IBBS-15 Abstract book. 15th International Biodeterioration and Biodegradation Symposium

15th International Biodeterioration &

Biodegradation Symposium

IBBS-15

Abstract Book

Katja Sterflinger & Guadalupe Piñar

19-24 September 2011

Vienna, Austria

IBBS-15 15th International Biodeterioration & Biodegradation Symposium

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PROGRAM & ABSTRACT BOOK

15th International Biodeterioration and Biodegradation Symposium

IBBS-15

19th – 24th September 2011

University of Natural Resources and Life Sciences

Vienna, Austria

Edited by: Katja Sterflinger & Guadalupe Piñar


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IBBS-15 was organized on behalf of the International Biodeterioration and Biodegradation Society (http://www.ibbsonline.org).

The symposium is hosted by the University of Natural Resources and Applied Life Sciences Vienna, and the Vienna Institute of Biotechnology.

We are very thankful to theUniversity for supporting IBBS-15 with technical facilities, access to lecture halls, poster boards and many servivces ranging from accounting to decoration.

We thank the Austrian Ministry of Education, Art and and Culture (bm:ukk), the Ministry of Agriculture, Forestry and Environment (Lebensministerium) and the Ministry of Science and Research (BMWF) for their geneous financial support.

We are very grateful to FEMS for supporting14 young scientists and 5 invited speakers of IBBS-15.


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Foreword by the organizers

This abstract book comprises 216 abstracts for the 15th International Biodeterioration and

Biodegradation Symposium held in Vienna, 19-23 September 2011 (http://ibbs-15.boku.ac.at). During the celebration of the IBBS-14 (2008) in the wonderful and sunny Sicily we were asked if we would be willing to organize the upcoming IBBS-15. Honestly, it took us just one – very short - Italian coffee to decide that we want to seize the chance and bring IBBS-15 to Vienna. And now here it is!

We have tried to organize IBBS-15 following the tradition of previous IBB-symposia, maintaining the family atmosphere created by the participants, some of them becoming good friends through the years. We want to encourage all new participants to mix in and become members of this IBB network-family. Therefore, we have particularly promoted the participation of young scientists, allowing them the opportunity to present their work to the scientific community, as well as to discuss topics in more detail with experts in topic-related working groups, creating a kind of “mentoring” which we think could be developed beyond this Symposium.

During IBBS-15 you will have the opportunity to present and to discuss your innovative research relating to different topics in the field of biodeterioration, biodegradation and bioremediation. In IBBS-15 - as is almost a tradition from the previous IBB-symposia - there is a strong focus on biodeterioration and bioconversion of various substrates such as wood, metals, rocks, building materials and especially materials of works of art and cultural heritage. However, we also decided to provide space for two large sessions on bioremediation of soil and water in order to signal the power of the so-called “white biotechnology”, and its significant potential to solve major ecological problems: these sessions are devoted to biodegradation of persistent pollutants and the bioremediation of polluted environments. Moreover, the IBBS-15 program also pays tribute to emerging technologies - genomics and proteomics - that can help us get a deeper understanding of organisms and their functions in biodeterioration and biodegradation.

We are proud that people from 41 different countries have registered for IBBS-15. We hope that you all feel comfortable in Vienna and at our University, and we are sure that you will benefit from your participation at IBBS-15.

We are very happy to welcome you to Vienna, Austria!

Katja Sterflinger & Guadalupe Piñar

Organizers of the 15th International Biodeterioration and

Biodegradation Symposium

Dep. Biotechnology, University of Natural Resources and Life Sciences Vienna, Austria.


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Foreword by the IBBS president

Dear colleagues,

The International Biodeterioration and Biodegradation Society (IBBS) is pleased to welcome delegates to its 15th International Symposium. This triennial series began in Southampton, UK, over 40 years ago. Initially concentrating on biodeterioration, the topics later came to include the increasingly important area of biodegradation, which today comprises around 50% of the submitted papers. This evolution has been mirrored in the Society Journal, International Biodeterioration and Biodegradation. From its beginnings, as the International Biodeterioration Bulletin, it progressed to its present title via “International Biodeterioration” and, in the first four issues of the current year, has published 26 papers on biodeterioration and 58 on biodegradation.

IBBS 15, held in the charming city of Vienna, continues in the tradition of previous Symposia, offering a forum for the presentation of review and research papers together with essential social and academic interaction between industry and higher education/research institutes. The success of any event depends on the dedication and assistance of many people and institutions. The Society thanks the University of Natural Resources and Applied Life Sciences in Vienna, members of the Symposium committees, and especially the two principal organisers, Drs. Katja Sterflinger and Guadelupe Piñar.

Christine Gaylarde

IBBS president


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Coordinators: Katja Sterflinger and Guadalupe Piñar (Austria)

Organizing committee: Advisory committee: Scientific committee: Ksenija Lopandic (A) Christine C. Gaylarde (UK) Cesáreo Saiz-Jimenez (ES) Christian Voitl (A) Joanna Verran (UK) Clara Urzí (IT) Jörg Ettenauer (A) John Gillat (UK) Diego A. Moreno (ES) Kristina Zakharova (A) Diego A. Moreno (ES) Flavia Pinzari (IT) Günther Ellersdorfer (A) Hans-Curt Flemming (GER) Hans-Curt Flemming (GER) Daniela Watzinger (A) James T. Walker (UK) James T. Walker (UK) Eva Maria Fritz (A) Geoffrey Michael Gadd (UK) Juan Luis Ramos (ES) Caroline Poyntner(A) Victor de Lorenzo (ES) Bärbel Kovarik (A) Guadalupe Piñar (A) Donatella Tesei (A) Katja Sterflinger (A)

Sessions:

IBBS15-1 Biofilms and Biofouling.

IBBS15-2 Biogenic transformations of rock, minerals, metals (MIC) and radionuclides.

IBBS15-3 Biodeterioration and bioconversion of lignin, cellulose and paper materials: biotechnological applications.

IBBS15-4 Biodeterioration of Cultural Heritage and building materials in indoor and outdoor environments: assessment and control, methods, treatments, prevention, cleaning.

IBBS15-5 Biodeterioration of miscellaneous materials: spacecraft materials, medical devices, glass, teflon, “nano” coatings.

IBBS15-6 Biodegradation of hydrocarbons and persistent pollutants: oil hydrocarbons, plastics, PAH, PCBs.

IBBS15-7 Bioremediation of contaminated soils and water, bio-filtration of industrial pollutants, development of GMOs and their use in bioremediation.

IBBS15-8 Biodiversity, ecophysiology and function of organisms involved in BBB

IBBS15-9 State of the art methods to study organisms and processes of BBB – genomics/proteomics/

IBBS15-10 Control of biodeterioration: new physical and chemical methods.

The text of the abstracts in this book was not modified by the editors. The authors are fully responsible for the submitted material. In exceptional cases we removed figures or citations because these were exceeding the maximum length announced in the guideline for authors.


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IBBS-15 program

Vienna, September 19th -24th 2011 University of Natural Resources and Applied Life Sciences Department of Biotechnology

Monday, 19th

16:00 – 19:00 Registration

Poster Set up

16:00 - Welcome cocktail “Come together”

Location:

BOKU, Muthgasse 11, 1190 Vienna,

Metro U4 station Heiligenstadt

Tuesday, 20th

9:00 – 9:30 Opening ceremony

Welcome words by Christine Gaylarde (President of IBBS), Gerold Stanek (FEMS delegate), Josef Glössl (Vice-Rector for Research and International Research Collaboration, BOKU) and Katja Sterflinger and Guadalupe Piñar (organizers)

Session

IBBS-15-1

Biofilms and Biofouling

(chair: Joanna Verran)

9:30 – 10:00 keynote: Hans-Curt Flemming (University Duisburg-Essen, Biofilm Center, Germany)

“Biofouling – Open questions, insufficient approaches and distant light”

10:00 -10:15 Pierangela Cristiani, Andrea Franzetti, Giuseppina Bestetti

“Bacterial profiles of biofilms operating in Microbial Fuel Cells”

10:15– 10:30 Carla C. C. R. de Carvalho “Bacterial adaptation to antineoplastic agents involve biofilm formation”

10:30– 10:45 Ruth Nithila , R.P. George, B.Anandkumar and U. Kamachi Mudali

“Effect of applied potential and ultrasonication on mircorbial adhesion of condenser materials of cooling water systems”


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10:45– 11:00 Sergey Dobretsov, M. Teplitzki, V. J. Paul, P. Proksch

“Bacterial quorum sensing inhibitors can prevent biofouling”

coffee break

Session

IBBS-15-5

Biodeterioration of miscellaneous materials: spacecraft materials, medical devices, glass, teflon, “nano” coatings (chair:Hans-Curt Flemming)

11:30– 12:00 keynote: Kasthuri Venkateswaran

(NASA-JPL, USA)

“Safeguarding the Crew and Engineering Systems for Human Missions”

12:00– 12:15 Jimmy Walker , Tom Pottage, Allan Bennett, Thomas Rohr, Chantal Fowler, Martin Wilson

“Effectiveness of gaseous decontamination technologies for use on spacecraft and their components”

12:15– 12:30 Gavin Bingley, Craig Banks, Gordon Craig, Mark Bodner, Joanna Verran

“Fungal colonisation and contamination of cinematographic film: implications for film and archivists”

12:30– 12:45 I. Vivar, S. Borrego, D.A. Moreno, A.M. García

“Fungal Biodeterioration of Cinematographic Films of the Cultural Heritage of Cuba”

12:45– 13:00 D.S.Rathore, S. Chauhan “Effect of climatic conditions on the succession of fungi on vegetable tanned sole leather (Buff) during storage”

lunch break

Posters are already available during lunch break


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Session

IBBS-15-3

Biodeterioration and bioconversion of lignin, cellulose and paper materials: biotechnological applications I

(chair: Kari Steffen)

14:00– 14:30 Keynote: Flavia Pinzari,

(ICPAL, Italy)

“From plant to book leaves: microbial exploitation of cellulose”

14:30– 14:45 N. Mesquita, A. Portugal, J. Loureiro, I. Nunes, S. Cabo-Verde, M.L. Botelho, G. Piñar

“Assessing sub-lethal gamma radiation effects on the physiology of fungi isolated from ancient documents: from CFU count to flow cytometry and fluorescence techniques”

14:45– 15:00 Valentina Raimondi, Giovanni Agati, Laura Berni, Giovanna Cecchi, Ioana Gomoiu, David Lognoli, Lorenzo Palombi

“Autofluorescence spectral features of fungal strains at different observational scales and their exploitation for the diagnostics of paper biodeterioration”

15:00– 15:15 Grit Kabiersch , Johanna Rajasärkkä, Susanna Hillebrand, Marko Virta, Annele hatakka, Marja Tuomela, Kari Steffen

“Bioluminescent yeast essays to follow the degradation of endocrine disrupting compounds after treatment with lignin-degrading fungi”

Coffee break

Session

IBBS-15-3 Biodeterioration and bioconversion of lignin, cellulose and paper materials: biotechnological applications II

(chair: Flavia Pinzari)

16:00– 16:15 Michael Schrems, Philipp Vejdovszky, Martin, Zeilinger, Sonja Bednarik, Emmerich Haimer, Falk Liebner, Thomas Rosenau, Antje Potthast

“Lignin-based soil improvers as key for profitable biomass conversion”

16:15– 16:30 Spérandio Mathieu, Hernandez-Raquet Guillermina, Paul Etienne, Lefebvre Xavier, Pommier Sebastien

“Analysis and modelling of cellulosic fibers degradation by bacterial consortia in mixed bioreactor”

16:30– 16:45 Daniel J. Yelle, Alexander N. Kapich, Kenneth E. Hammel

“White rot fungi selectively cleave threo-form of �-aryl ether linkages in spruce lignin”


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16:45– 17:00 GP Malyon, SD Streeter, SM Cragg

“Endogenous lignocellulosic digestive mechanism of the marine wood-boring isopod Limnoria quadripunctata”

17:00– 18:00 Poster session I

Topics IBBS-15-1 to IBBS-15-5

18:00 - open IBBS annual general meeting

Room will be announced.


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Wednesday 21st room: HS XX

Session

IBBS-15-4 Biodeterioration of cultural heritage and building materials I (chair: Cesareo Saiz-Jimenez)

09:00– 09:30 keynote: Christine Gaylarde (UK)

“Biodeterioration of cultural heritage buildings in tropical and sub-tropical climates”

09:30– 09:45 Pilar Bosch Roig, Rosa Montes Estellés, Jose Luis Regidor Ros, Pilar Soriano Sancho, Maria Teresa Doménech Carbó

“Biocleaning of wall paintings with bacteria”

09:45– 10:00 M. López-Miras, I. Martín-Sánchez, J. Romero-Noguera, F.C. Bolívar-Galiano, J. Ettenauer, K. Sterflinger, G. Piñar

“Analysis of microbial communities on a painting (oil on canvas): cultivation, molecular identification and evaluation of their biodegradative potential”

10:00– 10:15 P. Zalar, M. Lesar Kikelj, S. Kramar, A. Mladenovič, A. Padovnik, N. Gunde-Cimerman

“Investigations of fungi on wall paintings from a 16th century Lutheran cellar from Slovenia”

10:15– 10:30 Billeci N., Mancuso F. P.,

Pellegrino L., Palla F. “Investigation of biocenosis in a archaeological rock-cut settlements with relevant historic-artistic evidences. A case study in South-east of Sicily.”

10:30– 10:45 A. de los Ríos, B. Camara, Sergio Pérez-Ortega, M. Speranza, J. Wierzchos, C. Ascaso

“Problems met when trying to eliminate endolithic microorganisms damaging stone buildings using biocides”

10:45– 11:00 Elisabetta Zanardini, Kevin J.Purdy, Eric May, Helene Moussard, Simon Williams, Brian B.Oakley, J. Colin Murrell

“Investigation of archaeal and bacterial community structure and functional gene diversity in exfoliated sandstone at Porchester Castle (UK).”

Coffee break

11:30 - 11:45 Fadwa Jroundi, Carlos Rodriguez Navarro, Beatriz

“Bioconsolidation treatment at the Royal Chapel of Granada (Spain)”


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Martín Peinado, Javier Martín Peinado, Maria Teresa Gonzalez-Muñoz

11:45– 12:00 Jörg Ettenauer, Guadalupe Piñar, Katja Sterflinger, Maria Teresa Gonzalez-Muñoz and Fadwa Jroundi

“Molecular monitoring of the microbial dynamics occurring on historical limestone buildings during and after the in situ application of different bio-consolidation treatments”

12:00– 12:15 Laura Selbmann, Daniela Isola, Massimiliano Fenice, Laura Zucconi, Silvano Onofri

“Fungal cell wall degradation. New insights for removal of rock black fungi.”

12:15– 12:30 Črtomir Tavzes, Jernej Palčič, Karin Fackler, Franc Pohleven, Robert Koestler

“Biomimetic system for removal of fungal melanin staining on cultural heritage objects”

12:30– 12:45 O.A. Cuzman, D. Pinna, C. Riminesi, B. Sacchi, P.Tiano

“Evaluation of treatments efficiency against crustose lichens growing on monumental stone by electrical conductivity ”

12:45– 13:00 Gazzano C., Favero-Longo S.E., Piervittori R.

“Lichen secondary metabolites: potential biocides against rock-dwelling microcolonial fungi and cyanobacteria?”

Lunch break

Session IBBS-4

Biodeterioration of cultural heritage and building materials II (chair: Clara Urzi)

14:00– 14:15 A contribution by the Austrian artist Sofie Gangl

Biogenesis of cultural heritage: the beginning and “the end”!

14:15– 14:45 keynote: Frank Maixner

(Institute for mummies and the Iceman, Italy)

“Molecular investigations of microbial communities in mummified human remains.”

14:45– 15:00 Patrícia Guiamet, Miguel Crespo, Paola Lavin, Beatriz Ponce, Christine Gaylarde, Sandra Gómez de Saravia

“Biodeterioration of funeral sculptures in La Recoleta cemetery, Buenos Aires, Argentina. Pre- and post-interventions studies”

15:00– 15:15 Adamson, C.; Smith, B.J.; “The Influence of Aspect on the


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Warke, P.A.

Biological Colonization of Stone in Northern Ireland”

Coffee break

15:45– 16:00 Caroline Laffont , Yashiho Kikkawa

“Assessment of mould growth risk at the French national Library by monitoring and building simulations”

16:00– 16:15 Querner, Pasqual “Integrated Pest Management (IPM) and distribution of pest species in Berlin and Vienna – a comparison”

16:15– 16:30 J. von Werder, H. Venzmer

“The Potential of Pulse Amplitude Modulation Fluorometry for Evaluating the Resistance of Building Materials to Algal Growth”

20:00 Congress Dinner at the “Vienna Heurigen

Fuhrgassl-Huber”

Bus departure in front of the BOKU University at 19:30


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Wednesday, 21st room: HS XXI

Session IBBS-15-6

Biodegradation of hydrocarbons and persistent pollutants: oil hydrocarbons, plastics, PAH, PCBs I

(Chair: Abraham Esteve-Nùñez)

9:00 – 09:30 Keynote: Juan-Luis Ramos

(Consejo Superior de Investigaciones Cientificas-CSIC, Spain)

“Genomics and Post-Genomics of Solvent Tolerance in Pseudomonas putida”

09:30– 09:45 Hong Liu, Shu-Jun Wang, Jun-Jie Zhang, Hui Dai, Huiru Tang, and Ning-Yi Zhou

“Patchwork Assembly of nag-like Nitroarene Dioxygenase Genes and 3-Chlorocatechol Degradation Cluster for the Pathway Evolution of 2-Chloronitrobenzene Catabolism in Pseudomonas stutzeri ZWLR2-1”

09:45– 10:00 Irina Kosheleva, Sergei Sokolov, Alexander Boronin

“Ecology and Function of Catabolic Plasmids for Biodegradation of Petroleum Hydrocarbons”

10:00– 10:15 Noor Faizul Hadry Nordin , Amir Feisal Merican and Yusrizam Sharifudin

“Implication of rpoN loss-of-function in xenobiotic compounds degradation by Burkholderia xenovorans LB400”

10:15– 10:30 M. Afzal, S. Yousaf, T.G. Reichenauer, V. Andria and A. Sessitsch

“Efficient colonization, competitive ability and the expression of hydrocarbon degradation genes correlates with efficient phytoremediation of soil contaminated with diesel fuel”

10:30– 10:45 John Culhane, Nicholas Otieno, Kieran Germaine, Dina Brazil, David Ryan, David Dowling

“Development of endophytic and rhizospheric bacterial inoculants with remediation potential for practical application in the bio-energy crop industry”

Coffee break

11:15– 11:30 L. Santa Cruz-Calvo, J.I. Vílchez, J.J. Picazo-Espinosa, J. Narváez-Reinaldo, R. González-

“Isolation of desiccation tolerant microorganisms for the rhyzoremediation of soils subjected to seasonal drought”


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López, M. Manzanera

11:30- 11:45 Ondrej Uhlik , L. Musilova, M. Strejcek, J. Wald, C. Vlcek, J. Ridl, M. Mackova, T. Macek

“Aromatics-Utilizing Bacteria and Their Functional Genes in Long-Term PCB and PAH Contaminated Soil”

11:45– 12:00 K. Demnerová, J.Vrkoslavova, H. Stiborova , J.Purkrabová , J.Hajšlová

“Polybrominated flame retardans (BFRs) and their fate in the Czech Republic”

12:00– 12:15 Han Wang, Heli Xu, Anyi Hu, Qian Sun, Chang-Ping Yu

“Isolation and Characterization of Melamine (2, 4, 6-triamino-1, 3, 5-triazine)-degrading Bacteria ”

12:15– 12:30 Miriam Santo, Ronen Weitsman, Alex Sivan

“Biodegradation of polyethylene by bacterial laccase (phenol oxidase)”

Lunch break

Session IBBS-15-6

Biodegradation of hydrocarbons and persistent pollutants: oil hydrocarbons, plastics, PAH, PCBs II

(Chair: Victor de Lorenzo)

14:00– 14:15 Urooj Zafar , Geoff. D. Robson “Biodegradation of Polyurethane under compost at different temperatures”

14:15– 14:30 I.F. Ferreira , C.C.C.R. de Carvalho, D.I.C. Wang, M.R. Aires-Barros

“Crude desulfurization in organic:aqueous phase biocatalytic systems “

14:30– 14:45 H. Friman , A. Schechter, Y. Nitzan, R. Cahan

“Toluene Biodegradation in Bio electrochemical Cells (BEC) simulating microbial fuel cell (MFC)”

14:45– 15:00 Carolina Castro-Silva, V. M. Ruiz-Valdiviezo, S. G. Rivas-Rivera, A.R. Sosa-Trinidad, M. Luna-Guido, R. Marsch, L. Dendooven

“Bioavailability and dissipation of anthracene from soil with different alkalinity and salinity”

Coffee break

15:45– 16:00 Hugo Ribeiro, A. P. Mucha, C. Marisa R. Almeida, Adriano A. Bordalo

“Influence of rhizosediments characteristics on hydrocarbon degradation potential of Juncus maritimus plant-microorganisms


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Associations”

16:00– 16:15 Peressutti S.R, Herrera Seitz K., Pouyte K.1, Pedetta A., Babay P.A., Espinosa M., Costagliola M., Studdert C.

“Phenanthrene degradation and strategies to improve bioavailability by microorganisms isolated from sediments and marine waters in Argentine”

20:00 Congress Dinner at the “Vienna Heurigen

Fuhrgassl-Huber”

Bus departure in front of the BOKU University at 19:30


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Thursday, 22nd

Session IBBS-15-7

Bioremediation of contaminated soils and water, developmet of GMOs and their use in bioremediation of soils

(chair: Juan Luis Ramos)

9:00 – 09:30 Keynote: Victor de Lorenzo (Centro Nacional de Biotecnología, Spain)

“Capitalising microbial diversity for bioremediation: from systemic analyses to synthetic agents”

09:30-09:45 Akhilesh Tiwari , Anuradha Samyal, M.U. Charaya

“Rapid FTIR spectroscopic assessment of the efficiency of pretreated fungal biomass to adsorb lead”

09:45- 10:00 Mohd. Nizam Yusof, Siew-Moi Phang

“Biodegradation of Several Polycyclic Aromatic Hydrocarbons by Microalgae Isolated from Contaminated Site.”

10:00 -10:15 Satarupa Chakraborty, Abhishek Mukherjee, and Tapan K. Das

“Biochemical Characterization of a Lead-tolerant Strain of Aspergillus foetidus : An Implication of Bioremediation of Lead from Aqua- Enviroment”

10:15– 10:30 Maria S. Kuyukina , Irena B. Ivshina, Tatyana N. Kamenskikh, Maria V. Zhigalova

“Survival of poly(vinyl alcohol) cryogel-immobilized Rhodococcus cells in crude oil-contaminated soil and impact on biodegradation efficiency”

10:30– 10:45 María Markúsdóttir, Oddur Vilhelmsson

“Surfactant-degrader bioprospecting in sub-Arctic pristine and anthropogenically impacted seashore and freshwater environments”

Coffee break

11:15– 11:30 Lara Valentín , M. Tuomela, K. Steffen, E. Winquist, M. Moreira

and A. Hatakka

“Two fungal bioremediation methods for contaminated soil”


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11:30– 11:45 L. Zidkova, M. Otevrel, L.Wimmerova, J. Szokol, J. Nesvera, M. Patek

“Use of modified Rhodococcus strains for degradation of phenol in industrial wastewater.”

11:45– 12:00 R. Schroll, A. Krug, F. Wang, U. Dörfler, J.C. Munch

“Soil inoculation with specific microbial communities attached on carrier material - can this become a useful tool in soil remediation?”

12:00– 12:15 Olukanni Olumide D., Adekola Adejoke Adebimpe, Sunmola Damilola Banke, Osunmuyiwa Olufolabi

“Effect of redox mediators and various media on the decolourisation of the azo dye Methyl Red; and its biodegradation by Providencia rettgeri strain ODO”

12:15– 12:30 Subramanian Ramachandran, Marianne Saldanha, Irshad Ahmed, Pankaj Kumar Jain and Sanjay Kumar Verma2

“Application of recombinant Nostoc calcicola for the removal of Zinc(II) from Textile Industry Effluents”

12:30– 12:45 Abraham Esteve-Núñez, Karina Boltes, Alejandro Reija, Noemi Bragado, Ainara Dominguez, Jose Carpatos, Miriam Biel, Antonio Berná

“Microbial conversion of pollutants into electric current: strategies for monitoring and stimulating the bioremediation capacity of soil microbial communities.”

Lunch break

Session

IBBS-15-2

Biogenic transformations of rock, minerals, metals and radionuclides

(chair: Diego Moreno)

14:00– 14:30 Keynote: Geoff Gadd

(University of Dundee, UK)

“Biogenic transformation of rock, minerals, metals and radionuclides”

14:30– 14:45 Favero-Longo S.E., Turci F., Fubini B., Castelli D., Piervittori R.

“Lichen deterioration of asbestos and asbestiform minerals of serpentinite rocks in Western Alps”

14:45– 15:00 Ramanathan Baskar, Sushmitha Baskar, Joyanto Routh

“Moonmilk deposits in the Mawmluh Cave Ecosystem, Meghalaya, India: Some biogenic evidences”

15:00– 15:15 Jean Herisson, Pascal Taquet, Eric van Hullebusch, Thierry

“Development of a biodeterioration accelerated test to understand


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Chaussadent

behaviour of cementitious materials made with Calcium Aluminate Cement in sewer networks.”

15:15– 15:30 M. López-Fernández, O. Fernández Sanfranzisco, M. Martínez Garcia, M.L. Merroun

“Bacterial communities in clay formations and their interactions with radionuclides and heavy metals.”

coffee break

16:00– 17:00 Poster session II

topics IBBS-15-6 to IBBS-15-10

17:00-open Networking and mentoring sessions: topic related groups

Students meet teachers in topic-related working groups


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Friday, 23rd

Session

IBBS-15-10

Control of biodeterioration: new physical and chemical methods

(chair: Laura Selbmann)

9:00 – 09:30 keynote: Joanna Verran (Manchester Metropolitan University,UK)

“Antimicrobial surfaces: do we need them? Do they work?”

9:30 – 09:45 Mauro Tretiach , Fabio Candotto Carniel, Stefano Bertuzzi

“Biocide resistance in selected epi- and endolithic lichens after mild thermic pre-treatment: a case study from the Athenaion temple of Paestum (S Italy)”

9:45 – 10:00 Paul Hsieh, Jens Z. Pedersen, Patrizia Albertano

“Generation of Reactive Oxygen Species upon Red Light Exposure of Cyanobacteria from Roman Catacombs”

10:00– 10:15 Gladis, Franziska; Schumann, Rhena

“Testing the photocatalytic inactivation of aeroterrestrial algal growth”

Coffee break

Session

IBBS-15-8

Biodiversity, ecophysiology and function of organisms involved in BBB

(chair: Christine Gaylarde)

10:45– 11:15 Keynote: Clara Urzì ,

(University of Messina, Italy)

“Are we applying the right approach to study the microflora colonizing our built cultural heritage?”

11:15– 11:30 Rosa Margesin, Christoph Moertelmaier, De-Chao Zhang

“Bacterial diversity and biodegradation in hydrocarbon-contaminated soil from an alpine industrial site”

11:30– 11:45 Lydia Gustavs, Manuela Görs, Lubos Polerecky, Rhena Schumann, Ulf Karsten

“Algae ′aground′ – Biodiversity and ecophysiology of aeroterrestrial greenalgal biofilms ”

11:45– 12:00 S. Böck, K. M. Keiblinger, S. Zechmeister-Boltenstern, D.

“Fungal succession at early stages of beech litter decay”


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Bandian, A. Richter, J. Strauss, Markus Gorfer

12:00– 12:15 Adrian Langarica-Fuentes, Pauline S. Handley, Geoff D. Robson

“Biodiversity assessment and degradation potential of thermophilic fungi found in two different composting processes using traditional and molecular methods”

12:15 - 12:30 B. O. Ortega Morales “Biodeterioration processes and climate change: the Mayan cultural heritage as a case study”

Lunch

Session

IBBS-15-9

State of the art methods to study organisms and processes of biodeterioration and biodegradation – genomics / proteomics / transcriptomics

(chair: Geoffrey Gadd)

13:30– 14:00 keynote: Katrin Riedel

(TU Braunschweig & Helmholz Centre, Ger)

“Metaproteomics: Linking Microbial Diversity to Ecosystem Functioning”

14:00 - 14:15 Canalizo-Hernandez, Monica; A. Michael Enzien

“Evaluation of current microbial control technologies for biofilm dispersion using time-lapsed video microscopy”

14:15– 14:30 D.J. Beale, M.S. Dunn, P.D. Morrison, D.R. Marlow, D.C.O Marney

“Using a Metabolomic Approach to Identify Bio-Corrosion in Copper Pipes”

14:30 - 14:45 Lopandic Ksenija, Bond Ursula, Atanasova Lea, Druzhinina S. Irina, Sterflinger Katja

“Molecular characterisation of the interspecies yeast hybrids”


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14:45– 15:15 concluding ceremony:

Katja Sterflinger : “Phylogeny and evolution of biodeterioration research: let us boost horizontal knowledge transfer!"

Katja Sterflinger, Chris Gaylarde, Guadalupe Piñar

Coffee break

Departure

Saturday, 24th

8:30 – 18:00 Visit of the UNESCO World Cultural Heritage region “Wachau” at the Danube river and the monastery

“Stift Melk”.

Bus departure in front of the BOKU University Muthgasse 18 at 8:30


23

IBBS-15 List of presentations

Vienna, September 19th -24th 2011 University of Natural Resources and Applied Life Sciences Department of Biotechnology

O=oral / P=poster

Session IBBS-15-1

Biofilms and Biofouling: methods, treatments, prevention, cleaning

Authors Title Presentation No.

Page No.

Hans Curt Flemming Biofouling – open questions, insufficient approaches and distant light.

keynote lecture 46

Pierangela Cristiani, Andrea Franzetti, Giuseppina Bestetti

Bacterial profiles of biofilms operating in Microbial Fuel Cells.

IBBS-15-1-O01 47

Carla C. C. R. de Carvalho Bacterial adaptation to antineoplastic agents involved in biofilm formation.

IBBS-15-1-O02 48

Ruth Nithila , R.P. George, B.Anandkumar and U. Kamachi Mudali

Effect of applied potential and ultarsonication on microbial adhesion of condenser materials of coolong water systems.

IBBS-15-1-O03 49

Sergey Dobretsov, Max Teplitski, Valerie J Paul, Peter Proksch

Bacterial quorum sensing inhibitors can prevent biofouling.

IBBS-15-1-O04 50

Posters

I.R. Melo; S. L. Urtiga Filho; F. J. S. Oliveira; F. P de França

Biocossosion in pipeline during transport of diesel biodiesel mixture (B5).

IBBS-15-1-P01 51


24

Patricia Guiamet, Paola Lavin, Sandra Gómez de Saravia

Isolation of a Bacillus sp. strain from documents stored at La Plata Archive and evaluation of its biodeterioration ability.

IBBS-15-1-P02 52

Sonia Mihai, Octav Pantea, Andreea Bondarev, Dragos Ciuparua

Gold Shell Architectures and optoelectronic properties

IBBS-15-1-P03 53

B. Prieto, D. Vázquez-Nion, P. Sanmartín, B. Silva

Tracking pigment content variations using CIELAB color coordinates: a simple method of biofilm-forming organism assessment

IBBS-15-1-P04 54

Annika Vaksmaa, Sergey Dobretsov

Biofouling community changes due to the colour of substrate

IBBS-15-1-P05 55

Denise Cristina Moretti Vieira; Zilda Aparecida de Melo; Thereza Christina Vessoni Penna

Biofilm (biocellulose membrane) production by Glucoacetobacter xylinum from waste residues of fruits and tea leaves.

IBBS-15-1-P06 56

Jimmy Walker , Tom Pottage, Scott Hovland and Allan Bennet

A review of decontamination of microorganisms in space module habitats and on EVA suits

IBBS-15-1-P07 57

Andrea Steitz, Gerald Aschacher,Irmgard Matzinger

Microbial Monitoring of Biofilms in Paper Machines by Molecular Tools

IBBS-15-1-P08 58

Session IBBS-15-2

Biogenic transformations of rock, minerals, metals and radionuclides


Page No.

Geoff Gadd Biogenic transformation of rock, minerals, metals and radionuclides

keynote lecture 60

Favero-Longo S.E, Turci F., Fubini B., Castelli D., Piervittori R.

Lichen deterioration of asbestos and asbestiform minerals of serpentinite rocks in Western Alps

IBBS-15-2-O01 61

Ramanathan Baskar, Sushmitha Baskar, and Joyanto Routh

Moonmilk deposits in the Mawmluh Cave Ecosystem, Meghalaya, India: Some biogenic evidences

IBBS-15-2-O02 62

Margarita López Bacterial communities in clay IBBS-15-2-O03 63


25

Fernández, Omar Fernández Sanfrancisco, Marta Martínez García, Mohamed L. Merroun.

formations and their interactions with radionuclides and heavy metals.

Jean Herisson, Pascal Taquet, Eric van Hullebusch, Thierry Chaussadent

Development of a biodeterioration accelerated test to understand behaviour of cementitious materials made with Calcium Aluminate Cement in sewer networks.

IBBS-15-2-O04 64

Session IBBS-15-3

Biodeterioration and bioconversion of lignin, cellulose and paper materials: biotechnological applications


Page No.

Flavia Pinzari, Katja Sterflinger, Guadalupe Piñar

From plant to book leaves: microbial exploitation of cellulose

Keynote lecture 66

N. Mesquita; A. Portugal; J. Loureiro; I. Nunes; S. Cabo-Verde; M.L. Botelho; G. Piñar

Assessing sub-lethal gamma radiation effects on the physiology of fungi isolated from ancient documents: from CFU count to flow cytometry and fluorescence techniques

IBBS-15-3-O01 67

Valentina Raimondi, Giovanni Agati, Laura Berni, Giovanna Cecchi, Ioana Gomoiu, David Lognoli, Lorenzo Palombi

Autofluorescence spectral features of fungal strains at different observational scales and their exploitation for the diagnostics of paper biodeterioration

IBBS-15-3-O02 68

Grit Kabiersch , Johanna Rajasärkkä, Susanna Hillebrand, Marko Virta, Annele hatakka, Marja Tuomela, Kari Steffen

Bioluminescent yeast essays to follow the degradation of endocrine disrupting compounds after treatment with lignin-degrading fungi

IBBS-15-3-O03 69

Michael Schrems, Philipp Vejdovszky, Martin, Zeilinger, Sonja Bednarik, Emmerich Haimer, Falk Liebner, Thomas Rosenau, Antje Potthast

Lignin-based soil improvers as key for profitable biomass conversion

IBBS-15-3-O04 70

Spérandio Mathieu, Hernandez-Raquet Guillermina, Paul Etienne, Lefebvre Xavier, Pommier

Analysis and modelling of cellulosic fibers degradation by bacterial consortia in mixed bioreactor

IBBS-15-3-O05 71


26

Sebastien

Daniel J. Yelle, Alexander N. Kapich, Kenneth E. Hammel

White rot fungi selectively cleave threo-form of �-aryl ether linkages in spruce lignin

IBBS-15-3-O06 72

GP Malyon, SD Streeter, SM Cragg

Endogenous lignocellulosic digestive mechanism of the marine wood-boring isopod Limnoria quadripunctata

IBBS-15-3-O07 73

Posters

Eduardo Montero, Antonio Berna, David Yañez-Ruiz, Abraham Esteve-Núñez

Harvesting electrical energy from cellulose: microbial fuel cells hosting goat rumen bacteria in combination with Geobacter

IBBS-15-3-P01 74

Hrdinová Jitka , Jagošová Vanda, Kozumplíková Milena, Minařík Miroslav, Píštěk Vlastimil

Biologic pre-treatment of lignocellulosic wastes to improve biogas production

IBBS-15-3-P03 75

Parisa Mohammadi, Bentolhoda Sargazi, Shahla, Rodbar Mohammadi

Assay of Antifungal Effects of Walnut (Juglance nigra) and Pomegranate (Punica granatum) Extracts on Book Deteriorating Fungi

IBBS-15-3-P07 76

M. Nadi, Vahid Mozafari , A. Golchin and E. Sedaghati

The Effects of Vermicomposting on Organic Wastes

IBBS-15-3-P08 77

Astrid Michaelsen, Flavia Pinzari, Nicoletta Barbabietola, Guadalupe Piñar

Molecular monitoring of the effects of different conservation treatments on paper infecting fungi

IBBS-15-3-P10 78

Abdul Rehman and Shumaila shakoor

Isolation and characterization of cellulose degrading bacteria from local environment

IBBS-15-3-P11 79

Li Yongfeng, Liu Yixing, Dong Xiaoying, Wang Fenghu

Comparison of Decay Resistance of Wood and Wood-Polymer Composite Prepared by In-situ Polymerization of Monomers

IBBS-15-3-P12 80

Andrea Steitz, Birgit Schmöllerl, Notburga Pfabigan, Roland Gründlinger

Molecular and Visual Monitoring of Sapstaining Fungi and Moulds on Sawn Timber

IBBS-15-3-P13 81


27

Session IBBS-15-4

Biodeterioration of Cultural Heritage and building materials in indoor and outdoor environments: assessment and control, methods, treatments,

prevention, cleaning


Page No.

Christine Gaylarde Biodeterioration of cultural heritage buildings in tropical and sub-tropical climates

Keynote lecture 83

Frank Maixner , T. Rattei, G. Piñar, K. Sterflinger, D. Piombino-Mascali, G. Cipollini, A. Zink

Molecular investigations of microbial communities in mummified human remains

Keynote lecture 84

P. Bosch Roig, R. Montes Estellés, J. L. Regidor Ros, P. Soriano Sancho, M. T. Doménech Carbó

Biocleaning of wall paintings with bacteria

IBBS-15-4-O01 85

M. López-Miras, I. Martín-Sánchez, J. Romero-Noguera, F.C. Bolívar-Galiano, J. Ettenauer, K. Sterflinger, G. Piñar

Analysis of microbial communities on a painting (oil on canvas): cultivation, molecular identification and evaluation of their biodegradative potential.

IBBS-15-4-O02 86

P. Zalar, M. Lesar Kikelj, S. Kramar, A. Mladenovič, A. Padovnik and N. Gunde-Cimerman

Investigations of fungi on wall paintings from a 16th century Lutheran cellar from Sloveni

IBBS-15-4-O03 87

Billeci N., Mancuso F. P.,

Pellegrino L., Palla F. Investigation of biocenosis in a archaeological rock-cut settlements with relevant historic-artistic evidences. A case study in South-east of Sicily.

IBBS-15-4-O04 88

A. de los Ríos, B. Camara, Sergio Pérez-Ortega, M. Speranza, J. Wierzchos, C. Ascaso

Problems met when trying to eliminate endolithic microorganisms damaging stone buildings using biocides

IBBS-15-4-O05 89

Elisabetta Zanardini, Kevin J.Purdy , Eric May, Helene

Investigation of archaeal and bacterial community structure and functional

IBBS-15-4-O06 90


28

Moussard, Simon Williams, Brian B.Oakley, J. Colin Murrell

gene diversity in exfoliated sandstone at Porchester Castle (UK).

Fadwa Jroundi, C. Rodriguez Navarro, B. Martín Peinado, J. Martín Peinado, M. T. González-Muñoz

Bioconsolidation treatment at the Royal Chapel of Granada (Spain)

IBBS-15-4-O07 91

Jörg Ettenauer, G. Piñar, K. Sterflinger, M. T. Gonzalez-Muñoz and F. Jroundi

Molecular monitoring of the microbial dynamics occurring on historical limestone buildings during and after the in situ application of different bio-consolidation treatments.

IBBS-15-4-O08 92

Laura Selbmann, D. Isola, M. Fenice, L. Zucconi, S. Onofri

Fungal cell wall degradation. New insights for removal of rock black fungi.

IBBS-15-4-O09 93

Črtomir Tavzes, Jernej Palčič, Karin Fackler, Franc Pohleven, Robert Koestler

Biomimetic system for removal of fungal melanin staining on cultural heritage objects

IBBS-15-4-O10 94

O.A. Cuzman, D. Pinna, C. Riminesi, B. Sacchi and P.Tiano

Evaluation of treatments efficiency against crustose lichens growing on monumental stone by electrical conductivity

IBBS-15-4-O11 95

Gazzano C., Favero-Longo S.E., Piervittori R.

Lichen secondary metabolites: potential biocides against rock-dwelling microcolonial fungi and cyanobacteria?

IBBS-15-4-O12 96

Patrícia Guiamet., Miguel Crespo, Paola Lavin, Beatriz Ponce, Christine Gaylar, Sandra Gómez de Saravia

Biodeterioration of funeral sculptures in La Recoleta cemetery, Buenos Aires, Argentina. pre- and post-intervention studies.

IBBS-15-4-O13 97

Adamson C., Smith B.J., Warke P.A.

The influence of aspect on the Biological colonization of Stone in Northern Ireland

IBBS-15-4-O14 98

Caroline Laffont , Yashiho Kikkawa

Assessment of mould growth risk at the French national Library by monitoring and building simulations

IBBS-15-4-O15 99

Querner, Pascal

Integrated Pest Management (IPM) and distribution of pest species in Berlin and Vienna – a comparison

IBBS-15-4-O16 100


29

J. von Werder and H. Venzmer

The Potential of Pulse Amplitude Modulation Fluorometry for Evaluating the Resistance of Building Materials to Algal Growth

IBBS-15-4-O17 101

Posters

Hakan Sert Birds Cause Biodeterioration on Historical Monuments

IBBS-15-4-P03 102

Hacer Sert and K.

Sterflinger

Role of black fungi in biodeterioration processes of stone in historical monuments in Turkey

IBBS-15-4-P04 103

Gazzano C., Favero-Longo S.E., Matteucci E., Piervittori R.

Index of Lichen Potential Biodeteriogenic Activity: towards a statistical validation and the extension to other biodeteriogens

IBBS-15-4-P05 104

D. Haas, J. Habib, H. Galler, J. Posch, G. Zarfel, R. Schlacher, E. Marth, F.F. Reinthaler

Airborne fungal spores in moldy and non moldy homes

IBBS-15-4-P06 105

Katja Sterflinger, Laura Barnabei, Guadalupe Piñar, Frank Maixner and Dario Piombino-Mascali

The Capuchin Catacombs of Palermo, Italy: fungal deterioration of mummies and contamination of the indoor air

IBBS-15-4-P07 106

Asako Kusumi, Xian-Shu Li, Yu Osuga, Ji-Dong Gu, Yoko Kaayama

Bacterial Community Analyses of Biofilms on the Deteriorated Sandstone of Bayon Temple of Angkor Monuments in Cambodia

IBBS-15-4-P08 107

Katja Kavkler , Nina Gunde Cimerman, Polona Zalar, Andrej Demšar

Structural changes of artificially aged textiles after inoculation with the selected “museum” fungi

IBBS-15-4-P09 108

Lucia Kraková , Katarína Chovanová, Samy A. Selim, Alexandra Šimonovičová, Alena Maková, Domenico Pangallo

Evaluation of the microbial contamination risk of different historical documents conserved in the Slovak National Library

IBBS-15-4-P10 109

Asako Kusumi, Xian-Shu Li, Yoko Katayama

Isolation and distribution of facultative sulphur-oxidizing bacterium Mycobacterium sp. Strain THI503 on deteriorated sandstone of Bayon temple in Angkor monuments, Cambodia

IBBS-15-4-P11 110

T-P. Nguyen, T. Basset, S. Decagraph: Early detection of IBBS-15-4-P12 111


30

Moularat, F. Bousta, G. Orial, R. Berardo, A. Lama, Caroline Laffont , P. Ramond, E. Robine

biological and chemical contaminants of paper based cultural heritage.

A. Laudy, D. Ruszkowski, L. Rajpert, A. Jagielski and A. Skłodowska

The potential sensitivity to biodegradation processes of chosen natural dyes extracted from ancient silk decorations in Wilanow Palace – Museum

IBBS-15-4-P13 112

Sikha Mandal, Jnanendra Rath, Samit Ray

Algal colonization and deterioration of fine sculptures of terracotta temples of Bishnupur, West Bengal, India

IBBS-15-4-P14 113

S. Gutiérrez-Patricio, Ana Zélia Miller , M. L. Coutinho, M.l A. Rogerio-Candelera, M. Filomena Macedo, M. Hernandez-Mariné, C. Saiz-Jimenez

Characterization and quantification of phototrophic biofilms on artistic tiles

IBBS-15-4-P15

114

Irene Arroyo , M. Isabel Sarró, Julia Montero

Assessment of biodeterioration with different analytical techniques in El Castillo and Las Monedas caves. Cantabria, Spain

IBBS-15-4-P16 115

Julia Montero, Irene arroyo, José Vicente Navarro

Biodeterioration influence in the stone material: La Casa de las Conchas. Salamanca. Spain

IBBS-15-4-P17 116

Leone Montonen, Sanna Kettunen, Kari Steffen

Microbial flora of the 240 year old submerged wooden shipwreck Vrouw Maria

IBBS-15-4-P18 117

S. Gómez-Cornelio, Benjamin O. Ortega-Morales, S. De la Rosa-García, P. Quintana, J. Narváez-Zapata, A. Oliva-Hernández, H. Bullen, H. Urcia and W. Santiago

Fungal biogenic alteration of limestone monuments at Chichen Itza

IBBS-15-4-P19 118

Kalle Pilt , Uwe Noldt Measurement of mechanical properties in situ and modeling of timber structures in Ruhnu churches (Estonia) damaged by wood-destroying insects.

IBBS-15-4-P20 119

Valentina Raimondi, G. Cecchi, C. Cucci, O. Cuzman, C. Fornacelli, I. Gomoiu, D. Lognoli, D. Mohanu, L. Palombi, M.

Feasibility of fluorescence lidar remote sensing on frescoed surfaces: test studies

IBBS-15-4-P21 120


31

Picollo, P. Tiano

Jnanendra Rath, Sikha Mandal, Samit Ray

Survival strategies of desiccated Lyngbya corticcola in terracotta monuments of Bishnupur, India causing biodeterioration

IBBS-15-4-P22 121

Veridiana Reyes, A. Alvarez, C. Cano Canchola and J. Cervantes

Actinomycetes in welded tuffs IBBS-15-4-P23 122

M. Speranza, M. Sanz, M. Oujja, A. de los Ríos, S. Pérez-Ortega, M. Castillejo and C. Ascaso

Laser treatment as an effective tool to control epilithic and endolithic microbiota from stone

IBBS-15-4-P24 123

Katharina Wiegner, Oliver Hahn, Matthias Farke, Sabine Kalus and Wolfgang Horn

Determination of MVOCs like formic and acetic acid with LC-MS/MS

IBBS-15-4-P25 124

Manuela Gagliardi, Daniela Isola, Laura Selbmann, Maria Andaloro, Paola Pogliani, Silvano Onofri, Laura Zucconi,

Study on biodeteriorigenous agents in the Holy Saviour’s Cave (Vallerano, Italy)

IBBS-15-4-P26 125

Pilar Bosch Roig, Inés Martin Sanchez, Fernando Bolivar Galiano, Rosa Montes Estelles

Molecular biology identification of microorganisms presents in wall paintings

IBBS-15-4-P27 126

M. Garcia-Valles, G.Piñar, D. Gimeno-Torrente, J.L. Fernández-Turiel, J. Ettenauer, K. Sterflinger

Molecular investigation of the micro-biota asscociated to the decayed medieval stained window glasses of two Catalonian churches

IBBS-15-4-P28 127

Liam Morrison , Edward P. Lynch, Fabio Rindi

Biogenic weathering of calcareous building stone due to colonisation by subaerial algal assemblages

IBBS-15-4-P29 128


32

Session IBBS-15-5

Biodeterioration of miscellaneous materials: spacecraft materials, medical devices, glass,teflon,“nano” coatings


Page No.

Kasthuri Venkateswaran Safeguarding the Crew and Engineering Systems for Human Missions

Keynote lecture 130

Jimmy Walker , T. Pottage, Al. Bennett, T. Rohr, C. Fowler, M. Wilson

Effectiveness of gaseous decontamination technologies for use on spacecraft and their components

IBBS-15-5-O01 131

Gavin Bingley, Craig Banks, Dr Gordon Craig, Mark Bodner, Joanna Verran

Fungal colonisation and contamination of cinematographic film: implications for film and archivists

IBBS-15-5-O02 132

I. Vivar, S. Borrego, D.A. Moreno, A.M. García

Fungal Biodeterioration of Cinematographic Films of the Cultural Heritage of Cuba

IBBS-15-5-O03 133

D.S.Rathore, Shashi Chauhan

Effects of climatic conditions on the successions of fungi on vegetable tanned sole leather (buff) during storage

IBBS-15-5-O04 134

Posters

E. Damiano , F. Pinzari, D. Matè, M. C. Sclocchi

The microscopic effects of fungal biodeterioration on 20th century photographic documents revealed by vp sem-eds technique

IBBS-15-5-P01 135

D. Vellati, M. Adamo, D. Matè, F. Pinzari, D. Ruggiero, L. Botti, L. Residori, M.C. Sclocchi

Assays on the resistance of photographic paper treated with gamma rays to fungal biodeterioration

IBBS-15-5-P02 136

F. De Leo, G. Campanella, E. Proverbio, Clara Urzì

Assessment risk of fungal corrosion of unbonded lubricated post-tensioned tendons

IBBS-15-5-P03 137


33

Session IBBS-15-6

Biodegradation of hydrocarbons and persistent pollutants: oil hydrocarbons, plastics, PAH, PCBs


Page No.

Juan-Luis Ramos Genomics and Post-Genomics of Solvent Tolerance in Pseudomonas putida

Keynote lecture 139

Hong Liu, Shu-Jun Wang, Jun-Jie Zhang, Hui Dai, Huiru Tang, Ning-Yi Zhou

Patchwork Assembly of nag-like Nitroarene Dioxygenase Genes and 3-Chlorocatechol Degradation Cluster for the Pathway Evolution of 2 Chloronitrobenezene Cataloism in Pseudomonas stutzeri ZWLR2-1

IBBS-15-6-O01 140

Irina Kosheleva, Sergei Sokolov, Alecander Boronin

Ecology and function of Catabolic plasmids for Biodegradation of Petroleum Hydrocarbons

IBBS-15-6-O02 141

Noor Faizul Hadry Nordin , Amir Feisal Merican and Yusrizam Sharifudin

Implication of rpoN loss-of-function in xenobiotic compounds degradation by Burkholderia xenovorans LB400

IBBS-15-6-O03 142

M. Afzal, S. Yousaf, T.G. Reichenauer, V. Andria and A. Sessitsch

Efficient colonization, competitive ability and the expression of hydrocarbon degradation genes correlates with efficient phytoremediation of soil contaminated with diesel fuel

IBBS-15-6-O04 143

John Culhane, Nicholas Otieno, Kieran Germaine, Dina Brazil, David Ryan, David Dowling

Development of endophytic and rhizospheric bacterial inoculants with remediation potential for practical application in the bio-energy crop industry

IBBS-15-6-O05 144

L. SantaCruz-Calvo, J.I. Vílchez, J.J. Picazo-Espinosa, J. Narváez-Reinaldo, R. González-López, and M. Manzanera.

Isolation of desiccation tolerant microorganisms for the rhyzoremediation of soils subjected to seasonal drought

IBBS-15-6-O06 145


34

Ondrej Uhlik , L. Musilova, M. Strejcek, J. Wald, C. Vlcek, J. Ridl, M. Mackova, T. Macek

Aromatics-Utilizing Bacteria and Their Functional Genes in Long-Term PCB and PAH Contaminated Soil

IBBS-15-6-O07 146

K.Demnerová, J.Vrkoslavova, H. Stiborova , J.Purkrabová , J.Hajšlová

Polybrominated flame retardans (BFRs) and their fate in the Czech Republic

IBBS-15-6-O08 147

Han Wang, Heli Xu, Anyi Hu, Qian Sun, Chang-Ping Yu

Isolation and characterization of Melamine (2, 4, 6,-triamnion-1, 3, 5- triazine)-degreding Bacteria

IBBS-15-6-O09 148

Miriam Santo, Ronen Weitsman and Alex Sivan.

Biodegradation of polyethylene by bacterial laccase (phenol oxidase)

IBBS-15-6-O10 149

Peressutti S.R, Herrera Seitz K., Pouyte K., Pedetta A., Babay P.A., Espinosa M., Costagliola M., Studdert C.

Phenanthrene degradation and strategies to improve bioavailability by microorganisms isolated from sediments and marine waters in Argentine

IBBS-15-6-O11 150

Urooj Zafar , Geoff. D. Robson

Biodegradation of Polyurethane under compost at different temperatures

IBBS-15-6-O12 151

I.F. Ferreira , C.C.C.R. de Carvalho, D.I.C. Wang, M.R. Aires-Barros

Crude desulfurization in organic:aqueous phase biocatalytic systems

IBBS-15-6-O13 152

H. Friman , A. Schechter, Y. Nitzan and R. Cahan

Toluene Biodegradation in Bio electrochemical Cells (BEC) simulating microbial fuel cell (MFC)

IBBS-15-6-O14 153

Carolina Castro-Silva, V. M. Ruiz-Valdiviezo, S. G. Rivas-Rivera, A. Rosa Sosa-Trinidad, M. Luna-Guido, R. Marsch, L. Dendooven

Bioavailability and dissipation of anthracene from soil with different alkalinity and salinity

IBBS-15-6-O16 154

Hugo Ribeiro, Ana P. Mucha, C. Marisa R. Almeida, Adriano A. Bordalo

Influence of rhizosediments characteristics on hydrocarbon degradation potential of Juncus maritimus plant-microorganisms associations

IBBS-15-6-O19 155

Posters

Natalia González, M. Carmen Molina, Raquel Simarro , Cintia Casado, L.

Biodegradation of naproxene and carbamazepine by a bacterial consortium and particularly a single

IBBS-15-6-P01 156


35

Fernando Bautista, Emanuele Schiavi

strain identified as Pseudomonas fluorescens both isolated from waste water

G. Valducci Vecchi, M. Baratieri, S. Angeli, M. Grigiante and L. Brusetti

Laboratory-scale microbial bioremediation of pinewood-tar following a steam gasification process

IBBS-15-6-P03 157

Carla C. C. R. de Carvalho

Improving the bioremediation abilities of Rhodococcus erythropolis

IBBS-15-6-P04 158

Bappaditya Roy Rajat Banerjee, Sumana Chatterjee

Biodegradation of heat treated commercial Polyethylene by extracellular enzymes of Staphylococcus epidermis SPECIES BP/SU1.

IBBS-15-6-P06 159

Leonardo Jordão da Silva, Flávia Chaves Alves, Francisca Pessôa de França

Analysis of the Technologies Solutions applied to Oily Waste treatment from Oil Industry

IBBS-15-6-P07 160

Wael A. El Moslimany, Noura A. Alhamad, Wael S. El Sayed, Ashraf Al Nayal, Riyad Y. Hamzah

Emulsification and Biodegradation of Crude Oil by Bacteria Isolated from Petroleum-Contaminated Soil from Bahrain and Kuwait

IBBS-15-6-P09 161

Wael S. El-Sayed Characterization of Microbial Consortium Reductively Dechlorinating 2,3-dichlorophenol and 2,4,6-trichlorophenol

IBBS-15-6-P10 162

García-Díaz, C., Ponce-Noyola, T., Esparza-García, F.J., Rivera-Orduña, F., Ríos-Leal E. and Barrera-Cortés J.

Removal of Polycyclic Aromatic Hydrocarbons by Selected Strains Isolated from Organic Materials and Humic Acids

IBBS-15-6-P12 163

Azam Hassaninasab, Yoshiteru Hashimoto and Michihiko Kobayashi

Screening of microorganisms with curcumin-converting ability

IBBS-15-6-P13 164

Chang-Young Hong , Ki-Seob Gwak , Su-Yeon Lee, Seon-Hong Kim, In-Gyu Choi

Biodegradation of polychlorinated biphenyls by Ceriporia sp. ZLY-2010: enzymes production and degradation rate

IBBS-15-6-P14 165

Muhammad Ishtiaq Ali , Safia Ahmed and Abdul Hameed

Isolation and characterization of fungal strains capable of degrading Polyvinyl chloride (PVC)

IBBS-15-6-P15 166

Andrés Izquierdo, Joaquim Fuel component degradation and IBBS-15-6-P16 167


36

Vila, Lina Ortega, and Magdalena Grifoll

associated microbial community dynamics during the bioremediation of polluted soil in a biopile

Vladimir Jirku , Barbora Kosova , Alena Cejkova and Jan Masak

The Multiingredient Biosurfactant - enhanced Biodegradation of PAHs

IBBS-15-6-P17 168

Mehlika Karamanlioglu , Pauline Handley, Geoff Robson

The influence of temperature on degradation of the compostable plastic packaging material polylactic acid

IBBS-15-6-P18 169

Mphekgo P. Maila, Priscilla Randima and Thomas E Cloete

The removal of pahs from contaminated soil: effectiveness of seeding a root colonising bacteria

IBBS-15-6-P20 170

Igor Kviatkovski , Shay Zoharand and, Segula Masaphy

Degradation of high p-Nitrophenol concentrations Arthrobacter sp. isolated from agricultural soil.

IBBS-15-6-P21 171

Toru Matsui , Katsuya Kato, Tomoyuki Namihira, Naoya Shinzato and Hisashi Semba

Stereospecific degradation of phenylsuccinate by actinomycetes.

IBBS-15-6-P22 172

Dariush Minai-Tehrani , Z. Sobhani Damavandifar and S. Minoui

Biotreatment of oil sludge contaminated water by bacterial consortium

IBBS-15-6-P23 173

Mohd Naqiuddin Mohd Zairi , and Geoff Robson

Biodegradation of Polyurethane Plastics Exposed to the Environment

IBBS-15-6-P24 174

Somayeh Mollasalehi and Geoff Robson

Biodegradation of polyvinyl alcohol (PVA) by fungi

IBBS-15-6-P25 175

Sivaraman C, Anasuya Ganguly and Srikanth Mutnuri

Purification and Characterization of Cyclodextrin Glucanotransferase Enzyme and Its Role in Biodegradation of Diesel Oil Contaminated Soil

IBBS-15-6-P26 176

Wannarak Nopcharoenkul, Parichat Netsakulnee and Onruthai Pinyakong

Biodegradation of diesel by free and immobilized Pseudoxanthomonas sp. RN402

IBBS-15-6-P27 177

J. Harichová, E. Karelová, J. Godočíková, M. Bučková, A. Puškárová, B. Polek, P. Ferianc, Domenico Pangallo

Degradation of polycyclic aromatic hydrocarbons (PAHs) and long-chain hydrocarbons by bacterial strains isolated from polluted soil

IBBS-15-6-P28 178

Petra Prouzová, E. Hoskovcová, M. Holečková, T. Macek, K. Demnerová and

Study of microbes and plants interactions in PCB contaminated soil

IBBS-15-6-P29 179


37

M. Macková

Sidra Ilyas and Abdul Rehman

Decolourization and detoxification of Azo Dye, Synozol Red HF-6BN, by Aspergillus niger and Nigrospora sp.

IBBS-15-6-P30 180

Hugo Ribeiro, M.Teresa Borges, Ana Matos, Carlos M. Pereira, Paula M. L. Castro

Effects of biofouling on nitrification performance of a biofilter operating in a commercial marine aquaculture recirculation system

IBBS-15-6-P32 181

Agnieszka Richert, Maria Swiontek Brzezinska, Maciej Walczak

The influence of modified polyhexamethylene guanidine on the biodegradation of polylactide.

IBBS-15-6-P33 182

Rizzo C., Malavenda R., Lo Giudice A., Michaud L., Hoermann B., Gerce B., De Domenico M., De Domenico E., Syldatk C. , Hausmann R.

Biosurfactant-producing bacteria from Branchiomma luctuosum and Megalomma claparedei (Polychaeta, Sabellidae).

IBBS-15-6-P34 183

Safia Ahmed, Fazal-ur-Rehman, Zulfiqar A. Malik and Abdul Hameed

Biodegradation of Polycyclic Aromatic Hydrocarbons by newly isolated bacterial strain Bacillus cereus KWS2

IBBS-15-6-P35 184

A.-M. Tanase, I. Chiciudean, R. Ionescu, O. Csutak, D. Pelinescu, T. Vassu, I.Stoica

Characterization of hydrocarbon-degrading microbial strains isolated from oil polluted soil

IBBS-15-6-P37 185

Ajda Ulcnik, Lucija Zupancic-Kralij, Crtomir Tavzes, Franc Pohleven

Determination of remediation potentials of ligninolytic fungi for degradation of lindane

IBBS-15-6-P39 186

Session IBBS-15-7

Bioremediation of contaminated soils and water, bio-filtration of industrial pollutants, development of GMOs and their use in bioremediation, PAH, PCBs


Page No.

Victor de Lorenzo (Centro Nacional de biotecnologia, E)

Capitalising microbial diversity for biormediation : from systemic analyses to synthetic agents

Keynote lecture 188

Satarupa Chakraborty, Biochemical Characterization of a Lead- IBBS-15-7-O02 189


38

Abhishek Mukherjee, and Tapan K. Das

tolerant Strain of Aspergillus foetidus : An Implication of Bioremediation of Lead from Aqua- Enviroment

Mohd. Nizam Yusof, Siew-Moi Phang

Biodegradation of Several Polycyclic Aromatic Hydrocarbons by Microalgae Isolated from Contaminated Site.

IBBS-15-7-O03 190

Maria S. Kuyukina , Irena B. Ivshina, Tatyana N. Kamenskikh, Maria V. Zhigalova

Survival of poly(vinyl alcohol) cryogel-immobilized Rhodococcus cells in crude oil-contaminated soil and impact on biodegradation efficiency

IBBS-15-7-O04 191

María Markúsdóttir and Oddur Vilhelmsson

Surfactant-degrader bioprospecting in sub-Arctic pristine and anthropogenically impacted seashore and freshwater environments

IBBS-15-7-O05 192

Abraham Esteve-Núñez , K. Boltes A. Reija, N. Bragado, A. Dominguez, J. Carpatos, M. Biel and A. Berná

Microbial conversion of pollutants into electric current: strategies for monitoring and stimulating the bioremediation capacity of soil microbial communities

IBBS-15-7-O06 193

Lara Valentín, M. Tuomela, K. Steffen, E. Winquist, M. Moreira and A. Hatakka

Two fungal Bioremediation Methods for contaminated soil

IBBS-15-7-O07 194

R. Schroll, A. Krug, F. Wang, U. Dörfler, J.C. Munch

Soil inoculation with specific microbial communities attached on carrier material - can this become a useful tool in soil remediation?

IBBS-15-7-O08 195

L. Zidkova , M. Otevrel, L. Wimmerova, J. Szokol, J. Nesvera, M. Patek

Use of modified Rhodococcus strains for degradation of phenol in industrial wastewater

IBBS-15-7-O09 196

Olukanni Olumide David, A. Adejoke Adebimpe, S. Damilola Banke, O. Olufolabi

Effect of redox mediators and various media on the decolourisation of the azo dye Methyl Red; and its biodegradation by Providencia rettgeri strain ODO

IBBS-15-7-O10 197

Subramanian Ramachandran, M. Saldanha, I. Ahmed, P. Kumar Jain and S. Kumar Verma

Application of recombinant Nostoc calcicola for the removal of Zinc(II) from Textile Industry Effluents

IBBS-15-7-O11 198

Posters


39

Sina Azami, Pejman Rohanifar, Dariush Minai-Tehrani, Saeed Minoui

Effect of soy peptone as a nutrient on biodegradation of oil sludge in the soil

IBBS-15-7-P01 199

Andreea Bondarev, Sonia Mihai, Daniela Popovici and Octav Pantea

Biomaterials for Heavy Metals Adsorption

IBBS-15-7-P02 200

Ng Chow Goon, Miao Huang, Sze Chun Chau

Microbial dynamics in two-phase-partitioning bioreactors for hexane removal show partitioning of distinct groups into non-aqueous phase

IBBS-15-7-P03 201

Wafaa M. Abd El-Rahim, Magda A. El-Meleigy, Eman Refaat

Isolation and identification of fungi capable of textile dye degradation

IBBS-15-7-P05 202

E. Samuel Hartikainen, P. Lankinen, J. Rajasärkkä, H. Koponen, M. Virta, A. Hatakka, M. A. Kähkönen

Effects of Cu and Zn to the litter-decomposing fungi

IBBS-15-7-P08 203

Mohammad Ahmed Khiyami

Treatment of radium in groundwater of Saudi Arabia by PCS fungi biofilm

IBBS-15-7-P09 204

Lada Mathurasa, Ekawan Luepromchai, Chantra Tongcumpou, David A. Sabatini

Tributyltin Bioavailability and Biodegradation in Soil after Surfactant Flushing

IBBS-15-7-P12 205

Maria Laura Turino Mattos; Andre Andres; José Francisco da Silva Martins

Rhizoremediation of rice soils with residual clomazone herbicide

IBBS-15-7-P13 206

Elisabeth Mayer, Eva-Maria Gössl, Gonçalo A. Santos, Michaela Mohnl

Probiotics – Solution for bioremediation in fish and shrimp farming

IBBS-15-7-P14 207

Raquel Simarro, Natalia González, L. Fernando Bautista, M. Carmen Molina, Miguel Pérez, Luis Pérez

Assessment of bioremediation techniques (bio-stimulation, bio-

augmentation and natural attenuation) in a creosote polluted soil and restoration of the microbial community

IBBS-15-7-P15 208

Grzegorz Pasternak, Barbara Kolwzan

Biodegradation of carbazole by methylotrophic bacteria Methylobacterium sp. GPE1

IBBS-15-7-P16 209

Lenka Rucká, Michaela Šimčíková, Juraj Szőköl,

Construction of chromosomal deletions and gene-replacements in Rhodococcus

IBBS-15-7-P17 210


40

Petr Halada, Jan Nešvera and Miroslav Pátek

strains degrading aromatic compounds

Subramanian Ramachandran, M. Saldanha, I. Ahmed, P. Kumar Jain and S. Kumar Verma

Biosorption of Cr(VI) and Cr(III) by Nostoc calcicola immobilized on silica coated calcium alginate and silica gel

IBBS-15-7-P18 211

D. Guedes Ferreira, Márcia Teresa Soares Lutterbach, V. Oliveira, L. Silva Contador, M. Machado Galvão, D. Dias de Carvalho, E. Flávia Camporese Sérvulo

Bioremediation of clay soil contaminated with gasoline blended with different levels of ethanol

IBBS-15-7-P20 212

Moraes, D.A.; Anselmo, A.F; Souza, E.C ; Vessoni Penna, T.C.

Effect of broth composition and salinity in the volumetric mass transfer coefficient (kLa), for aerobic bioprocess to break organic gas residuals.

IBBS-15-7-P21 213

Tatoba R.Waghmode, Mayur B. Kurade, Sanjay P. Govindwar

A study on degradation of Remazol Red by Galactomyces geotrichum MTCC 1360 and effect of dye and degraded metabolites on iron uptake in Sorghum vulgare and Phaseolus mungo

IBBS-15-7-P23 214

Aakash Welgama, Christine Edwards, Linda A. Lawton

Biodegradation of natures own contaminants, microcystins and nodularin

IBBS-15-7-P24 215

Erika Winquist , Festus Anasonye, Kalle Salonen, Markus Räsänen, Kari Steffen and Marja Tuomela

Bioremediation of contaminated soil with fungi in aerobic conditions

IBBS-15-7-P25 216

Zhang Jing, LIN Xian-Gui, YIN Rui, LIU Wei-Wei

Enhance of cattle manure and sewage sludge on bioremediation of Aged Polycyclic Aromatic Hydrocarbons Contaminated Agricultural Soils

IBBS-15-7-P26 217

María Markúsdóttir, Oddur Vilhelmsson


IBBS-15-7-P27

218


41

Session IBBS-15-8

Biodiversity, ecophysiology and function of organisms involved in BBB


Page No.

Clara Urzi Are we applying the right approach to study the microflora colonizing our built cultural heritage

Keynote lecture 220

Benjamín Otto Ortega-Morales

Biodeterioration processes and climate change: the mayan cultural heritage as a case study

IBBS-15-8-O01 221

Rosa Margesin, Christoph Moertelmaier, De-Chao Zhang

Bacterial diversity and biodegradation in hydrocarbon-contaminated soil from an alpine industrial site

IBBS-15-8-O02 222

Lydia Gustavs, M. Görs, L. Polerecky, R. Schumann, U. Karsten

Algae ′aground′ – Biodiversity and ecophysiology of aeroterrestrial greenalgal biofilms

IBBS-15-8-O03 223

S. Moll, K. M. Keiblinger, S. Zechmeister-Boltenstern, D. Bandian, A. Richter, J. Strauss, and Markus Gorfer

Fungal succession at early stages of beech litter decay

IBBS-15-8-O04 224

Adrian Langarica-Fuentes, Pauline S. Handley, Geoff D. Robson

Biodiversity assessment and degradation potential of thermophilic fungi found in two different composting processes using traditional and molecular methods

IBBS-15-8-O05 225

Posters

Simona Ghita, Iusuf Loredana, Ioan I. Ardelean

Some microbiological parameters in marine microcosms supplemented with gasoline and gasoline-enriched marine populations

IBBS-15-8-P01 226

Margit Balazs, A. Szvetnik, A. Németh, A. Rónavári and

I. Kiss

Performance of DNA polymerases

IBBS-15-8-P03 227

Ortansa Csutak, C. Pavlusenco, I. Stoica, R.

Diversity and physiology analysis of new yeast strains isolated from oil-polluted

IBBS-15-8-P04 228


42

Ghindea, A. M. Tanase environment

L. Giaramida , L. Lawton,

C. Edwards and B. Singh

Trying to unravel the relation between natural bacterial communities’ diversity and bioremediation

IBBS-15-8-P05 229

Magdalena M. Majewska

Biodiversity of Cracow-Czestochowa Upland limestone-inhabiting fungi – summary of preliminary studies

IBBS-15-8-P06 230

Lydia Hemala, Rosa Margesin

Production of antimicrobial compounds by bacteria isolated from an alpine industrial site contaminated with hydrocarbons

IBBS-15-8-P07 231

Luiz Carlos Martins das Neves, M. Ishii, I. Conceição Roberto, T. Christina Vessoni Penna

Optimization of biosurfactant production by cultivation of Bacillus atrophaeus ATCC9372 in semi-defined olive oil and whey culture media

IBBS-15-8-P09 232

Dony Chacko Mathew and Chieh-Chen Huang

Isolation and Characterization of a potential mercury resistant endophytes, Photobacterium halotolerans , Meld1

IBBS-15-8-P10 233

Éva Mészáros, R. Sipos, R. Pál, C. Romsics, and K. Márialigeti

Stimulation of trichloroethylene biodegradation in anaerobic three-phase microcosms

IBBS-15-8-P11 234

Rita Sipos, É. Mészáros, G. Cebe, A. Mohr, R. Pál, C. Romsics, K. Márialigeti

Complex molecular biological investigation of TCE biodegradation with respect to electron donor amendments using anaerobic microcosms

IBBS-15-8-P12 235

Swiontek Brzezinska Maria , Jankiewicz Urszula, Walczak Maciej

Role of Streptomyces rimosus in degradation of chitin substances

IBBS-15-8-P13 236

Dong-Sheng Wei, Olaf Schmidt, Walter Liese

Fungal degradation studies on bamboo IBBS-15-8-P14 237

Session IBBS-15-9

State of the art methods to study organisms and processes of BBB – genomics/proteomics


Page No.


43

Katrin Riedel ( TU Braunschweig & Helmholz Centre, Ger)

Metaproteomics: Linking Microbial Diversity to Ecosystems Functioning

Keynote lecture 239

Monica A. Canalizo-Hernandez and Michael V. Enzien

Evaluation of current microbial control technologies for biofilm “dispersion” using time-lapsed video microscopy.

IBBS-15-9-O01 240

D.J. Beale, M.S. Dunn, P.D. Morrison, D.R. Marlow, and D.C.O Marney

Using a Metabolomic Approach to Identify Bio-Corrosion in Copper Pipes

IBBS-15-9-O02 241

Lopandic Ksenija, U. Bond, L. Atanasova, I. S. Druzhinina and K. Sterflinger

Molecular characterisation of the interspecies yeast hybrids

IBBS-15-9-O03 242

Posters

Zahra Sobhani Damavandifar, Manouchehr Mirshahi, Sina Sarikhani, Dariush Minai-Tehrani

A suitable method to detect polycyclic aromatic hydrocarbons (PAHs) as a water contaminant by monoclonal antibody

IBBS-15-9-P01 243

P. Sanmartín, D. Vázquez-Nion, B. Silva, B. Prieto

Advantages of contact-color techniques vs. improved chlorophyll extraction method for quantification of phototrophic biomass on stone surfaces

IBBS-15-9-P02 244

Session IBBS-15-10

Control of biodeterioration: new physical and chemical methods.


Page No.

Joanna Verran Antimicrobial surfaces: do we need them? Do they work?

Keynote lecture 246

Mauro Tretiach, Fabio Candotto Carniel, Stefano Bertuzzi

Biocide resistance in selected epi- and endolithic lichens after mild thermic pre-treatment: a case study from the Athenaion temple of Paestum (S Italy)

IBBS-15-10-O01 247

Paul Hsieh, Jens Z. Generation of Reactive Oxygen Species IBBS-15-10-O02 248


44

Pedersen, Patrizia Albertano upon Red Light

Exposure of Cyanobacteria from Roman Catacombs

Gladis, Franziska; Schumann, Rhena

Testing the photocatalytic inactivation of aeroterrestrial algal growth

IBBS-15-10-O03 249

Posters

Joanna Foksowicz-Flaczyk, Judyta Walentowska

Antimicrobial activity of ionic liquid applied to linen fabric

IBBS-15-10-P01 250

Chang Wook Jo, Jung Eun Choi, Young Hee Kim, Jin Young Hong, Mi Hwa Jung, Soo Ji Kim

Insecticidal activity of Asarumsieboldii against termites

IBBS-15-10-P03 251

Young-Hee Kim, Jung-Eun Choi, Jin-Young Hong, Mi-Hwa Jung, Chang-Wook Jo

Stability test of natural pesticides on materials for conservation of organic cultural heritage

IBBS-15-10-P04 252

P. Mohammadi, M. Ejadpanah-Saravi

Assessment of Antifungal Effect of Nano Metal Particles against Isolated Fungi from Ghajar Inscription

IBBS-15-10-P05 253

Bohuslav Uher One microbial bioassay for a method of cyanobacterial and algal removal

IBBS-15-10-P07 254

Judyta Walentowska, Joanna Foksowicz-Flaczyk

Natural essential oil for antimicrobial protection of natural textiles

IBBS-15-10-P08 255

Catalina Calin, Gina Vasile, Octav Pantea, Andreea Bondarev, Sonia Mihai

The influence of dithiocarbamates used for phytosanitary treatments of vineyard on the microelements contents from soil

IBBS-15-10-P09 256


45

Topic 01- Biofilms and Biofouling


46

KEYNOTE LECTURE Biofouling - unsolved problems, insufficient approaches and some light at the horizon Hans-Curt Flemming1

Biofilm Centre, University of Duisburg-Essen, Universitätsstr. 5, D-45141 Essen, [email protected] Microbial biofouling is a very costly problem. Five general reasons can be identified which continuously contribute to costs: i) Biofouling is detected by its effect on process performance or product quality and quantity. Early warning systems are very rare although they could save costly countermeasures necessary for removing established fouling. ii) For diagnosis, the common practice is to take water samples which give no information about site and extent of biofouling deposits. iii) When finally the diagnosis is established, biocides are employed which, in many cases, for the best kill microorganisms but do not remove them. Killing is not cleaning, and in many cases, the presence of biomass and not its physiological activity is the problem. iv) Nutrients are not considered as potential biomass. Biofouling is a biofilm phenomenon and based on the fact that biofilms grow at the expense of nutrients; oxidizing biocides can make things even worse by breaking recalcitrant molecules down into biodegradable fragments. v) Efficacy control is performed again by process performance or product quality only and not optimized by biofilm monitoring, verifying successful removal. Thus, further biofouling is inevitable. In order to overcome this vicious circle, an integrated strategy is suggested. It includes the selection of low-adhesion, easy-to-clean surfaces, good housekeeping, early warning systems, limitation of nutrients, improvement of cleaners, strategic cleaning and monitoring of deposits. The goal of a holistic anti-fouling strategy is: to learn how to live with biofilms and keep their effects below the level of interference in the most efficient way.


47

IBBS-15-1O01

Bacterial profiles of biofilms operating in Microbial Fuel Cells

Pierangela Cristiani1, Andrea Franzetti2, Giuseppina Bestetti2

1 RSE - Ricerca sul Sistema Energetico S.p.A., Dipartimento Ambente e Sviluppo Sostenibile, Milan, Italy. [email protected]

2 Dipartimento di Scienze dell’Ambiente e del Territorio, Università degli Studi di Milano Bicocca, Milan, Italy.

The microbial communities of operating membraneless microbial fuel cells has been analyzed sampling different sites, from the anaerobic anode to the aerobic area of the cathode, using the fingerprinting molecular technique, DGGE (Denaturing gradient gel electrophoresis).

The anode materials sampled were: carbon fibres of a brush, carbon cloth and stainless steel chips (AISI 304). The cathodes were carbon cloth based.

The microbial fuel cells were inoculated with row wastewater coming from a municipal plant of Milan city and weekly fed with sodium acetate 3 g/l.

DGGE profiles enabled to calculate the Jaccard similarity indexes for the bacterial communities that developed between the electrodes of the MFCs. In this way the degree of selection of microbial communities was defined in relation to the constructive characteristics, materials used and distance between the electrodes The cluster analysis based on similarity data, showed that the microbial populations, coming from the same inoculum, gradually diverged in relation to the structural characteristics of the cell, as well as to the material and the geometry of the anode. In addition, the excision and sequencing of some bands permitted the characterization of the different communities and bacteria groups.


48

IBBS-15-1O02

Bacterial adaptation to antineoplastic agents involve biofilm formation


Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Department of Bioengineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal ([email protected])

About 30% of patients receiving chemotherapy for non-hematologic neoplasms and 85% of patients receiving induction chemotherapy for acute leukemias develop potentially life-threatening infections (1). This high rate of infections in cancer patients has been ascribed to a lower ability of the host defense system, to invasive procedures (e.g. surgery and application of catheters) and chemotherapy they have to endure (2). However, bacterial adaptation to antineoplastic agents can also result in an increased resistance to antibiotics (3).

During the adaptation tests to the several antineoplastic agents, it was observed that Staphylococcus aureus, Rhodococcus erythropolis, Mycobacterium aurum and Escherichia coli cells adhered to surfaces and that the size of the adherent aggregates, during the first step of biofilm formation, reflected the concentration of the antineoplastic agent. The formation of biofilms has been related to the pathogenicity of some bacteria, such as staphylococci, and to the establishment of chronic infections (4).

In the present study, the adaptation to the antineoplastic agents resulted in a considerably increased resistance to antibiotics. One of the reasons was the existence of persisters, which could be observed by waiting 36h instead of the usual 16-24h for MIC determination. Other mechanisms involved modifications in the cellular membrane and cell wall composition, EPS production and changes in the physicochemical properties of the cell surface that resulted in cell aggregation and biofilm formation.

References:

(1) Sylvester RK (2006) Infections in patients with cancer. ACCP's Pharmacotherapy selfassessment program, Book 10: Hematology/ Oncology I and II. (Schumock GT, Brundage DM, eds) pp. 147-165, American College of Clinical Pharmacy, Kansas City, MO.

(2) Guinan JL, McGuckin M, Nowell PC (2003) Oncology (Williston Park) 17:415-420.

(3) de Carvalho C.C.C.R. (submitted)

(4) Costerton JW, Stewart PS, Greenberg EP (1999) Science 284:1318–1322.


49

IBBS-15-1-O03

Effect of applied potential and ultrasonication on microbial adhesion of condenser materials of cooling water systems

Ruth Nithila, R.P. George, B.Anandkumar and U. Kamachi Mudali

Corrosion Science and Technology Division, Indira Gandhi Center for Atomic Research, Kalpakkam 603 102, INDIA

Biofouling of corrosion resistant titanium reduces heat transfer efficiency in equipments such as titanium-tubed steam condensers of power plants. Conventional fouling control strategies do not guarantee a problem free cooling water system. Hence there is the necessity to develop non-conventional techniques to supplement the present treatment programmes and thereby effectively defeat biofouling. In the present study, the impact of applied potential and ultrasonication on biofilm formation was analyzed. Anodic and cathodic applied potentials of 600 mV (SCE) for the duration of one hour was tested in both biofilm forming environments and already formed biofilms on titanium. Potentiodynamic polarization, Open circuit potential and adhesion rate monitoring was done to select these test parameters. MTCC cultures of Gram-positive Bacillus sp., Micrococcus sp. and Gram negative Pseudomonas sp. and Flavobacterium sp. were used in this study. Total viable count and Direct Acridine orange Count, epifluorescence microscopy and Bacterial adhesion to Hydrocarbons assay was used to quantify and visualize microbial adhesion and biofilm formation. Results showed approximately 50% reduction in adhesion and biofilm formation. Control of Gram-positive species with cathodic applied potentials was faster compared to Gram-negative species. Effect of ultrasonication at a frequency of 24 kHz and power of 200 W was also studied. The ultrasonication time of 30 minutes to get maximum kill was standardized and intermittent cooling was done to differentiate the effect of heat. Present results showed good reduction in microbial density in water and epifluorescence micrographs showed absence of biofilm formation, reduction in microbial density and lesser fluorescence indicating lesser activity on titanium surfaces exposed to ultrasonicated water.


50

IBBS-15-1-O04

Bacterial quorum sensing inhibitors can prevent biofouling

Sergey Dobretsov1, Max Teplitski2, Valerie J Paul3, Peter Proksch4

1 Department of Marine Science and Fisheries, College of Agricultural and Marine Sciences, Sultan Qaboos University, Sultanate of Oman

2 Soil and Water Science Department, University of Florida, Gainesville, USA

3 Smithsonian Marine Station, Fort Pierce, Florida, USA

4 Institut für Pharmazeutische Biologie und Biotechnologie, Düsseldorf, Germany

Quorum sensing (QS) - a bacterial population density dependent gene regulatory mechanism in response to production, release and perception of low molecular weight signal molecules – is important for regulation of bacterial adhesion, biofilm formation and virulence. In this study, 78 natural products isolated from marine organisms (sponges, algae, fungi, tunicates and cyanobacteria) and terrestrial plants were screened for the inhibition of bacterial QS using a reporter strain Chromobacterium violaceum CV017. Forty two percent of the tested compounds inhibited QS of the reporter. QS inhibitory activities of the most active and abundant compounds were further investigated using the LuxR-based reporter E. coli pSB401 and the LasR-based reporter E. coli pSB1075. Demethoxy encecalin and hymenialdisin reduced QS dependent luminescence of the LasR-based reporter induced by N-3-oxo-dodecanoyl-L-homoserine lactone. Hymenialdisin, demethoxy encecalin and microcolins A and B inhibited response of the LuxR-based reporter induced by N-3-oxo-hexanoyl-L-homoserine lactone. Coatings with kojic acid (concentration = 0.5% w/w) - a QS inhibitor found in this study - reduced two-fold the densities of bacteria and diatoms and inhibited recruitment of macrofoulers on the plates in the field bioassays. Our study suggests that compounds that interfere with bacterial QS signals might be a good alternative to traditional antifouling compounds.


51

IBBS-15-1-P01

Biocorrosion in pipeline during transport of diesel-biodiesel mixture (B5) I.R. Melo1; S. L. Urtiga Filho2; F. J. S. Oliveira3; F. P de França1 1 Departamento de Engenharia Bioquímica, Escola de Química, UFRJ, Centro de Tecnologia, Bloco E, Ilha do Fundão, CEP- 21949-900, Rio de Janeiro - RJ – Brasil.

2 Federal University of Pernambuco/Recife/Brazil

3 Petróleo Brasileiro S.A. Av. Almirante Barroso, 81, 23 andar, Centro, CEP- 20031-004, Rio de Janeiro – RJ, Brasil.

The mixture is obtained of different portions of diesel and esters of vegetable oils, called biofuels. It emerges as an alternative for the cutback of oil derived fuel dependence, but it can suffer alteration in its properties throughout time, due to the hydrolytic, microbiological and oxidative reactions. The fuel corrosion and degradation processes may be induced and speeded due to the presence of water or microorganisms. This paper aims to assess the formation of biofilms in dynamic system using as circulating fluid sweet water with 10, 60 e 80% of B5 in the metallic surface of the carbonic steel AISI1020. A closed looping system was used built on stainless steel 316L. The process was conducted for 15 days, being monitored through the quantification of plankton and sessile microorganisms and mass loss of the metallic coupons. The findings have shown that there has been variation of the microbial throughout the experiment, the adherence of microorganisms over the metallic surface. In the biofilms significant aerobic, anaerobic, fungi, iron bacteria, aerobic and anaerobic acid producer bacteria, Pseudomonas aeruginosa and sulfate- reducing bacteria (SRB) were quantified. The corrosion rate as of 0,15± 0,03;0,18±0,01 and 0,14±0,005mm/year,respectively, with the 15-day-process.


52

IBBS-15-1-P02

Isolation of a Bacillus sp. strain from documents stored at La Plata Archive and evaluation of its biodeterioration ability Patricia Guiamet (1,2), Paola Lavin (1,3), Sandra Gómez de Saravia (1,4)

1Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas (INIFTA), Departamento de Química, Facultad de Ciencias Exactas, UNLP, CCT La Plata- CONICET, C.C. 16, Suc.4 (1900), La Plata. Tel: 54-221-4257430, Fax: 54-221-4254642. [email protected] 2Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, CONICET. 3Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CONICET 4Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, CICBA.

It is widely known that biofilms (complex microbial communities embedded in a matrix of extracellular polymeric substances -EPS-) may cause valuable economic damage in the industry. When these biofilms attach to materials belonging to documentary heritage e.g. paper, photos and magnetic and optical media, among others, social and cultural damage is irreparable due to loss of valuable information contained in these media. This process is known as biodeterioration. One of the genera causing biodeterioration is Bacillus. This genus has been the most frequently found in documents and environmental sampling carried out in three Archives from Argentina (Archivo Histórico del Museo de La Plata, Archivo del Colegio de Escribanos y Departamento de Investigación Histórica y Cartográfica de la Dirección de Geodesia). Bacillus spp can attack cellulose, parchment and glues causing biodeterioration of the documents due to the production of amylase, cellulase, N-acetyl ß-glucosaminidase, lactic acid, acid phosphatase. In this study, the ability of a strain identified by molecular biology techniques as Bacillus sp., isolated from documents of Archivo Histórico del Museo de La Plata to attack materials such as paper and optical media (CD-ROM) was evaluated. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed that this strain of Bacillus was able to attach to both substrates and form biofilms causing damage on them, which demonstrates the biodeterioration ability of the microorganism.

Acknowledgments We are very grateful to the Universidad Politécnica de Madrid (Spain) for its financial support (grants AL07-PID-020 and AL08-P (I+D)-08), and we acknowledge Ana M. García and Diego A. Moreno for their assistance with biomolecular techniques.


53

IBBS-15-1-P03

Gold Shell Architectures and optoelectronic properties

Sonia Mihai,1* Octav Pantea1, Andreea Bondarev1 and Dragos Ciuparu1

1 Petroleum and Gas University of Ploiesti, Ploiesti , Romania,. Fax: +40 - 244 - 575 847; Tel + 40 - 244 - 573 171; E-mail: [email protected]*

Synthesis, characterization, and functionalization of self-assembled, ligand-stabilized gold nanoparticles are studied in the chemistry of nanomaterials. Gold auto-assembled films were prepared by interaction of Schiff Base with tris(triphenylphosphinegold)oxonium tetrafloroborate. The gold cluster compounds were characterized by various techniques UV-vis, FT-IR spectroscopy, X-ray photoelectron (XPS), SEM and AFM. The AFM allowed the examination of the topography of the Schiff Base adsorbed films on gold auto-assembled films at the nanometric scale and confirmed that the Schiff Base forms a stable layer. For all Schiff Base was found to be adsorbed on gold surfaces and to form a stable layer. We have investigated the electrical and optical properties of gold films nanoparticles.

Acknowledgements:

Authors recognise financial support from the European Social Fund through POSDRU/89/1.5/S/54785 project : "Postdoctoral Program for Advanced Research in the field of nanomaterials”.


54

IBBS-15-1-P04

Tracking pigment content variations using CIELAB color coordinates: a simple method of biofilm-forming organism assessment

B. Prieto, D. Vázquez-Nion, P. Sanmartín, B. Silva

GEMAP-USC, Grupo de Estudios Medioambientales Aplicados al Patrimonio Natural y Cultural. Departamento de Edafología y Química Agrícola, Facultad de Farmacia, Universidad de Santiago de Compostela, 15782-Santiago de Compostela, Spain. [email protected] (B. Prieto)

Pigment content provides valuable information about the growth and physiological status of organisms, such as those found in a biofilm. The interest in monitoring the biofilm development on cultural heritage surfaces as part of their preventive control, explains the importance of checking a rapid, simple, cost-effective, on site and non-destructive method of organism assessment based on color measurements. Our objective was to investigate the relationship between reflectance color measurements, expressed by means of the CIELAB color coordinates, and pigment content (chlorophyll a, total carotenoids and phycobiliproteins) in response to variations in environmental conditions: nitrate and phosphate sources and light intensity. An aeroterrestrial biofilm-forming cyanobacterium of the genus Nostoc was chosen as test organism. Findings show that the CIELAB color coordinates are close correlated with pigment content, which highlights the relevance of the implementation of color techniques in biodeterioration studies in the cultural heritage field.


55

IBBS-15-1-P05

Biofouling community changes due to the colour of substrate

Annika Vaksmaa, Sergey Dobretsov

Department of Marine Science and Fisheries, College of Agricultural and Marine Sciences, Sultan Qaboos University

We hypothesized that substrate colour can affect not only the settlement of macro-fouling species but also the formation of microbial communities, which can directly or indirectly affect the formation of macro-fouling communities. Acrylic panels and slides painted with two contrast colours (black and white) and covered with transparent sheets were exposed to biofouling for the period of 40 days in the Marina Bandar al Rowdha (Sea of Oman). Micro-fouling communities were sampled after 5th, 10th and 20th days. Densities of different species of diatoms and bacteria were counted using an epiflourescence microscope. Development of macro-fouling communities was assessed with 10 day intervals using a dissecting microscope. Community structure and densities of Bacillus-like and Cocci-like bacteria changed over the time but no effect of colour was observed. Macro-fouling communities altered over time and formed two different clusters (after 20 and 30 days) on a MDS plot. Major species that contributed to the dissimilarities between communities were Ulva sp., Folliculina sp. and Balanus amphitrite. The species richness was higher on the white colour panels whereas the evenness of communities was higher on the black panels. The results of the present study emphasize the importance of substrate colour for the formation of fouling communities that need to be taken into account in the future antifouling studies.


56

IBBS-15-1-P06

Biofilm (biocellulose membrane) production by Glucoacetobacter xylinum from waste residues of fruits and tea leaves. Denise Cristina Moretti Vieira1; Zilda Aparecida de Melo1; Thereza Christina Vessoni Penna1

1 School of Pharmaceutical Science – Universidade de São Paulo. São Paulo. Brazil The biofilm (biocellulose membrane (C6H1005) n), produced over the static culture of Glucoacetobacter xylinum associated to Saccharomyces cerevisiae, was obtained from waste residues of fruits and tea leaves (for instance green tea). For green tea as substrate, the obtained biofim yield was 2.38 g fiber / day. Green tea added with orange juice increased the yield of biofilm up to 2.93 g fiber / day. For black and mate tea leaves, the highest productivity cellulose was 1.09 g fiber / day and 0.38 g fiber / day, respectively. The addition of (1.2%) collagen to the culture with mate tea increased productivity by 5 times. The addition of orange juice increased productivity by 3.7 times in tea media. It was concluded that the tea leaves and fruit juices used are suitable for the production of bacterial cellulose. DSC analysis for the obtained biofilm confirmed thermal degradation up to 130°C and a melting point of (-15°C).


57

IBBS-15-1-P07

A review of decontamination of microorganisms in space module habitats and on EVA suits

Jimmy Walker¹, Tom Pottage¹, Scott Hovland² and Allan Bennet¹ ¹HPA , Microbiological Services Division, Porton Down Salisbury. ² European Space Agency, ESTEC, Holland.

Biofilms on surfaces of the MIR space station caused material biodegradation that may have led to safety issues and there are concerns that biofilm microorganisms on future planetary missions may forward contaminate the target planet breaching international planetary protection treaties if not decontaminated sufficiently. Therefore, there is a requirement to decontaminate spacecraft module interiors during the mission and also Extra Vehicular Activity (EVA) space-suits whilst on Mars to reduce forwards or backwards contamination. A literature review was carried out of decontamination technologies that could be employed singularly or in combination for habitat module interiors on missions < 6 months and > 6 months using a scoring criterion and trade-off matrix to select appropriate technologies. For missions < 6 months, disinfectant wipes were recommended in combination with air disinfection/filtration systems. Missions > 6 months would also require additional measures like gaseous decontamination systems to periodically reduce the inaccessible bioburden. In addition, decontamination technologies for the inside and outside surfaces of EVA space-suits (on Lunar and Martian surfaces) were also reviewed. Only physical decontamination of the exterior EVA suit surface is required for Lunar missions, but for Martian missions this would be enhanced with gaseous disinfection. The interior of the suit could be decontaminated using passive antimicrobial fabrics and active cleaning using disinfectant wipes. Conclusions: A range of different decontamination technologies were recommended to prevent the colonisation of surfaces in the habitat module and on the EVA suits, and further research is required to develop these technologies for use in actual missions.


58

IBBS-15-1-P08

Microbial Monitoring of Biofilms in Paper Machines by Molecular Tools

Andrea Steitz1, Gerald Aschacher1,Irmgard Matzinger1

1Holzforschung Austria, Franz Grill Straße 7 A-1030 Vienna,

Two paper machines were investigated regarding the microbial composition of their biological slimes (biofilms) and the characteristics of their process waters. Biofilms from the wet end of the paper machines were examined by Denaturing Gradient Gel Electrophoresis (DGGE). Clone libraries were established for the identification of the microbial population of the biofilms. The performance of one biocide was intensively investigated by measuring adenosine triphosphate (ATP) values, colony forming units (CFU) and bacterial counts estimated by fluorescence microscopy. Highly significant differences between the process waters in the two paper machines were detected for pH values, temperature, redox potential, total organic carbon (TOC) and total nitrogen (TN). Chemical oxygen demands (COD) deviated significantly from each other, whereas the differences in phosphate concentration, phenol index and conductivity were insignificant. Bacteria were identified as prevalent colonizers of the biofilms whereas fungi were not found. The impact of the cleaning procedure (hydrogen peroxide) showed almost no influence on the bacterial population, the microbial fingerprints obtained at various time points showed only a variation in intensity of the charcteristic bands. The identification of the bacterial population in the two paper machines revealed two completely different ecosystems. The paper machine running with a significant higher temperature (41-68 °C) was populated mostly by thermophilic bacteria of the Deinococcus-Thermus-group and species number was smaller than in the paper machine operating at a lower temperature (44-50 °C). Members of the alpha- and beta-proteobacteria were dominating in this paper machine. The addition of the biocide showed only a temporary (15 min.) and local suppression on bacterial growth and their metabolism.


59

Topic 02–Biogenic transformations of rock, minerals, metals and

radionuclides


60

KEYNOTE LECTURE

Biogenic transformation of rock, minerals, metals and radionuclides

Geoffrey M. Gadd

Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, Scotland, UK

Microbes play key geoactive roles in the biosphere particularly regarding element biotransformations and biogeochemical cycling, metal and mineral transformations, decomposition, and bioweathering. All kinds of microbes, including prokaryotes and eukaryotes and their symbiotic associations with each other and “higher organisms”, can contribute actively to geological phenomena, and central to many processes are metal and mineral transformations. Microbes possess several properties that effect changes in metal speciation and mobility, as well as mineral formation or mineral dissolution. Such mechanisms are important components of natural biogeochemical cycles for metals as well as associated elements in rocks and minerals, e.g. sulfur and phosphorus, and metalloids, actinides and metal radionuclides. Apart from being important in natural biosphere processes, such metal-mineral transformations can have beneficial or detrimental consequences in a human context. Bioremediation refers to the application of biological systems to clean-up of pollution with bacteria and fungi being the most important organisms for reclamation, immobilization or detoxification of metallic pollutants. In contrast, metal and mineral transformations by microbes may result in spoilage and destruction of natural and synthetic materials, rock and mineral-based building materials, e.g. concrete, acid mine drainage and associated metal pollution, biocorrosion of metals, alloys, and related substances, and adverse effects on radionuclide speciation, mobility and containment. Our research seeks to understand mechanisms of metal and mineral biotransformations, and their environmental and applied significance in bioremediation, biodeterioration and corrosion. Some examples outlined in this presentation will include fungal degradation of rocks and metal-containing minerals, depleted uranium and uranium oxides, and fungal biodeterioration of concrete which may have implications for radioactive waste containment as well as being of general biodeteriorative significance.


61

IBBS-15-2-O01

Lichen deterioration of asbestos and asbestiform minerals of serpentinite rocks in Western Alps

Favero-Longo S.E.1,2, Turci F.1,3, Fubini B.1,3, Castelli D.1,4, Piervittori R.1,2

1, University of Torino, Interdepartmental Centre “G. Scansetti” for Studies on Asbestos and Other Toxic Particulates

2, University of Torino, Dipartimento di Biologia Vegetale - Viale Mattioli 25, 10125, Torino, Italy

3, University of Torino, Dipartimento di Chimica IFM - Via Giuria 7, 10125, Torino, Italy

4, University of Torino, Dipartimento di Scienze Mineralogiche e Petrologiche - Via Valperga Caluso 35, 10125, Torino, Italy

Serpentinites of Western Alps contain fibrous minerals as vein-filling minerals, including (a) chrysotile (serpentine) and tremolite (amphibole) asbestos, commercially used in the past, (b) minerals usually having fibrous habit (asbestiform minerals), as balangeroite (gageite group) and carlosturanite (serpentine), and (c) other minerals, usually non fibrous, which locally occur with a fibrous habit, such as antigorite (serpentine) and diopside (pyroxene). Fibres exposed at the surface of natural outcrops are frequently colonized by lichens, well known agents of physico-chemical deterioration. This may result in modulating also the toxicity-relevant properties of such fibres, e.g. surface induced free radical release. In particular, we showed that lichen colonization (in the field) and lichen metabolites (in vitro) affect the surface chemistry of chrysotile from the disused asbestos mine of Balangero (the largest in Western Europe), inducing a partial reduction of toxic properties. The mineral deterioration by Candelariella vitellina is here investigated on a wider set of natural outcrops exposing veins containing: (a) chrysotile-carlosturanite-fibrous diopside, (b) calcite-fibrous tremolite, (c) fibrous antigorite. All serpentine fibres contacted by lichen hyphae are significantly modified in their chemical composition, while tremolite and diopside are unaffected (SEM-EDS). In vitro all serpentine fibres were significantly depleted in magnesium by lichen metabolites, up to a complete conversion into amorphous silica upon incubation with oxalic acid 50 mM (SEM-EDS, ICP-AES). Minor leaching was observed for fibrous tremolite and diopside. On the other hand all incubations induced significant modification of the amount of free radicals released by all fibres (EPR), signalling a likely altered toxic potential.


62

IBBS-15-2-O02

Moonmilk deposits in the Mawmluh Cave Ecosystem, Meghalaya, India: Some biogenic evidences

Ramanathan Baskar 1, Sushmitha Baskar 1, and Joyanto Routh 2, 3

1Department of Environmental Science and Engineering, Guru Jambheshwar University of Science and Technology, Hisar 125001, Haryana, India E-mail: [email protected]

2Department of Earth Sciences, IISER-Kolkata, Mohanpur 741252, India

3Department of Natural Sciences and Technology, MTM, Örebro University, Örebro, 70182 Sweden

Abstract

Moonmilk deposits, actively form on the floor of Krem Mawmluh in Meghalaya, Northeastern India. The initial observations reveal abundant micrite and calcified microbial filaments and we hypothesize that these deposits form as a result of biogenic interactions. Electron microscopy (SEM) showed abundant needle calcite, fibre calcites (micro-fibre and nano-fibre calcite crystals), biofilm and microbial filaments. The total viable culturable microbes showed high population densities and finally in vitro culture experiments, confirmed the ability of many of the isolated strains (Bacillus sp. and Actinomycetes) to precipitate calcite These results support the biogenic nature of the deposits at Krem Mawmluh.

Keywords: Geomicrobiology, Caves, Moonmilk, Bacteria, Calcite precipitation


63

IBBS-15-2-O03

Bacterial communities in clay formations and their interactions with radionuclides and heavy metals. Margarita López Fernández1, Omar Fernández Sanfrancisco1, Marta Martínez García1, Mohamed L. Merroun1. 1 Dept of Microbiology, Universidad de Granada, Campus Fuentenueva, s/n, 18071 Granada, Spain.

Abstract Microbial occurrence and -processes in granitic and salt deposits as host rock candidate for the radioactive waste repositories has been previously investigated. In the case of clay formations, however, very few studies on the presence and viability of microbes in these formations were performed. This work describes the culture dependent bacterial diversity of two bentonite samples recovered from clay deposits of “Cortijo de Archidona” (Almeria, Spain). These samples were chosen as potential host rock model for the deep geological disposal of radioactive wastes. The mineralogy of these two samples (BI and BII) is dominated by clay mineral of smectite and differs by the presence of the iron sulphate mineral phase, jarosite (KFe3+

3(SO4)2(OH)6) for the sample BII. The evaluation of aerobic bacterial populations clearly indicated the presence of high numbers of cultivable bacteria in both samples. Bacteria belonging to Actinobacteria (Arthrobacter, Micrococcus, Amycolatopsis), Gammaproteobacteria (Pseudomonas), Bacilli (Bacillus), etc. were identified. The isolated bacterial strains were characterized biochemically and physiologically. The heavy metals tolerance study showed that a high fraction of the bacterial population was able to tolerate high concentrations of metals, i.e. up to 4 mM U, 3 mM Pb, etc. The bacterial uranium tolerance mechanisms were studied using transmission electron microscopy and infrared spectroscopy. The results obtained indicated that U precipitation as uranium phosphate mineral phases (e.g. meta-autunite) was involved in the bacterial tolerance to this radionuclide.


64

IBBS-15-2-O04

Development of a biodeterioration accelerated test to understand behaviour of cementitious materials made with Calcium Aluminate Cement in sewer networks.

Jean Herisson1, Pascal Taquet2, Eric van Hullebusch3, Thierry Chaussadent1

1 Paris-Est University, French Institute of Science and Technology for Transport, Development and Networks (IFSTTAR), 58 boulevard Lefebvre, 75732 Paris Cedex 15, France

2 France Kerneos,1 rue Le Chatelier, ZAC Parc Technologique, 38090 Vaulx-Milieu, France

3 Paris-Est University, Laboratoire Géomatériaux et Environnement (LGE), EA4508, 5 boulevard Descartes, 77454 Marne-la-vallée Cedex 2, France

Contact: [email protected], [email protected], [email protected]

Sewer networks contain many aggressive and corrosive agents for pipe materials. One type of damage can be attributed to concrete corrosion by biogenous sulfuric acid. Under particular conditions biogenic hydrogen sulfide may be released in the headspace. This latter sulfur species is biologically converted into sulfuric acid by the action of a succession of sulfur-oxidizing bacteria in the pipe headspace. According to field data, cementitious materials made with Calcium Aluminate Cement (CAC) offer far better performance on site than those made of Ordinary Portland Cement (OPC). The development of an accelerated and pertinent laboratory test is necessary to better understand the mechanisms involved in both cementitious materials.

Biodegradation mechanisms can be illustrated by four steps:

� Abiotic oxidation of H2S into elemental sulfur on the surface of cementitious material � Oxidation of sulfur to sulfuric acid by bacteria � Migration of sulfuric acid into gypsum layer � Acid attack of cementitious materials producing gypsum expansive layer.

The study of each step allows identifying elements necessary to establish an accelerated test reproducing the conditions found in the aerial part of sewer networks.

Such a test is nowadays developed in Ifsttar. Meanwhile, an in situ exposure campaign is also performed in France.

This study aim at comparing the results obtained on the different conditions of exposure on several materials as follows: OPC/sand (use as a reference), CAC/sand and CAC/CAC aggregate.


65

Topic 03–Biodeterioration and bioconversion of lignin, cellulose and

paper materials: biotechnological applications


66

KEYNOTE LECTURE

From plant to book leaves: microbial exploitation of cellulose

Flavia Pinzari1, Katja Sterflinger2, Guadalupe Piñar2

1 Istituto Centrale per la Conservazione ed il Restauro del Patrimonio Archivistico e Librario - Laboratorio di Biologia. Via Milano, 76, 00184 Rome. Italy.

2 Institute of Applied Microbiology, Department of Biotechnology, Institute of Bio Technology (VIBT), University of Natural Resources and Applied Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.

Cellulose is the main structural component of photosynthetic organisms and the most common organic compound on Earth. The fungal and bacterial breakdown of cellulose is a key process in the global carbon cycle. Estimates indicate that annually over 1011 tons of cellulose is synthesized by plants, and contemporaneously a comparable amount is degraded by microorganisms. In soil environments, this cellulosic matter provides a carbon and energy source to many heterotrophic microorganisms, which in turn produce CO2 via respiratory processes. The human use, or intended use, turns cellulose from “matter” to “material”, and its spoilage from a primary natural process to a problem to solve. The accumulation of books and archival materials inside buildings devoted to their conservation, created new environmental opportunities for cellulolytic fungal and microbial species. The fungal and bacterial consortia that establish on books are not too far from the communities of decomposers which, in natural environments, transform cellulose of lifeless organic matter into low molecular forms, and cellular energy. The development and sustained presence of a fungal assemblage on an individual book depend on the type of spore that reaches the material surface of the host, the particular microenvironment (temperature, relative humidity, and light), the water activity of the substrate, and the casual events which promote the colonisation of materials. The accurate assessment of the colonisation and breakdown of materials which has been caused by microorganisms and the successful distinguishing of deteriogens from associated non-deteriogens species still represent a key topic that is going to benefit from the new molecular and microscopy techniques.


67

IBBS-15-3-O01

Assessing sub-lethal gamma radiation effects on the physiology of fungi isolated from ancient documents: from CFU count to flow cytometry and fluorescence techniques

N. Mesquita1; A. Portugal1; J. Loureiro1; I. Nunes2; S. Cabo-Verde2; M.L. Botelho2; G. Piñar3

1. Centre for Functional Ecology, Department of Life Sciences, Faculty of Sciences and Technology, University of Coimbra, 3001-401 Coimbra, Portugal.

2. Unit of Physics and Accelerators, Nuclear and Technological Institute, Estrada Nacional 10, 2686-953 Sacavém, Portugal.

3. Universität für Bodenkultur, Vienna Institute of Biotechnology (VIBT), Muthgasse 11, A-1190, Vienna, Austria.

Recent research in paper biodeterioration aims at developing and optimizing methodologies that prevent the degradation of historical documents. Cladosporium cladosporioides is radio resistant, and was used as a reference to assess the radiation dose required to treat documents. This isolate was collected from an ancient document from the Archive of the University of Coimbra.

A batch of paper disks impregnated with spores was submitted to different radiation doses; the effects on fungal biomass, radial growth and viability were assessed using two separate methodologies: 1) colony radial growth and biomass, and 2) colony forming units count.

The determined D10 for the tested strain was of 2.7 kGy, and 15.4 kGy completely inhibited the culture growth. Results show that it is possible to disinfect documents colonized by fungi, by decreasing their biological activity when using radiation doses up to 14.4 kGy, a dose that has been tested and described as safe to paper supports in terms of physical resistance and color changes.

Furthermore, the effects of sub-lethal radiation doses on spores are also being analysed using different flow cytometry techniques; with the use of different fluorochromes it is possible to analyse the effects in spore growth, viability and even metabolism, in a more immediate time-frame. By allowing the analysis of several thousands of spores in a short period of time, we think that the further development of this technique, when applied to fungal spores, will offer better information on the effects of different disinfection techniques, with a physiological basis.


68

IBBS-15-3-O02

Autofluorescence spectral features of fungal strains at different observational scales and their exploitation for the diagnostics of paper biodeterioration

Valentina Raimondi1, Giovanni Agati1, Laura Berni2, Giovanna Cecchi1, Ioana Gomoiu3, David Lognoli1, Lorenzo Palombi1

1 'Nello Carrara' Applied Physics Institute - National Research Council, Via Madonna del Piano 10, I- 50019, Sesto Fiorentino, Firenze, Italy

2 Freelance paper conservator, Italy

3 National Arts University of Bucharest, Institute of Biology, Splaiul Independentei, 296, R-77700 Bucharest, Romania

The autofluorescence features of fungal strains are here investigated with several fluorescence techniques at different observational scales, from the microscopic scale, by using an epifluorescence microscope coupled to a microspectrofluorimeter, to the macro scale by using laser induced fluorescence imaging on biodeteriorated paper samples. Fluorescence measurements at microscopic scale were carried out with an optical epifluorescence microscope, coupled to a CCD camera, a standard webcam and a microspectrofluorimeter. This instrumentation was synergetically used to record in vivo real-time autofluorescence spectral features of spores and hyphae in Aspergillus niger while exposed to ultraviolet irradiation. The results showed a remarkable change in the autofluorescence features of the sample after only few minutes of exposure and can contribute to the interpretation of data obtained with other fluorescence techniques. Macro-scale fluorescence measurements were conducted both on isolated cultures and on biodeteriorated paper samples by means of the laser induced fluorescence hyperspectral imaging technique. The latter can be used to obtain fluorescence images of the target, each pixel of which contains information relevant to the whole visible spectrum with a spectral resolution better than 2 nm. The aim was to investigate the potential on biodeterioration detection on paper substrate at an early stage of development of the biological growth. Results are compared with biological sampling data and open good prospects for an early detection.


69

IBBS-15-3-O03

Bioluminescent yeast assays to follow the degradation of endocrine disrupting compounds after treatment with lignin-degrading fungi

Grit Kabiersch, Johanna Rajasärkkä, Susanna Hillebrand, Marko Virta, Annele Hatakka, Marja Tuomela, Kari Steffen

Department of Food and Environmental Sciences, University of Helsinki, P.O. Box 56, 00014 Helsinki, Finland

Endocrine disrupting compounds are substances that mimic the action of natural hormones and interfere with the hormone system. We are studying the degradation of two different compounds, namely (i) bisphenol A (BPA), which interacts with the estrogen receptor and is suspected to increase the risk of breast and prostate cancer, and (ii) tributyltin (TBT), which interacts with the retinoid X receptor and induces a phenomenon called “imposex” in female gastropods. Both compounds were treated by lignin-degrading fungal cultures, i.e. Stropharia coronilla, S. rugosoannulata or Kuehneromyces mutabilis, or with lignin-degrading enzyme solutions containing manganese peroxidase (MnP) and/or laccase. The successful removal of hormonal activity has been followed using bioluminescent yeast assays displaying the estrogen receptor α or the retinoid X receptor. In both assays, the receptor is expressed from a plasmid. After binding a ligand, it dimerizes and induces the expression of luciferase in a dose-dependent manner. Finally D-luciferin is added and the emitted light is detected. 7-Day old cultures of S. coronilla were able to remove the estrogenic activity of BPA within 24 hours completely. Treatment of BPA with purified neutral MnP from this fungus also resulted in 100% reduction of estrogenic activity within 6 hours. A supersaturated TBT solution was treated with concentrated culture liquid from K. mutabilis or a purified MnP. Removal of TBT was observed during a 24 hour treatment time. Supersaturated TBT solution mimicked TBT contaminated sediment suspension, which could be treated with a fungus or an enzyme.


70

IBBS-15-3-O04

Lignin-based soil improvers as key for profitable biomass conversion Michael Schrems, Philipp Vejdovszky, Martin, Zeilinger, Sonja Bednarik, Emmerich Haimer, Falk Liebner, Thomas Rosenau, Antje Potthast

Christian-Doppler-Laboratory “Advanced cellulose chemistry and analytics” Department of Chemistry, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austria [email protected]

Due to fading fossil resources a strong need for valorization of renewable resources is on the rise. Bioconversion of renewable resources by treatment with hot aqueous ethanol, known as the Organosolv process, is an old concept dating back to the early 1950s. With the beginning of the 1980s the interest in this research area increased remarkable focusing on the delignification of renewable materials, mainly wood. But in spite of overbalanced advantages existing production sites have been closed down by the end of the 1990s and the interest ebbed away. Herewith we want to present a new approach for a profitable bioconversion process utilizing Organosolv technology using the good accessibility of by-products, which has always been the strength of this concept compared to others like kraft or sulfite pulping. Lignin, the second most abundant natural polymer on earth, shows a great potential to make such a process economically feasible on the large scale. Until now rather neglected by simply using it as fuel, various more beneficial applications are possible. A very promising application is the use of Organosolv lignin as a source for ammonoxidized lignins which can be applied as soil improvers combating progressive desertification in parts of the world. At the conference we will present the development of an Organosolv process aligned for maximal lignin output using rye straw as source for bioconversion. We will also give insights into the mechanism of delignification through investigations of the reaction kinetics using Apocynol, a phenol derivative, as a simple model for lignin.


71

IBBS-15-3-O05

Analysis and modelling of cellulosic fibers degradation by bacterial consortia in mixed bioreactor Spérandio Mathieu, Hernandez-Raquet Guillermina, Paul Etienne, Lefebvre Xavier, Pommier Sebastien

1 Université de Toulouse; INSA,UPS,INP; LISBP ; 135 Avenue de Rangueil, F-31077 Toulouse, France 2 INRA, UMR792 Ingénierie des Systèmes Biologiques et des Procédés ; F-31400 Toulouse, France 3 CNRS, UMR5504 ; F-31400 Toulouse, France

Cellulose and lingo-cellulose are the most abundant renewable carbon resources for production of energy and bioraffinery. Cellulose from paper industry is also a significant component of domestic and industrial waste. The hydrolysis of such solid substrate by bacteria constitutes the limiting step for its transformation. Here, we proposed original biokinetic experiments and modelling approach to explore the cellulose bio-transformation. The hydrolysis of pure cellulose and waste-cellulose (from primary sludge) was studied in batch bioreactors using diverse bacterial inocula. Our results demonstrated that biofilm colonisation of cellulose was the limiting step of the process. Whatever the origin of the cellulose studied, respirometric analysis and microscopy observations showed that degradation preceded into two steps: an accelerating phase, which could be related to the colonisation of the substrate and, a slowing down phase, explained by a limited hydrolysis reaction due to the substrate surface. For waste cellulose, the maximal colonisation and hydrolysis rate occurred after 2-3 days while pure cellulose was colonised only after 5 days. Similarly, 70 % of the waste cellulose was degraded after 10 days whereas it took about 20 days to degrade the same amount of pure cellulose. These results suggest that the degradation rate was controlled by the initial level of colonisation of the cellulosic substrate by bacterial. Moreover, the addition of suspended micro-organisms has no significant effect on the degradation rate. A specific model was developed and calibrated to the experimental responses. This model accurately predicts the hydrolysis rate and bacterial growth in different conditions.


72

IBBS-15-3-O06

White rot fungi selectively cleave threo-form of -aryl ether linkages in spruce lignin Daniel J. Yelle1, Alexander N. Kapich1,2, Kenneth E. Hammel1

1 USDA Forest Products Laboratory, One Gifford Pinchot Drive, Madison, Wisconsin, 53726, USA

2 International Sakharov Environmental University, 23 Dolgobrodskaya Str, 220009 Minsk, Belarus

White rot fungi aggressively degrade all components of the wood cell wall, utilizing both enzymatic and non-enzymatic mechanistic pathways. Certain white-rotters are able to selectively cleave specific stereoisomers of the major lignin linkage in wood, the �-aryl ether, revealing the nature of specific oxidative enzymes produced by these fungi. Ceroporiopsis subvermispora and Phanerochaete chrysosporium degraded spruce wood (approx. 25% wt. loss) were extracted sequentially with n-hexane, chloroform, acetone, methanol, and 1,4-dioxane and precipitated (pH 3) to isolate a high molecular weight (HMW) fraction of the degraded lignin polymer. The HMW fraction was analyzed using one- and two-dimensional NMR spectroscopy experiments, including quantitative 13C, heteronuclear single quantum coherence (HSQC) and heteronuclear multiple bond correlation (HMBC). Integration of the specific C� peaks (i.e., erythro and threo stereoisomers) in the NMR spectra of the HMW 1,4-dioxane lignin extract revealed that the threo-form of �-aryl ethers is preferentially cleaved by both C. subvermispora and P. chrysosporium. These results suggest that the oxidative mechanisms involved in lignin degradation by these fungi involve preferential decay of lignin linkages via selective enzymatic pathways.


73

IBBS-15-3-O07

Endogenous lignocellulosic digestive mechanism of the marine wood-boring isopod Limnoria quadripunctata

GP Malyon, SD Streeter, SM Cragg

University of Portsmouth, Institute of Marine Sciences, Ferry Road, Portsmouth, PO4 9LY, UK

The marine wood-boring isopod Limnoria quadripunctata has previously been reported to possess a digestive tract free of resident flora, and our current work provides further evidence of this. Furthermore, extracts from both the secretory gland – the hepatopancreas – and the gut have been shown to inhibit microbial growth. Extracts of both the hepatopancreas and gut were proven to possess cellulolytic properties by means of a carboxymethyl cellulose/agar plate assay technique. In situ hybridisation assays have identified the hepatopancreas as the site of synthesis of these cellulolytic enzymes and Western blot assays have demonstrated their presence in both the hepatopancreas and the gut, strongly suggesting that L. quadripunctata is capable of cellulolytic activity independently of any resident gut symbionts. Indeed, analysis of expressed sequence tags derived from the hepatopancreas indicated that a significant proportion of the transcripts identified – greater than 20% – correspond to glycosyl hydrolases. Proteomic analysis has identified the presence of endogenously-produced haemocyanin in the gut. It is possible that the protein may be acting as a phenoloxidase with lignin-modifying properties, as has been suggested in other detrital feeders. Further studies are currently underway to test this hypothesis.


74

IBBS-15-3-P01

Harvesting electrical energy from cellulose: microbial fuel cells hosting goat rumen bacteria in combination with Geobacter

Eduardo Montero1, Antonio Berna2, David Yañez-Ruiz3, Abraham Esteve-Núñez1,4

1 Department of Analytical Chemistry and Chemical Engineering, University de Alcalá,

Alcalá, Madrid, Spain

2Department of Geology, University of Alcalá, Alcalá de Henares, Madrid, Spain

3Department of Animal Nutrition, Estación Experimental del Zaidín, CSIC, Granada, Spain

4IMDEA water, Parque Tecnológico de Alcalá,Alcalá de Henares, Madrid, Spain

The concept of renewable sources of energy can be altered by a recent and exciting finding: Fe(III)-reducing bacteria of the genus Geobacter (1) can directly transfer electrons to solid conductive surfaces as graphite so clean electricity can be harvested through devices as microbial fuel cells (MFC) . These types of fuel cells can be fuelled by a number of organic compounds that microorganism can oxidize (acetate, glucose, and more complex mixtures of organic matter). The aim of this work was to couple the oxidation of vegetal residues to electricity production by combining cellulose oxidizer bacteria and the electrogenic bacteria Geobacter.

For oxidizing cellulose we have used goat rumen bacteria with well known ability for fermenting cellulose (2) while producing acetate as dead-end metabolite. In the natural habitat of this bacteria, acetate is converted into methane by methanogens. In contrast, in our MFC system we have avoided methane formation by using Geobacter to succesfully oxidize acetate to CO2 using an anode as electron acceptor so current can be harvested.


75

IBBS-15-3-P03

Biologic pre-treatment of lignocellulosic wastes to improve biogas production Hrdinová Jitka, Jagošová Vanda, Kozumplíková Milena, Minařík Miroslav, Píštěk Vlastimil EPS, s.r.o., V Pastouškách 205, 686 04 Kunovice, Czech Republic [email protected] The extension of the raw material basis for biogas station was the main object of this research. The biological pre-treatment of lignocellulosic wastes (straw, waste paper, sawdust, and poppy-heads residues) improvement in biogas production was investigated. In the first stage, the lignocelluloses were hydrolyzed aerobically for 2-12 weeks, using three cellulolytic microorganisms: Trichoderma reesei, Trichosporon cutaneum and strain with operational name - Tur3. This stage was followed by anaerobic digestion at 55 °C for 40 days. The biogas volume and concentration of released methane were determined during the process. The biological pre-treatment of poppy-heads residues with Trichoderma reesei increased the production of methane by 6.5 % after four weeks; pre-treatment of straw by 26.4 % after 12 week, pre-treatment of paper by 9.7 % after 12 weeks and sawdust by 14.7-40.8 % after 2-4 weeks. Four week pre-treatment of poppy-heads residues with Trichosporon cutaneum improved methane production by 25.0 %, pre-treatment of straw by 10.4-28.9 % after 2-4 weeks, pre-treatment of paper by 3.3-16.5 % after 8-12 weeks, pre-treatment of sawdust by 21.0 % after 12 weeks. Strain Tur3 did not prove significant increase in methane production (straw 2.5 % 12 weeks and sawdust 9.3 % 2 weeks). In other cases, the biological pre-treatment had negative influence on methane production.


76

IBBS-15-3-P07

Assay of Antifungal Effects of Walnut (Juglance nigra) and Pomegranate (Punica granatum) Extracts on Book Deteriorating Fungi

Parisa Mohammadi 1, Bentolhoda Sargazi 1, Shahla. Rodbar Mohammadi 2

1) Department of Biology, Faculty of Science, ALzahra University, Tehran, IR of Iran 2) Tarbiat Modares University of Medical Science, Tehran, IR of Iran E-mail address: [email protected]

Paper and parchment in ancient books and documents are deteriorated by different microorganisms and represent a cause of great concern for libraries and archives in all over world. Many plant extracts such as extract of gallnut, mimosa bark, clove, thyme and marjoram have been proven to have antifungal properties. Some of these natural products such as walnut and pomegranate were formerly used to dye papers and textiles in Iran. The material which was dyed by these natural compounds seems to be more resistance to biodeteriorating damages as well this compound is more compatible to environment. For this reason in this study effect of green walnut and pomegranate peel extracts were investigated against fungi isolated from ancient paper materials. The methanol and ethyl acetate extracts of walnut and pomegranates peel with broth dilution method were tested on Alternaria sp, Ulocladium sp, Chaetomium sp and Penicillium sp according to NCCLS-M27A protocol. Then the minimum inhibitory concentrations of them were defined. The results show ethanolic extract of green walnut is most effective than pomegranate extract and was able to inhibit the growth of 50% of tested fungi. Ethanolic extract of pomegranate peel did not have any effect to remove these fungi. The efficient concentration of ethyl acetate and ethanol extract of walnut on Alternaria sp was 5 and 1 µg/ml, respectively. 10 µg/ml of this extract is necessary to inhibit the growth of Chaetomium sp. Further investigations are necessary to purify effective compounds of walnut extract, assay fungicidal impact of effective fractions and study antifungal mechanism of these compounds. This approach can be useful for paper materials from conservation and renovation aspect.

Keyword: paper, deteriorating fungi, natural products


77

IBBS-15-3-P08

The Effects of Vermicomposting on Organic Wastes

Marieh Nadi1, Vahid Mozafari2, Ahmad Golchin3 and Ebrahim Sedaghati4

1Department of Soil Science, Szent Istvan University, Godollo, Hungary.

2Department of Soil Science, Vali-e-Asr University of Rafsanjan, Rafsanjan, Iran.

3 Department of Soil Science, Zanjan University, Zanjan, Iran.

3Department of plant protection, Vali-e-Asr University of Rafsanjan, Rafsanjan, Iran.

Abstract

Composting and vermicomposting are efficient methods for converting organic wastes (OW) to products. Incorporation of composts and vermicomposts into potting and container media is a potential use for these materials and a suitable alternative for peat and other organic substrates. Samples of different solid organic wastes (SOW) including cow manure (CM), pistachio wastes (PW), cotton residues (CR) and date wastes (DW) were collected from different regions of Iran and converted to various vermicomposts. The physico-chemical characteristics of organic wastes and prepared vermicomposts were significantly different. The bulk densities of organic wastes ranged from 0.11 gm-3 in CR to 0.65 gm-3 in DW, their water holding capacities differed significantly. The highest organic carbon, total nitrogen and potassium content were measured in PW. The highest and the lowest C/N ratios of 97.3 and 42.7 were measured for DW and AM respectively. DW and PW were acidic in nature. AM had a higher concentration of micronutrients in comparison to other organic wastes. After vermicomposting, there was a substantial decrease in organic carbon content, total K and electrical conductivity. The contents of ash, N, P, Ca, Mg and micronutrients increased in vermicomposts. The pH of organic wastes shifted toward neutrality and their bulk densities balanced between 0.41 to 0.46 gm-3. Water retention ability of organic wastes decreased as a result of worms activities and this reduction was in the range of 17 to 33% at field capacity. Keywords: Vermicompost, Pistachio wastes, Cotton residues, Animal manure, Date wastes.


78

IBBS-15-3-P10

Molecular monitoring of the effects of different conservation treatments on paper infecting fungi

Astrid Michaelsen1, Flavia Pinzari2,, Nicoletta Barbabietola2, Guadalupe Piñar3

1 Department of Microbial Ecology, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria

2 ICRCPAL-Istituto Centrale per il Restauro e la Conservazione del Patrimonio Archivistico e Librario, Laboratorio di Biologia, Ministero per i Beni e le Attivita Culturali, Via Milano 76, 00184 Rome, Italy

3 Institute of Applied Microbiology, Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 11, 1190 Vienna, Austria

Fungi are the main organisms that induce the biodeterioration of paper-based items of cultural heritage. Appropriate measures to deal with fungal infections include mechanical, chemical and biological treatments, which also have to consider possible undesirable effects on the paper material itself and health hazards for humans.

Three different conservation treatments, namely freeze-drying, gamma rays and ethylene oxide (EtO) fumigation, were compared in their short- and long-term effectiveness to inhibit fungal growth on different paper samples. After inoculation with the common paper spoilers Chaetomium globosum, Trichoderma viride and Cladosporium cladosporioides, as single strains or in a mixed culture, paper samples were treated and screened with DGGE for the recovery of fungal DNA. Freeze-drying displayed no short- or long-term effect on DNA recovery when compared to non treated samples, whereas the application of gamma rays showed slight reduction of DNA recovery as well as DNA fragmentation shortly after treatment. However, no difference could be singled out as long-term effect.

Additionally, RNA was used as an indicator for long-term fungal viability. Minor differences in RNA recovery from freeze-drying or gamma rays treated samples were recorded in samples inoculated with mixed cultures, but not in those inoculated with single strains, contrary to most studies proving gamma rays effectiveness in fungal disinfection. Only treatment with EtO fumigation proved to be negative for both DNA and RNA recovery suggesting possible bias in subsequent DNA-based methods that are used to analyse former biological infestations on EtO treated samples.


79

IBBS-15-3-P11

Isolation and characterization of cellulose degrading bacteria from local environment

Abdul Rehman and Shumaila Shakoor

Department of Microbiology and Molecular Genetics, University of the Punjab, Lahore, Pakistan

Cellulose is a polymer of glucose the most abundant organic material in nature resistant to decomposition. A large number of bacteria, fungi and actinomycetes are known to degrade cellulose. Cellulases are inducible enzymes which are synthesized by microorganisms during their growth on cellulosic materials. Cellulases have attracted much interest because of the diversity of their application. The major industrial applications of cellulases are in textile industry for ‘bio-polishing’ of fabrics and producing stonewashed look of denims, as well as in household laundry detergents for improving fabric softness and brightness. A potential challenging area where cellulases would have a central role is the bioconversion of renewable cellulosic biomass to commodity chemicals.

In the present study thirty bacterial isolates were grown on CMC salt medium and only two were selected as cellulose degrading bacteria (S2-2 and S3-2) by Congo red dye test and enzyme assay. These two selected bacteria showed clear hallow zones in Congo red dye plates. In enzyme assay test both isolates have exhibited high extracellular cellulase activity i.e. 80%. The optimum temperature for bacterial growth was found 370C and optimum pH was 7. Bacterial isolates were characterized biochemical and as well as on the basis of 16 S rRNA. The optimum temperature and pH of the enzyme was also ascertained. SDS PAGE was also performed to determine the molecular weight of the protein responsible top degrade cellulose.


80

IBBS-15-3-P12

Comparison of Decay Resistance of Wood and Wood-Polymer Composite Prepared by In-situ Polymerization of Monomers

Li Yongfeng1, Liu Yixing1*, Dong Xiaoying1,2, Wang Fenghu1

1. Key Laboratory of Bio-based Material Science and Technology of Ministry of Education, Northeast Forestry University, Harbin 150040, P. R. China;

2. College of Science, Northeast Forestry University, Harbin 150040, P. R. China

*Corresponding Authors:

Liu Yixing, [email protected], Tel: +86 451 82190134; Fax: +86 451 82190134

Fast-growing Poplar Wood (PW), Populus ussuriensis Kom, was modified by formation of wood-polymer composite (WPC) to improve its decay resistance. Two functional monomers, glycidyl methacrylate (GMA) and poly(ethylene glycol) dimethacrylate (PEG(200)DMA), added with a few initiator, Azo-bis-isobutryonitrile(AIBN), and catalyst, maleic anhydride, (MAn), were first impregnated into wood cell lumen under a vacuum-pressure condition, and then in-situ polymerized into copolymers through a catalyst-thermal treatment. Both the decay resistances of untreated wood and wood-polymer composite were tested, and the difference was also analyzed by SEM and FTIR. The test results indicated that the in-situ polymerized copolymers fully filled up wood cell lumen and also grafted onto wood cell walls with a certain degree, resulting in the blockage of passages for microorganisms and moisture to wood cell walls, as well as the modification of wood chemical components unidentified as food for figus. Thus, the decay resistance of wood-polymer composite was reached 2.81% in terms of weight loss, improved 96.2% over untreated wood.

Keywords: Wood-Polymer Composite, Monomer, In-situ Polymerization, Decay Resistance


81

IBBS-15-3-P13

Molecular and Visual Monitoring of Sapstaining Fungi and Moulds on Sawn Timber

Andrea Steitz1, Birgit Schmöllerl1, Notburga Pfabigan1, Roland Gründlinger1

1Holzforschung Austria, Franz Grill-Straße 7, A-1030 Vienna

In a current research project, the site specific impact of sapstaining fungi and moulds on sawn timber was investigated. Stacks of untreated sawn timber of Scots pine and spruce as well as stacks of sawn timber (Scots pine) treated with three different preservatives were sampled for the molecular investigation and sapstain was stated by visual assessment at the saw mills. The results of the fingerprint technique Denaturing Gradient Gel Electrophoresis (DGGE) of the untreated sawn timber showed a distinct difference between Scots pine and spruce, indicating different fungal colonisation due to the wood species. Fingerprints of the treated timber showed the same two main bands for the preservatives X and Y, whereas one characteristic band lacked in the case of the third preservative Z, indicating its protection against this fungal genus/species. Based on the visual assessment, the evaluated average blue-stain showed more or less the same performance of preservatives X and Y. In contrast, preservative Z revealed a better performance at two saw mills.


82

Topic 04–Biodeterioration of Cultural Heritage and building materials in indoor and outdoor environments: assessment and control, methods, treatments, prevention, cleaning


83

KEYNOTE LECTURE

Biodeterioration of cultural heritage buildings in tropical and sub-tropical climates

Christine Gaylarde

Univ. of Portsmouth, UK, and Univ. of Sao Paulo, Brazil.

The biodeterioration of buildings, both ancient and modern, is accelerated under the conditions of high temperature and humidity found in tropical and sub-tropical climates. The mature biofilms (found on long-undisturbed surfaces) differ from those seen under less aggressive conditions. In Europe, for instance, algae are much more prominent than cyanobacteria, the reverse of situations found in Latin America. This is because of the higher resistance to dehydration and ultraviolet irradiation of cyanobacteria. The microorganisms detected on a wide variety of stone-built cultural heritage buildings in Latin America and Asia are discussed, together with their potentially damaging effects. The influence of environmental parameters, such as incident light and vegetation is noted. Buildings of cultural heritage are a particular maintenance problem, since it is normally required that they retain as much as possible their original appearance. Restorations such as repainting should use materials similar to the original, but interventions that involve rebuilding or re-surfacing are nowadays recommended to be readily distinguishable from the original, as well as being reversible. An example of such an intervention is given from Tiotihuacan, Mexico.


84

KEYNOTE LECTURE

Molecular investigations of microbial communities in mummified human remains

Frank Maixner1, Thomas Rattei2, Guadalupe Piñar3, Katja Sterflinger-Gleixner3, Dario Piombino-Mascali1, Giovanna Cipollini1, Albert Zink1

1 EURAC- Institute for Mummies and the Iceman, Viale Druso 1, 39100 Bolzano, Italy

2 Department of Computational Systems Biology, University of Vienna, Austria

3 Institute of Applied Microbiology, Dep. Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria

Mummified human remains can be regarded as an independent “ecosystem“ in which, on the one hand, highly specialized microorganism thrive, some of which are involved in degradation processes. On the other hand, traces of ancient DNA in these specimens could provide detailed insight into the past microbiome of this mummy consisting of beneficial and potentially pathogenic bacteria. Until recently, the vast majority of complex microbial communities were largely underestimated due to major technical, time and financial constraints. Therefore, our knowledge of mummified bodies as “ecosystems“ is limited. However, advancements in high-throughput next-generation sequencing technology have yielded powerful new tools in terms of cost effectiveness, time saving and data recovery. “In-depth“ sequencing of phylogenetic marker genes in combination with classical cultivation approaches will shed light into the spectacular microbial diversity in various habitats in and on mummies.


85

IBBS-15-4-O01

BIOCLEANINING OF WALL PAINTINGS WITH BACTERIA

1Pilar Bosch Roig, 2Rosa Montes Estellés, 1Jose Luis Regidor Ros, 1Pilar Soriano Sancho, 1Maria Teresa Doménech Carbó 1Institute of Restoration of Cultural Heritage of the Polytechnic University of Valencia. 2Microbiology area in the Biotechnology department of the Polytechnic University of Valencia. Microorganisms are important biodeterioration agents of art works, however, nowadays new methodologies are developing the use of microorganisms for cleaning residual organic compounds and salt crusts difficult to remove by traditional restoration methods. This paper develops the use of Pseudomonas stutzeri for the cleaning of mural paintings, with the goal to remove traces of organic matter from ancient restorations or insoluble salt efflorescence. Different strains of Pseudomonas stutzeri from international collections of microorganisms are tested and different application supports are experienced to facilitate direct application to real work. A new application support is introduced to this field, the Agar, a polysaccharide, obtained from marine red algae, getting very advantageous results, showing a more homogeneous and superficial cleaning of vertical wall paintings compared with conventional support mediums, like the cotton. The better experimental results obtained in previous laboratory tests are successfully applied on wall painting areas of a Spanish Church named “Iglesia de los Santos Juanes” in the city of Valencia.


86

IBBS-15-4-O02

Analysis of microbial communities on a painting (oil on canvas): cultivation, molecular identification and evaluation of their biodegradative potential.

M. López-Miras1, I. Martín-Sánchez2, J. Romero-Noguera1, F.C. Bolívar-Galiano1, J. Ettenauer3, K. Sterflinger3, G. Piñar3.

1 Dep. of Painting. Faculty of Fine Arts. University of Granada. Avda. Andalucía s/n, 18071, Granada, Spain. e-mail: [email protected]

2 Dep. of Microbiology. Faculty of Sciences. University of Granada. Avda. Fuentenueva, 18071, Granada, Spain.

3 Institute of Applied Microbiology, Dep. of Biotechnology, Vienna Institute of Bio Technology (VIBT). University of Natural Resources and Life Sciences, Muthgasse 11, A-1190 Vienna, Austria.

Paintings contain a wide range of organic and inorganic constituents which are a good source of nutrition and energy for microorganisms. Furthermore, dirt and dust deposited on the surface of the painting may also support the development of microorganisms on paintings, causing different kinds of alterations, as aesthetic and structural damage. Although the biodiversity of the microbial communities found on paintings has showed to be high, only some specialized members of the community may grow and damage these works, usually those microorganisms which have high enzymatic activity and low nutritional requirements.

In this study we evaluate the cultivable and non-cultivable members of the microbial community colonising the painting “Virgen de Guadalupe”, San Antón´s convent, Granada (Spain), showing visible sings of biodeterioration. A combination of culture-dependent and -independent techniques was selected, as DNA extraction, PCR-DGGE, RAPD and sequencing analyses. In addition, an enzymatic characterization on the cultivable bacteria and fungi was performed.

Comparative sequence analyses showed that the majority of isolated bacterial strains belong to related species of the phylum Firmicutes (72,7%) such as Bacillus sp. (54,5%) and Paucisalibacillus sp. (9,1%) and most of the isolated fungal strains belong to related species of the order Eurotiales (75%) such as Penicillium (50%) and Eurotium (12,5%). The majority of the non-cultivable members of the bacterial community belong to related species of the phylum Proteobacteria (60%) such as Stenotrophomonas sp. (10%); the non-cultivable members of the fungal community belong to related species of the order Pleosporales (50%) and Saccharomycetales (50%).

Concerning enzymatic activity test, the most widespread activities among bacterial isolates were esterase C4 and naphtol phosphohydrolase; among fungi were lipases (phosphatase alcaline and esterase C4) and phosphatases (phosphatase acide and naphtol phosphohydrolase).


87

IBBS-15-4-O03

Investigations of fungi on wall paintings from a 16th century Lutheran cellar from Slovenia

P. Zalar1, M. Lesar Kikelj2, S. Kramar2, A. Mladenovič2, A. Padovnik2 and N. Gunde-Cimerman1

1 University of Ljubljana, Biotechnical Faculty, Department of Biology, Večna pot 111, 1000 Ljubljana, Slovenia

2 Institute for the Protection of Cultural Heritage of Slovenia, Restoration Centre, Poljanska 40, 1000 Ljubljana, Slovenia

Due the high iconographic value and quality of the wall paintings, Lutheran cellar (Sevnica, Slovenia) is considered as one of the most important Slovenian historical monument. The wall paintings, which reflect rare Protestant spirit of late 16th c. in Slovenia, are made in for the wall paintings unusual tempera technique. This special technique represents quite difficult task for the conservation-restoration intervention. The wall paintings are highly deteriorated due to the various environmental factors. High humidity (capillary, condensation), soluble salt crystallization, insufficient ventilation and temperature variations over the season together with the inappropriate use of the cellar (wine cellar with alcohol vapour) results in high damaging affect on the wall paintings. First documented conservation-restoration works were performed in the years 1911-1912, then in 1965 and 1973. Current conservation-restoration interventions include architectural rehabilitation and establishment of appropriate environmental conditions of the monument, which are necessary for the preservation of the wall paintings. Due to the moldy appearance of the wall paintings we have taken samples for mycological analyses, in which we have detected fungi directly and indirectly. We have identified isolates according to valuable taxonomic standards, including sequencing of different DNA regions: ITS rDNA, parts of exons and introns of beta tubulin and actin gene. Representatives of different wood decaying fungi that might be seasonally airborne have been identified in adition to yet unidentified lichen, and mold species of genera Cladosporium and Penicillium. Because of specific nature of the wall paintings and of ubiquitous presence of fungi in different parts of the cellar the suggested method of sanitation could be fumigation.


88

IBBS-15-4-O04

Investigation of biocenosis in a archaeological rock-cut settlements with relevant historic-artistic evidences. A case study in South-east of Sicily.

1Billeci N., , 1Mancuso F. P., 2Pellegrino L., 1Palla F.

1University of Palermo, Italy, Dipartimento di Biologia Ambientale e Biodiversità, Via Archirafi 38, 90123 Palermo;

2Regione Siciliana, Italy, Centro Regionale per la Progettazione e il Restauro, Via dell’Arsenale 52, Palermo, Italy.

The aim of this study was to explore the biological communities colonizing a rupestrian cave and its artistic surfaces. The site, named Saints cave, belong to a biggest group of rock-cut settlements located in Licodia Eubea (CT). These semi-confined environments represent very peculiar habitat where microclimate strongly influences the development and complexity of biological colonization. In the Saints cave two antrums (chamber A and B) were distinguished and a different biological colonization, responsible for chemical and physical damages, was revealed. A complex biocenosis characterized by herbaceous species, bryophytes, insects and widespread microbial consortiums, was detected inside chamber B. In particular, were recognized macroscopic organisms as Adiantum capillus-veneris, Parietaria judaica and insects belonging to the species Diptera Brachycera. Combining optical and confocal laser scanning microscopy (O.M., C.L.S.M.) with cultural-independent techniques (DNA extraction, PCR in vitro amplification, Sequences Analysis), were revealed the Cyanobium as predominant species of cyanobacteria.

This study is a part of a project which aims to perform the conservative restoration and the sustainable fruition of this important archaeological site, that also provided the planning of environmental monitoring.


89

IBBS-15-4-O05

Problems met when trying to eliminate endolithic microorganisms damaging stone buildings using biocides

A. de los Ríos, B. Camara, Sergio Pérez-Ortega, M. Speranza, J. Wierzchos, C. Ascaso

Museo de Ciencias Naturales, CSIC, Serrano 115 dpdo, Madrid-28006, Spain.

The ineffective use of biocides to treat stone buildings damaged by biological growths is a cause of concern. In this study, several chemical treatments (Biotin T, Biotin R, Acticide LV706, Acticide CF, Acticide IOG, Preventol) followed or preceded by mechanical cleaning were tested and their efficacy examined using the SEM-BSE technique. The success of these treatments was found to depend on the methods and products chosen, but was especially conditioned by the microorganisms colonizing the monument and their localization in the stone. Biocide treatments applied on a small scale to monuments and quarry stone revealed the differential elimination of endolithic and epilithic microbial forms. Thus, treatments most effective at combating epilithic lichens were often insufficient to eliminate endolithic forms. The different accessibility of the biocides to the target microorganisms and their ability to persist in the stone and therefore exert their actions for longer periods emerged as important factors for their biocidal activity. In some treatments, epilithic lichens or their remains after prior mechanical cleaning behaved as a protective layer, preventing the agents penetrating into deeper substrate zones and their actions on endolithic microorganisms. These problems mean it is very difficult to totally eliminate endolithic fungi, and viable cells persisting after treatment are a potential source of recolonization. Our findings highlight the idea that candidate treatments need to be first assessed, preferably in situ, in terms of their short and long term effects on both the endolithic and epilithic microbiota damaging the monument.


90

IBBS-15-4-O06

Investigation of archaeal and bacterial community structure and functional gene diversity in exfoliated sandstone at Porchester Castle (UK).

Elisabetta Zanardini1,2, Kevin J.Purdy 1, Eric May3, Helene Moussard1, Simon Williams1, Brian B.Oakley4, J. Colin Murrell1

1 School of Life Sciences, University of Warwick, Coventry (UK)

2 Department of Chemical and Environmental Science, University of Insubria, Como (Italy)

3 School of Biological Sciences, University of Portsmouth, Portsmouth (UK)

4 RBRRC, Agricultural Research Service, Athens, GA (USA)

Diversity and function of the microbial communities involved in the biodeterioration processes on outdoor stoneworks of Cultural Heritage (CH) has not been extensively studied.

In this study, archaeal and bacterial diversity in exfoliated sandstone at Porchester Castle was examined using cultivation-independent methods including PCR-DGGE, clone library analysis and pyrosequencing.

The most predominant types of Bacteria and Archaea identified included Chloroflexi, Actinobacteria, Deinococcus, α- and β- proteobacteria, Cyanobacteria and Bacteroidetes and halophilic archaea related to the family Halobacteriaceae, respectively.

Functional gene analysis, performed on the same samples, revealed the presence of both archaeal and bacterial ammonia oxidizers-(AOA and AOB respectively; amoA genes) and denitrifying bacteria (nirK gene).

In particular for AOB, the majority of the environmental sequences retrieved from our samples were related to the genus Nitrosospira. This was also confirmed by the analyses of the sequences of green-like ribulose-1,5-bisphosphate carboxylase/oxygenase large-subunit genes (cbbL). These showed a very low level of diversity and were closely related to the cbbL genes of Nitrosospira and Nitrobacter.

We showed that pyrosequencing, already applied for the in-depth studies of the microbial diversity in various terrestrial and aquatic ecosystems, can also represent a promising and cost-effective method for the analysis of the prokaryotic community associated with biodeterioration of CH stoneworks, thus permitting a better evaluation of biodeterioration processes that can compromise our cultural and historical patrimony.

IBBS-15-4-O07


91

Bioconsolidation treatment at the Royal Chapel of Granada (Spain) Fadwa Jroundi1, Carlos Rodriguez Navarro2, Beatriz Martín Peinado3, Javier Martín Peinado3, Maria Teresa González-Muñoz1 1 Dept of Microbiology, 2 Dept of Mineralogy and Petrology, Sciences Faculty, University of Granada, Avda. Funtenueva, sn., 18002 Granada, Spain, 3 TARMA, SCA, Granada, Spain

Bioconservation of ornamental stones by means of bacterial carbonatogenesis has been developed to be an environmentally-friendly alternative approach, with which it is possible to induce a new formation of natural cementing products. A number of researchers are studying this biomineralization processes and their application on stone [1]. In this sense, we have developed a method based on the selective activation of the carbonatogenic microbiota that inhabits the stone by the application of a suitable nutritional solution. Experiments under laboratory conditions showed an effective stone consolidation without blocking of stone pores [2]. These results prompted to employ this method in small-scale-in situ bioconsolidation treatment performed at San Jeronimo Monastery in Granada, Spain. Carbonatogenic microorganisms are stimulated to grow and induced the in situ formation of new calcium carbonate cement [3]. All these positive results encouraged us to perform large-scale-in situ bioconsolidation applications at the Royal Chapel in Granada, Spain. We evaluate the effects of such a treatment on an extremely damaged stones which were at the same time subjected to extensive conservation Works using conventional methods. The treatment was applied by spraying a M-3P nutritional solution on the surface of the carved crestings [3]. The results showed the formation of new carbonate cement due to the fact that all of the activated culturable microbiota was carbonatogenic. The development of no dangerous microbial communities was detected after treatment. Furthermore, a clear reduction of the weight loss in comparison with the control (untreated) stone was observed and color measurements before and after treatment of the calcarenite stone blocks show no significant differences. [1] De Muynck et al (2010) Ecol. Eng. 36: 118–136 [2] Rodriguez-Navarro et al (2003) Appl Environ Microbiol 69:2182-2193 [3] Jroundi et al (2010) Microb Ecol 60:39–54


92

IBBS-15-4-O08

Molecular monitoring of the microbial dynamics occurring on historical limestone buildings during and after the in situ application of different bio-consolidation treatments. Jörg Ettenauer1, Guadalupe Piñar1, Katja Sterflinger1, Maria Teresa Gonzalez-Muñoz2 and Fadwa Jroundi2 1 Institute of Applied Microbiology, Department of Biotechnology, Vienna Institute of Bio Technology (VIBT). University of Natural Resources and Life Sciences, Muthgasse 11, A-1190 Vienna, Austria. [email protected]; [email protected]; [email protected] 2 Department of Microbiology, University of Granada, Fuentenueva s/n, 18071 Granada, Spain. [email protected], [email protected]

Microbial induced carbonate precipitation has been proposed as an environmental friendly method to protect decayed ornamental stone and introduced in the field of preservation of Cultural Heritage. Recent conservation studies performed under laboratory conditions on non-sterile calcarenite stones have successfully reported on the application of a suitable nutritional solution, inoculated and non-inoculated with Myxococcus xanthus, as bioconsolidation treatment. Furthermore, this procedure has been very recently applied in situ to selected historical buildings in Granada (Spain). For the first time, we evaluate the efficiency and risks of the in situ application of the above mentioned treatments onto two historical buildings in Granada. The evaluation consists of a detailed investigation of the micro-biota actively growing during the seven days of the treatments -short-term monitoring and of the one remaining in the stones after six and twelve months of the application –long-term monitoring. A molecular strategy, including DNA extraction, PCR amplification of 16S rRNA sequences, construction of clone libraries and fingerprinting by DGGE (Denaturing Gradient Gel Electrophoresis) analysis followed by sequencing was used to gain insight into the microbial diversity present on the differentially treated stones. The monitoring of M. xanthus was done by PCR using species-specific primers. Similar dynamics were triggered on both buildings by the application of the nutritional solution (inoculated or non-inoculated). 16S rDNA sequencing revealed the dominant occurrence of members belonging to the Firmicutes and Proteobacteria during the seven days of the treatment, whereas after one year microorganisms of the order Bacillales of the phylum Firmicutes were predominantly detected. M. xanthus could be detected only during the seven days of the treatment. The treatments seem to activate no dangerous microorganisms during the seven days of the treatment and furthermore, to select the remaining of a homogeneous group of carbonatogenic bacteria on the stones after a long period of time.


93

IBBS-15-4-O09

Fungal cell wall degradation. New insights for removal of rock black fungi.

Laura Selbmann, Daniela Isola, Massimiliano Fenice, Laura Zucconi, Silvano Onofri

DECOS, Università degli Studi della Tuscia, Largo dell’Università snc, Viterbo, Italy

Abstract

Rock black fungi are recognized as the main agent of biopitting, a phenomenon characterized by lesions in size ranging up to 2 cm in diameter and depth on stone. They are present epi- and endolitically both on monuments and natural rocks; their thick and melanised cell wall confers rigidity enough to dwell the rock. Black fungi are resistant to chemicals and are not easily removed by biocides or other anti-microbial treatments. It is of high interest to find new methods to remove black spots from monuments and control their growth and deteriorating activity.

Cryomyces spp. are rock black fungi adapted to the harshest Antarctic conditions, and successfully tested for their survival to space. Their cell wall structure is one of the main factors involved in their ability to survive different external stressors. Cells are embedded in a thick and strongly melanised cell wall encrusted with black rigid plaques giving a supplementary protection and making them practically impregnable being refractory even to many commercial enzymes as chytinases and glucanases. The Antarctic soil fungus Lecanicillium muscarium CCFEE 5003 is known for its ability to degrade the cell wall of different food spoiling and opportunistic fungi as well as plant pathogenic oomycetes. Microscopic observations of dual culture of Cryomyces spp. and the mycoparasitic fungus L. muscarium, both in liquid and solid media, revealed that the cell wall of Cryomyces was heavily damaged with production of protoplasts. This finding open new prospects for conservative interventions on stone monuments damaged by rock black fungi.


94

IBBS-15-4-O10

Biomimetic system for removal of fungal melanin staining on cultural heritage objects Črtomir Tavzes1, Jernej Palčič2, Karin Fackler4, Franc Pohleven2, Robert Koestler3 1The Institute for the Protection of Cultural Heritage of Slovenia, Conservation Centre, Research Institute, Poljanska cesta 40, SI-1000 Ljubljana, Slovenia 2University of Ljubljana, Biotechnical Faculty, Department of Wood Science and Technology, Rožna dolina, Cesta VIII/34, SI-1000 Ljubljana, Slovenia 3Smithsonian’s Museum Conservation Institute, 4210 Silver Hill Road, Suitland, MD 20746-2863, U.S.A. 4Vienna University of Technology, Institute of Chemical Engineering, Getreidemarkt 9/166 A-1060 Vienna, Austria Fungal-caused melanin-staining is a problem on many cultural objects from the French Palaeolithic cave at Lascaux to books and papers in museum collections. Melanin, because it is insoluble and resistant to bleaching, may leave behind undesirable stains on the cultural heritage object, monuments, or buildings long after the fungal infestation has been controlled. Research into removal of melanin stains from paper and other sensitive substrates using industrial biomimetic oxidizing systems has shown considerable success and is reported on in this presentation. The reaction kinetics were studied, as was the relative concentration of the reagents, to optimize the systems. Bleaching of melanin with the biomimetic copper-pyridine complex proved to be far superior to the effect of white-rot fungi oxidising enzymes, previously reported by this group.


95

IBBS-15-4-O11

Evaluation of treatments efficiency against crustose lichens growing on monumental stone by electrical conductivity O.A. Cuzman1, D. Pinna2, C. Riminesi1, B. Sacchi1 and P.Tiano1

1ICVBC-CNR, Via Madonna del Piano, Sesto Fiorentino, 50019, Italy 2OPD, Florence, Italy Crustose lichens frequently cover large areas of monumental stones and archeological areas. The topic related to their elimination is under debate, and usually their removal is carried out applying biocide formulates or physical stresses. The evaluation of the effective biocidal action of such treatments some methods can be applied such as epifluorescence and respirometry. Because a damaged organism changes its cell functionality, we propose a simple method based on the measurements of these changes at a physiological level. Fragments of crustose lichens sampled from monumental stone surfaces, were subjected to different stress degrees either by temperature heating and chemical treatment using the biocide Rocima 103. The vitality and membrane integrity of treated and non treated samples were evaluated by ions leakage measurements, ionic chromatography and optical microscopy. Preliminary results shown that electrical conductivity is a sensitive method for measuring the induced stress and for the assessment of the efficiency of control treatments applied.


96

IBBS-15-4-O12

Lichen secondary metabolites: potential biocides against rock-dwelling microcolonial fungi and cyanobacteria?

Gazzano C., Favero-Longo S.E., Piervittori R.

University of Torino, Dipartimento di Biologia Vegetale - Viale Mattioli 25, 10125, Torino, Italy

Antibiotic, antiviral and anti-proliferative functions of lichen secondary metabolites (LSM) have been recognized, suggesting their potential use for therapeutic applications. As LSM play a role in lichen-plant allelopathic interactions, their use as herbicide has been also recently suggested. The antimicrobial activity of LSM has been assessed against a wide set of bacteria and fungi, but researches poorly considered their effects against rock-dwelling organisms, thus preventing their potential use for the control of the biological colonization of Cultural Heritage. This work analyzes the effects of three LSM (parietin, norstictic and usnic acid), solubilised in acetone, against black yeasts (Sarcinomyces petricola), meristematic microcolonial fungi (Coniosporium apollinis, C. perforans, C. uncinatum, Phaeococcomyces-like sp.), coccoid and filamentous cyanobacteria (Chroococcus, Phormidium, Scytonema), which commonly occur in black crusts on stonework. In vitro, the inhibition of the colonial growth varied, depending on the treated species and the lichen metabolite, but the inhibition was always lower than that obtained with a water solution of benzalkonium-chloride at 1%. Higher inhibition effects were observed 35 days after the treatment, while no differences between treated and untreated colonies were observed in most cases after 3 months. The inhibitory effect of acetone alone cannot be neglected. The effects of LSM applications were finally monitored in the field on black crusts occurring on travertine blocks in the Roman theatre of Aosta (Italy).


97

IBBS-15-4-O13

Biodeterioration of funeral sculptures in La Recoleta cemetery, Buenos Aires, Argentina. pre- and post-intervention studies. Patrícia Guiamet.1,2, Miguel Crespo3, Paola Lavin1,4, Beatriz Ponce5, Christine Gaylarde6, Sandra Gómez de Saravia1,7

1INIFTA, Departamento de Química, Facultad de Ciencias Exactas, UNLP, CCT La Plata - CONICET. CC 16, Suc. 4 (1900), La Plata, Argentina. [email protected]

2Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, CONICET.

3Programa de Conservación y Restauración de Monumentos y Obras de Arte del Cementerio de La Recoleta. Subsecretaría de Patrimonio. Ministerio de Cultura G.C.B.A. Ciudad de Buenos Aires. Argentina. [email protected]

4Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CONICET.

5INTEMIN -SEGEMAR . San Martín. Buenos Aires. Argentina. [email protected]

6University of Portsmouth,Portsmouth, UK. [email protected]

7Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, CICBA.

Stone materials exposed to weather are subject to biological colonization and consequently to biofilm formation, causing biodeterioration. The colour changes on the stone substrates due to biogenic pigments, mechanical stress on the mineral structure due to extracellular polymeric substances (EPS) and the accumulation of atmospheric pollutants by the biofilm itself are some of the deteriogenic effects, which modify the esthetic appearance of the work. The aim of this work was to study the biodeterioration of marble tombstones from La Recoleta Cemetery: José C. Paz, Rufina Cambaceres, Carlos Brandsen and Carlos Pellegrini, as well as the biocide effect of benzalkonium chloride on biofilm formation, producing a chart of intervened tombstone. Sectorized monitoring was performed according to biodeterioration on the surface of tombstones and their orientation in the emplacement area. Pre- and post- intervention microbiological studies, scanning electron microscopy (SEM) and X-ray dispersion analysis (EDX), were carried out. Results showed a significant decrease of the biofilm flora after treatment. However, algae of the genus Trentepohlia were difficult to eradicate. These studies are a valuable contribution to determine restoration criteria against biofilm formation, to characterize chromatic variations of biological origin on the stone and to formulate conservation and restoration policies.


98

IBBS-15-4-O14

The Influence of Aspect on the Biological Colonization of Stone in Northern Ireland

Adamson, C.; Smith, B.J.; Warke, P.A.

Department of Geography, Archaeology and Palaeoecology, Queen’s University Belfast, BT7 1NN

The rate and type of biological colonization of stone is influenced by a wide array of climatic, micro-climatic and environmental controls. A series of experiments were designed to compare the rate and type of biological colonization of stone at varying locations over a 21-month time period. Exposure trials were set up at nine different sites across Northern Ireland, including coastal, inland, urban, rural, low-lying, and higher altitude locations, as well as sites in areas of high rainfall or rain-shadow. Additionally, further experiments were designed to compare colonization on limestone versus sandstone, smooth compared with tooled surfaces, and differences in aspect. To determine aspect-related differences in colonization, blocks were placed on the north and south facing sides of purpose-designed exposure racks. A series of analyses, including colorimetry and visual identification, were carried out on blocks exposed for increasing time intervals.

Results showed considerable differences in greening levels between north-facing and south-facing blocks for both sandstone and limestone, with significantly higher levels of greening on north facing blocks. This difference is likely to be representative of the fact that in Northern Ireland’s wet climate and northern-latitude position, the north face of a building will not receive direct sunlight. Therefore north-facing blocks, once wet, will remain damp for much longer than blocks on other façades. This slow-drying regime is much more hospitable for biological colonization and continued growth than the hostile environment of rapid wetting and drying cycles experienced on the south face.


99

IBBS-15-4-O15

Assessment of mould growth risk at the French national Library by monitoring and building simulations

Caroline LAFFONT 1, Yashiho KIKKAWA2

1. Bibliothèque nationale de France – Laboratoire du département de la conservation –5 rue Vivienne 75002 Paris France – [email protected] ;

2. Japan Society for the Promotion of Science, National Research Institute for Cultural Properties ; 13-40, Ueno Park, Taito-city Tokyo, JAPAN – [email protected]

Abstract

The French national Library in charge of the preservation of its cultural heritage implements a preventive risk management strategy to avoid biodeterioration on collections and building materials. When such a biocontamination occurs, the decontamination is time and staff consuming, costly and sometimes detrimental for collections.

Since this strategy (which was presented during the 14Th IBBS) is an iterative and continuous improvement process, the laboratory sets about a collaborative study in order to assess the mould growth risk by using building simulations. The aim of these simulations is to predict the indoor climate, essential parameter to estimate the mould germination probability.

Building simulations are perform with the Delphin simulations software which take into account the outdoor climatic records and the architectural parameters such as the potential of moisture and heat insulation of the walls materials.

To evaluate if these software give precise data and thus if they can widely use into the whole storage rooms, the outdoor and indoor climate with datalogger and outdoor and indoor aerobiocontamination are also monitored.

We propose to present this research methodology and the first outcomes of this one year research.


100

IBBS-15-4-O16

Integrated Pest Management (IPM) and distribution of pest species in Berlin and Vienna – a comparison

Querner, Pascal

University of Natural Resources and Life Sciences, Department of Integrated Biology and Biodiversity Research, Soil Ecology Group, Institute of Zoology, Gregor-Mendel-Straße 33, A-1180 Vienna – Austria

E-Mail: [email protected]

Integrated Pest Management (IPM) was developed by the food industry and later also applied in museums and archives. The main objective of an IPM program is to prevent damage of art, ethnological or natural history collections made of wood, textiles from animal fibers, natural history collections or other organic materials from insect pests or mould. IPM is part of preventive conservation and focuses on preventing an infestation and reduction of pesticide application with chemical free treatments like nitrogen fumigation, freezing or heating. Regular checking with insect tarps (monitoring) is an important part of IPM to find a pest infestations as early as possible, locate the infested objects, identify the pest species and evaluate the problem. Results from monitoring strategies in different museums in Vienna, Austria and Berlin, Germany are presented. In both cities the concept of IPM is not jet widely applied in all museums or collections. Most museums don’t apply pesticides anymore but preventive measures like regular cleaning of storage sites or sealing of the building are still lacking in both cities. Monitoring with sticky and pheromone traps showed that cloths moth Tineola bisselliella and varied carpet beetles Anthrenus verbasci are both widely occurring in both cities. Other pests like the brown carpet beetle (or vodka beetle) Attagenus smirnovi or the berlin beetle Trogoderma angustum were frequently found in museums in Berlin but not so far in Vienna’s art collections. On the contrary the black carpet beetle Attagenus unicolor, a commune pest in Viennese museums and homes, was not found in Berlin.


101

IBBS-15-4-O17

The Potential of Pulse Amplitude Modulation Fluorometry for Evaluating the Resistance of Building Materials to Algal Growth J. von Werder1 and H. Venzmer

1 Hochschule Wismar, University of Applied Sciences Technology, Business and Design, Wismar, Germany

The growth of algae on thermally insulated façades has developed to a serious, aesthetical problem over the last decade. Responsible for the boosted growth is the increased insulation level of building envelopes which has lead to a significant drop of the surface temperature and in its consequence to long lasting humidity films.

To avert guarantee claims manufacturers of facade and coating materials are looking for strategies to avoid or delay the establishment of biofilms.

An efficient product development is however prevented by the necessity of time consuming free weathering tests and the lack of objective measures to quantify the growth.

In several long term studies the potential of the pulse amplitude modulation fluorometry was tested for the quantification of algal growth on free weathered samples below the visual threshold.

By using an imaging PAM in combination with a scanning device the growth dynamics on the sample surfaces could be repeatedly visualized and analysed in detail.

It could be demonstrated that in case of water saturation algal growth can be quantified with sufficient sensitivity and accuracy by measuring the fluorescence yield. Already few months after the installation of the samples algal colonies could be detected by their fluorescence emission.

Unfortunately the representative growth pattern is not characterized by a linear accumulation of algal colonies. Instead a long phase of fluctuating biomass on a low level is followed by a steep increase. This way the measuring technique allows evaluation of the samples only 6-9 month before a first greenish shimmer becomes visible with the bare eye.


102

IBBS-15-4-P03

Birds Cause Biodeterioration on Historical Monuments

Hakan Sert

Akdeniz University, Faculty of Education,

Department of Elementary Science Education, TR 07058, Kampus-Antalya/Turkey

Corresponding Author: Hakan Sert ([email protected])

The alteration and weathering of stone is basically determined by natural and anthropogenic

impacts influencing various physical, chemical and biological damage factors at the object

site. The potential of animals (particularly birds and insects) to alter and destroy monuments

and works of art was shown in many studies in the last years. This study aims to determine

the deteriorations-effect of bird species on historical monuments in Side (Antalya/Turkey).

Field studies show that these organisms cause irreversible damage to surfaces resulting in the

scarring of monument fabric and damaging appearance of the rock surfaces in Side and thus

they have to be regarded as a serious threat to the antique cultural heritage in Turkey.

Keywords: Birds, Biodeterioration, Historical monuments, Side, Mediterranean Region


103

IBBS-15-4-P04

ROLE OF FUNGI IN BIODETERIORATION PROCESS OF STONE IN HISTORICAL

MONUMENTS IN ASPENDOS (TURKEY )

Hacer (Bakır) Sert1*, Katja Sterflinger2

1 Akdeniz University, Manavgat Vocational High School, TR-07600 Antalya, Turkey

2University of Natural Resources and Applied Life Sciences, Austrian Center of Biological Resources and Applied Mycology (ACBR), Muthgasse 18, A 1190, Vienna, Austria

*Corresponding Author, e-mail: [email protected]

Abstract

In this study the fungal diversity and deterioration on historical monuments were investigated. The Aspendos Ancient City (Antalya/Turkey) in Mediterranean Region was chosen as study area, because of its outstanding historic and artistic value and the large variety of monuments. The sample collection, isolation, morphological and molecular characterizations were done according to Sterflinger & Krumbein 1997 and Sterflinger & Prillinger 2001. From a total of around 108 samples 42 fungal strains were isolated, and could be assigned to 8 different genera(Coniosporium, Capnobotryella, Massarina, Mycocalicium, Phaeococcomyces, Phoma, Rhinocladiella, Sarcinomyces). The observations of the rock surfaces clearly demonstrate that there is a strong positive correlation between the fungi and the alteration of the rock surfaces.

Keywords: Biodeterioration, Black fungi, Aspendos, Turkey, Monument


104

IBBS-15-4-P05

Index of Lichen Potential Biodeteriogenic Activity: towards a statistical validation and the extension to other biodeteriogens

Gazzano C., Favero-Longo S.E., Matteucci E., Piervittori R.

University of Torino, Dipartimento di Biologia Vegetale - Viale Mattioli 25, 10125, Torino, Italy

An index of Lichen Potential Biodeteriogenic Activity (LPBA) has been proposed to quantify the impact of lichens on stonework on the basis of the extent of impact of each species, quantified both on the surface and within the substratum, and on other parameters related to reproduction, physicochemical action, and bioprotection (Gazzano et al. 2009). A scientific network is now under construction to statistically validate the LPBA index through (a) the elaboration of the index interpretative scale, by examining a wide set of study cases in terms of stonework types, lithotypes and climates, and (b) the calibration of each parameter scale by ad-hoc field and laboratory tests. A similar approach may also be followed to extend the index to other biodeteriogens of Cultural Heritage as free-fungi, cyanobacteria and green algae. Tentative applications of the original LPBA index to black crusts and green patinas on historical and culturally significant stone substrata of North-Western Italy preliminary address main difficulties and potential adjustments for the index extension. An index which covers the different groups involved in stonework deterioration may finally represent the practical tool needed by cultural heritage managers to plan and improve conservation by recognizing restoration priorities and identifying harmless colonizations.

Gazzano C., Favero-Longo S.E., Matteucci E., Roccardi A., Piervittori R. (2009) Int. Biodet. Biodegr. 63, 836-843.


105

IBBS-15-4-P06

Airborne fungal spores in moldy and non moldy homes D. Haas1, J. Habib1, H. Galler1, J. Posch1, G. Zarfel1, R. Schlacher2, E. Marth1, F.F. Reinthaler1 1 Institute of Hygiene, Microbiology and Environmental Medicine, Medical University Graz, Austria

2 Department of Environment Information of the Styrian Government, Austria

Quantitative and qualitative assessment of airborne fungal spores was carried out in 66 south-eastern Austrian dwellings in comparison with the ambient air over a one year period. The spore concentrations determined inside homes were related to the size of the area affected by mold. In a further step, the cultivable fungal species were identified in order to determine the most frequent fungi in indoor air.

For measuring the one-stage MAS-100® air sampler with an air stream of 100 litres per minute was used. This impactor was equipped with malt extract agar (MEA) and dichloran glycerol agar (DG18) plates; sampling time was one minute. Following the incubation of the plates for 7 to 10 days at 25°C, the total colony forming units per cubic meter (cfu/m³) air were calculated.

Homes without visible mold growth (n=29) showed concentrations similar to those in the outdoor air. The genus Penicillium was particularly high in spring with 44.4%, autumn with 70.3% and winter with 55.3%. Increased levels of Cladosporium sp. occurred in spring (37.0%) and summer (52.5%); in contradistinction, Aspergillus sp. occurred most frequently in the winter with 34.2% in the indoor air. In the indoor air of 37 homes with visible mold growth the median spore concentrations rose significantly (p<0.001) from 3x10² to 4x10³ spores per m³ with increasing size of moldy patches. The major indoor contaminants in moldy homes were Penicillium sp. and Aspergillus sp. In the outdoor air, Cladosporium sp. was more frequent in summer and fall than in other seasons. Penicillium sp. and Aspergillus sp. in the outdoor air were more strongly represented in spring (23,8% and 5,4%, respectively) and in winter (46,5% and 15,8%, respectively), as other fungal genera.

Increasing spore concentrations reflect an increasing problem of mold growth on surfaces. Season influences the fungal spore concentrations in the outdoor air as well as indoor air.


106

IBBS-15-4-P07

The Capuchin Catacombs of Palermo, Italy: fungal deterioration of mummies and contamination of the indoor air

Katja Sterflinger1, Laura Barnabei1, Guadalupe Piñar1, Frank Maixner2 and Dario Piombino-Mascali2

1 Institute of Applied Microbiology, Dep. Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria

2 EURAC- Institute for Mummies and the Iceman, Viale Druso 1, 39100 Bolzano, Italy

In the crypt of the Capuchin Convent of Palermo nearly two thousand mummies spanning the late 16th /mid-20th century are laying in their coffins or shelves, or standing in line at the walls of the crypt. Many of the mummies show serious signs of fast and recent biodeterioration. For this reason in 2010 a huge sampling campaign was carried out in order to investigate the micro-biological status of the mummies as a basis to prevent further biodeterioration. Samples were taken from different materials of the mummies including skin, bones, straw used as stuffing material, cloth, and leather. In order to investigate the impact of this microbial contamination on the indoor air quality, air samples were taken and analyzed both quantitatively and qualitatively. First results showed that the mummies are heavily contaminated with mold. Thirty-three fungal species were found on and inside the mummy materials, some of which are common airborne fungi and some of which originate from the material used as stuffing for the clothing of the mummies. In some areas of the crypt the amount of fungal spores in the air reached more than 2000 spores / m3. From a medical point of view this amount must already be classified as a potential health risk for visitors. We suggest a broad cleaning campaign carried out by experienced conservators with HEPA-filter equipped vacuum cleaners and a significant improvement of the indoor climate through creation of an airflow as immediate measures to prevent further contamination and deterioration of this precious biocultural heritage.


107

IBBS-15-4-P08

Bacterial Community Analyses of Biofilms on the Deteriorated Sandstone of Bayon Temple of Angkor Monuments in Cambodia

Asako Kusumi1, Xian-Shu Li1, 2, Yu Osuga1, Ji-Dong Gu3, Yoko Katayama1

1 Graduate School of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509, Japan

2 Department of Material Science and Engineering, Nagoya Institute of Technology, Nagoya, Aichi, 466-8555, Japan

3 School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong SAR, P. R. China

Bayon Temple stands in the center of Angkor Thom of Angkor Monuments in Cambodia. Deterioration of wall surfaces resulting from the colonization of microorganisms on the bas-reliefs of inner and outer gallery at Bayon Temple is a serious issue, however knowledge on microbial communities of the biofilm in this area is not available. We studied bacterial community structure by examining biofilms with different pigmentation by PCR amplification of 16S rRNA gene and sequences of the DNA bands after DGGE. Biofilms were collected by using sterilized adhesive sheets to avoid damaging to the bas-reliefs on the gallery wall. The results indicated that red-pigmented biofilms were characterized by the presence of bacteria in the genus Rubrobacter and genus Deinococcus, however homology indices of these microorganisms ranged around 90-95% suggesting unknown bacterial groups colonizing on the surface of the deteriorated sandstone. Bacteria belonged to the genus Rubrobacter are known to contain carotene and have radiation tolerance, which might have advantage to survive on stone surface exposed to sunlight in the tropic region. Green-pigmented biofilms were characterized by the presence of Cyanobacteria such as Anabaena, Spirullina, and Synechococcus in addition to Rubrobacter-related bacteria. Dark pigmented biofilms were found to contain Chloroflexi in addition to Cyanobacteria and Rubrobacter-related bacteria. Comparing three-dimensional structure of these biofilms indicated the distribution of stress tolerant bacteria of such Rubrobater-related microorganisms and phototrophic bacteria on the outermost layer of biofilms. These results suggest the formation of the unique microbial communities on the surface of sandstone in this region.


108

IBBS-15-4-P09

Structural changes of artificially aged textiles after inoculation with the selected “museum” fungi

Katja Kavkler 1,2, Nina Gunde Cimerman3, Polona Zalar3, Andrej Demšar4

1 Institute for the Protection of Cultural Heritage of Slovenia, Restoration Centre, Poljanska 40, 1000 Ljubljana, Slovenia 2 Gorenjska predilnica d.d., Kidričeva 75, 4220 Škofja Loka, Slovenia 3 University of Ljubljana, Biotechnical Faculty, Department of Biology, Večna pot 111, 1000 Ljubljana, Slovenia 4 University of Ljubljana, Faculty of Natural Sciences and Engineering, Department of Textiles, Sne_zni_ska 5, 1000 Ljubljana, Slovenia

Biodegradation of textile fibres of natural origin was investigated. First the "museum mycoflora" was determined by analyes of several visibly contaminated textile objects from different museums. Materials of textile substrates were determined and fungal strains isolated and identified from each object. Fungal strains belonging to six species, either the most often occurring or in a way significant, were selected and their enzymatic profiles studied. The selected species were: Aspergillus clavatus, Cladosporium cladosporioides, Fomes fomentarius, Hypoxylon fragiforme, Penicilluim chrysogenum and P. corylophilum. In the second part of the study three different materials, often found in museum environment (cotton, linen, wool) were choosen, and inoculated with the selected six fungal strains. Half of the specimens were artificially aged under extreme humid and warm conditions before the inoculation, and later compared to the other half of the samples, which represented the control group. Structural changes in the materials were investigated using FTIR spectroscopy and scanning electron microscopy. The results showed relatively significant changes already after 8 weeks of exposure, which were amplified with the prolonged exposure to biodegradation. However, due to different impacts of the fungi as well as different resistance properties of the textile materials, changes varied between samples.


109

IBBS-15-4-P10

Evaluation of the microbial contamination risk of different historical documents conserved in the Slovak National Library

Lucia Kraková1, Katarína Chovanová1, Samy A. Selim2, Alexandra Šimonovičová3, Alena Maková4, Domenico Pangallo1; 5

1 – Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, Slovakia

2 - Microbiology Section, Botany Department, Faculty of Sciences, Suez Canal University, Ismailia, Egypt

3 - Department of Soil Science, Faculty of Natural Sciences, Commenius University, Bratislava, Slovakia

4 – Slovak National Archives, Department of Archival Preservation, Bratislava, Slovakia

5 – Caravella, s.r.o., Bratislava, Slovakia

The microbial contamination of seventy historical documents (11 parchments and 6 paper letters) from the Slovak National Library was evaluated through the combination of cultural and molecular methods. The samples were recovered by swabs and after treated in two different ways: i) direct inoculation on casein and cellulose agar plates; ii) were brought to the laboratory suspended in physiological solution, decimal diluted and plated in specific plates for the growth bacteria and fungi. The microorganisms grew on different plates, prior identification, were selected by two different PCR-based methods the f-ITS and the f-CBH, for bacteria and fungi respectively. The f-ITS is based on the amplification of the internal transcribed sequence between the bacterial 16S and 23S rDNA. The f-CBH was applied in this study for the first time and is oriented to the fungal cellobiohydrolase gene. Both PCR selection methods produced typical profiles which consented to clusterize the isolates in order to reduce them for the consequent sequencing identification. The ITS fragment was utilized for the identification of fungi, while the 16S rDNA for bacteria. The cellulolytic and proteolytic abilities were screened through the use of two plate assays, the Ostazin Brilliant Red H-3B (OBR-HEC) and the gelatin agar respectively. Bacteria of the genus Massilia sp., Bacillus sp.; Microbacterium sp. Thermomonas dokdonensis and Curtobacterium sp. with the members of the fungal species Aspegillus fumigatus (all recovered from paper) displayed both biodegradative activities. The parchment isolates with an intensive proteolytic activity were members of the genus: Virgibacillus, Staphylococcus, Microbacterium, Penicillium and Phoma.

ACKNOWLEDGEMENTS

This work was mainly financed by the VEGA Agency: 2/0179/11 ‘‘Biodeterioration of natural and synthetic polymers in historical and contemporary art collections’’. We are grateful to company Caravella s.r.o. for its support that was necessary for finalization of our work.


110

IBBS-15-4-P11

Isolation and distribution of facultative sulfur-oxidizing bacterium Mycobacterium sp. strain THI503 on deteriorated sandstone of Bayon temple in Angkor monuments, Cambodia

Asako Kusumi1, Xian-Shu Li1, 2, Yoko Katayama1

1 Graduate School of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, 183-8509, Japan

2 Department of Material Science and Engineering, Nagoya Institute of Technology, Nagoya, Aichi, 466-8555, Japan

The ancient structures of Angkor monuments, Cambodia, represent one of the most famous cultural heritage sites in the world and are suffering serious biodeterioration. Sulfur-oxidizing bacteria have been reported as a dominant group involved in the biodeterioration due to the excretion of sulfuric acid. Thus, we enumerated sulfur-oxidizing microorganisms in samples collected from the deteriorated sandstone in Angkor monuments from 1998 to 2007. We report facultative chemolithoautotrophic characteristics of Mycobacterium spp. by using elemental sulfur as the energy source.

From positive culture of MPN for enumeration of sulfur-oxidizing microorganism in deteriorated sandstones of Angkor monuments, we obtained five strains of facultative chemolithoautotrophic sulfur-oxidizing bacteria. Phylogenetic analyses and some taxonomic characteristics indicated that they mostly related to rapid growing Mycobacterium spp. One of the isolates, strain THI503 decreased medium pH from pH4.7 to 3.6 and increased the concentration of thiosulfate and sulfate ions in a basal salt medium supplemented with elemental sulfur during 22 day’s incubation. Fluorescent microscopy of the DAPI stained cells showed that most bacterial cells are adhered on the surface of the granules of elemental sulfur. Growth by using elemental sulfur in the mineral salt medium was also confirmed by measuring amounts of ATP in the culture. The growth of Mycobacterium sp. on elemental sulfur is the first in the bacteria of the genus Mycobacterium. We will show the distribution of the facultative sulfur-oxidizing bacteria on deteriorated sandstone by quantitative-PCR and discuss contribution of these bacteria to stone deterioration in Angkor monuments.


111

IBBS-15-4-P12

DECAGRAPH : Early detection of biological and chemical contaminants of paper based cultural heritage.

Thi-Phuong Nguyen1, Tony Basset2, Stéphane Moularat3, Faisl Bousta4, Geneviève Orial4, Romain Berardo3, Anne Lama5, Caroline Laffont6, Patricia Ramond5, Enric Robine3

1 Bibliothèque nationale de France, Parc Gustave Eiffel, 14 avenue Gutenberg, 77600 Bussy Saint Georges, France, 2 Conservatoire National des Arts et Métiers, Réserves du musée, 11 rue du Landy, 93 000 Saint-Denis-la-Plaine, France, 3 Centre Scientifique et Technique du Bâtiment (CSTB), 84 avenue Jean-Jaurès, Champs sur Marne, 77447 Marne-la-Vallée Cedex 2, France, 4Laboratoire de Recherche des Monuments Historiques (LRMH), 29 rue de Paris, 77420 Champs-sur-Marne, France, 5Archives Nationales, 60 rue des Francs Bourgeois, 75141 Paris Cedex 03, France, 6 Bibliothèque nationale de France, 5 rue Vivienne, 75002 Paris, France

Contamination by fungi in storage facilities is a major and recurrent problem which not only concerns archives and libraries but every other institution in charge of the preservation of paper containing cultural heritage. Unfortunately such contaminations can still only be detected when they become visibly obvious on the documents or the building materials. At this stage of the fungi development, it is often too late and the decontamination, difficult and costly. The better way of avoiding such a contamination, is to detect it very soon, before it becomes invasive. That is the aim of DECAGRAPH project. The methodology presented here is based on the detection of organic volatiles emitted by fungi (MVOC’s) specifically found on paper based collections as soon as they start growing. The research program is divided into 3 main stages:

- definition of target tracers characteristic of a mould activity on paper,

- from these date, determination of a fungal contamination index which indicates the presence or absence of a mould activity,

- validation of the index by MVOC’s and classical biological air samplings in several book

storage facilities, contaminated or not.

The results of this project will be used to parameterize a sensor for continuous monitoring of biological indoor air quality in paper based collections storage facilities. Collection keepers could then be warned about a contamination from its very beginning and take rapidly the actions necessary to prevent it from being invasive and harmful to the collections.

DECAGRAPH which began in September 2009 is a 2 year’s national research program supported by the French ministry of Culture.


112

IBBS-15-4-P13

The potential sensitivity to biodegradation processes of chosen natural dyes extracted from ancient silk decorations in Wilanow Palace - Museum

A. Laudy 1, D. Ruszkowski 2, L. Rajpert 2, A. Jagielski 3 and A. Skłodowska 2

1) Wilanow Museum - Palace, Kostki Potockiego10/16 02-958Warsaw, Poland, http://www.wilanow-palac.pl/ 2) Laboratory of Environmental Pollution Analysis and 3) Department of Metabolic Regulation, Institute of Biochemistry, Faculty of Biology, University of Warsaw, Miecznikowa 1, 02-096 Warsaw Poland

e-mail of the corresponding author: [email protected]

Keywords: silk, natural dyes, biodegradation, bacteria

The Wilanow royal residence, property of King Jan III Sobieski, is one of the most beautiful Polish baroque palaces. The oldest apartments are decorated with dyed silk textiles, woven in Genoa in the beginning of XVIII century. The colorful floral patterns have been preserved in its original form until today. The goal of the research was to verify the biological agents affecting the degradation and potential fading processes of silk textiles. The natural dyes extracted from the ancient textiles within blue, green and flesh colors shown the presence of indigo and ellagic acid. The separation and identification of ancient dyestuffs’ constituents was performed by HPLC. Three proteolitic bacteria from genus Bacillus were tested for the potential ability to decolourization of dyes extracted from the silk threads.

Plant extracts, containing dyestuffs, was used as an only source of carbon and energy for isolates growth on solid media. Chemically pure standards of dyestuffs, identical with natural ones, were used to study the rate and efficiency of their biodegradation in liquid media.

Biodegradation of ellagic acid, indigocarmin and polish indigocarmin (from Isatis tinctoria) by bacteria was shown.


113

IBBS-15-4-P14

Algal colonization and deterioration of fine sculptures of terracotta temples of Bishnupur, West Bengal, India Sikha Mandal, Jnanendra Rath, Samit Ray Department of Botany, Visva-Bharati University, Santiniketan-731235, West Bengal, India ABSTRACT The blackish brown crust on the fine carvings of seven famous terracotta temples of Bishnupur was studied with regards to the presence of cyanobacteria and microalgae. Forty five taxa of micro algae belonging to 36 species of cyanobacteria, 5 species of chlorophyta and 4 species of Bacillariophyceae has been found from crusts of these monuments. The temples of Bishnupur were built in 17th century, crafted from the local lateritic soil and brick and are covered with terracotta tiles. However, due to colonisation by cyanobacteria and micro algae the fine sculptures are being damaged in course of time. Cyanobacterial taxa Aphanothece, Chroococcus, Cyanosarcina, Gloeothece, Lyngbya, Scytonema and Leptolyngbya occurs in almost all the temples and are the dominant species forming biofilm over the fine sculptures deteriorating its fine carvings. Though the temperature on the surface of these temples increased up to 500C during summer months, the cyanobacterial species survive forming black-brown crust. Aphanothece, Gloeocapsa, Gloeothece, Cyanosarcina, Chroococcus, Lyngbya and Nostoc species are surrounded by thick external envelope layers formed by extracellular polysaccharides that might be the cause of deterioration of the fine architecture of these beautiful temples.


114

IBBS-15-4-P15

Characterization and quantification of phototrophic biofilms on artistic tiles Sara Gutiérrez-Patricioa, Ana Zélia Millerb, Mathilda L. Coutinhoc, Miguel A. Rogerio-Candeleraa, Maria Filomena Macedod, Mariona Hernandez-Marinée, Cesareo Saiz-Jimeneza a Instituto de Recursos Naturales y Agrobiologia, CSIC, Av. Reina Mercedes 10, 41012 Sevilla, Spain b Centro de Petrologia e Geoquímica, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001, Lisboa, Portugal c REQUIMTE-CQFB - Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Monte de Caparica, Portugal d Vicarte, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Monte de Caparica, 2829-516 Caparica, Portugal e Universitat de Barcelona, Facultat de Farmacia, Av. Joan XXIII, s/n, E-08028 Barcelona, Spain Artistic Portuguese and Oriental tiles attributed to Wenceslau Cifka and dating from the nineteenth century were used as decorative building materials on outdoor walls of the Pena National Palace (Sintra, Portugal). The palace is located on the top of a hill surrounded by a forest, which produces characteristic microclimatic conditions that enhance biological colonization. Nowadays, these historic tiles present extended phototrophic biofilms, which are particularly apparent on the depression of the relieve motifs. In the course of a general survey concerning the conservation state of the historic tiles, an accurate campaign was performed in order to assess the main biodeterioration agents and subsequent biodeterioration effects. Bacteria, fungi, microalgae, and cyanobacteria obtained from these naturally colonized artistic tiles were characterized by DNA-based molecular analysis using the small subunit of the ribosomal RNA gene (16S and 18S for prokaryotes and eukaryotes, respectively), and complemented with cultivation techniques. Optical microscopy and confocal laser scanning microscopy were accomplished in order to assess the biofilm structure and cell organization. The amount of chlorophyll a was determined as an estimation of biofilm biomass present on the artistic tiles. This photosynthetic biomass quantification was complemented by digital image analysis by determining the surface areas covered by the biofilms. Our results unveiled complex microbial communities composed mainly by photoautotrophic microorganisms and fungi which growth resulted in dense biofilms formed on relieve motifs, crack formation patterns as well as on the slick tile surfaces. Regarding the ceramic glaze characterization, preliminary analyses were performed by energy dispersive X-ray fluorescence and X-ray electron probe micro-analysis. These techniques revealed the presence of a heterogeneous silica lead-tin glaze with tin dioxide (SnO2) crystals as opacifying and colouring crystalline pigment in the white glaze. Details on conservation and/or preservation works that need to be implemented are also presented in this work.


115

IBBS-15-4-P16

Assessment of biodeterioration with different analytical techniques in El Castillo and Las Monedas caves. Cantabria, Spain.

Irene Arroyo1, M. Isabel Sarró2 & Julia Montero1.

1 IPHE. Ministerio de Cultura.

2 CENIEH. Centro Nacional de Investigación sobre la Evolución Humana.

E-mail: [email protected], [email protected], [email protected]

Paleolithic paintings represent unique cultural assets that need to be preserved for future generations, due to these paintings represent the oldest artistic works created by humans. Previous reports have mentioned a generalized phenomenon of accelerated microbial colonization of rock art in caves. During the last years several studies have provided with valuable information on the composition of the microbial communities present in rock art and have highlighted the interest in understanding their complexity.

This work shows the biodeterioration of two important caves in Cantabria declared World Heritage by UNESCO. The caves of Las Monedas and El Castillo are located in Puente Viesgo, both are karstic cavities and they are an important tourist attraction, which receive a high number of visitors every year.

The aim of this study is to analyses the microbial populations present in the caves using different traditional and molecular microbiological techniques in order to obtain information about the different groups present and evaluate the effect of these populations in the support of the paintings and in the pigments used to elaborate the rock art.


116

IBBS-15-4-P17

Biodeterioration influence in the stone material: La Casa de las Conchas. Salamanca. Spain

Julia Montero1, Irene arroyo1, José Vicente Navarro1

1Instituto del Patrimonio Cultural de España

E-mail: [email protected], [email protected], [email protected]

The type of stone some monuments are made of may cause the emergence of different types of alteration, such as those generated by biological contamination and remains of other nature. Thus, the Villamayor sandstone, an ornamental rock used in the construction of La Casa de las Conchas (the House of Shells), shows this aspect.

It is a fine grained and brittle to the touch sandstone. It takes on a white-yellow and golden-brown coloration due to its content in iron, iron and titanium oxides and to surface oxidation.

Its porous nature makes it prone to attacks by different contaminants. In this case, a study has been made about the effect produced by biological agents.

Resistant species of lichens and microbial fauna, whose metabolism has altered the structure and the color of the stone, have been found.

In microbiology testing carried out, the Actynomicetes, the main attacker of the sandstone, are basically predominant, as well as several other species of both autotroph and heterotroph microorganisms.

It is also important to emphasize that microbiological and chemical analysis have been made on bird excrement samples found on the shells. They prove the existence of nitrifying bacteria that contribute to material degradation.

Moreover, studies with Scanning Electron Microscope (SEM) of patinas, that cover some areas of the building facade, have been carried out. They show the presence of calcium oxalates and plasters as products resulting from microbial action.


117

IBBS-15-4-P18

Microbial flora of the 240 year old submerged wooden shipwreck Vrouw Maria

Leone Montonen, Sanna Kettunen, Kari Steffen Department of Food and Microbiological Sciences, P.O.B. 56, 00014 University of Helsinki, Finland Degradation of wood is slow in the Baltic Sea, and shipwrecks can remain in good condition there for centuries. The water is brackish and, at the site of the wreck, constantly cold and dark with a low oxygen content. However, microbes are active and over time destroy wood also in this environment. We wanted to find out what microbes are present and what potential danger they constitute to the wreck, if left in situ or lifted.

The study was done by scanning for microbes by molecular methods, extracting DNA from approx. 0.2 g (wet weight) samples, amplifying, cloning and sequencing fungal, bacterial and archaeal 16/18S rDNA genes.

The major bacterial group clustered with uncultured Chloroflexi bacteria. Chloroflexi grow autotrophically with either H2S or H2 as electron donors, or heterotrophically in the dark. All cultured members of Chloroflexus are thermophilic but nonthermophilic marine Chloroflexi sequences are known. Several groups of planctomycetes sequences were also abundant. The third major group fell within the phylum Acidobacteria, which are widespread, but their ecological role is unknown. The archaeal sequences clustered with ubiquitous uncultured Crenarchaeota from cold marine environments. Their possible role in the destruction of submerged wood cannot yet be inferred.

Most of the fungal sequences belonged to ascomycetal groups with wood-degrading abilities. The majority clustered with an aquatic Acremonium-like sequence that has a mangane (II) oxidising enzyme with laccase activity. Sequences belonging to lignicolous clades of Lulworthia-like and Nais inornata-like fungi were also retrieved.


118

IBBS-15-4-P19

Fungal biogenic alteration of limestone monuments at Chichen Itza

Sergio Gómez-Cornelio1, Benjamin Otto Ortega-Morales1*, Susana De la Rosa-García1, Patricia Quintana2, José Narváez-Zapata3, Amanda Oliva-Hernández3, Heather Bullen4, Héctor Urcia4 and William Santiago2

1Departamento de Microbiología Ambiental y Biotecnología, Universidad Autónoma de Campeche, Av. A. Melgar s/n, C.P. 24039, Campeche, México, 2Departamento de Física Aplicada, CINVESTAV Unidad Mérida, C.P. 97310, Mérida, Yucatán, México, 3Centro de Biotecnología Genómica, Instituto Politécnico Nacional, Blvd Del Maestro esq. Elias Piña, Reynosa, 88710, Tamaulipas, Mexico, 4Department of Chemistry, Northern Kentucky University, Highland Heights, KY 41099, USA.

Chichen Itza is a large archaeological Mayan site in Mexico. Visual inspection has shown that extensive areas of microbial biofilm growth occur on most buildings at this site. Given that scant attention has been given to fungi in Mayan monuments, this work aimed at characterizing the biogeochemical consequences of metabolic activity on limestone by selected fungi isolated from biofilms at Chichen Itza. Our survey yielded 63 fungal isolates derived mainly from black epilithic black biofilms material inoculated using two different methods and two culture media. Three fungal isolates identified by ribosomal intergenic spacer analysis (RISA), Penicillium oxalicum TM1H52, Annulohypoxylon stygium TM1H19,

Rosellinia sp TM1H05 and shown to be very active calcium carbonate-dissolving activity with the subsequent formation of mygenic fabrics. Electron scanning microscopy (SEM), X-ray diffraction (XRD) and fourier transform infrared (ATR-FTIR) spectroscopy of crystalline precipitates showed they were composed of a mixture of calcite (CaCO3), calcium oxalates and calcium citrates. These results provide experimental evidence for the formation of secondary mycogenic minerals by naturally occurring fungi derived from microbial biofilms associated with Chichen Itza buildings. However, the absence of calcium oxalates and citrates on stone surfaces is noteworthy and is possible related to rapid turnover in the epilithic habitat.


119

IBBS-15-4-P20

Measurement of mechanical properties in situ and modeling of timber structures in Ruhnu churches (Estonia) damaged by wood-destroying insects. Kalle Pilt a , Uwe Noldt b a. Estonian University of Life Sciences, Department of Rural Building, Kreutzwaldi 1, Tartu, Estonia Estonia b. Johann Heinrich von Thünen-Institut (vTI) / Federal Research Institute for Rural Areas, Forestry and Fisheries, Institute of Wood Technology and Wood Biology (HTB), Leuschnerstrasse 91d, D-21031 Hamburg, Germany There are many factors which effect durability of wood in the existing timber structures. One of the most important factors effecting durability is biological, especially the wood-rotting fungi and wood-destroying insects. This paper gives overview about fieldworks on Ruhnu island and investigations on damages of House Longhorn beetle, Deathwatch beetle and Common furniture beetle in the churches built in 1642 and 1912 AD, respectively. Since both churches are architectural heritage only non-destructive methods in measuring mechanical properties of timber structures could be used, e.g. the resistograph IML-RESI F400. The modeling of timber structures was analyzed with modern computer design program ArchiCad 12 and engineering program Autodesk Robot Structural Analysis v. 22. Engineering calculations were taken from European Standards. The extent of various damages in two churches built at different time but located in the same environmental conditions was analyzed during discussions. The damages by insects differ by extent and by habitat. In the result the possibilities of preliminary work for the restoration of old timber structures in architectural heritages are described.

Keywords: architectural heritage, wood-destroying insects, beetles, non-destructive methods.


120

IBBS-15-4-P21

Feasibility of fluorescence lidar remote sensing on frescoed surfaces: test studies

Valentina Raimondi1, Giovanna Cecchi1, Costanza Cucci1, Oana Cuzman2, Cristina Fornacelli1, Ioana Gomoiu3, David Lognoli1, Dan Mohanu3, Lorenzo Palombi1, Marcello Picollo1, Piero Tiano2

1 'Nello Carrara' Applied Physics Institute - National Research Council, Via Madonna del Piano 10, I- 50019, Sesto Fiorentino, Firenze, Italy

2 Institute for the Conservation and Promotion of Cultural Heritage - National Research Council (CNR-ICVBC), Firenze, Italy

3 National Arts University of Bucharest, Institute of Biology, Splaiul Independentei, 296, R-77700 Bucharest, Romania

Fluorescence techniques are widely used for the detection of photoautotrophic biodeteriogens in several scientific fields, cultural heritage conservation included. In particular, fluorescence lidar remote sensing has already been applied to the diagnostics of biodeterioration on outdoor stone monuments for several years. The latter is a technique to carry out fluorescence measurements remotely in the field: it relies on the use of a pulsed laser as an excitation source and of a telescope to collect the emitted signal remotely. This technique offers several advantages for in-field operation, since it can be applied from a distance on extended surfaces and independently from external conditions like sunlight. In order to extend the applicability of this technique, we carried out a set of test studies to investigate the feasibility of non-invasive fluorescence lidar remote sensing of biological growth on frescoed surfaces. The study - conducted in the frame of the regional-funded TDTbioart project which addressed innovative technologies for the diagnostics and treatment of biodeterioration - consisted of fluorescence lidar and fiber optical reflectance spectroscopy (FORS) measurements on a fresco model samples with different pigment layers and inoculated with various concentrations of biodeteriogens, specifically a green alga (Apatococcus sp.) and a cyanobacterium (Chroococcus sp.). Results pointed out the feasibility of the remote detection of biodeterioration at a pre-visual stage of development from distances as far as tenths of meters and using very low laser fluence to prevent any damage to the cultural assets.


121

IBBS-15-4-P22

Survival strategies of desiccated Lyngbya corticcola in terracotta monuments of Bishnupur, India causing biodeterioration Jnanendra Rath, Sikha Mandal, Samit Ray Department of Botany, Visva-Bharati University, Santiniketan-731235, West Bengal, India Abstract: Lyngbya corticcolais is a dominant cyanobacteria occur as blackish crust in the seven terracotta archeologically important monuments of Bishnupur India. In summer months the temperature in the crust surface goes beyond 60 0C with extreme dryness RH of 88 %and high UV-B radiation of 0.42 W/m2 but it still survives in the monuments wall. The response of desiccation up to 6 months on the chlorophyll- a, carotenoid and mycosporine amino acid like substances of L. corticcola shows that though Chlorophyll-a was decreased continuously with the increase in the duration of desiccation, carotenoids and mycosporine content was progressively increased. The cellular carbohydrate and cell protein content of the organism was increased up to 4 months and 3 months of desiccation respectively and start decreasing then after. However Superoxide dismutase enzyme (SOD unit/mg protein) was increased with duration of desiccation. Induction of 32, 28 KDa protein and over expression of 68, 53 kDa protein probably provide protection to the organism. Morphological features shows the cells become compact, coiled and intertwined coiled and separated and finally breakage of the filament into small fragments with dense granulated and packed tightly and embedded in the mucilage sheath probably an adaptive strategies of this organism. Upon desiccation carbohydrate content of L. corticcolais increases and act as a primary coloniser and provide nutrients and moisture which supports and stimulate the growth of heterotrophic bacteria, unicellular green algae, fungi, mosses and small plant all of which reside within the microbial mats. Further the carbohydrate content produced by Lyngbya undergoes changes in volume during desiccation and rewetting cycles, thus loosening the terracotta tiles and acids excreted by certain microorganisms of the mats as metabolic end products cause severe corrosion of the substrate and deteriorate the fine carvings of century old Bishnupur terracotta monuments.


122

IBBS-15-4-P23

Actinomycetes in welded tuffs

Veridiana Reyes1, Aurelio Alvarez2 , Carmen Cano Canchola2 and Jorge Cervantes1

1Departamento de Química, 2 Departamento de Biología, Universidad de Guanajuato

This study reports the identification of Actinomycetes in degraded welded tuffs surface from the principal church of the city of Guanajuato, Mexico. A total of 25 isolates have been obtained from 30 samples taken before and after of restoration of the monument. The strains were identified by characterization of 16S rRNA sequences. Four genera grouped the strains identified: Streptomyces, Actinoplanes, Micrococcineae and Geodermatophilus. This project was supported by grants from CONACyT and Universidad de Guanajuato.


123

IBBS-15-4-P24

Laser treatment as an effective tool to control epilithic and endolithic microbiota from stone

M. Speranza1, M. Sanz2, M. Oujja2, A. de los Ríos1, S. Pérez-Ortega1, M. Castillejo2 and C. Ascaso1.

1 Museo Nacional de Ciencias Naturales, CSIC,Serrano 115, 28006 Madrid, Spain

2Instituto de Química Física Rocasolano, CSIC, Serrano 119, 28006 Madrid, Spain

In the last decades chemical biocides have been banned because of the environmental and health hazards associated with these toxic substances. Furthermore, the use of biocide, as well as UV radiation sterilization systems, is often restricted because of low substrate penetration power. In addition, previous work done by our group has shown the limited effect of biocide treatments in controlling endolithic microbiota. Although laser cleaning of stone is a well established technique in the field of cultural heritage, to our knowledge, this is the first study of its effect on endolithic microorganisms. In our study we tested different laser treatments with a Q-switched Nd:YAG laser, using infrared and ultraviolet nanosecond laser pulses to attack stone surface colonizers as well as the endolithic microbiota.The effect of the laser treatment on both ultra-structural epilithic and endolithic colonizers was assessed using Scanning Electron Microscopy using especially prepared samples (SEM-BSE microscopy). The EDS and micro-Raman spectroscopy were employed to detect structural and chemical changes in the stone. Not only was the ultrastructure of the photo- and mycobiont of Verrucaria located on the stone surface severely affected, but the endolithic microorganisms also suffered some ultrastructural changes in fungal cells. All the results indicated that this is a promising and effective treatment for the control of important internal biodeterioration processes produced by endolithic microorganisms. Field experiments are now in progress to formulate laser irradiation strategies, alone and in combination with low doses of biocides, thus allowing us to evaluate the biological recolonization of the irradiated surfaces.


124

IBBS-15-4-P25

Determination of MVOCs like formic and acetic acid with LC-MS/MS

Katharina Wiegner1, Oliver Hahn1, Matthias Farke2, Sabine Kalus1 and Wolfgang Horn1 1BAM Federal Institute for Materials Research and Testing, Berlin, Germany 2GLASBAU HAHN GmbH, Frankfurt am Main, Germany

Abstract:

Formic and acetic acids are often responsible for damage of cultural objects, e. g. glass and metal corrosion or changing and fading of colours. To minimize the risk of damage emissions must be quantified. Museums worldwide are equipped with different display cases. Exhibit display cases should protect cultural objects from dust as well as from mechanical / physical damage. Acetic and formic acids are typical metabolites of micro organisms and these emissions belong to MVOCs (microbial volatile organic compounds). Cultural objects based on organic matter, such as leather, paper and wood can emit formic and acetic acid into the indoor or display case air. Concentrations of harmful substances can decrease considerably in display cases because of their very high air tightness (air exchange rates are often smaller than 0.1 d-1). There is a need for a stable, robust, reproducible and comparable method with an easy local sampling and determination in laboratories. In this study different methods for determination of acetic and formic acid were compared to find the best suited method. Important factors were: in situ sampling, handling and preparation of loaded samples, limit of quantification and detection, recovery rate. Miniaturised emission test chambers and model display cases were used for these investigations. The first method is based on sampling on DNPH (2,4-Dinitrophenylhydrazin) cartridges, derivatisation, elution and quantification of the derivates with LC-MS/MS. The second one, which is only usable for the quantification of acetic acid, is done after sampling on TENAX® or CARBOTRAP® and thermal desorption-GC in accordance with ISO 16000-6 and ISO 16017-1.


125

IBBS-15-4-P26

Study on biodeteriorigenous agents in the Holy Saviour’s Cave (Vallerano, Italy)

Manuela Gagliardi1, Daniela Isola1, Laura Selbmann1, Maria Andaloro2, Paola Pogliani2,

Silvano Onofri1, Laura Zucconi1

1 Department of Ecology and Sustainable Economic Development, Università della Tuscia, Largo dell’Università snc, 01100 Viterbo, Italy 1 Department of Science of Cultural Heritages, Università della Tuscia, Largo dell’Università snc, 01100 Viterbo, Italy

Biodiversity on monuments and biodeterioration are strictly dependent to the environment. The first step to develop restoration strategies is the knowledge of the diversity and the destructive potential of biological agents involved. The most significant parameters affecting growth are represented by physical factors, mainly humidity, temperature, and light, as well as the chemical nature of the substratum. Mural paintings in rocky habitat are frequently subject to biological colonization causing different kinds of mechanical and chemical alterations such as discoloration, formation of crusts and loss of material.. This work aims to the restoration, valorization, and conservation of the Vallerano artistic rural area, in the frame of an interdisciplinary study between the Faculties of Science and Cultural Heritage Conservation of the University of Tuscia. The main biodeteriogenous organisms growing on the wall paintings of Holy Saviour’s Cave near Vallerano (Viterbo, Italy) were identified. The vault collapsed on 1888 and paints of the remaining vault and the northern and partially western lateral walls were exposed to the external conditions. Different kinds of biological alterations were found, mainly related to lichenic, fungal and algal colonization. Massive development of the lichen Dirina massiliensis f. sorediata, but also Caloplaca xantholyta and Lepraria sp. were observed. Alterations varying from brilliant to dark green and black gummy patinas were due to the presence of cyanobacteria (Chroococcus sp., Nostoc sp.) and green algae (Stichoccus sp., Trentepohlia sp.). Black fungi, here analyzed basing on ITS rDNA sequencing, were also present as dark spots.


126

IBBS-15-4-P27

Molecular biology identification of microorganisms presents in wall paintings

1Pilar Bosch Roig, 2Inés Martín Sanchez ,3Fernando Bolívar Galiano, 4Rosa Montes Estellés, 1Institute of Restoration of Cultural Heritage of the Polytechnic University of Valencia. (Spain) 2 Deparment of Microbiology. Faculty of Sciences. University of Granada. (Spain) 3Department of Paintings. Faculty of Fine Arts. University of Granada (Spain) 4Microbiology area in the Biotechnology department of the Polytechnic University of Valencia. (Spain) Wall paintings are made mostly by inorganic components but also by organic materials that can be altered by different types of microorganisms. Restoration processes and even the accumulation of dirt, air pollutants etc., on the wall paintings surfaces also provide new sources of nutrients for microorganisms. All these substrates can be used by microorganisms if given the appropriate environmental conditions like temperature, humidity and lighting. The growth of microorganisms on the surface of the wall paintings can cause physical, chemical and aesthetic damage, and that is why it is very important to study and control their presence on the murals surface.

In this work we carry out the identification by Molecular Biology Techniques of the microorganisms presents on the murals painted by A. Palomino (XVIII) in the vault of the Church of “Santos Juanes” in Valencia, Spain. Identifying fifteen different bacteria mainly from the genera Bacillus sp., and seven fungi mostly belonging to the genera Aspergillus sp. and Penicillium sp.

This investigation show that the wall paintings studied here have large numbers of microorganisms, that if there was a favorable change in the church environmental conditions may trigger serious problems of biodeterioration in these important murals.


127

IBBS-15-4-P28

Molecular investigation of the micro-biota associated to the decayed medieval stained window glasses of two Catalonian churches.

G. Piñar1, M. Garcia-Vallės2, D. Gimeno-Torrente2, J.L. Fernández-Turiel3, J. Ettenauer1, K. Sterflinger1

1 Institute of Applied Microbiology, Department of Biotechnology, Vienna Institute of Bio Technology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 11, A-1190 Vienna, Austria.

2 Facultat de Geologia, Universitat de Barcelona (UB), c/ Martí i Franquès s/n, 08028 Barcelona, Spain. 3 Institut de Ciències de la Terra Jaume Almera, CSIC, c/ Solé i Sabarís s/n, 08028 Barcelona, Spain. The decay of medieval stained glass is characterised by the presence of corrosion, patina development and mineral crust. Besides physico-chemical process, microorganisms as lichens, bacteria and fungi play a crucial role for the decay of stained glasses.

In the present study we investigate the micro-biota associated with decayed historical church window glasses located in two Catalonian churches: the Cathedral of Tarragona and the Church of Santa Maria del Mar in Barcelona, both under Mediterranean climate. Investigations were focused on glass with a K-rich composition, which is known to be more susceptible to biodeterioration, showing orange patinas and signals of bio-pitting. Biological activity in both studied sites showed to be fossil and only remains of this activity, as bio- pitting and mineral precipitation, was observed.

To overcome the limitations of culture-dependent techniques and with the goal to gain insight into the micro-biota associated with the observed decay, the biodiversity was investigated by the following molecular methods: DNA extraction from glass samples, amplification by PCR targeting the16S rRNA and ITS regions, and DNA fingerprint analyses by denaturing gradient gel electrophoresis (DGGE). In parallel clone libraries containing either PCR fragments of the 16S rDNA or the ITS1 region were screened by DGGE. Clone inserts were sequenced and compared with sequences listed in the EMBL database. Similarity values ranged from 89 % to 100 % to known bacteria and fungi.

Analyses revealed complex bacterial communities consisting of members and relatives of the phyla Proteobacteria, Bacteroidetes, Firmicutes and the dominance of Actinobacteria. Fungi showed lower diversity than bacteria and species of the genus Cladosporium were dominant. The detected Actinobacteria and fungi may be responsible for the observed bio-pitting phenomenon. Moreover, some of the detected bacteria are know for their capabilities of mineral precipitation. Sequence results also showed similarities with bacteria commonly found on deteriorated stone-monuments, what supports the idea that medieval stained glass biodeterioration in the Mediterranean area shows pattern comparable to those developed on stone.


128

IBBS-15-4-P29

Biogenic weathering of calcareous building stone due to colonisation by subaerial algal assemblages

Liam Morrison1, Edward P. Lynch1 and Fabio Rindi3

1 Department of Earth and Ocean Sciences, School of Natural Sciences, Ryan Institute, National University of Ireland, Galway, Ireland

2 Università Politecnica delle Marche, Ancona, Italy

Orange, red and pink patinas dominated by the filamentous green alga Trentepohlia umbrina (Kützing) are common features on limestone buildings in Galway City, Ireland. The presence of Trentepohlia tends to coincide with enhanced damage and decay of the limestone surface. Samples of the algae with fragments of their attached substrate were collected from two limestone buildings in the city; the 19th century Quadrangle Building, NUIG and the 14th century St. Nicholas’ Collegiate Church. The patinas have been analysed and characterised by Raman Spectroscopy, Scanning Electron Microscopy coupled with Energy Dispersive X-ray Spectrometry and image processing stereophotogrammetry software. This facilitated a microstructural and morphological characterisation. The combined application of SEM and stereomicroscopic techniques has allowed the first ever 3D reconstruction of the surface of the Trentepohlia dominated patina, enabling the investigation of topographic features, (texture and structure). The formation of these Trentepohlia assemblages as determined by SEM and Raman observations has lead to aesthetical and microstructural alterations in the building stone surface.


129

Topic 05-Biodeterioration of miscellaneous materials: spacecraft

materials, medical devices, glass, teflon, “nano” coatings


130

KEYNOTE lECTURE

Safeguarding the Crew and Engineering Systems for Human Missions

Kasthuri Venkateswaran

Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA 91109

Microbial detection and their mitigation systems to facilitate spacecraft/habitation cleaning, sterilization, and maintenance will be paramount to prolong the longevity of human habitation on other planets. Life-support processes (recycled resources, humidity, etc.) will promote the proliferation and colonization of microbes likely to play a role in subsequent bio-fouling/corrosion, and the pathogenesis of host organisms (e.g., crew members, plants). The objective of this concept paper is to understand whether spacecraft- and crew- associated microbiome can provide insight for a more efficient, lower cost, practical approach to specific challenges in space. In this regard, validated environmental monitoring systems and control strategies are crucial to (a) preserve acceptable microbial burden levels in human compartments and life-support components, (b) ensure negligible interference of false-positives with life-detection experiments, and (c) prevent the inadvertent exposure of humans to extraterrestrial materials.

Planetary protection policies derive from international treaties whose goal is “to preserve our ability to study other worlds as they exist in their natural states; to avoid contamination that would obscure our ability to find life elsewhere—if it exists; and to ensure that we take prudent precautions to protect Earth's biosphere in case it does.” Mandates are in place to minimize the likelihood of catastrophic outcomes as a result of human-associated cross-contamination between solar system bodies. To meet planetary protection obligations, NASA needs:

o Integrated system technologies to protect human life from alien microorganisms (should they exist) and to shield engineering systems from bio-corrosion.

o Assurance of compliance with evolving standards for planetary protection (both forward and backward contamination) relating to the human exploration of other celestial bodies.

o A sound technical basis to determine whether the inadvertent shedding of bio-contaminants from human explorers can be minimized to such a degree that the search for life can continue in an unobstructed, meaningful manner.

This concept paper identifies a body of work to address NASA needs relative to microbial monitoring and controlling the harmful impact of microbial corrosion. The key deliverable of this concept paper is an integrated microbial monitoring system, validated in a terrestrial Mars analog environment and ready for deployment on a human mission to Mars. Such a system is essential for human missions to comply with requirements to avoid harmful contamination and thereby facilitate the search for extraterrestrial life. The proposed integrated microbial monitoring system will bolster confidence in, and lend support to, planetary protection efforts, hardware reliability, and sustained crew health. By forewarning human explorers of any significant fluctuations in microbial burden, the system allows the crew to take immediate actions to significantly diminish any threat to crew health, or deterioration of the habitation module resulting from bio-corrosion. This approach will strive to directly integrate the technologies proposed herein with those being developed for robotic sample return missions, thereby providing a cradle-to-grave planetary protection implementation capability for human exploration.


131

IBBS-15-5-O01

Effectiveness of gaseous decontamination technologies for use on spacecraft and their components

Jimmy Walker¹, Tom Pottage¹, Allan Bennett¹, Thomas Rohr², Chantal Fowler³, Martin Wilson³

¹Biosafety Unit, Health Protection Agency, Porton Down, UK, ²European Space Agency, The Netherlands, ³Science and Technology Facilities Council, Rutherford Appleton Laboratory, UK.

The European Space Agency (ESA) and NASA currently use Dry Heat Microbial Reduction (DHMR) at >110°C for >30 hours to decontaminate whole spacecraft modules or components. However DHMR is a lengthy process that precludes the use of heat sensitive materials. Following an extensive literature review and selection process three gaseous decontamination technologies including vapour hydrogen peroxide (VHP, Steris Inc.), hydrogen peroxide vapour (HPV, Bioquell Ltd.) and Chlorine Dioxide (ClorDiSys Solutions Inc.) were tested for biological efficacy, material compatibility and residue formation at ambient pressure within a 20m3 environmental chamber. Following exposure at the highest concentrations both the VHP (Steris Inc) and HPV (Bioquell Ltd) technologies resulted in a 6 log reduction in commercially available biological indicators within 20 minutes. The ClorDiSys technology resulted in a >4 log microbial reduction after exposure for a 1 hour period. Three naturally occurring microorganisms typically found in cleanrooms used for spacecraft integration were also tested as biological indicators with Bacillus thuringiensis exhibiting similar survival rates to Geobacillus stearothermophilus after exposure to both VHP and HPV, but B. thuringiensis demonstrated greater resistance to chlorine dioxide. A range of 30 materials were exposed to the decontamination technologies. No change was witnessed with the hydrogen peroxide systems, whilst several materials showed signs of degradation after exposure to chlorine dioxide. Residue analysis carried out on exposed silicon wafers demonstrated that each decontamination system produced elemental and nitrogen containing hydrocarbon contamination whilst chlorine dioxide resulted in additional sulphate and hypochloride residues as well as an oxide layer. VHP has been recommended as an alternative to DHMR.


132

IBBS-15-5-O02

Fungal colonisation and contamination of cinematographic film: implications for film and archivists

Gavin Bingley, Dr Craig Banks, Dr Gordon Craig, Mark Bodner, Professor Joanna Verran

Studies were carried out on 19 cinematographic film reels contaminated with mould using air sampling, to quantify spores released from contaminated film during a simulated inspection process, in order to assess exposure of archivists to spores. Organisms present were identified and screened for gelatinase production, since gelatine is the major substrate for fungal growth on the film. The majority of fungi present were Aspergillus and Penicillium species, of which 18 out of 27 isolates produced gelatinase. For some films, fungi released spore numbers which exceeded the recommended safe exposure levels of 1000cfu/m3. Some films appeared contaminated, but no fungal growth was detected post-inspection. However, hyphal growth was evident across film frames, indicating that the damage may have taken place in the past. Culture of fungi may not be possible in archives but detection of the product of fungal growth may be possible. Detection of microbial volatile organic compounds (MVOCs) by gas chromatography mass spectrometry (GC-MS) coupled with solid phase microextraction (SPME) could be an alternative to culture methods to determine if active mould growth is occurring. 1-octen-3ol, 3-octanone and 3-octanol are all said to be general indicators of fungal growth and have been detected by GC. Conservation and storage is essential in archives. A knowledge of the amount of fungi, identification of key fungal contaminants, production of MVOCs and their gelatinolytic potential will help development of methods to enable early detection of actively growing mould.


133

IBBS-15-5-O03

Fungal Biodeterioration of Cinematographic Films of the Cultural Heritage of Cuba

I. Vivar1, S. Borrego1, D.A. Moreno2, A.M. García2

1 Archivo Nacional de la República de Cuba. Compostela 906 e/ San Isidro y Desamparados, Habana Vieja 10100, Ciudad de La Habana, Cuba

2 Universidad Politécnica de Madrid (UPM), Departamento de Ingeniería y Ciencia de los Materiales, Escuela Técnica Superior de Ingenieros Industriales, José Gutiérrez Abascal 2, Madrid 28006, Spain

At the Cuban Institute for Cinematographic Industry and Arts (known as ICAIC), samples were taken from several cinematographic films of cultural heritage affected by biofouling and biodeterioration problems. The films were analyzed by Environmental Electron Scanning Microscopy (ESEM) and Scanning Electron Microscopy (SEM) to determine the biodeterioration degree and the type of colonising microorganisms. To study fungal viability the films were dyed using fluorochromes and were then analysed using an epifluorescence microscope. Also, a protocol for the recovery of fungi from the surface of the films was developed. Their DNA was extracted and amplified using PCR to be automatically sequenced later, so as to identify which kinds of fungi were responsible for the biodeterioration of the cinematographic films.

A significant fungal colonisation was present on both sides of the films on all the samples, whereas on some films, there was a higher concentration of fungi on one of the two sides. The degree of colonisation also varied from one film to another, and on some of them, fungi could be seen to penetrate the film itself. We have detect some of the fungi are still active and their harmful effect on the films under study can, therefore, continue.

We are grateful to the UNESCO Fellowships Programme in support of Programme Priorities (2010-2011) – Request Nº 714-1 (CUBA) and to the Technical University of Madrid Aid Programme to carry out research activities with Latin America (Thematic Network on Biodeterioration of Historic and Cultural Heritage - Ref. AL11-RT-01)


134

IBBS-15-5-O04

Effect of climatic conditions on the succession of fungi on vegetable tanned sole leather ( buff ) during storage.

D.S.Rathore1 and Shashi Chauhan2

1.Department of Botany / Biotechnology, Govt. K.R.G.P.G.(Autonomous) College, Gwalior (M.P.) INDIA.

E.mail – [email protected]

2.S.O.S. in Botany, Jiwaji University, Gwalior (M.P.) INDIA

ABSTRACT

Bio-deterioration of leather by its definition, is concerned with the interaction of leather with microorganisms. Thus materials, organisms and environment, all intimately concerned in bio-deterioration. So, the climate of storage place is of great importance to the activity of the fungi of stored leather. Therefore, it is quite necessary to investigate various types of leather infesting fungi, their succession and ecological conditions which play an important role in the development of these organisms on such products. In the present study attempts have been made to study the succession of fungi on vegetable tanned sole leather ( Buff ) under varying storage conditions. The conducive factors which are taken into consideration include varying levels of the relative humidity and storage conditions at suitable temperature. Thus during the present studies three basic attempts were taken under considerations (a) Qualitatively-what kind, (b) Quantitatively - how many fungal forms inhabit the vegetable tanned sole leather (Buff) and (c) how do, relative humidity and duration of storage affect fungi.

Key Words –Bio-deterioration, Fungi, Succession, Climatic factors.


135

IBBS-15-5-P01

The microscopic effects of fungal biodeterioration on 20th century photographic documents revealed by vp sem-eds technique

E. Damiano1 , F. Pinzari1, D. Matè1, M. C. Sclocchi1

1Istituto Centrale per la Conservazione ed il Restauro del Patrimonio Archivistico e Librario, Via Milano, 76, 00184 Rome. Italy.

Microbial deterioration is a common problem in photographic collections and is considered a major cause of deterioration. However, few studies so far have been addressed to biological damage on these materials. There is the need for a better understanding of the interactions occurring between microorganisms, the environment, and the organic and inorganic substances composing photographic material. The aim of this work was the study of the damage caused by naturally occurring fungal infections on the gelatin silver emulsion of positive and negative 20th century photographic material. The study was conducted by observing photographic prints and films with different spots and damaged areas with a variable pressure scanning electron microscopy (VP-SEM) combined with electronic dispersion spectroscopy (EDS). The SEM allowed the observation of entire, unaltered samples without metallization thus with a not invasive approach. Morphological information could be obtained by means of a secondary electrons (SE) detector and chemical characteristics of surfaces were visualised by means of a backscattered electrons detector (BSE). Moreover, compositional data were obtained by collecting characteristic X-ray for each object visualised in SEM micrographs, with a topographic elemental analysis. SEM technique allowed visualizing several interesting aspects of fungal damage. The effects of the intimate interaction between the fungal hyphae and the organic and inorganic components of the substrates were described. In particular, the ability of some fungi to alter the distribution of silver crystals in the gelatin emulsion, and the selective degradation of some substances were documented.


136

IBBS-15-5-P02

Assays on the resistance of photographic paper treated with gamma rays to fungal biodeterioration

D. Vellati1, M. Adamo2, D. Matè1, F. Pinzari1, D. Ruggiero1, L. Botti1, L. Residori1 and M.C. Sclocchi1

1 ICRCPAL Istituto Centrale per il Restauro e la Conservazione del Patrimonio Archivistico e Librario, Laboratorio di Conservazione preventiva, Laboratorio di Biologia e Laboratorio di Fisica, Ministero per i Beni e le Attività Culturali, Via Milano, 76, 00184 Rome, Italy.

2 ENEA-UTTMAT-IRR CR. Casaccia -Via Anguillarese, 301 S. Maria di Galeria (Rome), Italy

Fungal and bacterial contamination is considered a dominant problem for the conservation of photographic material, and thus the development of appropriate treatment measures for contaminated objects is a challenge for restorers, curators and scientist involved.

The present study is aimed at the analysis of the effects of gamma radiation on the biodegradability of photographic documents. Samples of photographic paper containing baryte, and sensitized with gelatine-silver print were irradiated with gamma rays doses of 3, 5, 10, 100kGy. Both new and artificially aged (ISO 5630/3) photographic paper samples were considered for the treatments.

Assays on the resistance of irradiated and aged samples to fungal biodeterioration were performed. The growth of Penicillium chrysogenum Thom on photographic paper samples aged and/or irradiated was evaluated in different controlled microenvironments: (H.R. 100%, 92%, 82.5%, 72.5%, 63.3%, 56.2%). Moreover, chemical and surface alterations induced on gelatine and cellulose by irradiation and/or artificial ageing were evaluated by means of scanning electron microscopy, microanalysis (SEM-EDS) and FTIR-ATR spectroscopy. This last technique revealed that in the areas affected by fungal growth, in addition to gelatine chemical signals, there are also those of gelatine, baryte and paper. These results were observed in all samples, including control.

Several ageing effects on both gelatine and paper layers were documented. Moreover, the culturing results showed that P. chrysogenum developed only at 100% and 92% R.H. on both control and gamma-rays treated samples. A very peculiar interaction of the fungus with the barite layer was documented as well as a different spoiling attitude of the fungus at the two controlled microenvironments.


137

IBBS-15-5-P03

Assessment risk of fungal corrosion of unbonded lubricated post-tensioned tendons Filomena De Leoa, Giuseppe Campanellab, Edoardo Proverbiob, Clara Urzìa

aDepartment of Life Sciences “M. Malpighi”, Via F. Stagno D’Alcontres, 98166 Messina, Italy bDepartment of Industrial Chemistry and Materials Engineering C/da di Dio, 98166, Messina, Italy In unbonded post tensioned concrete used in buildings, parking structures, bridges, steel cable corrosion protection is provided by grease or plastic shield. Microbiologically influenced corrosion (MIC) of prestressing steel mainly due to bacteria has been reported in literature while only few paper report on the role of fungi in the corrosion failure of unbonded lubricated tendons.

Aim of this research was to evaluate a protocol for Stress Corrosion Tests (SCT) that includes ubiquitous fungi (A. flavus, Penicillium sp. Fusarium sp., Cladosporium sp.) and an extremophilic black yeast (Acidomyces acidophilum) as microorganisms test.

The ability of the chosen fungi to degrade a commercial calcium based grease (OVOLINE 71C), used as lubricant in post- tensioning tendons was evaluated, followed by small and full scale sample experiments. The extent of corrosion was studied by microscopy (LM and SEM) and by FTIR spectra to analyze the organic acid produced. The propagation of corrosion attack was measured by means of Acoustic Emission monitoring and by vibroacoustic analysis.

The results showed that all fungal strains were able to utilize the grease as sole carbon source and, through the production of organic acids they were able to produce diffused pits on steel surface. No Stress Corrosion Cracking (SCC) were noticed on the steel surface under tensile strength.

Results showed that the fungal action is carried out as summarized below:

1. decreasing of lubricating action of grease through its degradation;

2. steel corrosion through the release of organic acids;

3. possible mechanical weakening of tendons through the growth of fungal structures (hyphae and/or meristematic cells).


138

Topic 6-Biodegradation of hydrocarbons and persistent pollutants: oil hydrocarbons, plastics, PAH, PCBs


139

KEYNOTE LECTURE

Genomics and Post-Genomics of Solvent Tolerance in Pseudomonas putida

Juan-Luis Ramos, Estrella Duque, Craig Daniels, Jesús de la Torre, M. Antonia Molina-Henares, Lázaro Molina, Ana Segura, Jesús Lacal, Hortencia Silva-Jimenez and Tino Krell

Estación Experimental del Zaidín – CSIC, Granada, Spain

Pseudomonas putida DOT-T1E is a highly solvent-tolerant strain. When Pseudomonas putida cells encounter toluene in the growth medium, they perceive it simultaneously as a potential nutrient to be metabolized, as a membrane-damaging toxic drug to be extruded, and as a macromolecule-disrupting agent from which to protect proteins. Each of these efforts requires a dedicated transcriptional response that involves a large number of genes. We used DNA array technology to decipher the interplay between these responses. The resulting expression profiles suggest that the bulk of the available transcriptional machinery is redesigned to endure general stress, to remove toluene to the outer medium and reduce the permeability of the cell membranes. In addition, toluene affects motility functions in response to the aromatic compound. Toluene is mainly sensed by P. putida as a stressor rather than as a nutrient. Initially, in a very rapid adaptive response, fatty acids are converted to trans isomers and phospholipids headgroups are turned over with the aim of decreasing membrane fluidity. A number of transporters that extrude the solvent (TtgABC, DEF and GHI pumps) are induced in the presence of toluene. To determine the role of these transporters in solvent endurance we prepared mutants of the corresponding genes and carried out a wide series of phenomic analyses to assign a function to these transporters. A small share of the available machinery is redirected of the aromatic compound. Toluene induces the TOD pathways through a TCS system made of TodS/TodT.


140

IBBS-15-6-O01

Patchwork Assembly of nag-like Nitroarene Dioxygenase Genes and 3-Chlorocatechol Degradation Cluster for the Pathway Evolution of 2-Chloronitrobenzene Catabolism in Pseudomonas stutzeri ZWLR2-1

Hong Liu1, Shu-Jun Wang1, Jun-Jie Zhang1, Hui Dai2, Huiru Tang2, and Ning-Yi Zhou1*

1Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China

2State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Center for Magnetic Resonance, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan, 430071, China

*[email protected]

Pseudomonas stutzeri ZWLR2-1 utilizes 2-chloronitrobenzene (2CNB) as sole source of carbon, nitrogen and energy. To identify genes involved in this pathway, a 16.2-kb DNA fragment containing a conserved region of the ring-hydroxylating dioxygenase genes was cloned and sequenced. Of the products of the 19 open reading frames deduced from this fragment, CnbAc and CnbAd exhibited striking identities to the respective �- and �-subunits of the Nag-like ring-hydroxylating dioxygenases involved in the metabolism of nitrotoluene, nitrobenzene and naphthalene. Their encoding genes were flanked by two copies of insertion sequence IS6100. CnbAa and CnbAb are similar to the ferredoxin reductase and ferredoxin for anthranilate 1,2-dioxygenase from Burkholderia cepacia DBO1. E.coli cells expressing cnbAaAbAcAd converted 2CNB to 3-chlorocatechol with concomitant release of nitrite. Whereas the cnbCDEF gene cluster, homologous to a 3-chlorocatechol degradation cluster in Sphingomonas sp. strain TFD44, probably contains all the genes necessary for the conversion of 3-chlorocatechol to 3-oxoadipate. CnbC has been demonstrated to be a functional chlorocatechol 1,2-dioxygenase. The patchwork-like structure of this catabolic cluster suggests that the cnb cluster for 2CNB degradation has evolved by the recruitment of two catabolic clusters encoding a nitroarene dioxygenase and a chlorocatechol degradation pathway. This provides another paradigm to elucidate the bacterial evolution of catabolic pathways in response to xenobiotic chemicals.


141

IBBS-15-6-O02

Ecology and Function of Catabolic Plasmids for Biodegradation of Petroleum Hydrocarbons

Irina Kosheleva1,2, Sergei Sokolov1, Alexander Boronin1,2

1Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino 142290, Russia

2Pushchino State University, Pushchino 142290, Russia

The environment is constantly being polluted by various hydrocarbons originated from different sources. Petroleum hydrocarbons are known to be degraded by a variety of microorganisms. Great potential for developing and improving bioremediation technologies lies in improving our understanding of the metabolism and ecology of microorganisms capable of degrading oil and oil products.

Polycyclic aromatic hydrocarbons (PAH) are commonly found in crude oil and oil products. Degradation of PAH is often controlled by plasmids. Ecologically, plasmid-encoded pathways are advantageous because they provide genetically flexible systems and can be maintained in the population and transferred between bacterial species. Non homologous genetic systems involving in biodegradation of PAH and encoding isofunctional enzymes have been found in different species of bacteria. The IncP-9 and IncP-7 plasmids appear to be the common vehicle for degradative pathways and play an important role in dissemination of catabolic genes. Experiments on direct and exogenous isolation of plasmids show the prevalence of IncP-7 PAH degradation plasmids in heavily contaminated sites while IncP-9 plasmids seem to be ubiquitous. Direct evidence of dissemination of the naphthalene biodegradation plasmids in microbial populations in open soil under selective and nonselective conditions has been obtained.

Not only the type and amount of degradation plasmids but also the dynamics of plasmid transfer and thus, increased functional diversity within bacteria, should be used to assess the biodegradative potential of oil-contaminated sites.

This work was supported by ISTC #4033 and Analytical Program of Russian Ministry of Education and Science #2.1.1/10938.


142

IBBS-15-6-O03

Implication of rpoN loss-of-function in xenobiotic compounds degradation by Burkholderia xenovorans LB400

Noor Faizul Hadry Nordin1,3, Amir Feisal Merican1,2 and Yusrizam Sharifudin1,2

1. Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.

2. CRYSTAL, University of Malaya, 50603 Kuala Lumpur, Malaysia. 3. Department of Biotechnology, Kulliyyah of Science, International Islamic University Malaysia, 25200

Kuantan, Pahang, Malaysia. High genome plasticity of genus Burkholderia is likely responsible for their ability to occupy a broad spectrum of ecological niche by utilising a wide range of substrate including xenobiotic compounds. One of the factors involve in maintaining the fitness of bacteria such as Burkholderia is the alternative sigma subunit-54 (σ54). The core RNAP associated with sigma subunit forming holoenzyme (Eσ) complex can specifically recognise and initiate transcription from promoters. Alternative sigma subunit-54 enables specific sets of genes to be transcribed in response to environmental stimuli. Even though many of the characterised σ54-dependent promoters are involved in nitrogen metabolism, other previous studies clearly demonstrated that Eσ54 also control several other ancillary processes including degradation of xylene and toluene. Burkholderia xenovorans, a well-known degrader of polychlorinated biphenyls (PCBs) is also capable in degrading other xenobiotic compounds such as dioxins and polyaromatic hydrocarbons (PAHs). In this study, the loss-of-function mutants of the Burkholderia xenovorans LB400 rpoN genes were developed using pKNOCK suicide vector. Utilisation studies of dioxins and PAHs shows some significant differences between wild-type Burkholderia xenovorans and its rpoN mutants. This result suggests the utilisation and degradation of xenobiotic compounds by Burkholderia might be affected by rpoN loss-of-function. For further analysis, the degradation of dioxins and PAHs will be assessed using GC/MS.


143

IBBS-15-6-O04

Efficient colonization, competitive ability and the expression of hydrocarbon degradation genes correlates with efficient phytoremediation of soil contaminated with diesel fuel

M. Afzal, S. Yousaf, T.G. Reichenauer, V. Andria and A. Sessitsch

AIT Austrian Institute of Technology, Department of Health and Environment, A-2444 Seibersdorf, Austria

Plants in combination with their associated microflora can remediate soils that are contaminated with organic pollutants such as petroleum hydrocarbons. Inoculation of plants with efficient degrading bacteria is one approach to improve remediation processes. Several plant-associated bacteria, residing in the rhizo- or endosphere, have the capacity to degrade hydrocarbons, to promote plant growth or to alleviate plant stress. Although the potential of bacterial inoculation in phytoremediation applications has been demonstrated, the outcome of inoculation is sometimes not successful or leads to variable results. In many cases this is due to the fact that inoculant strains may insufficiently interact with or colonize plants used for phytoremediation and / or cannot compete with the resident microflora under certain environmental conditions. However, colonization and competitive ability of inoculant strains is generally rarely addressed, but is essential to deduce serious knowledge about the efficiency of an inoculation. We will present our results obtained from various experiments on the colonization capacity of several inoculants strains as well as catabolic gene expression in various plant environments (rhizosphere, root and shoot interior). Furthermore, we addressed the question how environmental parameters affect gene expression and colonization and to which extent inoculant strains compete with native bacteria. We will show how efficient colonization and expression of hydrocarbon degradation genes correlate with plant growth and efficient remediation of diesel fuel.


144

IBBS-15-6-O05

Development of endophytic and rhizospheric bacterial inoculants with remediation potential for practical application in the bio-energy crop industry

John Culhane, Nicholas Otieno, Kieran Germaine, Dina Brazil, David Ryan, David Dowling

Department of Science and Health, School of Science, Institute of Technology Carlow

Bacteria-assisted phytoremediation holds much promise for the sustainable removal of recalcitrant compounds from the soil environment. It involves the inoculation of plants with non-pathogenic bacteria that are proven xenobiotic degraders to assist in the accumulation and removal of these compounds that are otherwise toxic to their host. Previous research at Institute of Technology Carlow (ITC) has demonstrated that plants inoculated with bacteria capable of degrading compounds such as 2,4-Dichlorophenoxyacetic acid, a broad-leaf herbicide and naphthalene (PAH) can protect the host plant from the toxic effects of these chemicals and contribute to their removal from site. Endophytic and rhizospheric strains maintained and partially characterised at ITC have been selected and utilised in a ‘Smart Inoculation’ experiment. The experiment focused on introducing and colonising established bioenergy crops (Rye grass and Oilseed Rape) with “mastermixes” containing these strains with a view to investigating their possible benefits to the crops i.e. increased biomass or phytoremediation potential. Results have identified 2 mastermixes (20 strains) that have conferred increased growth in Oilseed Rape over a negative control (32%-36%). The strains comprised in these mixes are also being screened for resistance to heavy metals and, also, their ability to metabolise harmful or toxic organic compounds thus removing the threat they pose to the environment. Identifying and utilising strains with these traits coupled with their symbiotic growth promoting relationship with host plants could be of great benefit for the sustainable production of bioenergy crops.


145

IBBS-15-6-O06

Isolation of desiccation tolerant microorganisms for the rhyzoremediation of soils subjected to seasonal drought

L. SantaCruz-Calvo, J.I. Vílchez, J.J. Picazo-Espinosa, J. Narváez-Reinaldo, R. González-López, and M. Manzanera

Institute of Water Research, University of Granada, Granada, Spain

Microorganisms and plants establishment for the rhyzoremediation of recalcitrant pollutants such as polycyclic aromatic hydrocarbons (PAHs) in areas subjected to seasonal drought, is further impeded by additional physicochemical factors such as low precipitation, high evaporation, extreme temperatures, and high winds. These enviromental factors contribute to the extremely low water content in soil and reduce the survival of most of the isolated microbial strains with determined role in bioremediation of soils. We have set up a new technology in our laboratory for the isolation of desiccation tolerant microorganisms. To that end we have used the remarkable high stability that anhydrobiotic microorganisms present when dried to develop a selection method based on the use of organic solvents as selective agents. Since these solvents do not affect stable dried cells we can recover desiccation tolerant cells upon treatment. In this way we have used this tolerance to organic solvents as a selection method for the isolation of a collection of microbial isolates with increased desiccation-tolerance. We also determined that some of these microorganisms promote the growth of plants and therefore can be considered as PGPR under watering and drought conditions. Therefore we have developed biotechnological tools that could be useful for the rhyzoremediation of PAH-polluted soils in arid and semiarid regions.


146

IBBS-15-6-O07

Aromatics-Utilizing Bacteria and Their Functional Genes in Long-Term PCB and PAH Contaminated Soil

Ondrej Uhlik1,2, Lucie Musilova1, Michal Strejcek1, Jiri Wald1, Cestmir Vlcek3, Jakub Ridl3, Martina Mackova1,2, Tomas Macek1,2

1Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology Prague, Technicka 3, 166 28 Prague 6, Czech Republic

2Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, IOCB & ICT Joint Laboratory, Flemingovo n. 2, 166 10 Prague 6, Czech Republic

3Department of Genomics and Bioinformatics, Institute of Molecular Genetics, Czech Academy of Sciences, Videnska 1083, 142 20 Prague 4, Czech Republic

E-mail: [email protected], [email protected]

Certain autochthonous bacteria and fungi can degrade contaminants, such as polychlorinated biphenyls (PCB) or polycyclic aromatic hydrocarbons (PAH). The process of bioremediation is, however, under most environmental conditions slow. In order to make it more effective, researchers need to better understand microorganisms’ behavior directly in contaminated environments.

The aim of this study was (i) identification of metabolically active populations involved in intrinsic bioremediation, (ii) investigation of functional genes and linking their diversity to specificity towards different PCB congeners, (iii) tracking carbon flow from the molecules of (chloro)biphenyls, and (iv) evaluation of the influence of selected plants, their secondary metabolites or natural products on the diversity of metabolically active populations.

The main technique employed was stable isotope probing of DNA (DNA-SIP) which is considered one of the leading molecular tools for investigating the diversity of bacteria potentially responsible for ecologically relevant processes. Huge progress in application of DNA-SIP has been made by combining it with gene-targeted metagenomics, which is oriented to the analysis of amplicons of 16S rRNA genes or functional genes.

Results of this study contribute to our understanding of intrinsic bioremediation and diversity changes in response to the presence of plants or their secondary metabolites. They also confirm the importance of SIP as a method that allows for the identification of truly active members of the community, including those yet uncultured.

Acknowledgements: Work was supported by projects NPVII 2B08031, ME 10041, Z 40550506, MSM 6046137305 (Ministry of Education, Youth and Sports of the Czech Republic) and EU project FP7-KBBE-2010-4.


147

IBBS-15-6-O08 Polybrominated flame retardans (BFRs) and their fate in the Czech Republic K.Demnerová1, J.Vrkoslavova1, H. Stiborova 1, J.Purkrabová2 , J.Hajšlová2

1Department of Biochemistry and Microbiology, 2Department of Food Chemistry and Analysis Institute of Chemical Technology Prague, Technická 5, 166 28 Prague 6, Czech Republic [email protected]

Brominated Flame Retardants (BFRs) are used in various commercial and industrial applications to delay ignition and reduce the rate of combustion. They are commonly present in daily used products such as, computers, TVs, pipes, wires, cables, etc. Most of the BFRs are not covalently bound to polymeric materials and they are therefore over time released from the product into the environment, namely as vapour or dust. Due to their lipophilicity and persistence, they can be found in many environmental compartments, both biotic and abiotic. With regard to increasing concentration in human blood and breast milk, the production of commercial penta- and octa-BDEs technical mixtures was banned in the EU in 2004. However, due to their former wide usage, a release of lower brominated PBDE congeners into the environment still continues.

In this presentation, the data obtained within the national project BIOBROM will be introduced. One of the main tasks of this project was to investigate BFRs levels in the Czech ecosystem between 2006 and 2010 in various both abiotic and biotic matrices including river sediments, wastewater treatment plants sludge, human adipose tissue collected during the liposuction procedure, human breast milk, indoor household dust and car dust. The monitoring included PBDEs (polybrominated diphenyl ethers congeners 28, 47, 49, 66, 85, 99, 100, 153, 154, 183, 196, 197, 203, 206 and 209), HBCD (hexabromocyclodecane – isomers α, β, γ) and also six novel “emerging” non-PBDE BFRs including DBDPE (decabromodiphenyl ethane), BTBPE (bis-2,4,6-tribromophenoxy ethane), PBT (pentabromotoluene), PBEB (pentabromoethylbenzene), HBB (hexabromobenzene) and Br Indane (octabromo-1-phenyl-1,3,3-trimethylindane).


148

IBBS-15-6-O09

Isolation and Characterization of Melamine (2, 4, 6-triamino-1, 3, 5-triazine)-degrading Bacteria

Han Wang1, Heli Xu1, Anyi Hu1, Qian Sun1, Chang-Ping Yu1

1 Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen 361021, China

Melamine (2, 4, 6-triamino-1, 3, 5-triazine) is a nitrogen-containing heterocyclic compound belonging to the s-triazine family. Melamine is an important chemical intermediate to produce melamine resin, which has been broadly used in the manufacture of flame retardants and housewares. Melamine is considered as a contaminant on the horizon after the incidences of deliberate contamination of melamine and cyanuric acid in the infant formula, petfood and other animal feeds in several countries. China is the major exporter of melamine in the world. Our previous visit to a domestic melamine-producing factory (established more than 10 years) found milligrams per liter of melamine present in its final plant effluent. Concerns regarding the fate of melamine in the environment led us to find melamine-degrading bacteria in the vicinity of the factory. Currently, two strains of melamine-degrading bacteria were found from the gutter receiving treated wastewater inside the factory. Their most closely related bacterial species is Alicycliphilus denitrificans (accession number AJ418042) with less than 96% 16S rRNA gene similarity, suggesting these isolates might be novel melamine-degrading species. The isolates could use melamine concurrently as carbon and nitrogen sources to grow and ammeline, ammelide, cyanuric acid, allophanate and ammonia were identified as intermediates. Degradation ability of the isolates toward other s-triazine was investigated. Stable isotope fractionation was analyzed to reveal the kinetic isotope effect during melamine biodegradation. To our knowledge, this is the first report of isolation of melamine-degrading bacteria using melamine as both carbon and nitrogen sources from a long term melamine exposure site.


149

IBBS-15-6-O10

Biodegradation of polyethylene by bacterial laccase (phenol oxidase)

Miriam Santo, Ronen Weitsman and Alex Sivan.

Department of Biotechnology Engineering, Ben Gurion University of the Negev, Beer Sheva, Israel.

Polyethylene is considered one of the most durable plastic polymers. Practically non-biodegradable polyethylene accumulates in the environment posing an ecological threat to man and wildlife. We have previously isolated a strain of the actinomycete Rhodococcus ruber that was capable of utilizing and degrading polyethylene. Here, we report on the role of the bacterial copper-binding enzyme, laccase in the oxidation and degradation of polyethylene by this strain. Copper markedly affected the induction and activity of laccase, resulting in polyethylene degradation. RT-PCR, facilitating quantification of mRNA induction, showed a 13-fold increase in laccase mRNA levels as compared with the untreated control.Addition of copper to C208 cultures containing polyethylene enhanced the biodegradation of polyethylene by 75%, as compared with the non-amended control. Furthermore, when an extracellular isoform of laccase collected from the media of copper-induced cells was incubated with polyethylene, reductions of 20% and 15% were obtained in the Average Molecular Weight (Mw) and Average Molecular Number (Mn) with the polymer, respectively. FTIR analysis of similar polyethylene films incubated with the extracellular laccase exhibited an increase in the carbonyl peak, indicating that enzymatic oxidation by laccase plays a major in the biodegradation of polyethylene.


150

IBBS-15-6-O11 Phenanthrene degradation and strategies to improve bioavailability by microorganisms isolated from sediments and marine waters in Argentine Peressutti S.R1, Herrera Seitz K. 2, Pouyte K. 1, Pedetta A.2, Babay P.A.3, Espinosa M.3, Costagliola M.1, Studdert C.2

1Instituto Nacional de Investigación y Desarrollo Pesquero, Paseo Victoria Ocampo Nº1, (7600) Mar del Plata, Argentina. Tel.: +54-223-4862586; fax: +54-223-4861830. E-mail: [email protected] (S.R. Peressutti) 2Instituto de Investigaciones Biológicas (FCEyN, UNMdP), Funes 3250, 4to Nivel, UNMdP. 3Comisión Nacional de Energía Atómica, Centro Atómico Constituyentes, Av. Gral. Paz 1499, (1650) San Martín - Buenos Aires, Argentina. Tel.: +54-11-67727898; fax: +54-11-67727886. In the present work degradation of phenanthrene, a model PAH compound, by strains isolated from contaminated sediments and marine water was investigated. Eight strains were isolated from enrichment cultures; they belong to the genera Pseudomonas, Sphingomonas and Rhizobium as determined by 16S rRNA gene sequencing. All the strains were able to grow in mineral medium supplemented with phenanthrene as the sole carbon source, reaching counts of 109cell/ml. Phenanthrene disappearance was estimated by high performance liquid chromatography. Strains isolated in this work showed high PAH degradation potential, degrading between 90 and 100 of the phenanthrene added to the medium (200 mg/L -1) after 12 days. In order to study mechanisms that could influence PAH degradation, emulsifying activity and chemotaxis to phenanthrene were analyzed for all the strains. Several isolates showed clear chemotactic responses towards phenanthrene, as assessed by different methods. Also, various strains displayed emulsifying activity. Both chemotaxis and biosurfactants production may contribute to the enhancement of phenantrene bioavailability, and thus to the efficiency of in situ bioremediation of contaminated sites.


151

IBBS-15-6-O12

Biodegradation of Polyurethane under compost at different temperatures

Urooj Zafar 1, Geoff. D. Robson

Faculty of Life Sciences, Manchester University, Manchester, UK

Composting is a natural process involving the aerobic decomposition of organic wastes by a mixed microbial consortium that goes through a range of temperature changes due to the heat generated by microorganisms. Historically a major portion of municipal solid waste has been directed to landfills, resulting in the scarcity of available sites. Plastics are the major components of solid wastes and take a long time to degrade. A promising alternative could be composting of plastics rather directing them to landfill sites. Polyurethane, xenobiotic in nature, with a wide application has been taken as a model plastic to study the effects of a range of temperatures on degradation when buried under compost. Culture dependent technique was used to estimate the percentage of polyurethane degrading microorganisms in compost at 25, 45 and 55˚C. The isolated polyurethane degrading organisms were divided into different morphotypes and found that different groups of organisms dominate at different temperatures. Currently, the extent of degradation of polyurethane is being measured by assessing the loss in tensile strength at these temperatures. Culture dependent technique is used to quantify the microbial growth on solid polyurethane during degradation while culture independent techniques, such as DGGE, are being used to observe the succession of fungal community on polyurethane sheets at 25, 45 and 55˚C under compost.

This work is funded by HEC, Pakistan.


152

IBBS-15-6-O13

Crude desulfurization in organic:aqueous phase biocatalytic systems

I.F. Ferreira*1, C.C.C.R. de Carvalho1, D.I.C. Wang2, M.R. Aires-Barros1

1IBB- Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering. Instituto Superior Técnico, Avenida Rovisco Pais, 1049-001 Lisboa, Portugal.

2MIT- Massachusetts Institute of Technology. Chemical Engineering Department, 77 Massachusetts Avenue, 02139. Cambridge, USA.

In 2009, the crude oil demand in the world was c.a. 84 millions of barrels per day, generating around US$ 6 billion. During transformation and use of crude oil derivatives e.g. diesel and gasoline, aromatic and sulfur-containing toxic compounds are formed and released. Consequently regulatory agencies have been limiting the total amount of compounds such as sulfur. Petroleum companies are using hydrodesulfurization, an expensive chemical process, able to transform sulfur compounds into H2S but that is often ineffective when eliminating some complex organic compounds such as benzothiophenes. Biodesulfurization, BDS, offers a sustainable and environmentally friendly approach to eliminate sulfur containing compounds from crude oil.

In this work we evaluated the feasibility of using two different Rhodococcus strains for the desulfurization of crude oil using organic:aqueous phase biocatalytic systems. The effect of different phase ratios was also studied. The results showed that both strains were able to change their surface properties and membrane composition in fatty acids in order to adapt to both the organic phase and DBT. Moreover, it was observed that the cells migrated towards the organic:aqueous interface or were even in the organic phase itself. This allowed the cells to be in direct contact with the DBT-rich phase. The cells were capable of being in harsh conditions, maintaining both viability and high metabolic yields when n-alkanes were used as organic phase. When n-hexadecane was used as DBT reservoir, resting cells of R. erythropolis achieved a desulfurization yield of 100% and 60% in less than two hours for an initial DBT concentration of 0.1 mM and 1 mM, respectively.


153

IBBS-15-6-O14

Toluene Biodegradation in Bio electrochemical Cells (BEC) simulating microbial fuel cell (MFC)

H. Friman1,2, A. Schechter3, Y. Nitzan2 and R. Cahan1

1The Ariel University Center in Samaria, Ariel, ISRAEL, 2Bar-Ilan University, Ramt-Gan, ISRAEL,

3Department of Biological Chemistry, Ariel University Center of Samaria, Ariel 44837, Israel

The treatment of aromatic hydrocarbons in wastewater resulting from oil spills and chemical manufactories is becoming a key concern in many modern countries. Benzene, ethylbenzene, toluene and xylene (BETX) contaminate groundwater as well as soil. These compounds have an acute effect on human health and are known to be carcinogenic. Conventional removal of these toxic materials involves separation and burning of the wastes, however, the cost of chemical treatment is very high and energy consuming. This research focused on the effect of low voltage on Pseudomonas putida F1 grown in a Bio Electrochemical Cell (BEC) using toluene as a sole carbon and energy source. The BEC apparatus was designed as a typical two chambers microbial fuel-cell (MFC), but was operated under external constant voltage.

In this study we examined the effects of applied low external voltage on Pseudomonas putida F1 bacterial growth, electric transfer, toluene degradation, morphology and the bacterial proteome. Toluene degradation by Pseudomonas putida F1 in BEC under external voltage of 125, 250 and 500 mV (vs Ag/AgCl) led to increase of the bacterial cells growth 0.75, 0.6 and 0.5 OD respectively, compare to the control BEC (OD of 0.3). Examination of the current generated under external voltage of 125, 250 and 500 mV show that the maximal current was 24, 54 and 94 mA/m2 respectively. The majority of the current was received from the plankton bacterial cells. Toluene degradation is higher in BEC polarized to 125 mV than in the non-polarized control experiment. The residual toluene concentration after 139 h in BEC (125 mV) was 54 mg l-1 while in the control BEC the residual toluene was 75 mg l-1. Examination the proteome of bacterial cells growing in BEC (125 mV) shows the appearance of two groups of protein which are ascribed to electrons transfer to the electrode. Operating the BEC under external voltage of 500 mV has a detrimental effect on the bacterial cells however, operating under 125 mV enable bacterial growth and toluene degradation even in limited oxygen environment.

In summary, the BEC suggested in this research has a feasible potential for electro-bioremediation of toluene in industrial wastewater under limited dissolve oxygen.


154

IBBS-15-6-O16

Bioavailability and dissipation of anthracene from soil with different alkalinity and salinity

Carolina Castro-Silva, Víctor Manuel Ruiz-Valdiviezo, Sandra Gabriela Rivas-Rivera, Alma Rosa Sosa-Trinidad, Marco Luna-Guido, Rodolfo Marsch,Luc Dendooven*

Laboratory of Soil Ecology, GIB, Cinvestav, Av. I.P.N. 2508 C.P. 07360, México D. F., México. * Author for correspondence: Luc Dendooven, Tel: +52 55 5747 3319, Fax: +52 55 5747 3313, E-mail: [email protected]

Bioavailability of contaminants, such as anthracene (Anthra), a polycyclic aromatic hydrocarbon, has been related to their extractability with non-exhaustive techniques, such as hydroxypropyl-beta-cyclodextrin (HPCD) or n-butanol. Bioavailability of Anthra was determined with HPCD and n-butanol and its removal was monitored in sterilized and unsterilized soils of Texcoco, Mexico, with alkalinity ranging from pH 8.2 - 10.1 and electrolytic conductivities (EC) of 1.2, 3.2, 80.2 and 95.2 dS m-1 with an exhaustive ultrasonic extraction method during an aerobic incubation of 56 days. Biotic removal of Anthra, most accentuated in the first days, varied between 80 mg kg-1 and 169 mg kg-1 after 56 days and was not related to the measured soil characteristics. Bioavailability of Anthra, as measured by its extraction with n-butanol, nearly halved within the first 14 days with little change thereafter, and was not affected by soil characteristics and not related to the biotic dissipation. The amount of Anthra extracted with HPCD from soil was low and varied between 20 and 30% and it was not related to its biological dissipation. In soils with the highest EC and pH, more Anthra was removed than was available as defined with HPCD. It was found that the characteristics of alkaline saline soils did not affect the removal of Anthra from the Texcoco soil. The bioavailability of Anthra as defined with n-butanol and HPCD was not correlated to the biological removal of Anthra from soil.


155

IBBS-15-6-O19 Influence of rhizosediments characteristics on hydrocarbon degradation potential of Juncus maritimus plant-microorganisms associations Hugo Ribeiro1,2,3, Ana P. Mucha2, C. Marisa R. Almeida3, and Adriano A. Bordalo1,2

1 Laboratório de Hidrobiologia, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS-UP), Universidade do Porto, Largo Professor Abel Salazar, no. 2, 4099-003 Porto, Portugal 2 Laboratório de Hidrobiologia do CIMAR/CIIMAR – Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Porto, Portugal 3 Laboratório de Especiação Química e Biodisponibilidade (CS&B) do CIMAR/CIIMAR – Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto Porto, Portugal

Abstract

The influence of colonized sediments (rhizosediments) properties on hydrocarbon degradation potential of Juncus maritimus plant-microorganisms associations were investigated in four salt-marsh sites within the lower and middle stretches of a temperate estuary (Lima River, NW Portugal). Rhizosediments were sampled for hydrocarbon degrading microorganisms (HD, estimated using a modified most probable number protocol), total cell counts (TCC, obtained by DAPI direct count method) and total petroleum hydrocarbons (TPHs, determined by FTIR spectrophotometry after ultrasonic extraction) assessment. Rhizosediments were also characterized in terms of organic matter percentage (OM), water content, and particle size distribution. Additionally, evaluation of TPHs degradation potential was assessed from fortnight laboratory experiments carried out in 50 mL microcosms, where rhizosediments were mixed with Bushnell Haas medium with and without crude oil amendment. The OM, water and silt + clay percentages were significantly different among rhizosediments, increasing downstream. Generally, TPHs and TCC also followed the same trend. However, HD had a distinct behavior. Considering the three upper most sampling sites, there was a significant (p<0.05) negative correlation between HD and silt + clay content. Control experiments without amendment yielded different TPHs degradation rates, with higher rates in the upstream rhizosediments. Also, a significant (p<0.05) negative correlation between TPHs degradation rates and MO and silt + clay content, was found, suggesting the influence of rhizosediment characteristics on hydrocarbons bioavailability. In fact, in experiments with crude oil amendment, rhizosediment characteristics had no influence on degradation rates, probably because hydrocarbons were equally bioavailable. Our data suggest that sediments properties can, indeed, influence HD abundance and TPHs degradation rates, and should be considered on rhizoremediation strategies in estuaries.

Acknowledgements: This work was partially funded by Fundação para a Ciência e Tecnologia (FCT), Portugal, through the project PTDC/MAR/099140/2008, and the PhD fellowships awarded to Hugo Ribeiro (SFRH/BD/47631/2008).


156

IBBS-15-6-P01

Effect of surfactants on PAH biodegradation process by a bacterial consortium and dynamics of the bacterial community during the process

Natalia González1, Raquel Simarro1 , M. Carmen Molina1, L. Fernando Bautista2, Laura Delgado1, Juan A. Villa1

1 Department of Biology and Geology, 2 Department of Chemical and Environmental Technology. Universidad Rey Juan Carlos. C/ Tulipán s/n. E-28933 Móstoles, Madrid, Spain.

Abstract

The aim of this work was to evaluate the influence of the non-biodegradable and biodegradable surfactants (Tergitol NP-10 and Tween-80, respecitvely) on the growth, degradation rate and microbial dynamics of a polycylic aromatic hydrocarbon (PAH) degrading consortium (C2PL05) obtained from a petroleum polluted soil by applying culturable and non-culturable techniques. The consortium was incubated in BHB medium during 45 days at 25ºC in an orbital shaker with a mixture of naphthalene, phenanthrene and anthracene, and Tween-80 or Tergitol NP-10 as surfactant. With the non-biodegradable surfactant, growth and degradation rate of the microbial consortium were significantly lower than with the biodegradable surfactant. The medium toxicity did not show any significant reduction with the non biodegradable surfactant, whereas with the biodegradable surfactant, toxicity was almost eliminated (30%) after 40 days of cultivation. Regarding to the cultured bacteria, Pseudomonas and Stenotrophomonas groups were dominant during PAH degrading process with Tergitol NP-10 treatment, whereas Enterobacter and Stenotrophomonas were dominant with Tween-80. DGGE analyses (PRIMER and MDS) showed that bacteria composition was more similar between treatments when PAHs were consumed (end of the incubation time) than when PAHs concentration was still high (initial time). Community changes between treatments were a consequence of Pseudomonas sp., Sphingomonas sp., Sphingobium sp. and Agromonas sp.


157

IBBS-15-6-P03

Laboratory-scale microbial bioremediation of pinewood-tar following a steam gasification process G. Valducci Vecchia, M. Baratieria, S. Angelia, M. Grigianteb and L. Brusettia,* a Faculty of Science and Technology, Free University of Bolzano, piazza Università 5, 39100 Bolzano, Italy. b Department of Civil and Environmental Engineering, University of Trento, via Mesiano 77, 38100 Trento, Italy. * Corresponding author: [email protected] Biomass is one of the most attractive substitutes to conventional fossil fuels, for energy production through gasification. Since this process usually leads to significant tar concentrations in the synthesis gas, clean-up stages are foreseen in the plant layout. Tar originated by wood gasification consists of a mixture of organic compounds of high molecular mass. For this reason, wood-tar is of particular concern because of its carcinogenicity, toxicity and recalcitrance; and its disposal after the clean-up is still problematic. In this project microbial consortia were used to treat an experimental wood-tar, to study their microcosms evolution and their degradation activity. Pinewood sawdust was subjected to steam gasification at 800°C. The resulting wood-tar sample was suspended in water and inoculated in microcosms. A DGGE analysis and DNA band sequencing of the overall DNA extracted from each microcosm at the early and latter stage of treatment were performed in order to recognize the dominant species. The variations in the volatile chemical composition of tar were identified through direct headspace GC-MS analyses, in order to evaluate the actual effectiveness of catabolic action of bacteria and fungi. 2-butanone, diethoxy methane, furfural, ethyl acetate, 1,1-diethoxy ethane, 2-ethoxytetrahydrofuran, 1-methoxy 2-propyl acetate, 2,5-diethoxytetrahydrofuran, 2-methoxy-4-metyl phenol were tentatively identified in the headspace of untreated tar-water suspensions. After 20 days of incubation, a consistent reduction (even 100%) of some compounds was noted.


158

IBBS-15-6-P04

Improving the bioremediation abilities of Rhodococcus erythropolis


Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Department of Bioengineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal ([email protected])

Rhodococcus erythropolis cells present a considerable natural tolerance and adaptation ability to compounds usually toxic to other bacterial strains, including terpenes (1), hydrocarbons (2) and aromatic compounds such as toluene and xylene (3). Hydrocarbons could be metabolized both at 15 and 28 ºC even in the presence of 1.00%, 1.95% and 2.50% sodium chloride (4).

Most oil spills and soil contaminations occur under conditions far from the ideal conditions tested in the laboratory. The bacterial cells often have to deal with high/low temperature, high salt concentration and extreme pH values.

In the present study, R. erythropolis cells were adapted to both low and high temperatures and to high concentrations of sodium chloride, cupper sulfate and toluene. Biodegradation of alkanes and alcohols was used to evaluate the activity of the cells under extreme conditions.

The adaptation mechanisms presented by the cells involved: i) alterations in the cell wall and membrane composition; ii) modifications of the physicochemical properties of the cell surface; iii) increased degradation or bioconversion of the toxic compounds; iv) cell aggregation, and v) production of exopolymeric substances, including biosurfactants.

Using a stepwise adaptation strategy, the cells could degrade C6-C16 alkanes and C1-C12 alcohols at 4-37ºC, pH 3-11, in the presence of up to 7.50% NaCl and 1.00% CuSO4.

References:

(1) de Carvalho CCCR, Poretti A, da Fonseca MMR (2005) Applied Microbiology and Biotechnology 69:268-275.

(2) de Carvalho CCCR, Wick LY, Heipieper HJ (2009) Applied Microbiology and Biotechnology 82:311-320.

(3) de Carvalho CCCR, Fatal V, Alves SS, da Fonseca MMR (2007) Applied Microbiology and Biotechnology 76:1423-1430.

(4) de Carvalho CCCR, da Fonseca MMR (2005) FEMS Microbiology Ecology 51: 389-399.


159

IBBS-15-6-P06

Biodegradation of heat treated commercial Polyethylene by extracellular enzymes of Staphylococcus epidermis species bp/su1.

Bappaditya Roy Rajat Banerjee Sumana Chatterjee

Department of Chemistry , Basanti Devi College,147 /B Rashbehari Avenue, Kolkata 700029

India.

Abstract:

BP/SU1 can survive in a mineral media with shredded polyethylene as its only carbon source. During the process of its assimilation of carbon from polyethylene and its subsequent growth, it exudes some extracellular material into the mineral media. This extracellular supernatant is mainly composed of proteins and complex carbohydrates similar to the composition of the biofilm producing matrix in the same organism. This supernatant separated from the cells and incubated with pieces of polyethylene showed tangible evidence of polyethylene degradation by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The protein components of this mixture are separated by one dimensional sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS-PAGE) analysis. Tryptic digestion of the 38.5 kDa protein which is overexpressed when the mineral media is supplemented with shredded polyethylene is followed by matrix assisted laser desorption and ionization –mass spectroscopy (MALDI MS).Subsequent comparative proteomic analysis of the MS-MS data of this protein using MASCOT software showed almost 130 protein score with the Fructose bis phosphate (class I) aldolase group of enzyme from the same organism. It is highly probable that degradation of autoclaved shredded LDPE (low density polyethylene) occurs through a similar process like the enzymatic action of aldolase which catalyzes cleavage of C-C bond in hexose sugar.


160

IBBS-15-6-P07

Analysis of the Technologies Solutions applied to Oily Waste Treatment from Oil Industry Leonardo Jordão da Silva1, Flávia Chaves Alves2, Francisca Pessôa de França2

1Ministério da Ciência e Tecnologia, Esplanada dos Ministérios, Bloco E, Sala 589, 70067-900, Brasília, DF, Brasil Tel.: +55 61 3317-8663 Fax: +55 61 3317-7768 - E-mail: [email protected] 2Escola de Química, Universidade Federal do Rio de Janeiro, Av. Athos da Silveira Ramos, 149, 21941-909, Rio de Janeiro, RJ, Brasil Tel.: +55 21 2562-7621 Fax: +55 21 2562-7567 – E-mail: [email protected] The oil industry activities result several impacts on the environment due to large amounts of oily waste are generated. The oily sludge is a semi-solid material composed by a mixture of clay, silica and iron oxides contaminated with oil, produced water and chemicals used in the production of oil. Nowadays both the treatment and management of these wastes is essential to promote sustainable management of exploration and exploitation of natural resources. Biological, physical and chemical processes can be used single and/or combined in order to reduce environmental contamination by petroleum hydrocarbons into acceptable levels. The choice of treatment method depends on the waste physical-chemical properties as well as the availability of facilities to process these materials. Literature provides some operations and strategies for treatment and final disposal of oily waste, such as landfills, landfarming, incineration, co-processing (cement kiln), biopiles, centrifugation, destructive distillation, thermal oxidation, low temperature conversion, bioreactors, incorporation in ceramic materials, encapsulation, among others. The management of technology to be applied for the treatment of oily waste is essential to promote proper environmental management, prevailing alternatives to reduce, reuse and recycling the residue.


161

IBBS-15-6-P09

Emulsification and Biodegradation of Crude Oil by Bacteria Isolated from Petroleum-Contaminated Soil from Bahrain and Kuwait

Wael A. El Moslimany1, Noura A. Alhamad1, Wael S. El Sayed2, Ashraf Al Nayal1, Riyad Y. Hamzah1

1Biotechnology Program, College of Graduate Studies, Arabian Gulf University, Kingdom of Bahrain, Manama P.O. Box 26671, 2 Taiba University, Faculty of Science, Medina, Universities Avenue, P.O. Box 344

Several bacterial strains were isolated from petroleum-contaminated soil via enrichment in minimal medium containing 2% Arabian light oil as the sole carbon source. Based on metabolic fingerprinting using the API 50CH and API 20E (BIOMERIEUX) identification system, the isolated strains could be affiliated to different species of the genera Bacillus and Brevibacillus. The isolated strains produced stable oil-in-water emulsions within 1 to 7 days of incubation at 30 �C. The oil droplets of the produced emulsions had various sizes, indicating the production of different types of biosurfactants/bioemulsifiers. Cell-free culture supernatants produced clearing zones with different diameters in the semi-quantitative oil displacement assay. This suggests that emulsification was due to biosurfactants/bioemulsifiers that were released into the growth medium. The cells remained attached to the oil droplets in most of the cultures even after 6 weeks of incubation as indicated by the clear aqueous phase. In some cultures little turbidity was observed in the growth medium after two days of incubation, however, the turbidity remained constant afterwards. Gravimetric analysis of residual oil after 5 weeks of incubation at 30 �C revealed 20% to 36% loss of the original amount added to the shake flask cultures as compared to uninoculated controls. One isolate emulsified and utilized water-in-oil emulsions (26% degradation after 5 weeks), so called chocolate mousse, that are known to be microbiologically recalcitrant.


162

IBBS-15-6-P10

Characterization of Microbial Consortium Reductively Dechlorinating 2,3-dichlorophenol and 2,4,6-trichlorophenol

Wael S. El-Sayed1

1Microbiology Department, Faculty of Science, Ain Shams University, Cairo, Egypt

& Biology Department, Faculty of Science, Taibah University, Almadinah, KSA.

Abstract

A stable anaerobic microbial consortium reductively dechlorinating 2,3-dichlorophenol and 2,4,6-trichlorophenol was established from sedimentary environment. Defined sediment-free medium showing stable dechlorination activity was obtained in which dechlorination was restricted to the ortho chlorine in cultures enriched with hydrogen, acetate, and 2,3-DCP and to the para chlorine of 2,4,6-TCP cultures.

Chlororespiring species in established consortium were analyzed by PCR-denaturing gradient gel electrophoresis of 16S rRNA gene fragments, and at least 5 dominant bacteria were detected in dechlorinating cultures. Dominant fragments were sequenced and revealed their relation to members of Proteobacteria. Reductive dechlorination was confirmed by the stoichiometric dechlorination of di- and tri-CPs to lower ones and by the detection of chlorophenol RDase cprA gene fragments in chlorophenol-dechlorinating cultures. PCR recovered cprA gene fragments were cloned and sequenced and were found to cluster to cprA/pceA type of Dehalobacter restrictus. Only active dechlorinating cultures showed amplification of cprA gene confirming dehalorespiration on chlorophenols


163

IBBS-15-6-P12

Removal of Polycyclic Aromatic Hydrocarbons by Selected Strains Isolated from Organic Materials and Humic Acids García-Díaz, C., Ponce-Noyola, T., Esparza-García, F.J., Rivera-Orduña, F., Ríos-Leal E. and Barrera-Cortés J. Departamento de Biotecnología y Bioingeniería. Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN). Av. Instituto Politécnico Nacional No. 2508. Col. San Pedro Zacatenco. México 07360, D. F. Tel. (52-55) 5747-3800 Ext. 4380/4388. e-mail: [email protected] ABSTRACT: The purpose of this work was the isolation, characterization and isolation of hydrocarbonoclastic bacteria (HB) isolated from earthworm Eisenia andrei (EW), vermicompost (VC), leonardite extract as humic acids (LE) and sugar cane bagasse (SCB). Bacteria isolation was carried out by serial dilution and plated in nutritive agar, previous enrichment of the HB with kerosene. Seventy four HB were isolated: 15 from the LE, 15 from the VC, 18 from the EW and 26 from the SCB. The ability of these HB (and their mixtures) to remove 500 mg/kg of a PAHs mixture (phenanthrene, anthracene, fluoranthene, chrysene, and, benzo[a]pyrene; 100 mg/kg each PAH) was tested in liquid medium with and without kerosene added at a concentration of 5000 mg/kg. The highest PAH removal percentages (45%) were obtained with the trial prepared with LE and SCB. This trial was composed by 41 strains, that after identification by molecular biology methods, were classified within 15 species (Dietzia sp., Rhodococcus sp., Gordonia sp., Mycobacterium sp. Streptomyces sp., Kocuria sp., Microbacterium sp. Arthobacter sp., Brevibacterium sp. ensifer sp., Rhizobium sp. Naxibacter sp. Delftia sp., Stenotrophomonas sp. and Bacillus sp.) of the follow phylum: 9, Actinobacteria; 5, Proteobacteria and 1 Firmicutes. Considering the levels of PAHs removal obtained with the microflora extracted from these materials, we hope that this work will contributes to the microbiology of composting that uses low amounts of lignocellulosic materials for the treatment of soils highly contaminated with weathered hydrocarbons.


164

IBBS-15-6-P13

Screening of microorganisms with curcumin-converting ability

Azam Hassaninasab, Yoshiteru Hashimoto and Michihiko Kobayashi

Institute of Applied Biochemistry and Graduate School of Life and Environmental Sciences, The University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan

Curcumin, the major yellow coloring ingredient extracted from turmeric (Curcuma longa L., Zingiberaceae) has a variety of applications. It has been used for centuries as a yellow food dye and in traditional oriental medicine. It has a wide range of biological activities including antioxidant, anticancer, anti-inflammatory, antitumor and analgesic.

Despite the impressive applications of curcumin, basic investigations have lagged far behind those directed at biosynthesis, biotransformation and biodegradation.

To isolate the microorganisms with curcumin-converting ability, samples were inoculated in a test tube containing liquid medium with the addition of curcumin. After acclimatization culture, the culture solution was spread on agar plates and growing microorganisms were isolated. The curcumin bioconversion abilities of the isolates were analyzed using the cell-free extracts. The residual amount of curcumin was determined by HPLC, and one microorganism with the highest curcumin-converting activity was selected. To indicate whether degradation of curcumin is enzymatic or not, cell-free extracts prepared from the isolated microorganism were heat-treated by boiling for 10 min and used for the assay. The amount of curcumin did not decrease in the reaction mixture including treated cell-free extract, demonstrating the transformation is an enzyme reaction. These findings indicated that the strain contains an enzyme with curcumin-converting ability.


165

IBBS-15-6-P14

Biodegradation of polychlorinated biphenyls by Ceriporia sp. ZLY-2010: enzymes production and degradation rate

Chang-Young Hong 1, Ki-Seob Gwak 1, Su-Yeon Lee1, Seon-Hong Kim1, In-Gyu Choi1†

1 Dept. of Forest Sciences, College of agriculture and life sciences, Seoul National University, Korea

White rot fungi can be used for degradation of pollutants including polychlorinated biphenyls. In this study, Ceriporia sp. ZLY-2010 was selected to degrade Aroclor 1254 and 1260.

To evaluate degradation rates of Aroclor 1254 and 1260 by Ceriporia sp. ZLY-2010, liquid low-nitrogen medium was used and Aroclors were spiked 10 ppm in the medium. The results of biodegradation rate of Aroclors by Ceriporia sp. ZLY-2010 are as follows: the degradation rate of Aroclor 1254 reached 57.57% within 13 days, whereas the maximal degradation rate of Aroclor 1260 was 49.43% within 13 days. This result, compared with a previous study, indicated the higher degradation rate of PCB by Ceriporia sp. ZLY-2010.

To determine the ligninase activity related to the Aroclors degradation by Ceriporia sp. ZLY-2010, the protein contents and enzyme activities during cultivation were measured. So far, the ligninase activity of sample containing Aroclor 1254 was completed and that of Aroclor 1260 is ongoing. The extracellular protein contents were slightly affected by Aroclor 1254 in culture media. The total protein in the culture medium showed slight differences between the samples containing Aroclor 1254 and the control. Manganese-dependent peroxidase (MnP) activity rapidly increased with the addition to Aroclor 1254, indicating that MnP is related to Aroclor biodegradation. Laccase activity was lower than MnP activity when Aroclor 1254 was added, which implies that MnP had a greater influence on Aroclor 1254 biodegradation than Laccase. Furthermore, analysis of the genomic sequence of Ceriporia sp. ZLY-2010 related to degradation of PCBs is in preparation.


166

IBBS-15-6-P15

Isolation and characterization of fungal strains capable of degrading Polyvinyl chloride (PVC)

Author’s: Muhammad Ishtiaq Ali, Safia Ahmed and Abdul Hameed

Affiliation: Department of Microbiology, Quaid-i-Azam University, Islamabad Pakistan 45320

ABSTRACT

Polyvinyl chloride (PVC) has been included among those synthetic polymers which have got huge commercial application. Such chemical compounds due to their persistent chemical nature put a great stress on the environment where they are released. The present study was carried out to isolate and identify fungal strains capable of adherence and to degrade PVC . Low molecular weight thin films of PVC were buried in soil containing sewage waste. . A significant fungal growth and attachment was apparent on PVC film after 10 months of soil burial These fungal strains were identified on morphological and rRNA gene sequence basis as Phanerochaete chrysosporium PV1, Lentinus tigrinus PV2, Aspergillus niger PV3 and Aspergillus sydowii PV4. The biodegradation capabilities of fungal strains was further checked in shake flask experiments by taking thin films of PVC in Mineral Salt Medium (with no Carbon source) by taking as pure cultures. The biodegradation activity was analyzed by visual observation, plate assay, biomass quantification, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and gel permeation chromatography (GPC). PVC showed improved degradation pattern and changes in its physical properties due to fungal adhesion and activity.


167

IBBS-15-6-P16

Fuel component degradation and associated microbial community dynamics during the bioremediation of polluted soil in a biopile Andrés Izquierdo1, Joaquim Vila1, Lina Ortega2, and Magdalena Grifoll1

1Department of Microbiology, University of Barcelona, Barcelona, Spain 2Geotecnia 2000, Grupo Atisae, Tres Cantos-Madrid, Spain

The main interest of our group is to understand the microbial processes determining the fate of PAHs in soils, and their application in the optimization of bioremediation technologies and in risk assessment. We have a broad experience in the analysis of metabolic pathways involved in the degradation of single PAHs or PAH- environmental mixtures (creosote, crude oil) by single bacterial strains and enrichment cultures (consortia). Here we investigate the changes in hydrocarbon composition during the bioremediation of a fuel polluted soil and those in the microbial community structure in order to link the disappearance of specific substrates to key natural microbial populations.

The industrial soil under study was polluted due to repetitive accidental leaks of fuel containing underground storage tanks during fifty years. The preliminary risk assessment analysis recommended an aerobic biopile treatment. The homogenated soil was amended with nitrogen and phosphorous and treated in actively aerated biopiles during 90 days. Monthly soil samples were analysed for hydrocarbon composition (GC-MS) and microbial community structure using both molecular (PCR amplification of 16S rRNA genes followed by DGGE and clone library analysis) and culture-dependent methods. Chemical characterization of the residues will be used to investigate possible strategies to reduce the end-point concentrations.


168

IBBS-15-6-P17

The Multiingredient Biosurfactant - enhanced Biodegradation of PAHs

Vladimir Jirku, Barbora Kosova , Alena Cejkova and Jan Masak

Institute of Chemical Technology, 166 28 Prague 6, Czech Republic

Persistent pollutants are mostly C sources of relatively low solubility. To enhance their bioavailability, and accordingly the efficiency of their biodegradation, the use of chemically or biologically produced surfactants has been investigated. Synthetic surfactants, if applied above the critical micelle concentration, have been reported to be toxic toward water / soil mikroflora. Therefore, biosurfactants manifesting no inhibitory effects are in great demand. In this connection, different bacterial strains were used in the preliminary screening assays to select a multiingredient (rhamnolipid) biosurfactant producing strain. Different mixtures of carbon sources, as well as other modifications of cultivation media / conditions were used to affect reproducibly the yield and pattern of growth-associated production of stable mixture of these glycolipids. In the case of selected taxon : Pseudomonas aeruginosa , this study deals with optimization of rhamnolipid production, purification, identification, and evaluation of produced rhamnolipid mixture in a view of its biological activity, i.e., membrane permeabilization, protein leakage, changes in cell surface hydrophobicity and the effect on natural biofilm formation in bacterial and yeast populations exposed to obtained mixture of rhamnolipids. Cell populations of model biodegraders of naphthalene and phenanthrene exhibit significantly enhanced capability to utilize these compounds in technological cultures containing this multiingredient, not toxic biosurfactant.


169

IBBS-15-6-P18

The influence of temperature on degradation of the compostable plastic packaging material polylactic acid

Mehlika Karamanlioglu, Pauline Handley, Geoff Robson

Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester, M13 9PT, Manchester, UK

Poly(lactic acid) (PLA) is a synthetic aliphatic polyester with a hydrolysable backbone that is susceptible to degradation on the contrary of conventional plastics. As PLA short shelf-life products are thrown away after their use and they have the potential to replace conventional plastics, monitoring degradability of PLA in different environment conditions and understanding their degradation mechanisms are the main concerns of this study.

The effects of temperature, burial environment (soil and compost) and fungi on degradation of PLA were studied. PLA pieces were buried in compost and in soil at different temperatures (25-55ºC) and recovered over different time intervals. To determine degradation, gel permeation chromatography (GPC) and tensile strength measurements were performed. Results showed that molecular weight and tensile strength of PLA pieces are unaffected at 25° and 37°C, start decreasing at 45ºC and show the fastest rate of degradation at 55ºC. The degradation was not significantly different between soil and compost. Putative PLA degraders were isolated from recovered PLA pieces at each temperature and identified by rDNA sequencing. As the role of hydrolysis and microorganisms in PLA degradation are not clear, rate of hydrolysis in sterile medium was compared with the rate of degradation in compost and soil which suggested the first step of PLA degradation is hydrolysis.


170

IBBS-15-6-P20

The Removal of PAHs from Contaminated Soil: Effectiveness of Seeding a Root Colonising Bacteria

1Mphekgo P. Maila, 2Priscilla Randima and 2Thomas E Cloete

1Agricultural Research Council, Private Bag X79, Pretoria, 0001, South Africa (e-mail: [email protected]); 2Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, South Africa

Abstract

In this study, the potential of seeding a root colonising biodegrader in the soil was evaluated. Pseudomonas putida was seeded on the roots of Eleusine corocana in order to ascertain its feasibility to colonize the roots of the plant in different conditions. Scanning Electronic Microscopy (SEM) was useful in visualising the morphological structure of the bacteria that are able to colonize the roots of E. corocana. Examination by SEM and culture based techniques revealed that the roots that were inoculated and embedded with nutrients and Polycyclic aromatic Hydrocarbons (PAHs) had abundant colonized bacteria than other treatments. These results corroborated those of the rhizosphere enrichment cultures using optical density (OD) values and plate count technique, which shows that root colonizing bacteria, are predominant in the microcosms used. The high feasibility of colonizing the rhizosphere can be attributed to the affinity of the bacteria relative to the type of exudates secreted by the plant and the prevailing conducive environment that is favourable to the proliferation of the bacteria. The result obtained in this study also demonstrated a distinct ability of P. putida to colonize and grow differently with time, in different conditions. The results suggest that rhizoremediation can be improved by seeding the biodegraders in the root zone of the plants.


171

IBBS-15-6-P21

Degradation of high p-Nitrophenol concentrations Arthrobacter sp. isolated from agricultural soil.

Igor Kviatkovski1,2 , Shay Zohar1and and Segula Masaphy1,2,*

1Applied Microbiology and Mycology Department, MIGAL, P.O. Box 831, Kiryat Shmona 11016, Israel, 2 Tel Hai Academic College, Upper Galilee 12210, Israel *Author for correspondence; E-mail: [email protected], Tel: 972-4-6953519; FAX: 972-4-6944980

p-Nitrophenol (PNP) is one of the most important environmental contaminants among nitroaromatic compounds. It is toxic to many living organisms and may accumulate in the food chain. Whereas in the past, studies focused on low PNP concentrations, more recently, attention has turned to the isolation of bacteria capable of degrading PNP at high concentrations which may persist in industrial wastewater or in different technologies aimed at PNP removal. We have isolated a new Arthrobacter sp. capable of utilizing high PNP concentration as sole carbon, nitrogen and energy source, from local agricultural soil. The isolate was tolerant up to 800 mg/L PNP, and was able to degrade up to 600 mg/L PNP completely.


172

IBBS-15-6-P22

Stereospecific degradation of phenylsuccinate by actinomycetes.

Toru Matsui,1* Katsuya Kato,2 Tomoyuki Namihira,1 Naoya Shinzato,1 and Hisashi Semba3

1Center of Molecular Biosciences, University of the Ryukyus, Okinawa 903-0213, Japan

2National Inst. Advanced Industrial Science and Technology (AIST), Nagoya, Aichi 463-8560, Japan

3Strategic Technology Research Center, Nippon Shokubai Co., Tsukuba, Ibaraki 305-0856, Japan

Presenting author; Toru MATSUI, [email protected]

Carboxylate-substituted aromatic or heterocyclic compounds with high optical purity are interesting targets for either biodegradation or bioconversion. Degradation of phenylmalonate by Alcaligenes bronchisepticus KU 1201 has been extensively studied (Miyamoto and Ohta, 1992). The bacterium enantioselectively decarboxylates alpha-aryl-alpha-methylmalonates to optically active alpha-arylpropionates. The chemical process involved in the optical resolution of racemic phenylsuccinate (PS) by recrystallization has also been reported (Shiraiwa et al., 1994; Cesare and Stephani, 1997) The kinetic resolution of racemic compounds by microorganisms capable of stereospecific degradation is a simple method for the production of optically pure compounds (Suzuki and Kasai, 2003). While screening for aromatic dicarboxylate- degrading organisms, we isolated 2 actinomycetes that stereoselectively degrade PS although each preferentially degraded a different enantiomer. Here, we report the stereospecific degradation of PS by the actinomycetes.

Racemic phenylsuccinate was stereospecifically degraded by the actinomycetes PS9 and PS17 isolated from soil obtained from Okinawa Island, Japan. Strain PS9, identified as a Citricoccus sp., preferentially degraded the R-form, while strain PS17, identified as a Microbacterium sp., preferentially degraded the S-form of phenylsuccinate. Analysis of the culture broths of these species with phenylsuccinate as the sole carbon source revealed that benzoic acid was produced as a metabolic intermediate. Benzoic acid was further degraded by strain PS9 with m- and/or p-hydroxybenzoic acid but not o-hydroxybenzoic acid as possible intermediates.


173

IBBS-15-6-P23

Biotreatment of oil sludge contaminated water by bacterial consortium

Dariush Minai-Tehrani, Zahra Sobhani Damavandifar, Saeed Minoui

Faculty of Biological Sciences, Shahid Beheshti University, Tehran, IRAN

Most of the oil storage tanks and other vessels in most refineries contain bottom sediments which accumulate over time. These residues, called oil sludge, are composed of oil, water, solids. Their characteristics, such as high content of resin and asphaltene, make them highly recalcitrant and very difficult to reutilize. Oil sludge in the bottom of tank can accelerate corrosion, reduce storage capacity and disrupt operations. During cleaning operations, the oil sludge is removed and dumped in nearby lands, which may enter to the surface or sub surface water. In this experiment, biotreatment of oil sludge contaminated water by a bacterial consortium was studied. An oil degrading bacterial consortium was isolated form soil near reservoir oil tank, including three types of bacteria. The consortium was transferred to oil sludge contaminated water and adopted in oil containing medium with several passages. The growth curve showed a short Lag period (about 6 hours) with a logarithmic phase about 24 hours which indicated the high efficiency of the consortium for oil consumption. Our results showed that 35% of oil sludge reduced in culture medium after 140 hours of treatment. The yield of the cells (7g/L) and the amount of protein (17 mg/ml) increased while the oil sludge reduced in the medium. Analysis of aromatic and aliphatic fractions of oil sludge by GC and HPLC measurement showed that the highest reduction belongs to aliphatic (70%) followed by aromatic (40%) fractions, while the reduction of resins and asphaltine were not significant.

Keywords: Biodegradation, Oil sludge, Bacteria, contamination


174

IBBS-15-6-P24

Biodegradation of Polyurethane Plastics Exposed to the Environment Mohd Naqiuddin Mohd Zairi1, Geoff D. Robson1

1The University of Manchester, U.K. Polyurethanes (PU) are biodegradable synthetic polymers that are manufactured into many different products ranging from soft elastomers to rigid plastics. Consequently, PU plastics in particular have contributed significantly to the solid waste accumulation at landfill sites and are pollutants of terrestrial ecosystems. In previous studies, PU biodegradation were investigated in buried conditions for optimal microorganism action. However, in many cases, PU plastics are not successfully buried in soil but instead are left exposed to the natural air environment which offers different conditions for PU biodegradation. In this study, we aim to investigate the colonisation and development of microbial communities on PU plastics exposed to the environment over time. PU plastic coupons have been exposed to the environment on a rig system and were subjected to microbial recovery at 12 week time intervals. Air samples were also obtained at multiple occasions throughout the experiment to count and identify potential PU-degrading microorganisms that are readily found in the surrounding air. The extent of PU degradation was also determined through tensile strength tests. The amount of microorganisms scrapped off the plastic’s surfaces was found to be dependent on the weather where there are more microbial counts during the warmer summer months. The air samples also provided corresponding results. However, the dominantly found isolates from the air were not found dominantly colonising the coupons. Light microscopy recorded visual evidence of microorganisms colonising and penetrating the coupons’ surfaces. From stretching results, PU coupons have reduced in tensile strength due to degradation from environmental exposure.


175

IBBS-15-6-P25

Biodegradation of polyvinyl alcohol (PVA) by fungi Authors; Somayeh Mollasalehi, Dr Geoff Robson

Faculty of Life Sciences, University of Manchester, UK

Currently, large volumes of synthetic plastics are produced annually worldwide .As they are generally highly resistant to microbial degradation they are major pollutants of terrestrial and aquatic ecosystems. For this reason, during the last thirty years extensive research has been conducted to develop biodegradable plastics and the mechanisms by which they are degraded. Polyvinyl alcohol is a water soluble synthetic polymer which can be used to form plastics as well as being used extensively in paper and textile manufacture, adhesives and paints. Despite its growing use, relatively little is known about its degradation and in particular the role of fungi in this process. The aims of this study were;

• To isolate and characterizes fungi with the ability to degrade the water soluble polymer polyvinyl alcohol (PVA). Putative PVA degrading fungi were isolated from different environmental sites .The polymeric material tested were PVA’s of different molecular weights and degree of hydrolysis.

• To compare the growth curves of these putative PVA degraders were grown in minimal medium contain PVA (13-23 KD, 98% hydrolysed) as a sole carbon source.

• To investigate the fungal communities were present at studied soils Denaturing gradient gel electrophoresis (DGGE) profiles were used to compare the fungal community within the soils.

• In order to quantify the concentration of PVA during growth, a spectrophotometric assay was developed


176

IBBS-15-6-P26

Purification and Characterization of Cyclodextrin Glucanotransferase Enzyme and Its Role in Biodegradation of Diesel Oil Contaminated Soil

Sivaraman C, Anasuya Ganguly and Srikanth Mutnuri

Applied and Environmental Biotechnology Laboratory, Dept. of Biological Sciences, BITS Pilani - K.K Birla Goa Campus, Zuari Nagar, Goa 403726, India.

e-mail: [email protected], Telephone : 0091 832 2580125, Fax : 0091 832 2557033

Abstract

Microbial degradation of hydrocarbons is found to be attractive process for remediation of contaminated habitats. However the poor bioavailability of hydrocarbons result in low biodegradation rates. Cyclodextrins produced by Cyclodextrin Glucanotransferase (CGTase) enzyme on starch are shown to increase the bioavailability of variety of hydrophobic compounds. In the present work we purified the CGTase enzyme and studied its role on biodegradation of diesel oil contaminated soil. Purification of CGTase from E. cloacae was done which resulted in 6 fold increase in enzyme activity. The enzyme was active at neutral pH, 60 ˚C and molecular weight of 66 kDa. Addition of purified CGTase showed enhanced biodegradation of diesel oil (57 + 1.37%) but less significant when compared to treatment setup which was added with P. mendocina and E. cloacae (52.7 + 6.51%). Immobilization of P. mendocina on alginate resulted in better survival which resulted in biodegradation of hydrocarbons in diesel oil at 336 hours.

Keywords: diesel oil, bioavailability, cyclodextrin


177

IBBS-15-6-P27

Biodegradation of diesel by free and immobilized Pseudoxanthomonas sp. RN402

Wannarak Nopcharoenkul,1 Parichat Netsakulnee2 and Onruthai Pinyakong2, 3

1 Inter-department of Environmental Science, Graduate School, Chulalongkorn University, Bangkok 10330, Thailand 2 Bioremediation Research Unit, Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand 3 National Center of Excellence for Environmental and Hazardous Waste Management (NCE-EHWM), Chulalongkorn University, Bangkok 10330, Thailand

The use of immobilized cells in wastewater treatment has many advantages such as tolerance to pollutant concentration and possibility to reuse. This research aimed at the application of immobilized Pseudoxanthomonas sp. RN402 to clean-up diesel-contaminated water. The diesel biodegradability of RN402 was investigated in batch experiment and analyzed by TLC-FID. Free cells of RN402 at initial density of 109 MPN/treatment were capable of removing 100 and 200 mg/l diesel in carbon free minimal medium by 74% and 69%, respectively, within 3 days. The RN402 was immobilized on the surface of high-density polyethylene plastic pellets with maximum cell density of 108 MPN/g plastic pellets. The diesel biodegradation efficacy of immobilized cells (108 MPN/treatment) was compared with those of free cells (109 MPN/treatment) in a semi-continuous experiment by adding of 200 mg/l of diesel to the medium every 3 days for 15 days. The results revealed that the biodegradation efficiency of free cells was reduced after 6-day incubation, while that of the immobilized cells was maintained throughout the experiments. Moreover, no significant diesel reduction was observed when only plastic pellets were used. The results indicated that the immobilized cells gave better performance than free cells. Further study in batch experiment showed that about 64 to 33 % removals of diesel (300-2000 mg/l) were achieved within 3 days when the immobilized cells (108 MPN/treatment) were used. Moreover, the reusability test of immobilized cells revealed the stable activity after 12 cycles. This study suggested that immobilized RN402 has potential application for diesel-contaminated water treatment.


178

IBBS-15-6-P28

Degradation of polycyclic aromatic hydrocarbons (PAHs) and long-chain hydrocarbons by bacterial strains isolated from polluted soil

Jana Harichová, Edita Karelová, Jana Godočíková, Mária Bučková, Andrea Puškárová, Bystrík Polek, Peter Ferianc, Domenico Pangallo

Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, Slovakia

The microbial diversity of two polluted soils from a former chemical park in Bratislava was investigated. The analysis regarded the isolation of bacterial strains which potentially were able to degrade polycyclic aromatic hydrocarbons (PAHs) and long-chain hydrocarbons. The bacterial strains were isolated by Minimal Medium agar plates supplemented with a PAHs mixture and by the MST medium supplemented with oil powder and triphenyltetrazolium chloride (TTC). The microorganism able to grow on MST agar acquired a red colorization attesting the production of dehydrogenases. The bacterial strains able to grow in this two media were clusterized by the utilization of f-ITS. The internal transcribed spacer between 16S and 23S rRNA gene was amplified, followed by separation of fluorescently labeled PCR products by capillary electrophoresis. In a second step isolates showing characteristic f-ITS profile were identified by the sequencing of the 16S rRNA gene. All the isolates were screened for their ability to grow on minimal medium broth supplemented with a PAH mixture and to degrade long-chain hydrocarbons (hexadecane and engine oil) by the utilization of the 2,6-dichlorophenol indophenol assay (2,6-DCPIP). After 36 days of cultivation the unique bacterium able to grow on broth medium was an Arthrobacter strain, after 56 days of cultivation two Arthrobacter and two Pseudomonas strains. By the 2,6-DCPIP test were positive members of the genus Pseudomonas and other kinds of isolates such as Serratia marcescens, Acinetobacter sp., Ochrabactrum sp. and Microbacterium sp.

ACKNOWLEDGEMENTS

This work was supported by the the Research & Development Operational Program funded by the ERDF no. 26240220010


179

IBBS-15-6-P29

Study of microbes and plants interactions in PCB contaminated soil

Petra Prouzová1, Eva Hoskovcová1, Marcela Holečková2, Tomáš Macek2, Kateřina Demnerová1 a Martina Macková1

1Dept. of Biochemistry and Microbiology, ICT Prague, Technicka 3, Prague 6, Czech Republic, 166 28

2Inst. of Organic Chemistry and Biochemistry CAS, Flemingovo n. 2, Prague 6, Czech Republic, 166 10

Polychlorinated biphenyls (PCB) are one of the most toxic and persistent contaminants in the environment, which were spread due to nonregulated use. Using conventional methods for cleaning of PCB-contaminated sites is very expensive. Bioremediation, the use of biological organisms to remove pollutants, can provide an inexpensive and considerate solution to environmental contamination.

This work is focused on the following the fate of the selected microorganisms, Achromobacter xylosoxidans S12a and Pseudomonas putida KF, which were added to the contaminated soil planted by tobacco (Nicotiana tabacum) and horseradish (Armoracia rusticana). Achromobacter xylosoxidans S12a and Pseudomonas putida KF were both previously isolated from PCB contaminated sludge and they showed PCD degrading abilities. Samples were collected after 1 day, 3 and 4 months. DNA and bacteria isolates were isolated from each plant’s rhizosphere. Primers for specific segment of 16S rRNA sequences of Achromobacter xylosoxidans S12a and Pseudomonas putida KF were used to prove their occurrence in the samples. Bacteria were identified by MALDI-TOFF MS and screened for ability to metabolize PCB. As a model PCB system a recalcitrant mixture of PCB congeners with different degree of chlorination – Delor 103 was used. Residual content of PCB congeners was measured by GC analysis. P. putida and A. xylosoxidans were both identified by MALDI-TOFF MS in soil samples planted by horseradish and tobacco. The presence of both bacteria was confirmed by 16S rRNA analysis after 3 and 4 months cultivation.

ACKNOWLEDGEMENTS

The work was supported by NPV II 2B08031, FP7-KBBE-2010-4 and MSM 6046137305


180

IBBS-15-6-P30

Decolourization and detoxification of Azo Dye, Synozol Red HF-6BN, by Aspergillus niger and Nigrospora sp.

Sidra Ilyas and Abdul Rehman

Department of Microbiology and Molecular Genetics, University of the Punjab, New Campus, Lahore 54590, Pakistan

Abstract

In the present investigation the fungi, Aspergillus niger and Nigrospora sp. were employed for decolourization of Synazol Red HF-6BN. Decolourization study showed that A. niger and Nigrospora sp. were able to decolorize 88% and 96% Synazol Red 6BN, respectively in 24 days. It was also studied that 86% and 90% Synazol Red containing dye effluent was decolorized by A. niger and Nigrospora sp. after 28 days of incubation at room temperature. A protein with relative molecular mass of 70 kDa was partially purified and examined for enzymatic characteristics. The enzyme exhibited highest activity at temperature ranging from 40-50°C and at pH 6.0. The enzyme activity was enhanced in the presence of metal cations. High performance liquid chromatography analysis confirmed that these fungal strains are capable to degrade Synazol Red dye into metabolites. No zones of inhibition on agar plates and growth of Vigna radiata in the presence of dye extracted sample indicate that the fungal degraded dye metabolites are nontoxic to beneficial micro-flora and plant growth. A. niger and Nigrospora sp. have promising potential in colour removal from textile wastewater containing azo dyes.


181

IBBS-15-6-P32

Effects of biofouling on nitrification performance of a biofilter operating in a commercial marine aquaculture recirculation system Hugo Ribeiro1, Maria-Teresa Borges1,2, Ana Matos1, Carlos M. Pereira3 and Paula M. L. Castro4 1CIIMAR - Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Porto, Portugal 2Departamento de Biologia, Faculdade de Ciências da Universidade do Porto, Porto, Portugal 3Departamento de Química, Centro de Investigação em Química, Faculdade de Ciências da Universidade do Porto, Porto, Portugal 4CBQF- Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Porto, Portugal

Abstract

In Aquaculture facilities using water recycling there is a build-up of ammonia and nitrite, which are harmful to fish health. The elimination of these pollutants is achieved by biofiltration, via nitrification by autotrophic bacteria. However, the excessive colonization through time of the biofilter carriers by a diverse array of undesirable organisms (“biofouling”), including heterotrophic bacteria, can negatively affect the process. Studies on biofouling of biological treatment systems for saline effluents are rare. Having in mind this background, the present work addresses the influence of biofouling on nitrification efficiency in a Biofilter (MBBR) operating in a commercial fish farm.

Biofilter inlet and outlet water and carriers were sampled during a fish production cycle. Water physical-chemical analyses were done for total ammonia nitrogen (TAN), nitrite, and nitrate. Heterotrophic bacteria attached to the carriers were assessed by culturable counts using standard spread plating procedures. A unusual biofouling come into view and several microorganisms were identified, including marine cyanobacteria. Controlled nitrification experiments with biofilter carriers were carried out in laboratory microcosms to assess biofouling effects on nitrification.

The results show that the nitrification substrate concentration (TAN and Nitrite) available was 0.4 % of the minimum levels needed for nitrifying bacteria optimal development. The number of heterotrophic bacteria was ca. 10 times superior to those described in the literature for similar systems, suggesting a strong competition with nitrifying bacteria. Besides, controlled nitrification experiments demonstrated that biofouling negatively affected the nitrification process by removing the available substrate at a rate four times higher than nitrifying bacteria.

Given the results obtained, it is of outmost importance the study and development of more performing wastewater treatment processes in aquaculture recirculation systems.

We acknowledge the Fish farm manager and the Fish farm staff for permission and assistance. This work was funded by EC project ‘Raceways - a hyperintensive fish farming concept for lasting competitiveness and superior production’ (COOP-CT/016869/2006-2008).


182

IBBS-15-6-P33

The influence of modified polyhexamethylene guanidine on the biodegradation of polylactide.

Agnieszka Richert1, Maria Swiontek Brzezinska2, Maciej Walczak2

1 - Institute for Engineering of Polimer Materials and Dyes, Sklodowskiej-Curie 55, 87-100 Torun,

phone +48 (056) 650 00 44, e-mail: [email protected]

2 – Department of Environmental Microbiology and Biotechnology, Nicolaus Copernicus University, Gagarina 9, 87-100 Torun, Poland

Abstract

Degradability of polymeric materials is a function of the structures of polymeric materials, the presence of degradative microbial population, the environmental conditions and inhibiting substances. In this paper we present results of studies on impact of modified polyhexamethylene guanidine (PHMG) on biodegradation efficient of polylactide (PLA) and the biofilm formation on surface on this polymer. The degradation process was monitored by respiration activity of microorganisms in presence of PLA. The mechanical strength has been determined by static drawing standard procedure, the structural properties have been determined by SEM, the rheological properties have been determined by MFR (melt flow ratio value). The lowest concentrations of modified PHMG have no significant impact on investigated properties. In concentration modified PHMG higher than 0.4% (w/w) degradation efficient and changes in physical properties of PLA were significantly lower and in the biofilm structure many bacterial cells in the biofilm were dead.


183

IBBS-15-6-P34

Biosurfactant-producing bacteria from Branchiomma luctuosum and Megalomma claparedei (Polychaeta, Sabellidae).

Rizzo C.1,2, Malavenda R.1,2, Lo Giudice A.1, Michaud L.1, Hoermann B.2, Gerce B.2, De Domenico M.1, De Domenico E.1, Syldatk C. 2, Hausmann R.2

1) Dpt. of Animal Biology and Marine Ecology, University of Messina (Italy) – 2) Karlsruher Institut für Technologie (KIT) Institut für Bio- und Lebensmitteltechnik Bereich II: Technische Biologie, Universität des Landes Baden-Württemberg und nationales Forschungszentrum in der Helmholtz-Gemeinschaft (Germany).

To date, the isolation and characterization of BS-producing bacteria from biotic matrices has been never reported. In the present work, three bacterial strains were isolated from enrichment cultures performed by seeding crude oil-supplemented Bushnell Haas (BH) (1%) with homogenates (9 ml) of two filter-feeding Polychaetes species (i.e., Branchiomma luctuosum and Megalomma claparedei), and incubating at 28°C for one month. To analyse the production of BS molecules, isolates were preliminarily grown in a liquid medium supplemented with tretradecane (2%), and incubated at 28°C. After two and five days of incubation OD570 measure, C-TAB and blood agar (BA) plate assays, E24 index detection, surface tension measure, BS extraction and TLC were performed. Moreover, all tests except BS extraction and TLC were carried out also after growth on BH (with tetradecane and phenanthrene as carbon source) after ten and fifteen days of incubation. The strains showed different responses depending on the culture medium and the substrate added. The 16S rDNA sequencing showed that strains were related to Pseudovribio sp. (strain A27), and Alcanivorax spp. (strains A52 and A53). In conclusion, the isolation of BS-producing strains from filter-feeding organisms seems to be a promising approach, and for the first time we report about members of the genus Pseudovibrio as biosurfactant producer.


184

IBBS-15-6-P35

Biodegradation of Polycyclic Aromatic Hydrocarbons by newly isolated bacterial strain Bacillus cereus KWS2

Safia Ahmed, Fazal-ur-Rehman, Zulfiqar A. Malik and Abdul Hameed.

Department of Microbiology, Quaid-i-Azam University, Islamabad, Pakistan

Abstract

Polycyclic aromatic hydrocarbons (PAHs) are a ubiquitous class of organic compounds present in the environment. Microbial degradation of PAHs by bacteria and fungi is considered a safe and environment friendly method to remove such toxic contaminants. In the present study, bacterial isolates utilizing the PAHs (naphthalene, anthracene and phenanthrene as sole carbon and energy were characterized taxonomically and for their degradation potential for the respective PAH.

Bacterial samples were isolated from crude oil contaminated soil and checked for growth on PNR medium containing anthracene and pyrene. One bacterial isolate was selected on the basis of their best growth on PAHs and identified as Bacillus cereus KWS2. The optimum pH and temperature were 7 and 30ºC respectively, for degradation and growth by Bacillus cereus KWS2 where 45 % of reduction in anthracene concentration was observed after seven days of incubation.

Resting cell biotransformation studies of naphthalene, phenanthrene and anthracene were carried out by Bacillus cereus KWS2 and analyzed by GC/MS. From naphthalene biotransformation four metabolites, 2-naphthol, benzeneacetic acid, benzoic acid and benzaldehyde were detected and identified. From phenanthrene biotransformation, 9-phenanthrenol, 9,10-Dihydro,9,10-dihydroxyphenanthrene, bezeneacetic acid, 4-hydroxy-benzeneacetic acid and 2-hydroxy benzoic acid were detected and identified. Benzenecarboxylic acid, benzeneacetic acid and 4-hydroxy-benzeneacetic acid were detected and identified from anthracene biotransformation by B. cereus KWS2. The detection of both mono and dihydroxylated compounds suggested that B. cereus strains KWS2 harbor both mono and dioxygenase genes.


185

IBBS-15-6-P37

Characterization of hydrocarbon-degrading microbial strains isolated from oil polluted soil

Author(s):

A.-M. Tanase, I. Chiciudean, R. Ionescu, O. Csutak, D. Pelinescu, T. Vassu, I.Stoica

Institute(s):

Genetics Departament, Bucharest University, Bucharest, Romania

Microbial biodegradation is known to be an efficient process in the decontamination of oil-polluted environments. First step in establishing in situ bioremediation potential includes search for oil degrading indigenous microorganisms. To investigate the petroleum hydrocarbon (HC)-degrading potential of indigenous micro-organisms in oil polluted soil nearby an extraction field, a number of bacteria have been isolated and characterized for their ability to utilize n-alkanes as growth substrates.

Using culturable methods, a population of bacterial strains was detected in the contaminated soil. The isolated strains were able to degrade medium-chain n-alkanes as assesed by growth assays, using n-alkanes as sole carbon and energy source. Results showed a high diversity between strains at molecular level, but also in the degradation profiles.

Physiological and biochemical tests showed a similarity between a group of five strains that was confirmed by Biolog MicroLog analysis and identified as Pseudomonas aeruginosa. Other isolated strains showed similar physiological traits with Rhodococcus, Chriseobacterium or Acinetobacter species.

These preliminary results indicate that our strains could potentially be useful for the remediation of hydrocarbon pollution in natural environments.


186

IBBS-15-6-P39

Determination of remediation potentials of ligninolytic fungi for degradation of lindane

Ajda Ulčnik1, Lucija Zupančič-Kralj2, Črtomir Tavzes3, Franc Pohleven1

1Department of Wood Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia

2Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva cesta 5, 1000 Ljubljana, Slovenia

3Institute of Wood Science and Technology, and Sustainable Development, Celovška cesta 268, 1000 Ljubljana, Slovenia

The ability of four white-rot fungi (Trametes versicolor, Hypoxylon fragiforme, Chondrostereum purpureum, and Pleurotus ostreatus) and one brown-rot fungus (Gloeophyllum trabeum) to degrade an organochlorine insecticide, lindane, in liquid cultures was studied. Lindane was exposed to fungal liquid cultures for various lengths of time. In order to extract lindane from fungal liquid cultures, two different extraction procedures were used: extraction of lindane from filtrates, and from homogenised fungal cultures. Both procedures were examined for their appropriateness in determining the amount of lindane in liquid cultures. White-rots were able to degrade lindane. The amount of degraded lindane increased with its incubation period in the liquid cultures of all white rot fungi used, except C. purpureum. After 21 days of incubation, over 90 % of lindane was degraded by T. versicolor, H. fragiforme, and P. ostreatus. Degradation of lindane by a brown rot G. trabeum did not occur. The selection of extraction procedure, when liquid cultures of T. versicolor, H. fragiforme, and P. ostreatus were used, had no noticeable effect on the determined degradation. On the other hand, the amount of lindane in liquid cultures of C. purpureum and G. trabeum depended strongly on the extraction procedure. Lindane removal from the culture media, inoculated with C. purpureum and G. trabeum, presumably occurred via adsorption onto the mycelial biomass.


187

Topic 7-Bioremediation of contaminated soils and water, bio-filtration of industrial pollutants,

development of GMOs and their use in bioremediation


188

KEYNOTE LECTURE

Capitalising microbial diversity for bioremediation : from systemic analyses to synthetic agents

Víctor de Lorenzo,

Centro Nacional de Biotecnología, CSIC Madrid 28049 (Spain), [email protected]

According to some of the latest compilations we have information on the microbial degradation of about 1000 compounds, 700 catabolic enzymes and about 800 reactions of environmental interest (1). Yet, we are still unable to predict the fate and effects of many toxic compounds, let alone to rationally design bacteria able to metabolize many tough chemicals in granted ecosystems (2). In this context, two complementary ways to survey and exploit the catabolic gene pool available in the metagenome will be addressed. The first involves the application a Systems Biology approach to the global biodegradation network which is formed by integrating all known metabolic reactions exerted by microorganisms on unusual, mostly man-made chemical compounds (3,4). A number of genetic tools have been developed for mining desired activities out of such a catalytic landscape by coupling biotransformations to activation of a selectable property (5). With these tools, given reactions can be reclaimed directly from environmental DNA based on the biological activity instead of DNA sequence similarity. The second approach is framed in what has been recently denominate Synthetic Biology (SB). The basic notion behind SB is that any biological can be deconstructed in a limited number of components and reconstructed in an entirely different configuration for the sake of modifying existing properties or creating altogether new ones (6). Forward design of biological agents for biosensing nitro-compounds in soil (7) and for in situ biodegradation of lindane (hexachlorocyclohexane, HCH) will be discussed.

1. Nucleic Acids Res 31, 262-265. 2. Int Microbiol 8, 213-22. 3. Nucleic Acids Res 33, D588-592. 4. EMBO Rep 4, 994-999. 5. Environ Microbiol 8, 546-555. 6. Ann Rev Microbiol 64, 257-275. 7. Env Microbiol 10: 3305-3316.


189

IBBS-15-7-O02

Biochemical Characterization of a Lead-tolerant Strain of Aspergillus foetidus : An Implication of Bioremediation of Lead from Aqua- Enviroment.

Satarupa Chakraborty, Abhishek Mukherjee, and Tapan K. Das

Department of Biochemistry and Biophysics, University of Kalyani, Kalyani, West Bengal, India, [email protected] 919475944365(M) ABSTRACT The fungus ,Aspergillus foetidus isolated from the waste water treatment center, Kalyani has been identified as a Pb+2 tolerant strain by the supplementation experiment with the use of high concentration of Pb+2 . Growth studies of the strain in Czapex Dox (CD) media supplemented with different concentration of Pb+2 indicated that initially growth increased up to 200 mg/L of Pb+2 and then decreased gradually in increasing of Pb+2 concentration; but no significant growth was found at 2000 mg/L of Pb+2. Leakage of electrolyte & protein and presence of thiol groups of the strain grown in CD broth containing Pb+2 indicated disturbance in membrane structure as well as efflux pumping of lead outside the cell. The significant change of proline & thiol content while the strain was grown in CD medium supplemented with Pb+2 could be considered as the mechanism of lead tolerance shown by the strain. The different concentrations of Pb+2 affected the extent of lipid peroxidation, catalase & alkaline phosphatase activities in cell free extract of the PdR strain . Activities of guaiacol peroxidase ,glutathione reductase(GR) , and acid phosphatase were also assayed for further clarification of the characteristics of the lead tolerant strain of Aspergillus foetidus. The electron microscopic study was also carried out to show the deposition of salt of lead crystal to the cell surface. The preliminary studies on removal of lead ion from lead contaminated water indicated that the strain has the capacity to bioremediate the lead from aqua-environment.


190

IBBS-15-7-O03

Biodegradation of Several Polycyclic Aromatic Hydrocarbons by Microalgae Isolated from Contaminated Site.

Mohd. Nizam Yusof1, Siew-Moi Phang2

1Faculty of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam, Selanor, MALAYSIA.

2Institute of Biological Sciences, Universiti Malaya, 50603 Kuala Lumpur, MALAYSIA

Several species of microalgae have been isolated from the wastewater treatment pond of an oil refinery in Malaysia. These microalgae have been grown and enriched in solid Bold’s basal medium (BBM) containing selected PAHs namely naphthalene, phenanthrene and pyrene as their sole carbon sources. Several subculture processes have been conducted to obtain pure culture of dominant species. Selected species namely Chlorella and Scenedesmus were then subjected to PAHs biodegradation assays in 250 ml flasks containing liquid BBM and selected PAHs at different concentrations on an orbital shaker under fluorescence light. The medium then were centrifuged and subject to solid-phase extraction. Analytes were analysed using Gas Chromatography coupled with Mass Spectrometry detector (GC-MSD). Results showed reduction in peaks of PAHs parent compounds. These indicate that microalgae are capable to degrade the two-, three- and four-rings PAHs into less harmful compounds. Results obtained from this study can be used to design a proper bioremediation program for wastewater containing PAHs.


191

IBBS-15-7-O04

Survival of poly(vinyl alcohol) cryogel-immobilized Rhodococcus cells in crude oil-contaminated soil and impact on biodegradation efficiency

Maria S. Kuyukina1,2, Irena B. Ivshina1,2, Tatyana N. Kamenskikh1, Maria V. Zhigalova2

1 Institute of Ecology and Genetics of Microorganisms, Russian Academy of Sciences, 13 Golev Street, 614081, Perm, Russia. E-mail: [email protected].

2 Microbiology & Immunology Department, Perm State University, 15 Bukhirev Street, 614990, Perm, Russia

Bioremediation has a great potential to restore oil-contaminated soils by using biodegradative activities of indigenous or introduced microorganisms. However, bioaugmentation strategy could fail if oil-degrading strains are poor survivors or lose their catabolic activities when inoculated into natural ecosystems. Therefore, bioaugmentation requires inocula that, besides degrading pollutants in pure culture, can survive in oil-contaminated soil for a long time. The genus Rhodococcus is promising for biodegradation of recalcitrant contaminants, like petroleum hydrocarbons. Rhodococcus species are ubiquitous in hydrocarbon-contaminated environments, tolerate harsh conditions, compete successfully in complex bacterial populations and therefore could be efficiently used in bioaugmentation inocula [1]. Immobilization of Rhodococcus cells into poly(vinyl alcohol) cryogel improves their viability and catabolic potential [2]. In laboratory microcosms, we studied the survival and catabolic activity of immobilized and free Rhodococcus cells introduced into loam soil freshly contaminated with crude oil, mimicking spill situation. Inoculated R. erythropolis and R. ruber strains were monitored in soil and cryogel granules by selective agar plating and species-specific PCR. Introduced Rhodococcus cultures survived successively in oil-contaminated soil for 14 months. Moreover, immobilized cells demonstrated higher viability under soil desiccation conditions compared to free cells. Soil microcosms with immobilized Rhodococcus cells respired more actively than control (non-inoculated) and liquid culture-amended soils; that was in a good correlation with greater oil degradation rates in immobilized systems.

Research was supported by MCB Russian Academy of Sciences Presidium Program.

References

1. Kuyukina M.S., Ivshina I.B., 2010. In: Microbiology Monographs, 16, 231-262.

2. Kuyukina et al., 2006. J. Microbiol. Methods, 65, 596-603.


192

IBBS-15-7-O05



The University of Akureyri Department of Natural Resource Sciences. Akureyri, Iceland.

Although generally reputed pure and uncontaminated, most Icelandic aquatic environments are to some degree anthropogenically impacted. We surveyed two types of aquatic environments; three wastewater-exposed seashore environments and a freshwater system comprising the river Glerá and its watershed. The widespread and copious usage of several types of cleaning detergents and other surfactants results in their relatively high concentrations in both domestic and industrial sewage, which enters the aquatic environment untreated in most Icelandic localities. The river Glerá in Northern Iceland presents a convenient system for evaluating the effects of anthropogenic activities on microbial communities. The river originates in pristine surroundings and then flows past a local landfill and through an urban area, receiving inflows from various small-scale industries. Samples from the two environmental types were screened for surfactant-degrading activity and surfactant biodegraders isolated. All of the degrading isolates were found to belong to the class Gammaproteobacteria. In the freshwater system, surfactant degraders solely comprised pseudomonads, while the species composition was more diverse in the seashore samples. The surfactant-degrading freshwater pseudomonads mostly comprised members of the Pseudomonas fluorescens and P. syringae species groups. The seashore was generally found to be a poor source of surfactants degraders. The river environment was also screened for the appearance of mercury-tolerant bacteria, as an indicator of the degree of environmental pollution. It was found that the landfill leachate had an impact on the river microbiota downstream. Mercury resistance was markedly increased, whereas dominant species diversity, as determined by DGGE, was only modestly affected.


193

IBBS-15-7-O06

Microbial conversion of pollutants into electric current: strategies for monitoring and stimulating the bioremediation capacity of soil microbial communities.

Abraham Esteve-Núñez1,2, Karina Boltes1,2, Alejandro Reija2, Noemi Bragado1, Ainara Dominguez1, Jose Carpatos1, Miriam Biel1, and Antonio Berná3 1Departamento de Analítica e Ingeniería Química, Universidad de Alcalá, E-28871,

Alcalá de Henares, Madrid, Spain

2IMDEA water, Parque Tecnológico de Alcalá,Alcalá de Henares, Madrid, Spain

3Departamento de Geología, Universidad de Alcalá, Alcalá de Henares, Madrid, España

[email protected]

Abstract

Sedimentary microbial fuel cells (SMFC) are electrochemical devices that harvest electrical energy from the potential differences across sediment/water interfaces in the environment. Our aim is to apply this concept to the bioremediation of polluted soils where the organic contaminant will play the fuel role for natural microbial communities.

In this work, we introduce the concept of electrogenic microcosm as a novel tool for investigating and monitoring the in situ biodegradation microbial activity in polluted soils and sediments. For this purpose we have design a especial anode set up which allow to assay polluted soil samples and use chronovoltametry and chronoamperometry for measuring electrons released from the pollutant microbial oxidation that are directly transferred to the anode. The effective bioremediation task was also confirmed by soil extraction followed by HPLC-DAD analysis and ecotoxicological assays. Benzoate, and the herbicides Diuron and atrazine, N-(3,4-dichlorophenyl)N,N-dimethylurea, were used as representative model for aromatic pollutants. A serie of different concentrations benzoate, diuron and atrazine pulses were carried out in order to show the accuracy of the methodology. Interestingly, the response of the microbial population was faster (5-fold) when electrogenic populations were pre-exposed to benzoate versus non-exposed ones, proving an induction phenomenon. In addtion, polluted soils were bioremediated 10 times faster (3 days) using electrogenic approach in contrast with the natural biodegradation (30 days).

This novel methodology offers the possibility of a) monitoring the in situ pollutant biodegradation using environmentally-friendly tools, and b) evaluation of the natural biodegradation profile of a microbial population in the environment.


194

IBBS-15-7-O07

Two fungal bioremediation methods for contaminated soil

Lara Valentín1, Marja Tuomela1, Kari Steffen1, Erika Winquist2, Maite Moreira3 and Annele Hatakka1

1Department of Food and Environmental Sciences, University of Helsinki, Finland.

2Department of Biotechnology and Chemical Technology, Aalto University, Finland.

3Department of Chemical Engineering, University of Santiago de Compostela, Spain.

Corresponding author: [email protected]

A project for the evaluation of fungi to be used in bioremediation of contaminated soil is presented. Two ex situ technologies, a slurry bioreactor and a solid-phase pre-treatment of contaminated soil, were developed and scaled-up.

The first part was related to the fuel oil spill from the tanker Prestige that affected 1,900 km of northwest Spanish coastal areas in November 2002. A project was then initiated to develop a system for treating contaminated saline marshes based on the use of white-rot fungi in a slurry bioreactor focusing on PAHs. Considering that agitation and saline conditions may affect PAH degradability and fungal growth, fungi were screened for their halotolerance. The process was later scaled-up to a 5 l reactor with Bjerkandera adusta to determine the best PAH degradation efficiency under different conditions.

The second technology was related to the dioxin-contaminated soil in sawmills in Finland. Currently, the only method applied to these soils is thermal treatment. However, the large content of soil organic matter (SOM) hinders the combustion. Thus, a fungal pre-treatment of sawmill soils was developed to enhance the effectiveness of the subsequent combustion. The preliminary screening of 146 fungal strains showed that 34 strains extensively colonized non-sterile soil. Later, 18 selected strains were characterized by their production of enzymes during cultivation on Scots pine bark. Of six fungi selected, three were the most active SOM degraders (estimated SOM loss was 3.5 - 9.5% during 6 months). The process was scaled-up to 0.56 m3 with Stropharia rugosoannulata resulting in a SOM loss of 30.5 kg.


195

IBBS-15-7-O08

Soil inoculation with specific microbial communities attached on carrier material - can this become a useful tool in soil remediation?

R. Schroll, A. Krug, F. Wang, U. Dörfler, J.C. Munch

Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Institute of Soil Ecology, 85764 Neuherberg, Germany

[email protected]

Different types of agricultural soils were loaded with 14C-labelled model chemicals, and subsequently different microbial communities as well as isolated strains were inoculated to enhance the mineralization of such chemicals. Inocula were introduced in soils by different approaches: (i) soil inocula, (ii) application of isolated strain as well as microbial community via liquid media, (iii) isolated strain as well as microbial community attached to a carrier material. Most of the inoculation experiments were conducted in laboratory but one of these approaches was also tested under real environmental conditions in lysimeters and it could be shown that this approach was very successful.

The inoculation of soils with microbial communities attached on a specific carrier material showed the highest mineralization effectiveness and also the highest sustainability. Microbes attached on these carrier particles preserved their function over a long time period even if the specific microbial substrate was already degraded or at least not detectable any more. We already got some hints that the microbial communities which we inoculate to soils are “interacting communities” and that these interactions are the reason for the very high sustainability of our approach. Additionally, we could show that in specific cases some soil parameters might reduce the effectiveness of such an approach.

Results on isoproturon as a model for phenylurea-herbicides and 1,2,4-trichlorobenzene as an example for an industrially used chemical as well as the corresponding chemicals` degrading microbial communities and isolated strain will be presented.


196

IBBS-15-7-O09

Use of modified Rhodococcus strains for degradation of phenol in industrial wastewater

L. Zidkova1,2, M. Otevrel1, L. Wimmerova1, J. Szokol3, J. Nesvera3, M. Patek3

1DEKONTA, a.s., Dretovice 109, 273 42 Stehelceves, Czech Republic, www.dekonta.cz, [email protected]

2Institute of Chemical Technology, Department of Fermentation Chemistry and Bioengineering, Technicka 5, 166 28 Praha 6, Czech Republic

3Institute of Microbiology, AV CR, v. v. i., Vídeňska 1083, 142 20 Praha 4, Czech Republic

The strain Rhodococcus erythropolis CCM2595 was used for the construction of strains containing the genes for phenol degradation in the multicopy plasmid vector pSRK21. The constructs carried some of the genes pheA1+pheA2 (phenol hydroxylase), pheR (AraC-type regulator), catABC (catechol 1,2-dioxygenase, cis,cis-muconate cycloisomerase, muconolactone isomerase) and catR (IclR-type regulator) in various combinations and organization. Potential of the wilde-type strain and its seven recombinant derivatives to degrade phenol was first tested in a synthetic medium with phenol (300 mg/L). The strains were then applied in biodegradation of phenol in wastewater effluent from a chemical plant producing Ionex dian (Bisphenol-A) in a laboratory scale. The Rhodococcus strains were cultivated in 500 mL flasks in the wastewater containing phenol (600 mg/L) and decrease of phenol concentration in the medium was determined during the cultivation. Phenol in the wastewater was completely degraded within 18 days in the cultures inoculated by R. erythropolis (pSRK21phe), which carried the pheA1, pheA2 and pheR genes. The other strains degraded 38 – 97% of phenol during the same period. Plasmid DNA of the expected size was isolated from the R. erythropolis cells after completing the cultivation. This indicated that the plasmids were stably maintained in the cells during the cultivation. The results suggest that phenol hydroxylase activity was limiting for the phenol degradation under the conditions used and phenol was degraded faster due to the higher pheA1+pheA2 gene dosage.

The work was supported by grant 2B08062 AROMAGEN from the Czech Ministry of Education.


197

IBBS-15-7-O10

Effect of redox mediators and various media on the decolourisation of the azo dye Methyl Red; and its biodegradation by Providencia rettgeri strain ODO

Olukanni Olumide David *, Adekola Adejoke Adebimpe, Sunmola Damilola Banke, Osunmuyiwa Olufolabi

Environmental Biotechnology Laboratory, Department of Chemical Sciences, College of Natural Sciences, Redeemer’s University. PMB 3005 Redemption City, Ogun State, Nigeria.

*Author for Correspondence: Tel: +2348059360929, E-mail: [email protected],

Abstract

The discharge of coloured wastewaters into water-bodies is a serious environmental and public health problem. Developing efficient, effective and low cost procedure for wastewater treatment thus becomes expedient. A bacterium isolated from textile dye was identified using 16S rDNA as Providencia rettgeri. The organism decolourised various textile and non-textile dyes within 6 h at 100 mg/L. The prospect of redox mediators: quinol, nicotinamide adenine dinucleotide (NAD), reduced NAD (NADH), nicotinamide adenine dinucleotide phosphate (NADP) and reduced NADP (NADPH) to influence the decolourisation of the azo dye Methyl Red was investigated. Effects of various media on the decolourisation were also examined. The possible biodegradation of the model dye was investigated by subjecting the decolourised dye to UV-visible, high performance liquid chromatography (HPLC) and fouier transform infrared (FTIR) analyses. The organism preferred quinol to other redox mediators and nutrient broth to yeast, glucose, starch and peptone as media. The decolourisation time was reduced to 5 h. The results of the UV-visible and HPLC analyses suggested the biodegradation of the dye. This was confirmed by the removal of signature peaks of aromatic C-H bending (617, 704 and 834 cm -1) and the N=N peak at 1621 cm-1 observed when the FTIR of the dye was compared to that of its metabolite. The high Methyl Red decolourisation and biodegradation ability of Providencia rettgeri would enable this bacterium to be used in the biological treatment of industrial effluents containing azo dyes.

Keywords: Decolourisation, Biodegradation, Redox mediators, Methyl Red, Providencia rettgeri


198

IBBS-15-7-O11

Application of recombinant Nostoc calcicola for the removal of Zinc(II) from Textile Industry Effluents Subramanian Ramachandran1, Marianne Saldanha1, Irshad Ahmed1, Pankaj Kumar Jain2 and Sanjay Kumar Verma2

1Department of Biotechnology, Birla Institute of Technology and Science, Pilani, Dubai campus, UAE, e-mail: [email protected] 2Center for Biotechnology, Biological Sciences Group, Birla Institute of Technology and Science, Pilani, Rajasthan-333031, India

Viscose fibre is formed by the regeneration of cellulose treatment with a solution of electrolytes. Due to the environmental concern, Zinc(II) which act as catalyst in a vital process and provide the required strength to the fibre is being replaced with alum as less preferred alternative. The cellulosic fibre produced by this process is both fragile and low in quality. The quality and the market value of the product can be increased by many folds if an effective, environmental friendly technique is developed and employed to remove Zn from the effluent. We tested the wild type and the recombinant Nostoc calcicola expressing mouse metallothionein-I for the removal of Zn from the effluent. It was found that the recombinant N. calcicola could accumulate higher amount of metals including Zn. Although, the recombinant N. calcicola was sensitive to the effluent, it grew in effluent supplemented with culture medium. The growth of the cells was rapid at lowest concentrations of the effluent followed by a decrease upon increase in waste water in the culture medium. The growing cultures of recombinant N. calcicola in bioremediation of Zn from the effluent could avoid the need for a separate biomass production, in addition, the nitrogen-fixing N. calcicola release beneficial substances of plant growth promoting properties and fixed nitrogen into the culture. Hence, the treated water can be safely disposed into the environment or used for applications like plantations.

Key words: textile effluent, Zinc(II) bioremediation, recombinant Nostoc calcicola

Acknowledgements: The authors are thankful to the Director, BITS Pilani, Dubai campus for providing laboratory facilities and the sophisticated instrumentation facility (supported by the DST, India) at the All India Institute of Medical Sciences, New Delhi.


199

IBBS-15-7-P01

Effect of soy peptone as a nutrient on biodegradation of oil sludge in the soil

Sina Azami, Pejman Rohanifar, Dariush Minai-Tehrani, Saeed Minoui

Faculty of Biological Sciences, Shahid Beheshti University, Tehran, IRAN

Abstract

Oil sludge is a viscous material consisted of resin, asphaltine, sand and water. The asphaltine and resin content of oil sludge make it more resistant to biodegradation. The discharge of oil sludge into the soil causes damage to environment. Oil sludge is recalcitrant and can reside in the soil for long period. Some microorganisms are able to use hydrocarbon content of oil sludge as carbon source and can help to clean up pollution from soil. In this report the biodegradation of oil sludge in the soil was investigated for six months. The oil sludge was mixed with soil in different concentrations (0- 10% w/w) and the nutrients such phosphate and nitrate salts were added to the soil to help microorganism growth. On the other hand soy peptone was added to the samples to increase microorganism population and the rate of biodegradation was compared with samples without soy peptone (control). Our analysis showed that in peptone content samples, both total bacteria and oil degrading bacteria were increased in comparison with control. Increasing oil sludge concentration increased the microorganisms' population in the sample. TPH measurement in the soil after six months of treatment showed that in both peptone containing samples and the samples without peptone the amount TPH have decreased. However in peptone containing samples the reduction were higher than the control. Maximum reduction was observed in 1% sample (39%) while the minimum was seen in 10% sample (25%). Our result showed that peptone could increase biodegradation of oil sludge in soil.

Keywords: Biodegradation, oil sludge, soy peptone, nutrient


200

IBBS-15-7-P02

Biomaterials for Heavy Metals Adsorption

Andreea Bondarev1, Sonia Mihai1, Daniela Popovici1 and Octav Pantea1

aPetroleum-Gas University of Ploiesti, Bucharest Av. 39, Ploiesti, Romania

Naturally abundant biosorbants such as chitin and chitosan are recognized as excellent metal ligands, forming stable complexes with many metal ions. Incorporation of zeolites in different biopolymers membranes is an effective method to control the diffusion outside the zeolite crystals and these composite systems can find many opportunities in wastewater treatment. In short, the use of biopolymers and their composites as adsorbent materials for heavy metal removal from aqueous contaminated wastes is an environmentally friendly method.

In this paper, the results of heavy metal removal capacity of some chitosan-zeolite microspheres are compared to the adsorption capacity of other sorbents: chitosan, ethyl cellulose and clinoptillolite. The chitosan-zeolite composites were prepared by spray drying method and used as an adsorbent for some heavy metals in aqueous solution. The chitosan microspheres were characterized by FTIR and scanning electron microscopy. The concentration of heavy metal ions in supernatant was determined by the atomic absorption spectrophotometric method. Adsorptive experiments were carried out in a steady state regime, with initial concentration ranged from 30 to 100 mg/L. The adsorption behavior was described by Langmuir and Freundlich adsorption equations. The effect of several experimental parameters on the metal retention capacity of the chitosan microspheres, such as stirring system, time, amount of sorbent and temperature were investigated.


201

IBBS-15-7-P03

Microbial dynamics in two-phase-partitioning bioreactors for hexane removal show partitioning of distinct groups into non-aqueous phase Ng Chow Goon1, Miao Huang2, Sze Chun Chau1

1School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive Singapore 637551, Singapore 2School of Civil and Environmental Engineering, Nanyang Technological University, 50 Nanyang Avenue, Sinapore 639798, Singapore

Two-phase-partitioning bioreactors (TPPB) have been found to improve the biodegradation of toxic volatile organic compounds (VOC) by enhancing their bioavailability and/or reducing their toxicity on the microbial community. However, to date, there has been limited knowledge on the microbial dynamics within TPPB over long-term operation. This must be elucidated to improve its uses in bioremediation. A study was carried out to characterize the microbial dynamics in a model TPPB system for hexane removal over 66 weeks. This system consists of an aqueous phase that carries the microbial community and minimal nutrients, and a water-immiscible organic phase of silicon oil which exhibits high solubilisation capacity for the target VOC, hexane. The TPPB system monitored over 66 weeks was found to support better microbial growth, exhibit higher surfactant production and showed greater efficiency in hexane removal compared to the conventional monophasic bioreactors. Microbial community dynamics was analysed by a strategy combining 16S rDNA sequencing with PCR amplified ribosomal DNA restriction analysis (ARDRA), followed by denaturing gradient gel electrophoresis (DGGE). Interpretation of DGGE profiles across time was facilitated by a DGGE ladder, constructed using culturable and identified isolates from the bioreactors. Data showed difference in the microbial compositions between the two types of bioreactors, suggesting divergent development of microbial community due to the presence of the non-aquoues phase. More interestingly, microbial groups within the aqueous and non-aqueous phases within TPPBs showed considerable difference, suggesting partitioning of specific groups, most likely based on their cell surface hydrophobicity.


202

IBBS-15-7-P05

Isolation and identification of fungi capable of textile dye degradation

Wafaa M. Abd El-Rahim**, Magda A. El-Meleigy*, Eman Refaat*

* Department of Microbiology, Faculty of Science, Al-Azhar University (Girls' Branch)

**Department of Agriculture Microbiology, National Research Centre, Dokki, Cairo, Egypt

Abstract

In this study a trial was made to isolate fungal species efficient in biodegradation of textile dyes from the wastes of industrial textile factories and the surrounding environment. Ten wastewater samples were collected from industrial waste dumping sites at El-Mahalla El-Kubra, Shubra El-Khima, Kafr El-Dawar and Borg El-Arab Factories. The samples were then analyzed for their TSS, COD, pH, EC and BOD. Eleven fungal isolates were obtained by plating the factory effluents on mineral-basal media containing dye as the sole source of carbon and nitrogen. These isolates were identified as follows: Aspergillus terreus, Cladosporium sphaerospermum, two Fusarim sp., Cladosporium cladosporioides, Asperigillus flavus, Fusarium oxysporium, Aspergillus fumigatus, Aspergillus niger, Penicillium chrysogenum and Aspergillus versicolor . The fungal isolates were tested to demonstrate their abilities in removing Direct Violet dye. Three isolates Aspergillus fumigatus, Aspergillus terreus and Cladosporium cladosporioides were the most efficient in decolorization of Direct Violet dye. The decolorization % for Aspergillus fumigatus isolate amounted 58.69, Aspergillus terreus isolate 35.28 and Cladosporium cladosporioides isolate 24.68 after 4 days incubation. These decolorization % were increased by time until it reached about 91.21, 89.5 and 82.89 for these isolates respectively after 16 days of incubation. Since the biodegradation of Direct Violet dye can be achieved by these fungal isolates.

Keywords: Fungi identification, bioremoval, Direct Dye.


203

IBBS-15-7-P08

Effects of Cu and Zn to the litter-decomposing fungi

E. Samuel Hartikainen, Pauliina Lankinen, Johanna Rajasärkkä, Hilkka Koponen, Marko Virta, Annele Hatakka, Mika A. Kähkönen

Department of Food and Environmental Sciences, Division of Microbiology, P.O. Box 56, Biocenter 1, FIN 00014 University of Helsinki, Finland, [email protected]

Effects of Cu and Zn were tested on the growth of twenty litter-decomposing saprotrophic fungi and their production of extracellular enzymes, laccases and peroxidases, with different Cu (0, 100, 200, 400 mg dm-3) and Zn (0, 100, 200, 400 mg dm-3) concentrations in the ABTS indicator colour plates. Bioavailability and toxicity of Cu and Zn were tested with bioluminescent Saccharomyces cerevisiae yeast and Escherichia coli bacteria. Coniothyrium sp. was the best growing ascomycete both in the Cu and Zn containing indicator colour ABTS (2, 2’-azino-bis (3-ethylbentzthiazoline-6-sulfonic acid) plates. The most sensitive ascomycete was Sordaria sp. both in the Zn and Cu containing ABTS plates. Cu (200 mg dm-

3) inhibited (22-100 %) the growth of all eight basidiomycetous fungi in the ABTS plates compared to the non Cu contaminated plates. The growth of Agrocybe praecox, Gymnopus peronatus, Gymnopilus sapineus, Mycena galecurilata, Stropharia aeruginosa and Stropharia rugosoannulata was higher (2 – 272 %) with 100 mg Zn dm-3 compared to the control plates. The growth of the tested fungi decreased 70 - 100 % in the 400 mg Cu kg-1 and 39 -100 % in the 400 mg Zn kg-1 compared to the 0 mg kg-1 Cu or Zn containing plates, respectively. Bioluminescent S. cerevisiae yeast tests indicates that Zn was 2-20 times less toxic than Cu. Bioluminescent E. coli bacteria were four times more sensitive to Cu than Zn. Tested ascomycetous, zygomycetous and basidiomycetous fungi showed that Cu was more toxic than Zn to the growth and enzyme production of fungi.


204

IBBS-15-7-P09

Treatment of radium in groundwater of Saudi Arabia by PCS fungi biofilm

Mohammad Ahmed Khiyami

King Abdulaziz city for Science and Technology; [email protected]; Saudi Arabia, P. O. Box 6086, Riyadh 11442

Radium is a highly radioactive element that is chemically similar to calcium. Thus it has the potential to replace bone calcium causing great harm. Further, inhalation, injection, ingestion or body exposure to radium can cause cancer and other disorders.

Saudi Arabia relies on both desalinated sea water and treated ground waters. Studies on Saudi Arabian water assured the presence of 222 Radium, 226 Radium and 228 Radium in ground water samples and scale samples taken from pipes used in water wells. Although some fungi, such as mushrooms possess potential ability to accumulate long-lived fallout radionuclides and heavy metal species, only limited researches have been conducted.

In this study, water and soil samples were collected from local wells in Riyadh region. The samples were analyzed for chemical elements and radium concentration. Isolated fungi were identified morphology and genetically. Since low pH value is the most important water parameter linked to high radium concentration, all isolates were evaluated to grow at low pH ≤ 2.5. The ability of isolates to develop PCS biofilm and remove the radium from water samples were evaluated in 250ml Erlenmeyer flasks. Each flask was supplied with 100 ml media with known concentration of radium. The flasks were inoculate with isolated fungi and incubated for 14 days. After the incubation period, the dry mycelium, pH, K+ and Ca++ were determined. Also, radium reduction will be determined in the media and in the mycelium. Finally, a bioassay was developed to measure the safety of using the treated water.


205

IBBS-15-7-P12

Tributyltin Bioavailability and Biodegradation in S oil after Surfactant Flushing

Lada Mathurasa1,2, Ekawan Luepromchai2,3, Chantra Tongcumpou2,4,

David A. Sabatini5

1National Center of Excellence for Environmental and Hazardous Waste Management (NCE-EHWM), Chulalongkorn University, Bangkok, Thailand

2International Postgraduate Programs in Environmental Management, Graduate School, Chulalongkorn University, Bangkok, Thailand

3Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand

4Environmental Research Institute, Chulalongkorn University, Bangkok, Thailand

5School of Civil Engineering and Environmental Science, The University of Oklahoma, OK, USA

Biodegradation of tributyltin (TBT) in soil is known to be very slow. However, surfactant flushing might increase TBT biodegradation through improving its desorption and bioavailability. To prove this hypothesis, soil slurry microcosms were conducted by mixing 2 g sandy clay loam soil with 20 mL anionic surfactant, sodium dihexylsulphosuccinate (SDHS) and incubated for 2 months. The concentrations of TBT were 0.05 and 0.10 mg Sn of TBT g-1 soil, while SDHS were 0, 5, and 40 mM. The low concentrations of SDHS were selected to avoid soil bacterial cell lysis and simulate the remaining surfactant after soil flushing. TBT was rapidly desorped from soil. The increasing concentrations of SDHS increased TBT desorption, however the amounts of desorbed TBT from soil with different initial TBT concentrations were the same when similar SDHS concentration was used. At the end of experiment, the percentage of TBT removal compared to sterilized soil microcosms was around 50 and 80% in the microcosm with 5 and 40 mM SDHS, respectively. TBT-degrading metabolites, dibutyltin and monobutyltin were detected within 4 week. Consequently, TBT biodegradation was controlled by desorption mechanism. The numbers of total bacteria and TBT-degraders in soil containing both TBT and SDHS decreased during the first 3 weeks, while bacterial numbers in soil with either TBT or SDHS were almost constant. Beside, the PCR-DGGE profile showed that the soil bacterial community was majorly affected by the presence of TBT and its metabolites. The toxicity of desorbed TBT and its degrading metabolites were similar to other studies.

Keywords: Tributyltin, Surfactant, Biodegradation, Bioavailability


206

IBBS-15-7-P13

Rhizoremediation of rice soils with residual clomazone herbicide

Maria Laura Turino Mattos1; Andre Andres1; José Francisco da Silva Martins1

1Embrapa Temperature Climate, BR 392, km 78, 96001-970, Pelotas, RS, Brazil

[email protected]

Rhizoremediation is a beneficial interaction between plants and microorganisms to increase the degradation efficiency of pesticides. This combination is especially important in the remediation of soils contaminated with the herbicide clomazone of the technology Permit (TP). TP aims at weed control of rice cultures, consisting in the treatment of seeds that contain a protector that allows for greater flexibility in the dosages of this herbicide. However, it also generates residues that can cause phytotoxicity of sensitive cultures seeded post-utilization of this technology. The objective of this work was to evaluate species of forage grass for the phytoremediation of soils with residual clomazone, and isolating rhizobacteria associated with these forages. Soil samples were collected from experimental plots cultivated with forage sorghum during summer and perennial ryegrass during winter, after four and three consecutive years of irrigated rice culture with TP, respectively. The rhizobacteria were recovered in slides containing Thorton’s medium, totalling 26 isolates, which were purified and deposited in the Multifunctional Microorganisms Collection of Embrapa Temperate Agricultural. The taxonomic characterization tests of the isolates demonstrated biochemical diversity in the utilization of sugars. The technical basis for identification of the isolates will be generated with the continuity of tests at the biochemical and molecular levels. The rhizoremediation processes with forage sorghum and perennial ryegrass demonstrated efficiency in the degration of clomazone, without production of residues or incidence of phytotoxic effects. The inoculation with clomazone degrading rhizobacteria of seeds of the species evaluated may be an important additive to improve the efficiency of phytoremediation. In addition, it offers low cost when compared to conventional methods of remediation of soils contaminated with pesticides.


207

IBBS-15-7-P14

Probiotics – Solution for bioremediation in fish and shrimp farming Elisabeth Mayer, Eva-Maria Gössl, Gonçalo A. Santos, Michaela Mohnl BIOMIN Holding GmbH, Industriestrasse 21, A-3130 Herzogenburg Intensive aquaculture produces large amounts of organic wastes. Oxidation of these waste compounds facilitates the formation of toxic metabolites (H2S, NH3, NO2

-) causing deteriorated water quality. This also changes bacterial composition in water and soil of ponds, increasing the presence of pathogenic bacteria, thus contributing to the occurrence of diseases in fish and shrimp. The application of beneficial bacteria, probiotics and biodegrading microorganisms, to the ponds (water and soil) is a sustainable approach to minimize the environmental impact of aquaculture (bioremediation). Beneficial bacteria as biological control agents in aquaculture

Biodegrading strains

Gram-positive Gram-negative

Bacillus Paracoccus

Lactobacillus Thiobacillus

Enterococcus Nitrobacter

Propionibacterium Nitrosomonas

The heterotrophic Paracoccus impaired the nitrogen cycle by using NO3

- and other oxidized nitrogen compounds instead of oxygen and drastically reduced the amount of nitrogen compounds. The strain was able to degrade almost the total amount of 60 mg/L NO3

- in the test solutions within 24 hours. Furthermore, sodium acetate in the medium led to decrease of 16 mg/L NH4

+ to 7 mg/L within 48 hours. It has been shown that Paracoccus is able to control the above mentioned processes and reduce undesirable waste compounds.

NH4+- or NO2

-- removal is essential for the pond water quality and can be carried out by denitrifying bacteria such as Thiobacillus and Paracoccus. An in vitro study was conducted to test the biodegrading effect of Paracoccus pantotrophus 768 in two media: an aerobic buffer with 1 g/l yeast extract and 1 g/l sodium acetate as an additional carbon source, and without sodium acetate.


208

IBBS-15-7-P15

Assessment of bioremediation techniques (biostimulation, bioaugmentation and natural attenuation) in a creosote polluted soil and restoration of the microbial community

Raquel Simarro1, Natalia González1, L. Fernando Bautista2, M. Carmen Molina1, Miguel Pérez1, Luis Pérez2

1Department of Biology and Geology, 2Department of Chemical and Environmental Technology. Universidad Rey Juan Carlos. C/ Tulipán s/n. E-28933 Móstoles, Madrid, Spain

Abstract

Besides the changes undergone in the composition and structure of soils after a pollution spill, natural microbial population is usually significantly altered. Although it is well-known that physicochemical and thermal remediation techniques have a great impact in soil microbiology, bioremediation treatments can also affect soil biodiversity and function.

The aim of the present work was to assess different bioremediation treatments (bioaugmentation, biostimulation, combination of both and natural attenuation) applied to a creosote polluted soil with a two-fold purpose: effectiveness in removal of pollutants and degree of restoration of the microbial soil community.

Results showed that for all treatment creosote was degraded through time. However, no treatment performed significantly better than others when the total amount of organic compounds was considered. Analyzing specific polycyclic aromatic hydrocarbons, biostimulation achieved better biodegradation extent. In this way, naphthalene was completely removed and degradation of anthracene was deeper than that for phenanthrene and pyrene in all treatments. Toxicity (measured using bioluminescent bacterium Vibrio fischeri) raised and reached a maximum value at the end of the experiment due to the formation of smaller and more soluble compounds as a result of creosote oxidation. Biostimulated soils showed larger microbial growth rate for creosote-degrader bacteria once temperature had risen. Statistical analysis on DGGE results revealed that biostimulated soils displayed the highest microbial biodiversity. However, after the treatment, the most similar bacterial community to that of the unpolluted soil appeared in the soil where natural attenuation was applied.


209

IBBS-15-7-P16

Biodegradation of carbazole by methylotrophic bacteria Methylobacterium sp. GPE1

Grzegorz Pasternak, Barbara Kołwzan

Abstract

Heterocyclic compounds are the group of chemicals produced widely by the industry. Their toxic properties cause several environmental problems. This research was focused on utilization of carbazole – a model nitro-substituted heterocyclic compound. The bacterial strain used in this study was derived from soil contaminated by gasworks activity. The biodegradation tests were carried out for 20 days. During the test period bacterial growth was accompanied with significant decrease of surface tension. The decrease in carbazole concentration was 88 % and growth rate 0,29 h-1. Due to limited solubility of carbazole investigated strain was relatively not affected by its concentration. The genus of Methylobacterium sp. is widely present in the environment and known to stimulate the growth of plants. Therefore strain described in this study could be a promising component of bioremediation techniques.


210

IBBS-15-7-P17

Construction of chromosomal deletions and gene-replacements in Rhodococcus strains degrading aromatic compounds

Lenka Rucká, Michaela Šimčíková, Juraj Szőköl, Petr Halada, Jan Nešvera and Miroslav Pátek

Institute of Microbiology, AS CR, v. v. i., Prague 4, Czech Republic;

tel: (+420)241062398, fax: (+420)241722257, [email protected]

Rhodococcus strains are able to grow on various aromatic pollutants like phenol, p-chlorophenol, benzoate, hydroxybenzoate and aniline. Genes responsible for phenol degradation in R. erythropolis CCM2595 and R. jostii RHA1 were cloned and regulation of their expression was analysed. The pheA1+pheA2 genes (coding for subunits of phenol hydroxylase) were found to be regulated by the PheR activator (AraC-type protein) and the genes of the catABC operon (catechol 1,2-dioxygenase, muconate cycloisomerase and muconolactone isomerase, respectively) are controlled by the CatR repressor (IclR-type protein). Promoters of the studied genes were localized by transcription start site determination and binding of PheR, CRP and PcaR regulators within the regulatory regions of the genes were proved by protein-DNA interaction chromatography and mass spectrometry (MALDI-TOF). Rhodococcus strains harbouring multicopy plasmid vectors carrying some of the phenol degradation genes in various combinations and organization were shown to degrade phenol more efficiently than the wild-type strain. Based on the acquired knowledge, deletions, insertions and rearrangements within the genes and regulatory regions aimed at increasing expression of phenol degradation genes were introduced into the chromosome using the double homologous recombination events. The constructed modifications included deletion of the CatR repressor gene, insertion of an additional constitutive promoter upstream of the pheA2A1 operon and rearrangement of the common regulatory region of divergently transcribed catR and catABC genes. The developed Rhodococcus strains were evaluated in terms of ability to resist higher phenol concentrations and to degrade phenol efficiently.


211

IBBS-15-7-P18

Biosorption of Cr(VI) and Cr(III) by Nostoc calcicola immobilized on silica coated calcium alginate and silica gel

Subramanian Ramachandran1, Marianne Saldanha1, Irshad Ahmed1, Pankaj Kumar Jain2 and Sanjay Kumar Verma2

1Department of Biotechnology, Birla Institute of Technology and Science, Pilani, Dubai campus, UAE, e-mail: [email protected] 2Center for Biotechnology, Biological Sciences Group, Birla Institute of Technology and Science, Pilani, Rajasthan-333031, India

Chromium(VI) compounds exhibit carcinogenic and mutagenic effects on living systems due to their strong oxidizing nature. The remediation of residual chromium from industrial effluents is essential and the biological chromium removal has distinct advantages over conventional technologies. The present investigation was aimed at exploiting the cyanobacterium Nostoc calcicola for the removal of Cr(VI) and Cr(III). The optimum biosorption of Cr(VI), Cr(III) by N. calcicola occurs at pH 2.0 and pH 5.0 at 30oC. The lyophilized N. calcicola cells could accumulate a maximum of 159.5mg g-1 dry wt of cells within 12 hours. The maximum Cr(III) uptake was found to be 187.6 mg g-1 dry wt biomass and attains equilibrium at 2 hours. The N. calcicola cells were immobilized on silica gel by using sodium silicate and colloidal silica. The silica-immobilized cells were dried, powdered and sieved to pass 20-40 mesh size were used in the subsequent experiments. The silica-immobilized cells were found to be superior in terms of metal removal, metal loading capacity and reusability in multiple cycles. We also optimized a process for the reduction of Cr(VI) to less toxic Cr(III) in 24 hours with wet biomass. We identified the role of phycobiliproteins in the reduction of hexavalent chromium in acidic conditions. N. calcicola have potential applications not only for the remediation of hexavalent chromium but also as a cheap reducing agent for detoxification electroplating industry effluents.

Key words: Chromium compounds, biosorption, Nostoc calcicola, silica gel

Acknowledgements: The authors are thankful to the Director, BITS Pilani, Dubai campus for providing laboratory facilities and the sophisticated instrumentation facility (supported by the DST, India) at the All India Institute of Medical Sciences, New Delhi.


212

IBBS-15-7-P20

Bioremediation of clay soil contaminated with gasoline blended with different levels of ethanol Douglas Guedes Ferreira1,2,3, Márcia Teresa Soares Lutterbach2, Viviane Oliveira2, Luciana Silva Contador2, Mariana Machado Galvão2, Denize Dias de Carvalho3, Eliana Flávia Camporese Sérvulo1

1Laboratório de Microbiologia Industrial, Escola de Química, Universidade Federal Rural do Rio de Janeiro, Brasil. 2Laboratório de Biocorrosão e Biodegradação, Divisão de Corrosão e Degradação, Instituto Nacional de Tecnologia, Brasil. 3Laboratório de Tecnologia Ambiental, Escola de Química, Universidade Federal Rural do Rio de Janeiro, Brasil. Soil is a rich ecosystem in terms of biodiversity, but human activities constantly threaten the dynamic and even the maintenance of these species in the environment. Among these human activities is the contamination of terrestrial and aquatic ecosystems by petroleum and its derivatives, particularly gasoline due to its high toxicity to the environment and human health. The negative effects of gasoline can be enhanced by ethanol-blending, such as in the Brazilian gasoline. The damage caused by these accidents should be restricted and treated to recover soil natural conditions. An interesting alternative for environmental recovery is the use of biological treatment methods such as bioremediation, which relies on the ability of microorganisms to metabolize contaminants compounds. This work aimed to study bioremediation of gasoline blended with different levels of ethanol in clay soil, according to the nutritional condition, moisture and soil depth. This study was divided into two parts, the first part was conducted in microcosms with different conditions organized by a full factorial design and aimed to evaluate the interference of nutritional condition and soil moisture on bioremediation. The second part was performed in cylindrical columns with 60 cm high and 10 cm radius in order to monitor the bioremediation at different depths. The results obtained from these tests indicated that ethanol at the highest concentration tested (25%) did not interfere in the bioremediation; however it accelerates the percolation of the contaminant through the soil. On the other hand, the adjustment of the nutritional status to 50 ppm of phosphorus and moisture allowed more than 50% of the contaminant to be removed within 60 days. This fact indicates that the adjustment of the nutrient concentration and water content in the soil promote a better yield of microbial metabolism and consequently a more effective bioremediation.


213

IBBS-15-7-P21

Effect of broth composition and salinity in the volumetric mass transfer coefficient (kLa), for aerobic bioprocess to break organic gas residuals.

Moraes1, D.A.; Anselmo, A.F1; Souza, E.C. 1; Vessoni Penna, T.C. 1

1School of Pharmaceutical Science of University of São Paulo (USP). São Paulo, Brazil.

Correspondent author: [email protected]

Aeration and agitation are important variables to ensure effective oxygen transfer rate during aerobic bioprocesses to break the organic residuals (for instance: toluene) leaked into soils from the gas station; therefore, the knowledge of the volumetric mass transfer coefficient (kLa) is required. In view of selecting the optimum oxygen requirements for the fermentation of toluene in different broths, the kLa for the systems studied at agitation rate of 300 rpm and aeration from 1.0 (0.06 m/s) to 1.5 vvm (0.09 m/s) O2, were: sterile water (kLa= 1455 s-1); saline rejected residue (11 mOsm) obtained from reverse osmosis system (kLa = 749,3 s-1), rejected residue added with 1% w/v glucose (kLa = 727 s-1), rejected residue added with 5% w/v yeast extract (KLa = 356 s-1); saline (104 mOsm) broth added with 1% w/v glucose (kLa = 1288 s-1), saline broth added with 5% w/v yeast extract (kLa = 438 s-1); brain and heart (BH, 0.110 mOsm) medium (kLa = 645 s-1), BH added with 1% w/v glucose (kLa = 679,4 s-

1), BH added with 5% w/v yeast extract (kLa = 309 s-1); sea (0.660 mOsm) water (kLa = 767 s-1), sea water added with 1% w/v glucose (kLa = 408 s-1), and, sea water added with 5% w/v yeast extract (kLa = 442 s-1). It was observed that the combination of toluene added to BH or rejected residue or sea water media (without yeast extract) presented similarity of kLa (around 506-516 s-1), which was shown adequate for aerobic bioprocesses.


214

IBBS-15-7-P23

A study on degradation of Remazol Red by Galactomyces geotrichum MTCC 1360 and effect of dye and degraded metabolites on iron uptake in Sorghum vulgare and Phaseolus mungo

Tatoba R.Waghmode1, Mayur B. Kurade2, Sanjay P. Govindwar1

1Department of Biochemistry, Shivaji University Kolhapur-416004, India.

2Department of Biotechnology, Shivaji University Kolhapur-416004, India.

Abstract

Galactomyces geotrichum MTCC 1360, a yeast species showed 96.16% decolorization of Remazol red (50 mg l-1) within 36 h at 30oC and pH 11.0 under static condition with significant reduction in COD (Chemical oxygen demand) (62%) and TOC (Total organic carbon) (41%). Peptone (0.5%), Rice husk (5ml of 1% extract) and ammonium chloride (0.5%) were found to be more significant within other carbon and nitrogen sources used. The presence of tyrosinase, NADH-DCIP reductase, Riboflavin reductase and induction in azo reductase and laccase activity during decolorization indicated their role in degradation. HPTLC (High performance thin layer chromatography) analysis revealed the degradation of Remazol Red into different less toxic metabolite. FTIR (Fourier transform infrared spectroscopy) and HPLC (High performance liquid chromatography) analysis of samples before and after decolorization confirmed the biotransformation of dye. Atomic Absorption Spectroscopy (AAS) analysis revealed less toxic effect of degraded metabolite on iron uptake of Sorghum vulgare and Phaseolus mungo than Remazol Red dye. There were significant negative relationship between the uptake of iron by crop and the concentration of Remazol Red added to soil and positive relationship between the uptake of iron by crop and degraded metabolite. According to the extraction with 1.0 M ammonium acetate and concentrated nitric acid, the Remazol Red showed adverse effect on iron upatake by chelation and immobilization of iron, whereas degraded metabolite showed no chelation as well as immobilization of iron. Phytotoxicity study indicated the conversion of complex dye molecules into simpler oxidizable products having less toxic nature.


215

IBBS-15-7-P24

Biodegradation of natures own contaminants, microcystins and nodularin

Aakash Welgama, Christine Edwards, Linda A. Lawton

Institute for Innovation, Design, and Sustainability Research, Robert Gordon University, Aberdeen, AB25 1HG, UK

Cyanobacterial secondary metabolites, microcystins (MC) and nodularin have become common contaminants in most aquatic ecosystems over the recent years presenting a hazard to animal and human health. Unfortunately, these chemically diverse peptide hepatotoxins remain a challenge to most conventional water treatment due to their stable cyclic structures. Over the recent years, bioremediation of microcystins and nodularin has become one of the most exciting areas that hold promise for a successful and cost effective solution for water treatment process. The current work presents the biodegradation of microcystins and nodularin by bacterial isolates from three different bacteria genus Arthrobacter, Brevibacterium and Rhodococcus belonging to Actinobacteria. A total of five isolates representing the three genera were found to degrade MC-LR, -LF, -LY, -LW, -RR and nodularin in sterile water from their origin. The bacterial degradation rate of above cyanobacterial peptides were found to decrease with the multiple culturing of bacteria. However, a rapid degradation was discovered when the bacteria were pre-exposed to MC or other prokaryotic peptides demonstrating an inducible bacterial biodegradation involved. Elucidating genes behind the degradative mechanism would enhance the use of these bacteria in future water remediation strategies.


216

IBBS-15-7-P25

Bioremediation of contaminated soil with fungi in aerobic conditions

Erika Winquist1, Festus Anasonye2, Kalle Salonen1, Markus Räsänen1, Kari Steffen2 and Marja Tuomela2

1Department of Biotechnology and Chemical Technology, Aalto University, Finland

2Department of Food and Environmental Sciences, University of Helsinki, Finland

Corresponding author: [email protected]

Fungal treatment methods could provide new sustainable ways to treat contaminated soil. Especially new treatment methods are needed for high molecular mass organic contaminants that cannot be degraded by composting, e.g. polychlorinated dibenzo-p-dioxins and –furans (PCDD/F). In our studies, we used various soils contaminated with 1) oil hydrocarbons (44 000 mg/kg), 2) polyaromatic hydrocarbons (PAH) (5000 mg/kg), 3) PCDD/F (up to 50 000 ng/kg TEQ) and 4) trinitrotoluene (TNT) (1000 mg/kg). These contaminated soils originated from an oil refinery site, sawmill or military areas.

White-rot (WRF) and litter-decomposing fungi (LDF) produce extracellular oxidizing enzymes, such as laccase and manganese peroxidase (MnP), These enzymes are non-specific, and in addition to lignin they can oxidize a wide range of organic contaminants. We studied the capability of selected WRF and LDF to grow in the soil, produce lignin modifying enzymes and degrade toxic compounds.

Gymnopilus luteofolius, Kuehneromyces mutabilis, Phanerochaete velutina and Stropharia rugosoannulata were selected for laboratory experiments after extensive screening (Valentin et al., 2009). Fungi were grown in contaminated soil and aerated with moist air. For some cultivations fungal activity was measured by following CO2 production by the fungus on-line and for the other cultivations laccase and MnP activity in soil. Oil hydrocarbons, PAH PCDD/F and TNT were analysed at the beginning and after fungal treatment.

Valentin, L., Kluczek-Turpeinen, B., Oivanen, P., Hatakka, A., Steffen, K. and Tuomela, M., Evaluation of basidiomycetous fungi for pretreatment of contaminated soil, J Chem Technol Biotechnol 84 (2009) 851-858.


217

IBBS-15-7-P26

Enhance of cattle manure and sewage sludge on bioremediation of Aged Polycyclic Aromatic Hydrocarbons Contaminated Agricultural Soils

Zhang Jing1,2, Lin Xian-gui1,2* , Yin Rui1,2, Liu Wei-Wei3

1. State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008, China E-mail: [email protected]

2. Joint Open Laboratory of Soil and Environment, Institute of Soil Science and Hong Kong Baptist University, Nanjing 210008, China

3. College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, China

Abstract: The effectiveness of in-situ bioremediation of polycyclic aromatic hydrocarbons (PAHs) may be inhibited by low nutrient and organic carbon. The aim of this research was to evaluate the effect of organic wastes on the efficiency of bioremediation for removal of aged PAHs. The contaminated agricultural soils were amended with different contents of cattle manure and sewage sludge when alfalfa (Medicago sativa L.) were planted and inoculated with PAHs-degrading bacteria (Bacillus sp.and Flavobacterium sp) at the same time. The results indicated that organic wastes addition promoted plant growth and soil PAHs degradation, especially in those treatments with low content organic waste addition. After 60 days, removal percentage of PAHs was 25.8%, while with sewage sludge and cattle manure addition, the removal ratio reached to 32%~34.9% and 21.9%~43.7%, respectively, but only 5.6% for control. Especially the degradation of high-molecular-weight PAHs were more than 40%. Soil dehydrogenase activity and number of PAHs degrading bacteria significantly correlated with the removal ratio of PAHs, and were stimulated by addition of organic wastes. Therefore, organic wastes application on soil PAHs removal, to a great extent, dependent on the combined impact of the content and properties of waste organic matter and dissolved organic matter in compost soil.


218

IBBS-15-7-P27



The University of Akureyri Department of Natural Resource Sciences. Akureyri, Iceland.

Although generally reputed pure and uncontaminated, most Icelandic aquatic environments are to some degree anthropogenically impacted. We surveyed two types of aquatic environments; three wastewater-exposed seashore environments and a freshwater system comprising the river Glerá and its watershed. The widespread and copious usage of several types of cleaning detergents and other surfactants results in their relatively high concentrations in both domestic and industrial sewage, which enters the aquatic environment untreated in most Icelandic localities. The river Glerá in Northern Iceland presents a convenient system for evaluating the effects of anthropogenic activities on microbial communities. The river originates in pristine surroundings and then flows past a local landfill and through an urban area, receiving inflows from various small-scale industries. Samples from the two environmental types were screened for surfactant-degrading activity and surfactant biodegraders isolated. All of the degrading isolates were found to belong to the class Gammaproteobacteria. In the freshwater system, surfactant degraders solely comprised pseudomonads, while the species composition was more diverse in the seashore samples. The surfactant-degrading freshwater pseudomonads mostly comprised members of the Pseudomonas fluorescens and P. syringae species groups. The seashore was generally found to be a poor source of surfactants degraders. The river environment was also screened for the appearance of mercury-tolerant bacteria, as an indicator of the degree of environmental pollution. It was found that the landfill leachate had an impact on the river microbiota downstream. Mercury resistance was markedly increased, whereas dominant species diversity, as determined by DGGE, was only modestly affected.


219

Topic 8-Biodiversity, ecophysiology and function of organisms involved in

biodeterioration and biodegradation


220

KEYNOTE LECTURE

ARE WE APPLYING THE RIGHT APPROACH TO STUDY THE MICROFLORA COLONIZING OUR BUILT CULTURAL HERITAGE?

Clara Urzì1*, Filomena De Leo1, Laura Bruno2 and Patrizia Albertano2

1 University of Messina, Department of Life Sciences,, Viale F. Stagno d’Alcontres 31, 98166 Messina, Italy

*[email protected]

2 University of Rome “Tor Vergata”, LBA - Laboratory of Biology of Algae, Department of Biology, via della Ricerca scientifica snc, 00133 Rome, Italy

The methodological approach to assess the decay of monuments must be finalized to their conservation, and standardized procedures for cultural heritage used as already recommended by National and European committees. However, in the past years microbiologists dealing with the study of the numerous and diverse habitats present in the “cultural heritage environment” have often neglected the conservative purpose and privileged the floristic and biological aspects of these subaerial microbial communities. In this context, a three-phases multi-step interdisciplinary approach to study biofilm-associated alterations in archaeological subterranean sites and outdoor monuments is proposed.

The first phase consists of an in situ study of the geo-referenced site to characterize microclimate, substratum and physical, chemical and biological parameters. Digital images and descriptions of alterations are periodically collected, while data recording and sampling are conducted using non-invasive (NIT) and non-destructive (NDT) techniques. The second phase is carried out in laboratory thanks to a variety of microscopy investigations combined with culture-independent and culture-based techniques to identify microorganisms and to assess their biodeteriorative potential. The third phase includes the testing of conventional and innovative techniques to prevent and control the growth of biodeteriogens under laboratory conditions and the subsequent experimental application in field conditions. Preliminary examination of the methods and/or products applied are necessarily performed to avoid additional alteration of the structure and colour of the colonized substrata.

Further monitoring of the site with NIT and NDT techniques is recommended to confirm the effectiveness of the adopted measures and to provide prompt prevention of future microbial attacks.


221

IBBS-15-8-O01

Biodeterioration processes and climate change: the mayan cultural heritage as a case study

Benjamín Otto Ortega-Morales

Departamento de Microbiología Ambiental y Biotecnología, Universidad Autónoma de Campeche, Melgar, Col. Buenavista, CP 24039, Campeche, México.

Much research effort has been invested into understanding the influence of climate change on plant and animal communities yet relatively little is known about its potential impact on microorganisms. This is of paramount importance since microbes actively participate in the distribution of mineral resources, landscape evolution, soil development, water quality, agriculture along with the conservation and deterioration of cultural heritage. Mayan monuments on the Yucatan Peninsula of Mexico represent a natural laboratory to assess potential climate change impacts on microbial communities (biofilms) and building stone in tropical latitudes. Current climate models show that coastal sites such as Tulum, and Jaina, among others are endangered by sea level rise and by forecasted increase in the frequency and intensity of hurricanes. Conversely, Mayan ruins located inland of the Peninsula (Chicanna, Becan, Calakmul and Kohunlich) are expected to suffer of periods of drought leading to desiccation of surfaces and the subsequent thermal and salt-induced deterioration. The extent to which microbial biofilms may react towards this change, exacerbating, or perhaps protecting stone surfaces is not known. Baseline studies during the last 15 years suggest shifts in the extent of colonization, the composition of biofilms and the nature of ongoing deterioration processes. Ongoing interdisciplinary research aiming at providing a better understanding of the potential climate change impacts on subaerial biofilms on Mayan monuments will be briefly discussed.


222

IBBS-15-8-O02

Bacterial diversity and biodegradation in hydrocarbon-contaminated soil from an alpine industrial site

Rosa Margesin, Christoph Moertelmaier, De-Chao Zhang,

Institute of Microbiology, University of Innsbruck, Technikerstrasse 25, A-6020 Innsbruck, Austria

A combination of culture-dependent and culture-independent methods was used to investigate the microbial diversity in soil samples collected from an alpine industrial site containing high amounts of petroleum hydrocarbons. Forty-eight culturable bacterial strains were identified as potentially unique. The majority of these strains (71%) belonged to the phylum Proteobacteria, with a predominance of the two classes Alphaproteobacteria and Gammaproteobacteria; 19% of the strains were related to Actinobacteria. Only few representatives of the phyla Cytophaga–Flavobacterium–Bacteroides group (6%) and Firmicutes (4%) were detected. The number of sequences representing novel species (8 culturable novel species have been recently described) was remarkably high. All 48 culturable bacterial strains were examined for their ability to utilize a number of aliphatic, monoaromatic and polycyclic aromatic hydrocarbons as the sole source of carbon and energy at 15°C. Actinobacteria were the most versatile and efficient hydrocarbon utilizers. Selected strains were tested for their ability to degrade high amounts of phenol, anthracene and pyrene.

The bacterial 16S rRNA gene clone library was composed of 390 clones that grouped into 68 phylotypes and could be classified into four phyla: Proteobacteria (85% of the total clones), Bacteroidetes (9%), Actinobacteria (2%), Spirochaetes (1%). A number of phylotypes shared high similarities with strains previously described to be involved in hydrocarbon biodegradation.


223

IBBS-15-8-O03

Algae ′aground′ – Biodiversity and ecophysiology of aeroterrestrial greenalgal biofilms

Lydia Gustavs1, Manuela Görs1, Lubos Polerecky2, Rhena Schumann1, Ulf Karsten1

1Applied Ecology, University of Rostock, Albert-Einstein Str. 3, 18055 Rostock, Germany

2 Max-Planck Institute for Marine Microbiology,

Aeroterrestrial green algae typically colonize the interface between lithosphere and atmosphere, forming dense biofilms on natural and artificial substrata. These algal biofilms along with co-existing bacteria and fungi are a widespread and sometimes unwelcome phenomenon, as they cause discolorations and accelerate weathering of anthropogenic surfaces such as building facades. However, they are also promising communities to investigate algal adaptation strategies to the terrestrial environment. The dominant species have been identified by culture-dependent and independent methods. Further, optimum growth curves concerning various environmental factors were recorded, characterizing aeroterrestrial green algae as poikilo-tolerant. The high tolerance of the investigated strains against low water availability is partly based on the biochemical capability to synthesize and accumulate certain organic osmolytes. The discrepancy between the dominance of Apatococcus lobatus in aeroterrestrial biofilms and its conspicuously low growth rate unter controlled conditions led to investigations about mixotrophy in this species. Using a microsensor, the strong attenuation of ambient light was measured inside natural and artificial Apatococcus biofilms. As radiation is a limiting factor in deeper biofilm layers, mixotrophy is assumed as a competitive advantage against faster dividing algal taxa in the top layer.


224

IBBS-15-8-O04

Fungal succession at early stages of beech litter decay

Sandra Molla, Katharina M. Keiblingerb, Sophie Zechmeister-Boltensternb, Dragana Bandianc, Andreas Richterd, Joseph Straussa,c, and Markus Gorfera,c

a Fungal Genetics and Genomics Unit, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, A-1190 Vienna, AUSTRIA

b Federal Research and Training Centre for Forests, Natural Hazards and Landscape (BFW), Department of Forest Ecology and Soils, Unit of Soil Biology, A-1131 Vienna, AUSTRIA

c AIT Austrian Institute of Technology GmbH, Bioresources Unit, 2444 Seibersdorf, AUSTRIA

d Department of Chemical Ecology and Ecosystem Research, Faculty Center for Ecology, Faculty of Life Sciences, University of Vienna, A-1090 Vienna, AUSTRIA

Litter decomposition in beech forests (Fagus sylvatica) is a sequential, dynamic process driven by microbial succession. Initially present endo- and epiphytic phyllosphere fungi are rapidly replaced by saprophytic litter decomposers.

Although phyllosphere fungi cause little weight loss in the initial phase of litter decomposition, they strongly influence the subsequent steps. The presence of phyllosphere fungi can positively or negatively influence secondary invaders thereby shaping the succeeding community.

The objective of the current study was an evaluation of fungal succession during the early stages of beech litter decay when mass loss is negligible. It was hypothesised that the primary colonizers are epiphytic and endophytic phyllosphere fungi already present during leaf senescence. Community shifts were expected, where fungi better adapted to living leaves are continuously replaced by fungi better adapted to dead leaves. To this end the most prominent members of the fungal community in freshly fallen beech litter were identified by sequencing of cloned PCR products comprising the ITS- and partial LSU-region. Beech litter from four different sites in Austria was analyzed immediately after sampling and after incubation in the laboratory for two days and two weeks. Aquatic hyphomycetes represent the most prominent fungal early colonizers on all four different litter types. Diversity was increasing with longer incubation time on all samples. The obtained dataset gives information on the fate of phyllosphere fungi after leaf fall.


225

IBBS-15-8-O05

Biodiversity assessment and degradation potential of thermophilic fungi found in two different composting processes using traditional and molecular methods

Adrian Langarica-Fuentes1, Pauline S. Handley1, Geoff D. Robson1

1 Faculty of Life Sciences, University of Manchester, Michael Smith Bldg., Oxford Road, Manchester M13 9PT, United Kingdom

Thermophilic fungi are small group of organisms that have a minimum temperature for growth at or above 20˚C and a maximum above 50˚C that can in some cases be as high as 62˚C. From a biotechnological perspective, thermophilic fungi are of great interest due to their important role in the composting process and their ability to degrade recalcitrant materials. In this study, the biodiversity of thermophilic fungi in two different commercial composts was investigated by using traditional culture-based methods and two culture-independent techniques: denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP). Compost A was produced using forestry cuttings, while compost B used garden and municipal solid waste. Five and eight thermophilous species were isolated and identified from compost A and B, respectively, using culture-based techniques. However, DGGE and TRFLP analysis of these composts revealed a more complex fungal diversity suggesting that the culturable fraction of thermophilic fungi is only a small fraction of the total thermophilic community. To study the degradation ability of thermophilic fungi and the impact of adding different substrates on their community, DGGE analysis was performed to compost A that was mixed with two feedstocks (seaweed and barley straw) and incubated at 50 ˚C over a 6-week period. Visual evidence of fungal growth and high levels of degradation were observed in both substrates, suggesting there exist great potential for thermophilic fungi to degrade them. DGGE banding patterns throughout the experiments demonstrated that each substrate enriched for different fungi involved in the degrading process.

This research was supported by CONACyT and University of Manchester.


226

IBBS-15-8-P01

Some microbiological parameters in marine microcosms supplemented with gasoline and gasoline-enriched marine populations

Simona Ghita1,3, Iusuf Loredana4, Ioan I. Ardelean2,3

1Constanta Maritime University, 900663 Constanta, email [email protected]

2Institute of Biology, 060031 Bucharest, email [email protected] 3Ovidius University of Constanta, 900552 Constanta

4 NC "Maritime Ports Administration" SA Constanta, email [email protected]

In this paper we present the results concerning some microbial parameters [total cell count, dividing cells, permeabilised cells (propidium iodide positive cells), biomass and percentage growth rate] in marine microcosms supplemented with gasoline and gasoline-enriched marine populations. At the beginning of experiment, total cell population(labelled with either acridine orange or SYBR Green I) density was higher in microcosm supplemented with gasoline-enriched marine population and carbon / nitrogen source (gasoline (1% v/w) and ammonium nitrate (0.005% w/w) as compared with the other microcosms, then a significant decrease in total cell population density was observed. These results , mainly the density of dividing cells, the cells with intact plasma membrane [ total cell count minus permeabilised cells (propidium iodide positive cells] and percentage growth rate are discussed in corelation with the BOD5 in the atempt to take into account the intensity of aerobic respiration of microorganisms .The time evolution of cell densities are due to gasoline consumption and, probably, to the significant bacteriolithic activity (litic bacteriophages and/or predatory bacteria). Our results argue that gasoline can sustain the growth of endogenous micorbiota as well as of previously gasoline-enriched marine populations, thus sustaining the further research on the use of gasoline-enriched marine populations for bioremediation of polluted sites (bioaugmentation).


227

IBBS-15-8-P03

Performance of DNA polymerases

Margit Balazs*, Attila Szvetnik, Alexandra Németh, Andrea Rónavári, István Kiss

Institute for Biotechnology, Bay Zoltán Foundation for Applied Research,

Derkovits fasor 2., H-6726 Szeged, Hungary

* E-mail: [email protected], Phone: +36-62-432252, Fax: +36-62-432250

The polymerase chain reaction coupled with denaturing gradient gel electrophoresis (PCR-DGGE) has been used widely to determine the structure of microbial communities. DGGE and other PCR-based fingerprinting methods (TGGE, SSCP, T-RFLP, ARISA) brought a remarkable advancement in community analysis compared to traditional laboratory cultivation approaches that capture only about 1% of the population diversity. PCR-DGGE analysis is a multi-step procedure containing the extraction of total DNA from soil, PCR amplification with specific primers, the separation of amplicons based on sequence differences and the analysis of the patterns. The effect of the DNA preparation method and bias caused by the amplification steps on the outcome of PCR-DGGE analysis have been extensively studied. However, the DNA polymerase itself may affect the obtained patterns due to differences in processivity, inhibitor resistance and accuracy. Based on this consideration, the performance of two highly processive and accurate DNA polymerases was compared to the generally utilized Taq polymerase in PCR-DGGE analysis of three complex DNA samples isolated from polluted soils.


228

IBBS-15-8-P04

Diversity and physiology analysis of new yeast strains isolated from oil-polluted environment

Ortansa Csutak1, Camelia Pavlusenco2, Ileana Stoica1, Raluca Ghindea3, Ana-Maria Tanase1, Tatiana Vassu1

1 – Department of Genetics, Faculty of Biology, University of Bucharest, 1-3 Aleea Portocalelor, 060101 Bucharest, Romania, E-mail: [email protected]

2 – Master of Microbial Biotechnology and Genetics, Faculty of Biology, University of Bucharest, 1-3 Aleea Portocalelor, 060101 Bucharest, Romania

3 – MICROGEN, Faculty of Biology, University of Bucharest, 1-3 Aleea Portocalelor, 060101 Bucharest, Romania

Abstract

Microorganisms are subjects of many bioremediation studies due to their ability of assimilating a large range of organic compounds, such as hydrocarbons, main components of oil. The international interest regarding the possibility to use yeasts in biodegradation processes, led to intensive studies for the identification of new yeast species/strains with degradative abilities.

Six yeast strains (named DP1, DP2, DP3, DP4, DP5, DP6) isolated from oil-polluted environment, from Pitesti area, Romania, were identified using polyphasic ID. Strains DP1, DP2, DP4 and DP5 belong most probably to Rhodotorula glutinis, DP3 to Issatchenkia orientalis and DP6 to Candida rugosa.

All six strains grew on media with n-decane (C10), n-dodecane (C12), n-tetradecane (C14), n-hexadecane (C16) as sole carbon source. Best results after seven days were obtained on C12

and C16. Faint emulsification activity has been observed so far, mainly on n-hexadecane. Antimicrobial tests were performed for DP strains grown on n-alkanes plated against potential pathogen strains of Candida (C. albicans, C. parapsilosis, C. tropicalis, C. guilliermondii, C. parapsilosis) and Hansenula anomala. The results were recorded for almost 2 weeks. Halos were observed especially in the case of H. anomala - DP1, 2, 4, 5 on C16 and DP3 on C12, C. tropicalis - DP1 on C16 and DP2 on C12, C. krusei – most DP strains on C16. Cells prelevated from the halos did not show altered colonies or growth rate, but microscopic analysis revealed modifications of cell form or transition to hyphae, within the first two days.


229

IBBS-15-8-P05

Trying to unravel the relation between natural bacterial communities’ diversity and bioremediation

L. Giaramida1,2, L. Lawton1, C. Edwards1 and B. Singh3

1IDEAS Research institute – The Robert Gordon University, UK

2The Macaulay Land Use Research Institute – Aberdeen, UK

3Centre for Plants and Environment - University of Western Sydney, Australia

Biodiversity supports the vast range of ecosystem services that vitally contribute to human well-being. We are in the middle of the sixth mass extinction with biodiversity in decline in most of the ecosystems studied. Microorganisms diversity, directly or indirectly, is essential for most of the ecosystem services. Bacteria are the most diverse and abundant organisms on Earth and are essential for life on earth. They play a key role in nutrient cycling in both terrestrial and aquatic ecosystems. The relationship between bacterial diversity and ecosystem functioning remain poorly understood, representing a major research challenge. Biodegradation is one of the major functions of natural microbial community and in most cases determines the fate and persistence of man made and natural chemicals in the natural environment. Although there is some evidence of a positive relationship between microbial biomass and biodegradation rate, the relationship between community diversity and biodegradation in the environment remains to be confirmed. Here, in a dilution to extinction experiment, we used molecular biological (i.e. DNA fingerprinting and quantitative PCR) and analytical (i.e. HPLC) approaches to unravel the potential relationship between microbial diversity and biodegradation. Both natural and man made toxic compounds were tested in a system including freshwater from both pristine and polluted environments.


230

IBBS-15-8-P06

Biodiversity of Cracow-Czestochowa Upland limestone-inhabiting fungi – summary of preliminary studies

Magdalena M. Majewska

Department of Mycology, W. Szafer Institute of Botany, Polish Academy of Sciences

The aim of the studies was to define biodiversity of fungal species inhabiting inner surfaces of limestone formations in Cracow-Czestochowa Upland. Limestone is widely spread in this region of Poland. What is more, many castles and historical ruins made of this stone material are located around the Upland.

Samples from six localities in the middle part of the Cracow-Czestochowa Upland (from Olkusz up to Ogrodzieniec) were collected. 163 strains of fungi were isolated from this material during the three months length period of cultivation.

Among identified species were found Rhizopus sp., Aspergillus sp., Penicilium sp., Cladosporium sp., and some representatives of black yeast-like fungi - Aureobasidium pullulans (de Bary) Arnaud, Coniosporium perforans Sterflinger.

Actually, identification of other isolated strains is preformed. Genetical and phylogenetical analyses are scheduled.


231

IBBS-15-8-P07

Production of antimicrobial compounds by bacteria isolated from an alpine industrial site contaminated with hydrocarbons

Lydia Hemala, Rosa Margesin

Institute of Microbiology, University of Innsbruck, Technikerstrasse 25, A-6020 Innsbruck, Austria

We isolated 47 bacterial strains from an alpine industrial site contaminated with petroleum hydrocarbons and investigated the ability of these strains to inhibit growth of pathogenic bacteria and yeasts (E. coli, Sh. flexneri, S. enterica, P. aeruginosa, S. aureus, C. albicans, Cr. neoformans). The antimicrobial activities of the test strains were determined by an agar diffusion assay. Ten of the strains investigated were able to inhibit growth of at least one of the indicator strains, five of them produced sufficient biomass in liquid culture at 10°C and were also able to utilize petroleum hydrocarbons as the sole source of energy. Three strains belonging to the genus Pseudomonas inhibited growth of both bacterial and yeast indicators. Two strains belonging to the genus Serratia sp. were able to inhibit growth of Sh. flexneri, S. enterica, P. aeruginosa and S. aureus, while growth of the yeast indicators was not affected. None of the strains inhibited growth of E. coli. The chemical composition of the cell-extracts was analyzed by using thin-layer chromatography (TLC) and HPLC. The properties of the antimicrobial components produced by four test strains were similar to those of tetracycline, while one strain produced a compound with high similarity to the chemical properties of rifampicine.


232

IBBS-15-8-P09

Optimization of biosurfactant production by cultivation of Bacillus atrophaeus ATCC9372 in semi-defined olive oil and whey culture media

Luiz Carlos Martins das Neves1,2, Marina Ishii1, Inês Conceição Roberto2, Thereza Christina Vessoni Penna1

1Department of Pharmaceutical Technology, School of Pharmaceutical Science. 2Biotechnology Department, School of Engineering of Lorena/USP. University of São Paulo, Brazil.

High concentrations of environmental pollutants have represented serious risks for human health. Therefore, growing attention has been paying to their destruction by different competent public institutions. Biosurfactants are proteins with detergent, emulsifier and antimicrobial actions that have potential environmental applications. Bacillus atrophaeus strains are a non- pathogenic and suitable source of biosurfactants, including surfactin. The capability of B. atrophaeus strain to adapt its own metabolism to use several nutrients as energy sources and to preserve high levels of biosurfactants in the medium during the stationary phase is a promising feature for its possible application in biological treatments. The aim of this work is to establish a culture medium composition able to stimulate biosurfactants production by B. atrophaeus ATCC 9372. Batch cultivations were carried out at 150 rpm and 35°C for 24 h on lactose/casein/olive oil -based semi-defined culture media also containing mineral salts (NaCl, MgSO4, (NH4)H2PO4, KH2PO4, CaCl2, FeSO4.7H2O and MnSO4) and Whey culture media. Cell and Biosurfactant yields were compared. Kinetic study demonstrated a positive influence of lactose in the biosurfaction production (KS=0.739 g/L). Results indicated that whey are an interesting alternative culture media to biosurfactant production according cell and biosurfactant yields (PX = 0.3860 g/L.h; PB = 0.00198 g/L.h) when compared with semi-defined culture media (PX = 0.1427 g/L.h; PB = 0.00220 g/L.h).


233

IBBS-15-8-P10

Isolation and Characterization of a potential mercury resistant endophytes, Photobacterium halotolerans , Meld1 .

Dony Chacko Mathew and Chieh-Chen Huang

Department of Life Sciences, National Chung Hsing University, Taiwan.

A number of toxic synthetic organic compounds can contaminate environmental soil through either local (e.g., industrial) or diffuse (e.g., agricultural) contamination. Plant associated bacteria, such as endophytic bacteria (non –pathogenic bacteria that occur naturally in plant) have been shown to contribute to biodegradation of toxic organic compounds in contaminated soil and could have potential for improving phytoremediation.

Photobacterium halotolerans bacteria (1) is a gram negative, rod –shaped, found in salty marshes. This bacteria forms a distinct lineage with Photobacterium rosenbergii (2) and Photobacterium ganghwense (3) (showing 96.9 and 96.2% similarity respectively).

The current study was focused on the bioremediation of the toxic metal, methyl mercury into elemental mercury, which is a lesser toxic form of mercury. MerA gene specific primers A1s-n.F and A5- n.R (4) were used to detect merA gene in P.halotolerans. The merA gene was identified and showed 98% similarity to Vibrio shilonii AK1. Further studies were done to check the mercury tolerance level of this strain. It was observed that P. halotolerans were able to grow upto a concentration of 25 µM HgCl. It was found that this strain was salt tolerant (upto 6%) and had crude oil emulsification activity. In conclusion, the overall study demonstrates that Photobacterium halotolerans is a novel isolate for the bioremediation of mercury and oil contaminated sites. The role of Photobacterium halotolerans , as an endophytic bacterium will be analyzed in future.


234

IBBS-15-8-P11

Stimulation of trichloroethylene biodegradation in anaerobic three-phase microcosms

Éva Mészáros1*, Rita Sipos1, Róbert Pál2, Csaba Romsics1, Károly Márialigeti1

1Eötvös Loránd University. Department of Microbiology, Hungary, 1117-Budapest, Pázmány Péter stny. 1/C.

2Today Science Ltd, Hungary, 2083-Solymár, Cserje u. 13.

*[email protected]

Chlorinated ethenes, such as trichloroethylene (TCE), are among the most common groundwater contaminants worldwide. Careless handling and leakage from storage tanks resulted in large accumulation of halogenated compounds in the environment. Pump-and-treat technologies have limited effectiveness in the disposal of pollution, whereas stimulation of dechlorinating microorganisms is potentially the most promising and cost-effective technology for remediating contaminated sites.

To identify the most promising electron donor to be applied for in-situ bioremediation, model laboratory experiments are needed. Microcosm experiments were set up to enhance TCE biodegradation. Acetate as carbon source and hydrogen as electron donor were tested in a three-phase system. Previous studies showed only partial dechlorination of TCE in a two-phase system due to the lack of adhesion surface and the difficulty of biofilm formation, thus soil and synthetic carrier were used to ensure adequate surface for the settlement of bacteria. 2-bromoethane sulfonate (BES), an inhibitor of methanogens, was also tested to justify if these microorganisms play a role in TCE biodegradation.

The dynamics of biodegradation was monitored by gas-chromatography. Microbial community structure and function were characterized by molecular biological methods (TRFLP, clone library) and group specific detection of key taxa (group specific PCR, FISH) as well as key metabolic genes.

The results of the microcosm experiments are: (i) fast TCE degradation in the amended microcosms (ii) similar dynamics with a short log phase in the inhibitory systems (iii) very slow TCE degradation in the biotic control microcosms (no amendment) (iiii) key community members and (iiiii) dechlorinating bacteria were detected.

This work was supported by the Hungarian Scientific Research Fund (OTKA 75790) and the Fellowship of The European Union and The European Social Fund (TÁMOP 4.2.1./B-09/1/KMR-2010-0003)


235

IBBS-15-8-P12

Complex molecular biological investigation of TCE biodegradation with respect to electron donor amendments using anaerobic microcosms

Rita Sipos1*, Éva Mészáros1, Gábor Cebe1, Anita Mohr1, Róbert Pál2, Csaba Romsics1, Károly Márialigeti1

1Eötvös Loránd University. Department of Microbiology, Hungary, 1117-Budapest, Pázmány Péter stny. 1/C.

2Today Science Ltd, Hungary, 2083-Solymár, Cserje u. 13.

*[email protected]

Chlorinated ethene contamination is a serious problem worldwide, including Hungary. Biological remediation technologies seem to be the preferred method of choice, as they are more cost-effective than traditional technologies.

We conducted an anaerobic microcosm experiment consisting of TCE-contaminated groundwater with the addition of different electron donors to investigate an enhanced reductive dechlorination. The groundwater sample originated from a site where previously in situ bioremediation treatment was applied. The two-phase microcosms were monitored for VOCs and water chemical parameters, moreover the microbial community of the initial and the day 210 samples were thoroughly explored using 16S rRNA gene based T-RFLP and clone library construction. Group specific detection of dehalogenating bacteria and examination of catabolic genes of the degradation pathway were carried out.

First, we observed little bio-degradative activity difference owing to the application of different amendments, the degradation of TCE was quite slow and completed only to VC as of day 210. However, based on 16S rRNA-based molecular fingerprinting, the effect of the donors on the initial microbial community was quite remarkable. Later, we modified the setup to improve upon the initial redox conditions within the microcosms, and used H2 as primary or secondary electron donor. The prepared synthetic culture media was then inoculated with concentrated groundwater sample from the same site. The microbial community structure was examined with molecular biological methods, along with VOC monitoring. Preliminary results show a much faster TCE-to-VC transformation and a remarkable difference due to the various amendments.

This work was supported by the Hungarian Scientific Research Fund (OTKA 75790)


236

IBBS-15-8-P13

Role of Streptomyces rimosus in degradation of chitin substances

1Swiontek Brzezinska Maria, 2Jankiewicz Urszula, 1Walczak Maciej

1Department of Environmental Microbiology and Biotechnology, Institute of Ecology and Environmental Protection, Nicolaus Copernicus University, Gagarina 9, Toruń, Poland

2 Department of Biochemistry, Warsaw University of Life Science, Nowoursynowska 159, Warsaw, Poland

Chitinolytic actinomycetes was isolated from the agricultural land soils of the Central Poland. It was identified as Streptomyces rimosus. The identification was based on standard biochemical tests and 16SrRNA. The degradation of chitin substances (shrimp shell waste, chitin powder from crab and chitosan) was examined on the ground of oxygen consumption with application of OxiTop (WTW). The studies included purifying of chitinases by affinity chromatography. Purified proteins produced by Streptomyces rimosus were subjected to identification by mass spectrometry. Based on the analysis of resultant fragments of amino acid sequence of chitinase produced by examined bacteria, degenerate primers for PCR were designed. Our research results proved that Streptomyces rimosus degraded chitin substances on various levels. The best metabolised by Streptomtces rimosus substance is chitosan. However, this actinomycetes was equally efficient in hydrolysed of shrimp shell waste. Chitin powder from crab was the most poorly used source of carbon and nitrogen. Nucleotide sequence of amplified gene fragment proved similar to the sequence of chitinase encoding genes of the family 18 of glycoside hydrolases. Chitinolytic activity examination showed that shrimp shell waste was the best inducer for chitinases' synthesis.


237

IBBS-15-8-P14

Fungal degradation studies on bamboo

Dong-Sheng Wei, Olaf Schmidt, Walter Liese

Center for Wood Science, Department of Biology, University of Hamburg, Hamburg, Germany

It is well known that bamboo is a cheap and fast growing sustainable material with increasing interest for constructions and industrial utilization. However, bamboo has a low natural durability and is easily attacked by fungi. Little is known on its degradation by fungi. Therefore, studies were performed with several bamboo species. First, to evaluate in the laboratory conditions, mass loss of bamboos with small samples (3 x 1 cm) was measured in preserving jars after 1, 3, 12 month of incubation with white-rot, brown-rot and soft-rot fungi. Second, to imitate natural environmental conditions, bigger bamboo samples (25 cm long) were investigated in the so-called Fungus cellar-test whereby the bamboo is inoculated with pure cultures, however, incubated in large metal tube containing unsterile garden soil. The mass loss of bamboos was rather low after 1 month and increased within 1 year. The white-rot fungus Trametes versicolor and the soft-rot species Chaetomium globosum revealed considerable decay, whereas the brown-rot fungi Coniophora puteana and Gloeophyllum trabeum yielded less degradation. Fungal degradation in Fungus cellar-test was significantly affected by the water content of bamboos: C. puteana was aggressive in dry samples (about 30% moisture content). In contrast, the white-rot species Schizophyllum commune produced most decay in wet substrates (180 %u). The results improve the basic knowledge on the degradation of bamboo by fungi, which is needed for a better way to evaluate degradation of bamboo by fungi and for suitable protection measures of bamboo.


238

Topic 9-State of the art methods to study organisms and processes of BBB

– genomics/proteomics


239

KEYNOTE LECTURE

Metaproteomics: Linking Microbial Diversity to Ecosystem Functioning

Kathrin Riedel Institute of Microbiology, TU Braunschweig & Helmholtz-Centre of Infection Research, Braunschweig, Germany In contrast to nucleic acids, most proteins have an intrinsic metabolic function and can therefore be used to relate specific microbial activities to the identity of defined organisms. The assignment of proteins of a given habitat to specific phylogenetic and functional groups (referred to as metaproteomics) provides new insights into the role of microbial diversity in ecosystem functioning. The enormous potential of this emerging technology will be exemplified by two studies on (I) symbiotic partners present in lichens and (II) microbial communities involved in leaf litter decomposition.

(I) In order to link structure and function of microbes present in the lung lichen Lobaria pulmonaria metaproteomics was combined with fluorescence in situ hybridization and confocal laser scanning microscopy. The lung lichen harbours proteins of myco- and photobiont, but also of a highly diverse prokaryotic community, which is dominated by Proteobacteria. Proteins of the photobiont were found to be mainly associated with energy and carbohydrate metabolism; while fungal and bacterial proteins were involved in similar functions, e.g. stress response or secondary metabolite biosynthesis.

(II) To monitor spatial and temporal variations of protein composition and functioning during leaf litter decomposition we investigated litter biogeochemistry and community structure, quantified enzymatic activities, and analysed the protein complement by a semi-quantitative proteomics approach. Our results suggest that fungal and bacterial abundance positively correlates with the amount of certain macro- and micronutrients, e.g. P and Mn. The finding that almost all identified hydrolytic enzymes were of fungal origin suggests a prominent role of fungi during aerobic litter decomposition.


240

IBBS-15-9-O01

Evaluation of current microbial control technologies for biofilm “dispersion” using time-lapsed video microscopy.

Monica A. Canalizo-Hernandez1 and Michael V. Enzien1

1Dow Microbial Control. The Dow Chemical Company. Bacteria prefer to reside in biofilms where they are protected by a complex, permeable resistant matrix formed by extracellular polymer substances (EPS) produced by the bacteria. Biofilms pose significant challenges to many industrial processes. In gas and oil pipelines, bacterial growth and its consequent biofilm formation contribute to several deleterious processes such as reservoir souring, scale formation, pipeline clogging and microbiologically-influenced corrosion (MIC). The effects of current treatments for the control of bacterial growth on planktonic and sessile bacteria have been extensively studied, however, the efficacy of these technologies on biofilm removal and the mechanism by which the removal occurs are not well established. Therefore, understanding the effects that treatments widely used in the industry have on the mechanism of biofilm dispersion is imperative, not only to identify the most adequate treatment, but also to develop novel, superior technologies for biofilm dissolution. Here, we report on studies undertaken to examine the impact of common industrial biocide treatments on biofilm removal and grow-back using a novel, real-time video microscopy technique. Our results allow for the identification of the most promising technologies for the dispersive removal and grow-back prevention of biofilms.


241

IBBS-15-9-O02

Using a Metabolomic Approach to Identify Bio-Corrosion in Copper Pipes D.J. Beale1, M.S. Dunn2, P.D. Morrison2, D.R. Marlow1, and D.C.O Marney1

1CSIRO Land and Water, PO Box 56, Highett VIC 3190, Australia. 2RMIT, School of Applied Sciences, PO Box 2047, Melbourne VIC 3001, Australia. Copper is commonly used in residential, commercial and industrial pipe systems because of its relative un-reactivity and its ability to be easily manufactured and shaped. However, it does undergo corrosion as a result of microbial activity on its surface, even though it is traditionally viewed as a material with antimicrobial properties. This paper presents the application of metobolomic profiling of water samples taken from copper piping exposed to corrosion influencing bacteria. The samples were taken from accelerated copper microbially influenced corrosion (MIC) trials, extracted and derivitized with acyl, alkyl and silyl functional groups using microwave assisted techniques. Funcitonalisation using these derivitisation techniques enabled effective separation and identification of the metabolites from bacteria that cause MIC using gas chromatography in combination with mass spectrometry. Further classification of the identified metabolite compounds was achieved using chemometric Partial Least Squares Discrimination Analysis (PLS-DA), with the correlation of metabolites and substrates between each sample used to identify the consumption of fatty acid compounds in samples exposed to MIC. This approach can be used to identify metabolomic biomarkers (i.e., metabolites or substrates) that are indicative or otherwise of microbial activity within water distribution networks that can be attributed to MIC. It is also anticipated that the methodology described here-in can be used to identify biofilm related odor and taste problems within the networks.


242

IBBS-15-9-O04

Molecular characterisation of the interspecies yeast hybrids

Lopandic Ksenija1, Bond Ursula2, Atanasova Lea3, Druzhinina S. Irina3, Sterflinger Katja1

1Austrian Center of Biological Resources and Applied Mycology, Institute of Applied Microbiology, University of Natural Resources and Life Sciences, Vienna, Austria 2Moyne Institute for Preventive Medicine, Microbiology Department, Trinity College, University Dublin, Dublin, Ireland 3Division of Gene Technology and Applied Biochemistry, Institute of Chemical Engineering, Vienna University of Technology, Vienna, Austria

The whole genome comparisons have demonstrated that acquisition of foreign genes is not a rare event among eukaryotic microorganisms. Moreover genome duplication, introgression and interspecies hybridization are considered to be the main molecular mechanisms of yeast genome evolution (Dujon 2010). Several studies on natural yeast populations have shown that yeasts isolated from vineyard ecosystem and from the beer and vine fermentations may recombine their genomes and generate stable interspecies hybrids (e.g., González et al. 2006, 2008; Lopandic et al. 2007). Characterisation of such yeast hybrids can help us understand adaptive evolution and mechanisms involved in genomic and physiological responses of the cells to specific environments. The aim of this presentation is to provide an overview of the study on the native hybrid yeasts between species of the Saccharomyces sensu stricto complex. In order to achieve a reliable characterisation of the naturally occurring yeasts, a polyphasic approach that relied on different genotypic and phenotypic methods was used. The characterisation of the interspecies hybrids between S. cerevisiae and S. bayanus, or S. cerevisiae and S. kudriavzevii was approached by the amplified fragment length polymorphism (AFLP) technique, microarray based comparative genomic hybridization (CGH) and phenotype microarrays. The obtained results generated complex information on the hybrid genomes between native yeast isolates, on their chromosomal composition and structure, as well as on the new physiological properties that resulted from the genome merging.

González SS, Barrio E, Gafner J et al. (2006) Natural hybrids from Saccharomyces cerevisiae, Saccharomyces bayanus and Saccharomyces kudriavzevii in wine fermentations. FEMS Yeast Res 6:1221-1234.

Lopandic K, Gangl H, Wallner E, et. al. (2007) Genetically different wine yeasts isolated from Austrian vine-growing regions influence wine aroma differently and contain putative hybrids between Saccharomyces cerevisiae and Saccharomyces kudriavzevii. FEMS Yeast Res 7: 953-965.

González SS, Barrio E, Querol A. (2008) Molecular characterization of new natural hybrids of Saccharomyces cerevisiae and S. kudriavzevii in brewing. Appl Environ Microbiol 74: 2314-2320.

Dujon B. (2010) Yeast evolutionary genomics. Nature 11: 512-524.


243

IBBS-15-9-P01

A suitable method to detect polycyclic aromatic hydrocarbons (PAHs) as a water contaminant by monoclonal antibody

Zahra Sobhani Damavandifar1, Manouchehr Mirshahi1, Sina Sarikhani1, Dariush Minai-Tehrani2

1- Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IRAN

2- Faculty of Biological Sciences, Shahid Beheshti University, Tehran, IRAN

Abstract

Crude oil contains different kinds of polycyclic aromatic hydrocarbons (PAHs) from two rings to more complex four and five rings molecules. The spillage of crude oil in soil and water can damage the environment. Some PAHs are toxic and may be mutagenic and carcinogenic for living organisms. Detection of PAHs in the environment especially in water can help for preventing its harmful effect and also biotreatment of these toxic components from water. The chemical methods for detection of PAHs are not so sensitive and need laboratory materials and equipments. In this study a monoclonal antibody was prepared which can use for a sensitive and easy method for detection of PAHs in water. Pyrenyl butyric acid (PBA) was used as a PAH model for immunization of mouse to induce an antibody against PBA. Subcutaneous injection of PBA was done three times and the production of antibody against PBA in mouse serum was monitored by ELISA method. The ELISA showed that high affinity antibody was generated in mouse serum. Monoclonal antibody was made using mouse spleen lymphocytes and myeloma cells. Competitive ELISA showed high affinity of monoclonal antibody against PBA. This antibody will use for production a lateral flow strip kit to rapid, easy and sensitive detection of PAHs.

Key words: PAH, contamination, antibody,


244

IBBS-15-9-P02

Advantages of contact-color techniques vs. improved chlorophyll extraction method for quantification of phototrophic biomass on stone surfaces

P. Sanmartín, D. Vázquez-Nion, B. Silva, B. Prieto

GEMAP-USC, Grupo de Estudios Medioambientales Aplicados al Patrimonio Natural y Cultural. Departamento de Edafología y Química Agrícola, Facultad de Farmacia, Universidad de Santiago de Compostela, 15782-Santiago de Compostela, Spain. [email protected] (P. Sanmartín)

This work outlines two suitable methods for quantification of cyanobacterial and algal biomass on stone surfaces, (1) a new and improved chlorophyll a extraction method and (2) a non-destructive contact-color technique. Several known amounts of cyanobacteria solutions were inoculated on the top surface of leucogranite samples and their real biomass was compared with the biomass estimated using both methods. Also, the intensity and coverage of organisms on the surface of each sample was assessed by eight different observers endowed with normal color vision, in order to differentiate into two sets: perceptible and imperceptible presence of organisms. A similar accuracy between methods was found when the whole set of samples was considered together, regardless of visual evaluation. However, taking into account the samples with an imperceptible presence of organisms, only the colorimetric method was valid. Thus, the quantification in the non perceptible range together with the non-destructive nature of the procedure, allowing further analyses in the same sample, highlight contact-color techniques as an effective tool for cultural heritage studies.


245

Topic 10-Control of biodeterioration: new physical and chemical methods


246

KEYNOTE LECTURE

Antimicrobial surfaces: do we need them? Do they work?

Joanna Verran,

School of Health Care Science, Manchester Metropolitan University, Manchester, UK

One approach for reducing microbial contamination or colonisation of surfaces is to modify the surface so that it acquires ‘anti-microbial’ properties. These properties could encompass anti-adhesion characteristics, where the surface resists microbial attachment, and is therefore easy to clean; or anti-microbial properties, where approaching/attaching/attached microorganisms are inhibited, or, ideally, killed (and easily removed) by some component in or on the surface. In this latter case, the mode of action of the antimicrobial agent is key: does it diffuse from the surface? Is moisture required for optimum activity? Is an external trigger, such as irradiation, required? Is ‘contact kill’ the aim? All of these issues impact on the method used to demonstrate the point-of-use effectiveness of antimicrobial material.

This presentation will describe some of the work that has been done on putative antimicrobial surfaces, including those with photocatalytic properties, thin film coatings with antimicrobial metal alloys, encapsulated biocides that promote delayed release of antimicrobial, and others.

Finally, what is the intended use of the surface? Will the proposed antimicrobial activity be effective at point of use? Is there any added value in the surface being antimicrobial? Some attention will also be given to the actual or perceived value of antimicrobial surfaces, and specific cases will be described.


247

IBBS-15-10-O01

Biocide resistance in selected epi- and endolithic lichens after mild thermic pre-treatment: a case study from the Athenaion temple of Paestum (S Italy)

Mauro Tretiach, Fabio Candotto Carniel, Stefano Bertuzzi

Dipartimento di Scienze della Vita, Università degli Studi, Via L. Giorgieri 10, I-34127 Trieste, Italy

Verrucaria nigrescens Pers. and Bagliettoa parmigera (J.Steiner) Vězda & Poelt are two deleterious biodeteriogenous lichens of lithic substrata (limestone, travertine, marble etc.) which often occur together forming a mosaic of black and white thalli. Both species have a green alga as photobiont, but the former is epilithic, the latter endolithic. In this study we discuss the results of field and laboratory measurements on thalli treated with three commercial biocides, in order to elucidate the phenomena underlying a fast re-colonization observed on the pavements of the Athenaion temple in Paestum (S Italy) after the cleaning of the surfaces fourteen years ago. The efficacy of the three biocides was monitored with a portable pulse-amplitude-modulated (PAM) fluorimeter. For comparison, epifluorescence microscope observations were carried out in the laboratory on cross-sections of treated and untreated thalli, whereas confocal laser scanning microscopy, in combination with the use of specific fluorescent probes, allowed for the assessment of the physiological state of the cells. The two lichens showed different resistance to the biocides, probably related to the different affinity of the active principles with the constituents of the external layers. Mild thermal pre-treatments (45° C for 30-60 min.), totally inefficacious on dry thalli, greatly enhanced the negative effects of the three biocides when applied to fully hydrated thalli. These treatments might represent an interesting alternative method for removing lichens from open-air stone artefacts, without negative effects on the environment.


248

IBBS-15-10-O02

Generation of Reactive Oxygen Species upon Red Light

Exposure of Cyanobacteria from Roman Catacombs

Paul Hsieh, Jens Z. Pedersen, Patrizia Albertano*

Department of Biology, University of Rome ‘Tor Vergata’, Rome, Italy

Correspondents: [email protected] * [email protected] Fax: +39 06 2023500

Roman catacombs are threatened by phototrophic biofilms predominated by cyanobacteria and associated microorganisms. They grow on frescos, mortar, marble and tufa rock wherever artificial lighting is installed. Although light is inevitable for the biofilms, excessive light may endanger their survival.

Red light (620-650 nm) can drastically inhibit cyanobacterial growth. Within 25-hr irradiation, the photosystem II quantum yield of Leptolyngbya red species in culture is reduced to only 0.08 from original 0.41, and to 0.15 from 0.46 in Leptolyngbya green species. Instead, blue light (480 nm) promotes their photosystem activity to up to 0.53-0.61 in the same conditions.

Further, 5-aminolevulinic acid (D-ALA) is introduced to shorten the treatment. D-ALA is a colorless, non-toxic compound that can be transformed to protochlorophyllide inside cyanobacterial cells. Red light is switched on at the biofilm after overnight D-ALA incubation. Their quantum yield is lowered down by the extent of 0.1 than the untreated cells.

Cyanobacterial cells are abundant in phycobilisomes that act as antenna for photosystems by capturing a wide spectrum of dim light. The bilin pigments in the phycobilisomes, and chlorophyll in the photosystems may produce reactive oxygen species (ROS) such as 1O2

, O2- , H2O2 , and OH upon irradiation. When strong red light is applied, high amount of

ROS is generated beyond the quenching capacity of the organism. Electron spin resonance spectroscopy confirms that treatment with D-ALA causes the formation of ROS; spin trap experiments indicate that H2O2 has been produced in the system. Further tests will confirm the usefulness of the treatment under long-term exposure of biofilms to the applied monochromatic light.


249

IBBS-15-10-O03

Testing the photocatalytic inactivation of aeroterrestrial algal growth

Gladis, Franziska; Schumann, Rhena

Biocides against aeroterrestrial algal growth that impacts the appearance of man-made surfaces shall be replaced by environmental-friendly self-cleaning materials, e.g. photocatalysis. The efficiency of photocatalysis to degrade various chemical compounds and inactivate bacteria has already been shown, while their activity to inhibit biofilm formation is still not verified. The aim of this study was to develop a laboratory method to investigate the efficiency of photocatalytic materials against algal growth. Two algal isolates (“Chlorella” luteoviridis and Coccomyxa sp.) functioned well as model organisms because they grew on almost all test specimens at 100 % humidity and low UVA radiation. The newly developed experimental design may be useful for assessing the biological function of photocatalytic materials or other functional surfaces.

In this study, no growth inhibiting effects of photocatalytically coated glass were detected. Since aeroterrestrial algae are adapted to adverse conditions, cells may be protected well against photocatalytically generated hydroxyl radicals.


250

IBBS-15-10-P01

Antimicrobial activity of ionic liquid applied to l inen fabric

Joanna Foksowicz-Flaczyk, Judyta Walentowska

Institute of Natural Fibres & Medicinal Plants;e-mail: [email protected]

ABSTRACT

Linen fabrics, due to their special properties i.e. high hygroscopicity, cool and pleasant touch, excellent air permeability and low attraction of electrostatic charges on the surface, provide optimum comfort of using and has positive effect on human organism. Linen products create the best microclimate for the skin.

The studies were aimed at improving the functionality of the linen fabric by antimicrobial finishing using ionic liquid.

The literature on the subject proved that ionic liquids, belonging to quaternary ammonium salts, are characterized by antimicrobial activity. These compounds were successfully used in protection of cellulose materials, like wood and paper, against biodeterioration. Antimicrobial activity of ionic liquids strongly depends on their structure, especially of the anion type ones. [NO3]

- and [NO2]- salts are very effective agents against bacteria and fungi. Here we report on

applying ammonium based ionic liquid with nitrate anion in finishing process of linen fabrics to improve their antimicrobial functionality.

Linen fabrics, protected with ionic liquids, were tested to action of 5 mildew mixtures and bacteria species present on human skin.

The antimicrobial effectiveness of ionic liquid applied to linen fabric was determined by the use of Scanning Electron Microscopy.

Key words: biodeterioration, antimicrobial activity, ionic liquid, linen fabric.


251

IBBS-15-10-P03

Insecticidal activity of Asarumsieboldii againsttermites Chang Wook Jo, Jung Eun Choi, Young Hee Kim, Jin Young Hong, Mi Hwa Jung, Soo Ji Kim Conservation Science Division, National Research Institute of Cultural Heritage, Munji-dong, Yuseong-gu, Daejeon, 305-380, Korea

Termite cause very serious damage to wooden cultural heritages and wooden structures. Because they have wood-eating habit, control of termites is very important. To control of termites, we used Asarum sieboldii. Insecticidal component of A. sieboldii is known very effectively. Extracted a solution was filtered out impurities by filter paper and concentrated reduction of pressure with the rotary vacuum condenser. We used worker termite cause many damage in wooden cultural properties for experiment. The insecticidal activity against termites was investigated by fumigation method on the filter paper (55 ) applied to 250㎕ extracts and solvent fractions of the A. sieboldii. We observed and recorded after 24hr,48hr.

We used methanol extracts and solvent fractions of A. sieboldii for insecticidal bioassay. After 24hr · 48hr, methanol extract of A. sieboldii wasn't shown very effectively mortality but, after 72hr, mortality was risen rapidly and 0.005g/㎕ appeared over 80% mortality. Insecticidal activity of A. sieboldii hexane fractions were more effectively than methanol extraction solution after 24hr. Even 0.005g/㎕ concentration were insecticidal activity over 80% and after 48hr, almost reached insecticidal activity 100%. Overall, A. sieboldii was effectively against termite. Key word: Asarum sieboldii, insecticide, termite, wooden cultural heritages


252

IBBS-15-10-P04

Stability test of natural pesticides on materials for conservation of organic cultural heritage

Young-Hee Kim*, Jung-Eun Choi, Jin-Young Hong, Mi-Hwa Jung, Chang-Wook Jo

Conservation Science Division, National Research Institute of Cultural Heritage, 472 Munji-dong, Yuseong-gu, Daejoen, 305-380, Korea

Two kinds of natural pesticides extracted from plants were evaluated effects on materials of organic cultural heritage in showcase. Two kinds of natural pesticides are wood vinegar and Asarum sieboldii. Wood vinegar and Asarum sieboldii were showed the most significant insecticidal activity against adult of Lasioderma serricorne and termite. First, we used cotton fabrics and silk fabrics undyed and dyed as test samples of cultural property. Madder and indigo fabrics were used sappanwood and Polygonum tinctorium as material of naturally dyed. Undyed and dyed fabrics were exposed to natural pesticides of various concentrations for six months in relative humidity 55±1% and temperature 24±2oC. After exposure of pesticides, color difference (∆E*) of fabric samples were investigated for six months at two weeks intervals. As a results, dyed cotton, silk and undyed silk fabrics exposed wood vinegar were not nearly changed in their colors, but color of only undyed cotton fabric was clearly changed by wood vinegar. Especially color difference by wood vinegar on undyed cotton fabric was most distinct as the concentration increased. On the other hand, all of dyed and undyed fabrics exposed Asarum sieboldii were not nearly changed in their colors for six months. Therefore, this study first suggests that wood vinegar and Asarum sieboldii as natural pesticides could be used to conserve for cloth cultural properties from insects and microorganism, but wood vinegar couldn’t use the high concentration on undyed cotton fabric.

Keyword: natural pesticides, wood vinegar, Asarum sieboldii, conservation agents


253

IBBS-15-10-P05

Assessment of Antifungal Effect of Nano Metal Particles against Isolated Fungi from Ghajar Inscription

P. Mohammadi, M. Ejadpanah-Saravi Department of Biology, Faculty of Science, Alzahra University, Tehran, IR of Iran E-mail Address: [email protected]

Cheshme Ali calcareous inscription in Shahr e Ray expose to widespread of biodeteriorating microorganims. In the mountains above this spring, a large boulder can be seen with relief of Fath Ali shah Ghajar, some princes and chiefs on the rocks. Biodeterioration is a phenomenon that affects all materials including those used in buildings, monuments as well metals, pigments, frescoes, and fungi play an important role in this decay. Different type of fungal strains was isolated from this monument. Removing of this biological agent is important for conservation aspect of cultural heritage. Thus, inhibitory effect of nano metal powders was determined by microdilution method in RPMI 1640 medium according to CLSI protocol. The most frequently of common isolated fungal species were Ulocladium spp., Alternaria spp., Aurebasidium spp. and Fusarium spp. In this work nano particle of Tio2, Ag, Cu, Zno and Al was used to assay fungicidal effect. Minimum fungicidal concentration (MFC) of these nano particles were determined which was ≥12.5 µg/ml, ≥ 8µg/ml, ≥ 15 µg/ml, ≥10 µg/ml, 17µg/ml, respectively. This study shows that nano metal particles have appropriate antifungal properties against this species. Further investigation must be done to eliminate these types of fungi on laboratory model before applying this particle on carbonate cultural heritage.

Key word: biodeterioration, cultural heritage, nano metal particle, minimum fungicidal concentration


254

IBBS-15-10-P07

One microbial bioassay for a method of cyanobacterial and algal removal

Bohuslav Uher

Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlarska 2, 611 00 Brno, Czech Republic; [email protected]

Methods of cyanobacterial and algal removal have been offered in the literature, but with no supportive research as to their effect on the substrate, length of effectiveness with time or miniaturized microbial bioassays (biotests). Most of the methods are devised for the removal from buildings. The use of soft brushing after dilute ammonium, household bleach (hypochlorite), 1-2% orthophenylphenate, 4% zinc or magnesium silico fluorite, 5% formaldehyde and others chemical eradicators were applied in many building preservation and conservation processes. Before any cyanobacteria or algae removal methods are used on stone, their reaction on the substrate and effectiveness as algal inhibitors or eradicators must be assessed for each specific site. The environmental control of stone-monument sites should be looked at as a logical method of algal control such as the reduction of humidity, protective coating, drainage, impregnation, etc. The main aim was to develop the techniques of new grow inhibition test used on subaerial autophototrophic microorganisms (autochtonous, isolated from buildings and monuments). Financial support for this study was provided by the Funding of Ministry of Education of Czech Republic project No. MSM0021622416.


255

IBBS-15-10-P08

Natural essential oil for antimicrobial protection of natural textiles Judyta Walentowska1, Joanna Foksowicz-Flaczyk1

1Institute of Natural Fibres & Medicinal Plants, Poznan, Poland e-mail: [email protected] Textiles containing natural fibres e.g. flax and hemp non-wovens, linen and cotton fabrics, find application both as finishing and insulation materials in automotive and construction industries and for medical, shoe lining and packaging products. Problems occur when they are exposed to harmful external factors. High humidity and temperature, insufficient air circulation result in enhanced growth of micro-organisms, especially mildew. Uncontrolled mildew growth leads to complete degradation of natural textiles. The study was aimed at increasing the resistance of linen-cotton fabric to bacteria and mildew action. The studies were carried out on antimicrobial protection with the use of biocide of plant origin. A biocide characterized with low toxicity for humans and environment (complying with the requirements of UE Directive 98/8/EC on releasing on the market biocidal products) was selected for the tests. The biocide used in the study was thyme essential oil. The paper presents the values representing antimicrobial efficiency of thyme essential oil applied to linen-cotton fabric. Antibacterial and antifungal activity evaluation was done by determining the bacterial growth, degree of mildew growth, and their impact on the textiles strength. Microscopic evaluation of the tested fabrics was also made with the use of Scanning Electron Microscope. Applying the biocide in finishing processes of fabrics containing natural fibres allows for obtaining antimicrobial barrier properties. The achieved results will enable to continue the studies further in order to determine the durability of the applied essential oil for application of the treated natural fabrics in medical, shoe lining and packaging products.

Key words: antimicrobial protection, essential oil, natural textiles


256

IBBS-15-10-P09

The influence of dithiocarbamates used for phytosanitary treatments of vineyard on the microelements contents from soil

Catalina Calin1, Gina Vasile2, Octav Pantea1, Andreea Bondarev1, Sonia Mihai1

1 Petroleum- Gas University of Ploiesti, Bucharest Av. 39, Ploiesti, Romania

2University of Agronomic Science and Veterinary Medicine from Bucharest, Marasti Av. 59, Bucharest, Romania

Dithiocarbamates are included in the class of fungicides and are widely used because display high broad-spectrum activity against fungal plant diseases. These compounds influence the content of microelements from soil because they readily react with metallic ions forming very stable complexes. Dithiocarbamates decompose in soil and this behavior could influence the content of available metals.

The aim of our research was to study the evolution of microelements from soil before and after phytosanitary treatments with dithiocarbamates, during and after remanence period. In order to fulfill our objectives, soil samples were collected from Tohani vineyard, 20 and 40 cm depth.


257

LIST OF PARTICIPANTS


258

Name Affiliation Tel, Fax, E-mail

Adamson, Catherine

Department of Geograhy, Archaelogy and Palaeoecology, Queen’s University , University Road, Belfast, BT7 1NN, UK

Tel: 07761115891

[email protected]

Allsopp, Dennis Westbrook, II the Drive, Northampton NNI4RZ, UK Tel: +44 1604 792354

Fax: +44 1604 792016

[email protected]

Ardelean, Ioan Institute of Biology, Splaint Independentei 296, 060031 Bucharest, Romania

Tel: 0728129144

[email protected]

Arroyo, Irene Instituo del Patrimonio Cultural de Espana, Ministero de Cultury, c/el Greco n°4 28040 Madrid, Spain

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Azami, Sina Faculty of Biological Sciences, Shahid Beheshti University, Tehran, IRAN

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Barnabei, Laura University of Tuscia, Largo dell’universita’Viterbo, 01100, Italy

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Barrera Cortés, Josefina

Cinvestav-IPN, Departamento de Biotecnologia, Av. Instituto Politecnico Nacional 2508, Rd. San Pedro Zacatenco, 07360 Mexico, DF

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Ext:4380

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Baskar, Ramanathan

Department of Environmental Science and Engineering, Guru Jambheshwar University of Science and Technology, Hisar 125001, Haryana, India

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Österreichische Nationalbibliothek, Institut für Restaurierung, Josefsplatz 1, 1015 Wien, Austria

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Bertuzzi, Stefano

Universita degli studi di trieste, Dipartimento di scienze della vita, via giogieri 10, Trieste, 34127, Italy

Tel: +390405583885

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Billeci, Noemi University of Palermo, Italy, Dipartimento di Biologia Ambientale e Biodiversità, Via Archirafi 38, 90123 Palermo

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Bingley, Gavin Manchester Metropolitan Universit, Dept. Computing and Maths, John Dalton Building Chester Street, Manchester,MiSGD UK

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Bondarev, Andreea

Petroleum-Gas University of Ploiesti, Bucharest Av. 39, Ploiesti,100680 Bucharest, Romania

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Boronin, Alexander

Institute of Biochemistry and physiology of microorganisms RAS, Prospect Nauki 5, 142240 Russia

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Bosch Roig, Pilar

Institute of Restoration of Cultural Heritage of the Polytechnic University of Valencia, c/ Camino de vera 13 Valencia, 46022 Spain

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Brusetti, Lorenzo

Faculty of Science and Technology, Free University of Bolzano, piazza Università 5, 39100 Bolzano, Italy

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Calin, Catalina Petroleum-Gas University of Ploiesti,chemistry Department, Bucharest Av. No. 39, Ploiesti 100680 Romania

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Canalizo-Hernandez, Monica A.

Dow Microbial Control. The Dow Chemical Company, 727 Norristown Rd., spring House PA, 19477 USA

Tel: +1 847 6440227

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Carvalho, Carla C. C. R.

Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Department of Bioengineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal

Tel: +351 21 841 9189

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Castro-Silva, Carolina

Centro de investigacion y de studios avanzados del instituto politecnico nacional, av. instituto politecnico nacional 2508 san pedro zacatenco calz ticoman y acueducto gustavo a madero D.F. 07360 Mexico

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Chang-Ping Yu Key Laboratory of Urban Environment and Health, Tel: 865926190768


260

Institute of Urban Environment, Chinese Academy of Sciences,1799 Jimei Road, Xiamen 361021, China

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Chatterjee, Sumana

Department of Chemistry , Basanti Devi College, 147 /B Rashbehari Avenue, Kolkata 700029, India.

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Chow Goon, Ng

School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 05N-23, Singapore 637551,

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Ciccazzo, Sonia Distan, Maae, Via Celoria 2, 20133 Milano, Italy Tel: + 39 3201758177

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RSE – Ricerca sulSistema nergetico S.p.A., Dipartimento Ambente e Sviluppo Sostenibile, V. rubatti No 54, 20134 Milan, Italy

Tel: +390239924655

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Csutak, Ortansa Elisabeth

Department of Genetics, Faculty of Biology, University of Bucharest, Bd. Mihail Kogalniceanu, 36-46,Sector 5, Bucharest 050107, Romania

Tel:+40-213118077

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Culhane, John Department of Science and Health, School of Science, Institute of Technology Carlow, IT Carlow, Kilkenny Road, Carlow, Co-Carlow, Ireland

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Cuzman, Oana. A.

ICVBC-CNR, Via Madonna del Piano, Sesto Fiorentino, 50019, Italy

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Faculty of Biological Sciences, Tarbiat Modares University, Al-E-Ahmad Highway, Tehran, IRAN

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Das, Tapan K. Department of biochemistry and biophysics, University of Kalyani, Kalyani, West Bengal, India

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De Bolle, Miguel

Janssen Pharmaceutica NV- Preservation and material Protection-Janssen PMP, Turnhoutseweg 30, 2340 Beerse, Belgium

Tel: +32 14 60 7580

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De Carvalho, Carla C. C. R.

Institute for biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Department of bioengineering, Instituto Superior Technico, Av. Rovisco pais, 1049-001 Lisboa, Portugal

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261

de Franca, Francisca Pessoa

Escola de Quimica, Universidade Federal do Rio de Janeiro, Av. Athos da Silveira Ramos, 149, 21941-909, Rio de Janeiro, RJ, Brasil

Tel: 552125627621

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De Lorenzo, Victor

Centro Nacional de Biotecnologia, CSIC C/ Darwin, 3 (Campus de Cantoblanco) Madrid 28049, Spain http://www.cnb.csic.es/~meml

Tel +34 91585 4536

Lab +34 91585 4573.

Fax +34 91585 4506 Cellular phone (+34) 609 062 062.

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de los Ríos, Asunción

Museo de Ciencias Naturales, CSIC, Serrano 115 dpdo, Madrid-28006, Spain

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Demnnerova, Katerina

Department of Biochemistry and Microbiology, Department of Food Chemistry and Analysis

Institute of Chemical Technology Prague, Technická 5, 166 28 Prague 6, Czech Republic

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Dobretsov, Sergey

Department of Marine Science and Fisheries, College of Agricultural and Marine Sciences, Sultan Qaboos University, Al-Khoud 123, PO Box34,123 Oman

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Dossi, Michael Biomin Gmbh, Biomin Research Center, Technopark 1, 3430 Tulln an der Donau, Austria

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Dumont, Elisabeth

European Patent Office, Grasserstr. 9, 80339, Munich, Germany

Tel: +49 8923997704

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Duque, Estrella Estacion Experimental deel Zaidin, Consejo Superior de Investigaciones Cientificas, Profesor Albareda n°1, Granada, Spain

Tel:+34 958 181600-292

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El Moslimany, Wael A.

Biotechnology Program, College of Graduate Studies, Arabian Gulf University, Kingdom of Bahrain, Manama P.O. Box 26671 & Taiba University, Faculty of Science, Medina, Universities Avenue, P.O. Box 344

Tel: +9733 36146948

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El-Rahim, Wafaa M. Abd

Department of Agriculture Microbiology, National Research Centre, 32 Tahrir st.Dokki, Cairo, Egypt

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El-Sayed, Wael S.

Microbiology Department, Faculty of Science, Ain Shams University, Cairo, Egypt

Tel: 00966-568582670

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Ellersdorfer, Günther

Institute of Applied Microbiology, Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 11, 1190 Vienna, Austria

Tel: + 43 1 47654 6935

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Esteve-Nuñez, Abraham

Departamento de Analítica e Ingeniería Química, Universidad de Alcalá, E-28871, Alcalá de Henares, Madrid, Spain

Tel: +34 918 854950

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Ettenauer, Jörg Institute of Applied Microbiology, Department of Biotechnology, Vienna Institute of Bio Technology (VIBT). University of Natural Resources and Life Sciences, Muthgasse 11, A-1190 Vienna, Austria

Tel: +43 1 47654 6935

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Favero-Longo, Sergio Enrico

University of Torino, Interdepartmental Centre “G. Scansetti” for Studies on Asbestos and Other Toxic Particulates; University of Torino, Dipartimento di Biologia Vegetale - Viale Mattioli 25, 10125, Torino, Italy

Tel: +39 0116705972

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Ferreira, Isabel Filipa

IBB- Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering. Instituto Superior Técnico, Avenida Rovisco Pais, 1049-001 Lisboa, Portugal.

+351 218419668

+351 218419062

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Flaczyk, Joanna Foksowicz

Institute of Natural Fibres & Medicinal Plants, vL. Wojska Polskiego 71B, Poznan, PL60630 Poland

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Flemming, Hans Curt

University of Duisburg-Essen; Faculty of Chemistry; Biofilm Centre;Universitätsstrasse 5;D-45141 Essen.

Phone: +49-201-1836601 Fax: +49-201-1836603 Cell phone: +49-172-9066119 [email protected]

Friman, Hen The Ariel University Center in Samaria, Ariel, Israel & Bar-Ilan University,PO Box 3 Ariel, 40700 Ramt-Gan, Israel

Tel: 97239066606

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Gadd, Geoffrey

(keynote)

Division of Molecular Microbiology College of Life Sciences;University of Dundee;Dundee;DD1 5EH;Scotland;UK

Tel: +44 (0)1382 384767 Fax: +44 (0)1382 388216

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Garcia, Ana Universidad Politecnica de Madrid (UPM), Departamento de Ingenieria y Ciencia de los Materiales, Escuela Tecnica Superior de Ingenieros Industriales, Jose Gutierrez Abascal 2, madrid 28006, Spain

Tel: 34913363164

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Garcia-Diaz Departamento de Biotecnologia y Bioingenieria. Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional (CINVESTAV-IPN). Av. Instituto Politecnico Nacional No. 2508. Col San Pedro Zacatenco. Mexico 07360, D.F.

Tel. (52-55) 5747-3800 Ext. 4380/4388 [email protected]


263

Garcia-Vallés, Maite

Facultat de Geologia, Universitat de Barcelona (UB), c/ Martí i Franquès s/n, 08028 Barcelona, Spain

Tel:934021348

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Gasser, Nora Technisches Museum Wien, Mariahilferstraße 212, 1140 Wien, Austria

Tel: 01 899982830

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Gaylarde, Christine

Univ. of Portsmouth, UK, and Univ. of Sao Paulo, Brazil.

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Gazzano, Claudia

Universita di Torino, Dept. Plant Biology, viale mattioli 25, Torino 10125, Italy

Tel: 00390116705933

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Giaramida, Luca

IDEAS Research institute – The Robert Gordon University,Central Services Building, Garthoee Road, Aberdeen, ABIO7FY UK & The Macaulay Land Use Research Institute – Aberdeen, UK

Tel:(+44)(0)7965026127

Fax: 00441224262473

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Giebeler, Katharina

European Patent Office, Grasserstraße 9, 80339 Munich, Germany

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Gillatt, John Thor Specialities (UK) Limited, Wincham Avenue, Northwich,Cheshire, CW9 6GB, UK

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Gladis, Franziska

University of Rostock, Institute of biology, Applied Ecology, Einstein-Str. 3, 18051 Rostock, Germany

Tel: 00493814986095

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González Benítez, Natalia

Universidad Rey Juan Carlos, Departamento Biología y Geología

Tel:0034645587325

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Gorfer, Markus Fungal Genetics and Genomics Unit, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences,UFT, Konrad-Lorenz Str.24, 3430 Tulln, AUSTRIA

Tel: 0664 8251444

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Guiamet, Patricia

Instituto de Investigaciones Fisicoquimicas Teoricas y Aplicadas (INFTA), Departamento de Quimica, Facultad de Ciencias Exactas, UNLP, CCT La Plata – CONICET, C.C. 16, Suc.4 (1900), La Plata; Facultad de ciencias Veterinarias, Universidasd nacional de La Plata, Conicet

Tel: 54-221-4257430

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Gustavs, Lydia Applied Ecology, University of Rostock, Albert-Einstein lydia.gustavs@uni-


264

Str. 3, 18055 Rostock, Germany rostock.de

Haas, Doris Institute of Hygiene, Microbiology and Environmental Medicine, Medical University Graz, Universitätsplatz 4, 8010 Graz,Austria

Tel: +43 3163807716

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Hajj Chehade, Mohamad

Cerib, rue des longs Reages- Epernon, 28231 France Tel:0237184800

Fax: 0237186266

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Hartikainen, e. Samuel

Department of Food and Environmental Sciences, Division of Microbiology, P.O. Box 56, Biocenter 1, Viikinkaari 9,FIN 00014 University of Helsinki, Finland,

Tel:+358 0919159332

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Hassaninasab, Azam

Institute of Applied Biochemistry and Graduate School of Life and Environmental Sciences, The University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan

Tel:+81(29)8563565

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Herisson, Jean Paris-Est University, French Institute of Science and Technology for Transport, Development and Networks (IFSTTAR), 58 boulevard Lefebvre, 75732 Paris Cedex 15, France

Tel: +33140435418

Fay: +33140436514

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Hong, Chang-Young

Dept. of Forest Sciences, College of agriculture and life sciences, Seoul National University, Lab of wood chemistry, Program in Environmental Materials Science, Department of Forest Sciences, CALS, San 56-1 Sillimdong, Gwanak-gu,151-921 Seoul, Korea

Tel: 82 2 880 4795

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Hrdinová, Jitka EPS, s.r.o., V Pastouškách 205, 686 04 Kunovice, Czech Republic

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Hsieh, Paul Dipartimento di Scienze della Vita, Università degli Studi, Via L. Giorgieri 10, I-34127 Trieste, Italy

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Ishtiaq Ali, Muhammad

Department of Microbiology, Quaid-i-Azam University, Islamabad Pakistan 45320

Tel: +925190643196

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Izquierdo, Andrés

Universitat de barcelona, Department of Microbiology, Avda. Diagonal 645, 08028 Barcelona, Spain

Tel: +34648924832

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Jirku, Vladimir Institute of Chemical Technology, Technicka 5, 166 28 Prague 6, Czech Republic

Tel:+420 602 973 879

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265

Jo, Chang Wook

Conservation Science Division, National Research Institute of Cultural Heritage, 472 Munji-dong, Yuseong-gu, Daejeon, 305-380, Korea

Tel: +82428609415

Fax:+82428614928

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Jroundi, Fadwa Dept. of Microbiology, Sciences Faculty, University of Granada, Avda. Funtenueva, sn., 18002 Granada, Spain

Tel: 0034958249331

Fax: 0034958249486

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Kabiersch, Grit Department of Food and Environmental Sciences, University of Helsinki, P.O. Box 56, 00014 Helsinki, Finland

Tel:+358 45 2322275

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Karamanlioglu, Mehlika

Faculty of Life Sciences, University of Manchester, Oxford Road,1800 Stopford Building, Manchester, M13 9PT, Manchester, UK

Tel: +4407810020375

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Kari Steffen Department of Food and Microbiological Sciences, P.O.B. 56, 00014 University of Helsinki, Finland

Tel:+358 405509115

Fax:+358 519159322

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Katayama, Yoko

Graduate School of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509, Japan

Tel:+81 0423675732

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Kavkler, Katja Institute for the Protection of Cultural Heritage of Slovenia, Restoration Centre, Poljanska 40, 1000 Ljubljana, Slovenia & Gorenjska predilnica d.d., Kidričeva 75, 4220 Škofja Loka, Slovenia

Tel: +386 12343120

Fax:+386 12343176

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Kesserü, Peter Bay Zoltan Foundation for applied Research Institute for Biotechnology, Szeged, Derkovits Fasor 2, 6726 Hungary

Tel:+36309327746

Fax: +3662432250

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Khiyami, Mohammad Ahmed

King Abdulaziz city for Science and Technology

Saudi Arabia, P. O. Box 6086, Riyadh 11442

Tel:96614813772

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Kikkawa, Yashiho

Imperial Household Agency/Shoison Treasure House, 127, Zoshicha, Naza city, 630-8211 Japan

Tel:+81 742262811

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Kim, Young-Hee

Conservation Science Division, National Research Institute of Cultural Heritage, 472 Munji-dong, Yuseong-gu, Daejoen, 305-380, Korea

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Koos, Akos Bay Zoltan Foundation for applied Research Instiute for Biotechnology, Szeged, Derkovits Fasor 2, 6726 Hungary

Tel: 0662432248

Fax: 0662432250


266

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Kosheleva, Irina

Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences,prospect Nauki, 5, Pushchino 142290, Russia & Pushchino State University, Pushchino 142290, Russia

Tel: +791 63887704

Fax: 8 (495)9563370

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Kraková, Lucia Institute of Molecular Biology, Slovak Academy of Sciences, Dubravska Cesta 21, 84551 Bratislava, Slovakia

Tel:+421259307439

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Kusumi, Asako Graduate School of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8, Saiwai-Cho, Fuchu, Tokyo, 183-8509, Japan

Tel:+81 0422675732

Fax:+81 0423675732

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Kuyukina, Maria S.

Institute of Ecology and Genetics of Microorganisms, Russian Academy of Sciences, 13 Golev Street, 614081, Perm, Russia.

&Microbiology & Immunology Department, Perm State University, 15 Bukhirev Street, 614990, Perm, Russia

Tel: +7(342)2808114

Fax: +7(342)2809211

[email protected].

Laffont, Caroline

Bibliothèque nationale de France, 5 rue Vivienne, 75002 Paris, France

Tel : +330153798598

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Langarica-Fuentes, Adrian

Faculty of Life Sciences, University of Manchester, Michael Smith Bldg., Oxford Road, Manchester M13 9PT, United Kingdom

Tel: 447512076392

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Laudy,A. Museum - Palace, Kostki Potockiego10/16 02-958Warsaw, Poland, http://www.wilanow-palac.pl/

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Li, Yongfeng Key Laboratory of Bio-based Material Science and Technology of Ministry of Education, Northeast Forestry University, Harbin 150040, P. R. China;

Tel:+86 45182190134

Fax:+86 45182190134

[email protected],

Lopandic, Ksenija

Austrian Center of Biological Resources and Applied Mycology, Institute of Applied Microbiology, University of Natural Resources and Life Sciences, Vienna, Austria

Tel: +43 1 47654 6943 [email protected]

López-Fernandez, Margarita

Institute of Microbiology, Universidad de Granada, Campus Fuentenueva, s/n, 18071 Granada, Spain

Tel:0034958249331

Fax:0034958249486

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López-Miras, Maria del Mar

Dep. of Painting. Faculty of Fine Arts. University of Granada. Avda. Andalucía s/n, 18071, Granada, Spain

Tel: 616052138

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Maila, Mphekgo P.

Agricultural Research Council, Private Bag X79, Pretoria, 0001, South Africa

Tel: +27 0123102648


267

Fax: +27 (0)12-323 1157

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Maixner, Frank EURAC- Institute for Mummies and the Iceman, Viale Druso 1, 39100 Bolzano, Italy

Tel: +39 0471 055 567 Fax: +39 0471 055 579

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Majewska, Magdalena M.

Department of Mycology, W. Szafer Institute of Botany, Polish Academy of Sciences, Lubicz Str. 46, 31512 Krakow, Poland

Tel: +48 509907697

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Mandal, Sikha Department of Botany, Visva-Bharati University, Santiniketan-731235, West Bengal, India

Tel:+919232316615

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Margesin, Rosa Institute of Microbiology, University of Innsbruck, Technikerstrasse 25, A-6020 Innsbruck, Austria.

Tel: 0512 5076021

Fax: 0512 5072929

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Martins das Nevey, Luis Carlos

Department of Pharmaceutical Technology, School of Pharmaceutical Science.& Biotechnology Department, School of Engineering of Lorena/USP. University of São Paulo, Brazil.

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Masaphy, Segula

Applied Microbiology and Mycology Department, MIGAL, P.O. Box 831, Kiryat Shmona 11016, Israel & Tel Hai Academic College, Upper Galilee 12210, Israel

Tel: 972-4-6953519;

FAX: 972-4-6944980

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Mathew, Dony Chacko

Department of Life Sciences, National Chung Hsing University, Taichung, 40227, Taiwan

Tel:+8864 22840416

Fax: + 886422874740

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Mathurasa, Lada

National Center of Excellence for Environmental and Hazardous Waste Management (NCE-EHWM), Chulalongkorn University, Bangkok, Thailand

Tel: (6689) 776 0964

Fax: (662) 218 4227

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Matsui, Toru Center of Molecular Biosciences, Tropical ioshere Research Center, University of the Ryukyus,1 Senbaru Nishihara-cho, Okinawa 903-0213, Japan.

Tel: +81 98 895 8943

Fax: +81 98 895 8944

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Mattos, Maria Laura Turino

Embrapa Temperature Climate, BR 392, km 78, 96001-970, Pelotas, RS, Brazil

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Mayer, Elisabeth

BIOMIN Holding GmbH, Industriestrasse 21, A-3130 Herzogenburg, Austria

Tel: 02782 8030

Fax: 02782 80330

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268

Mesquita, Nuno Centre for Functional Ecology, Department of Life Sciences, Faculty of Sciences and Technology, University of Coimbra,POBOX 3046, 3001-401 Coimbra, Portugal

Tel: +351 969830668

Fax: +351 239855241

[email protected] [email protected]

Meszaros, Éva Eötvös Lorand University, Department of Microbiology, Hungary, 1117-Budapest, Pazmany Peter stny. 1/C.

Tel: +36 1 372250008384

Fax: +361 381 2178

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Mihai, Sonia Petroleum and Gas University of Ploiesti,B-Aul, 100680 Ploiesti, Romania

Fax: +40 244-575847

Tel: +40-244-573-171

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Miller, Ana Zélia

Centro de Petrologia e Geoquímica, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001, Lisboa, Portugal

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Minai-Tehrani, Dariush

Faculty of Biological Sciences, Shahid Beheshti University, Chamran Highway, Evin, Tehran, IRAN

Tel:+98 2188022039

Fax:+98 2122431664

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Mohammadi, Parisa

Department of Biology, Faculty of Science, Alzahra University, Vanak Street, Tehran, IR of Iran

Tel: +9821 88009088

Fax:+982188058912

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Mohd Zairi, Mohd Naqiuddin

The University of Manchester, 22 Chelford Close, Manchester, M13ODB, UK

Tel: +447 593081492

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Mollasalehi, Somayeh

Faculty of Life Sciences, University of Manchester, 19 Stadium Drive, Beswick,MII3NB Manchester, UK

Tel: 00447958153891

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Montero, Julia Instituto del Patrimonio Cultural de España, C/Pintor el Greco, 4, Madrid, 28040 Spain

Tel: +34915504486

Fax:+34915504444

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Montes, Rosa Universidad Politecnica de Valencia, Etsiamn-Biotecnologica, Camino de Vera sin Valencia, 46022 Spain

Tel:+34 619138550

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Moreno, Diego Universidad Politécnica de Madrid (UPM) ;Dep. de Ingeniería y Ciencia de los Materiales ; Escuela Técnica Superior de Ingenieros Industriales ; José Gutiérrez Abascal, 2 ; 28006 Madrid (SPAIN)

Tel: 679 158 424

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Morrison, Liam Department of Earth and Ocean Sciences, School of Natural Sciences, Ryan Institute, National University of

Tel: +353 877741806


269

Ireland,University Road, Galway, Ireland [email protected]

Mozafari, Vahid

Department of Soil Science, Vali-e-Asr University of Rafsanjan, 518 Rafsanjan, Iran

Tel:+983413202046

Fax:+983913202042

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Mutnuri Applied and Environmental Biotechnology Laboratory, Dept. of Biological Sciences, BITS Pilani - K.K Birla Goa Campus, Zuari Nagar, Goa 403726, India

Telephone : 0091 832 2580125,

Fax : 0091 832 2557033

[email protected]

Nemeth, Alexandra

Bay Zoltan Foundation for applied Researc,h Institute for Biotechnology, Szeged, Derkovits Fasor 2, 6726 Hungary

Tel: +3662432248

Fax.+3662432250

[email protected]

Ning-Yi Zhou Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44, Xiao Hong Shan,Wuhan, 430071, China

Tel: 0086 2787197655

Fax:0086 2787197655

[email protected]

Nir, Irit Moshav TAad, Misgav, 20155 Israel Tel: 97249909342

Fax: 97249909342

[email protected]

Nithila, Ruth Corrosion Science and Technology division, Indira Gandhi Centre for Research,Kalpakkam 603 102, India

Tel: +91 4427480500 extn.:22072

Fax: +91 4427840121

[email protected]

Nopcharoenkul, Wannarak

Inter-department of Environmental Science, Graduate School, Chulalongkorn University, Phayathai Rd., Pathumwan, Bangkok 10330, Thailand

Tel: (6681)5822861

Fax: (662)2187668

[email protected]

Nordin, Noor Faizul Hardy

Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. & Department of Biotechnology, Kulliyyah of Science, International Islamic University Malaysia,Jalan Sultan Ahmad Shan, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia

Tel: 60193903696

[email protected]

Oliveira, Ana Fundacas de ciencia, Aplicacoes e Tecnologia, Espaciasis – Funcate, Av. Dr. Soas Guilhermino, 429-11-Sao Jose Dos Campos-SP, 12210-131, Brazil

Tel:552121231201

Fax:552121271194

[email protected]

Olukanni, Olumide David

Environmental Biotechnology Laboratory, Department of Chemical Sciences, College of Natural Sciences,BLK E FLAT 3 Run Quaters, Redemtion Camp, LAG-Ibadan Exp

Tel: +2348059360929, olukanni_olumide@yahoo.


270

Redeemer’s University. PMB 3005 Redemption City, Ogun State, Nigeria

com

Ortega-Morales, Benjamin Otto

Departamento de Microbiología Ambiental y Biotecnología, Universidad Autónoma de Campeche, Melgar, Col. Buenavista, CP 24039, Campeche, México.

Tel: 52 981 8119800

[email protected]

Pangallo, Domenico

Institute of Molecular Biology, Slovak Academy of Sciences, Dubravska cesta 21, 84551, Bratislava, Slovakia

Tel: +421903311652

Fax. +421259307416

[email protected]

Pasternak, Grzegorz

Wroclaw University of Technology, 1-15, Institute of Environmental Engineering, WYB Wyspianskiego 27, 50-370 Wroclaw, Poland

Tel:506245451

[email protected]

Patek, Miroslav Institute of Microbiology, AS CR, v. v. i., Prague 4, Videnska 1083, 14220 Czech Republic;

Tel: (+420)241062398,

Fax: (+420)241722257, [email protected]

Peressutti, Silvia Rita.

Instituto Nacional de Investigación y Desarrollo Pesquero, Paseo Victoria Ocampo Nº1, (7600) Mar del Plata, Argentina.

Tel.: +54-223-4862586; fax: +54-223-4861830. [email protected]

Pilt, Kalle Estonian University of Life Sciences, Department of Rural Building, Kreutzwaldi 1, Tartu,51014 Estonia

Tel: +372 566606663

[email protected]

Piñar, Guadalupe


Tel: + 43 1 47654 6943

[email protected]

Pinzari, Flavia ICPAL - Istituto Centrale per il Restauro e la Conservazione del Patrimonio Archivistico e Librario; Laboratorio di Biologia; Via Milano 76, I-00184 Rome, Italy

Tel ++3906 48291215 Fax. ++3906 4814968

[email protected]

Prieto, Beatriz GEMAP-USC, Grupo de Estudiaos Medioambientales Aplicados al Patrimonio natural y Cultural. Departamento de Edafologia y Quimica Agricola, Facultad de –Farmacia, Universidad de Santiago de Compostela, 15782-Santiago de Compostela, Spain

[email protected]

Prouzova, Petra Dept. of Biochemistry and Microbiology, ICT Prague, Technicka 3, Prague 6, Czech Republic, 166 28

Tel:+420 220443634

[email protected]

Querner, Pascal University of Natural Resources and Life Sciences

Department of Integrated Biology and Biodiversity Research. Soil Ecology Group, Institute of Zoology, Gregor-Mendel-Straße 33, A-1180 Vienna - Austria

[email protected]

Raimondi, 'Nello Carrara' Applied Physics Institute - National Research Council, Via Madonna del Piano 10, I- 50019,

Tel:+39555226379


271

Valentina Sesto Fiorentino, Firenze, Italy Fax:+39555226328

[email protected]

Ramos, Juan-Luis

EEZ-CSIC;C/Profesor Albareda 1;E- 18008 Granada, Spain

Tel: +34 958 181608 Fax: +34 958 135740

[email protected]

Rath, Jnanendra Department of Botany, Visva-Bharati University, Santiniketan-731235, West Bengal, India

Tel: +919474766362

[email protected]

Rathore Department of Botany / Biotechnology, Govt. K.R.G.P.G.(Autonomous) College, Gwalior (M.P.) INDIA.

[email protected]

Regidor, Jose Luis

Universidad politecnica de Valencia – IRP, Camino de Vera S/N, 16022 Spain

Tel: +34656887931

[email protected]

Rehman, Abdul Department of Microbiology and Molecular Genetics, University of the Punjab, 54590 Lahore, Pakistan

Tel: 0924239238531

[email protected]

Reyes, Veridiana

Departamento de Química, Departamento de Biología, Universidad de Guanajuato, Lascurain de Retana 5, Guanajuto, GTO 36000, Mexico

Tel: +524737312936

Fax:+524737320006

[email protected]

Ribeiro, Hugo Laboratório de Hidrobiologia, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS-UP), Universidade do Porto, Largo Professor Abel Salazar, no. 2, 4099-003 Porto, Portugal

Tel: +351 222062285

Fax: + 352 222063384

[email protected]

Richert, Agnieszka

Institute for Engineering of Polimer Materials and Dyes, Sklodowskiej-Curie 55, 87-100 torun, Poland

Phone +48 (056) 650 00 44 [email protected]

Riedel, Kathrin Department of Microbial Proteomics. Institute of Microbiology, TU Braunschweig

& Helmholtz Centre of Infection Research, Inhoffenstrasse 7, D-38124 Braunschweig

Tel. +49 531 6181 3041 (office HZI) +49 531 391 5800 (secretary) Fax. +49 531 391 5854

[email protected]

Rizzo, Carmen Dpt. of Animal Biology and Marine Ecology, University of Messina, viale ferdinando stagno d’alcontres (ex Sauta sperone) 31, 98166 Messina Italy

Tel:+39 3289046925

Fax:090 393409

[email protected]

Romero-Noguera, Julio

Dept. of Painting Fac. Fine Arts, University of Granada, Avda. Andalucia s/n Granada, 18071 Spain

Tel.:+54697940516

[email protected]

Safia, Ahmed Department of Microbiology, Quaid-i-Azam University, Islamabad, Pakistan

Saiz-Jimenez, Cesareo

Instituto de Recursos Naturales y Agrobiologia, Avda Reina Mercedes 10, 41012 Sevilla, Spain

[email protected]


272

Sanmartin, Patricia

GEMAP-USC, Grupo de Estudios Medioambientales Aplicados al Patrimonio Natural y Cultural. Departamento de Edafología y Química Agrícola, Facultad de Farmacia, Universidad de Santiago de Compostela, 15782-Santiago de Compostela, Spain

[email protected]

Santa Cruz Calvo, Lucia

Universidad de Granada, Institudo del Agua, Ramon y cajal No. 4, 18071 Granada Spain

Tel: 626321099

Fax 958

[email protected]

Schrems, Michael

Christian-Doppler-Laboratory “Advanced cellulose chemistry an analytics”, Department of Chemistry, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austria

[email protected]

Schroll, Reiner Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Institute of Soil Ecology, Ingulstaedter Landstr.1, 85764 Neuherberg, Germany

Tel: +49 893187 3319

Fax:+49 893187 3376

[email protected]

Sclocchi, Maria Carla

ICRCPAL Istituto Centrale per il Restauro e la Conservazione del Patrimonio Archivistico e Librario, Laboratorio di Conservazione preventiva, Laboratorio di Biologia e Laboratorio di Fisica, Ministero per i Beni e le Attività Culturali, Via Milano, 76, 00184 Rome, Italy

Tel: 0648291215

[email protected]

Selbmann, Laura

DECOS, Università degli Studi della Tuscia, Largo dell’Università snc, Viterbo,01100 Italy

Tel: +390761357012

Fax:+390761357751

[email protected]

Sert, Hacer Akdeniz Universitiesi manavgat meslek Yuksekokulu, Emek Mah. 3049 Sok No.1 Manavgat Antalaya, 07600 Turkey

Tel:+905054564987

[email protected]

Sert, Hakan Akdeniz Univeritesi egitim fakultesi Ilkogretim Bolumu fen bilgisi ogretmenligi, kampus Antalya 07058 Turkey

Tel:+905363224067

[email protected]

Sessitsch, Angela

AIT Austrian Institute of Technology, Department of Health and Environment, Donau-Citx-Straße 1 A-2444 Seibersdorf, Austria

Tel: 050 5503509

Fax: 050 550 2037

[email protected]

Sipos, Rita Eötvös Lorand University, Department of Microbiology, Hungary , Pazmany Peter stny. 1/C, 1117-Budapest

Tel: +36 13812177

Fax: +36 1 3812178

[email protected]

Sivan, Alex Department of Biotechnology Engineering, Ben Gurion University of the Negev, p.O.B.653 Beer Sheva, 84105 Beer Sheva, Israel

Tel: 972 86461961

Fax: 97286479035

[email protected]


273

Soares-Lutterbach, Marcia Teresa

Laboratório de Biocorrosão e Biodegradação, Divisão de Corrosão e Degradação, Instituto Nacional de Tecnologia, Dr. Joan Guichermino, 429-11, Sao Jose Jos Campos 5P, 12210-131 Brasil.

Tel:552121232703

Fax:552121271194

[email protected].

[email protected]

Spérandio, Mathieu

Université de Toulouse; INSA,UPS,INP; LISBP ; 135 Avenue de Rangueil, F-31077 Toulouse, France

[email protected]

Stehrer, Thomas

Fronius International GmbH, Günter Fronius Straße 1, Thalheim, 4609, Austria

Tel:+43 72422418504

[email protected]

Steitz, Andrea Holfzforschung Austria, Franz Grill-Str. 7, 1030 Wien, Austria

Tel: +43 (0)1 798262337

[email protected]

Sterflinger, Katja


Tel: + 43 1 47654 6260

[email protected]

Stoica, Ileana University of Bucharest, Faculöty of Biology, Genetics Dept.,Bd. Miahil Kogälniceanu nr. 36-46, sector 5, 050107, Bucharest, Romania

Tel: +40213118977

Fax:+40213118077

[email protected]

Streeter, Simon University of Portsmouth, Institute of Marine Sciences, Ferry Road, Portsmouth, PO4 9LY, UK

Tel: 44 (0) 2392845807

Fax:44(0) 2392845800 [email protected]

Subramian, Ramachandran

Department of Biotechnology, Birla Institute of Technology and Science, Pilani, Dubai campus,POBox 345055, Dubai International Academic city, UAE

Tel:+97144200700

Fax:+97144200844

[email protected]

Swiontek, Brzezinska Maria

Department of Environmental Microbiology and Biotechnology, Institute of Ecology and Environmental Protection, Nicolaus Copernicus University, Gagarina 9,87100 Toruń, Poland

Tel:+48566112522

Fax:+48566114443

[email protected]

Szvetnik, Attila Bay Zoltan Foundation for applied Research Instiute for Biotechnology, Szeged, Derkovits Fasor 2, 6726 Hungary

Tel:36 305288614

Fax:3662432250

[email protected]

Tanase Ana-Maria

Genetics Department, Faculty of Biology, Bucharest University, Bd. Miahail Kogälniceanu nr. 36-46, sector 5, 050107,Bucharest, Romania

Tel:+40 213118077

Fax:+40213118077

[email protected]


274

Taran, Mojtaba Department of Biology, Faculty of Science, Razi University, Kermanshah, Iran

[email protected]

Tavzes, Crtomir The Institute for the Protection of Cultural Heritage of Slovenia, Conservation Centre, Research Institute, Metelkova 6, SI-1000 Ljubljana, Slovenia

Tel: ++386 051369311

Fax:++386 012343176

[email protected]

Tesei, Donatella


Tel: + 43 1 47654 6935

Tiano, Piero CNR-ICNBC, Via Madonna Del Piano 10, 50019 Sesto Fiorentino, Italy

Tel: +390555225415

Fax.:+390555228403

[email protected]

Tretiach, Mauro Dipartimento di Scienze della Vita, Università degli Studi, Via L. Giorgieri 10, I-34127 Trieste, Italy

Tel: +39 0405583886

Fax:+39040568855

[email protected]

Tuomela, Marja University of Helsinki/ Dept. Of Food and Environmental Science, P.O. Box 56, 00014 Helsinki, Finland

Tel: +358 919159321

Fax:+358 919159322

[email protected]

Uher, Bohuslav Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlarska 2, 611 00 Brno, Czech Republic;

Tel: +420532146113

Fax:+420532146213

[email protected]

Uhlik, Ondrej Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology Prague, Technicka 3, 166 28 Prague 6, Czech Republic

[email protected], [email protected]

Ulcnik, Ajda Department of Wood Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia

Tel: +38613203640

Fax:+38612572297

[email protected]

Urzi, Clara (keynote)

Dept. of Life Sciences, Sect. of Microbiological, Genetic and Molecular Sciences Viale F. Stagno d'Alcontres, 31 98166 Messina, Italy

[email protected] [email protected]

Vaksmaa, Annika

Department of Marine Science and fisheries, college of Agriculture and Marine Sciences, Sultan Qaboos University, Oman

[email protected]

Valentin, Lara Department of Food and Environmental Sciences, University of Helsinki, Aurinkotuulenkatu 14 A 6,00990 Helsinki Finland.

Tel:+3580440669484

[email protected]

Vassu-Dimov, University of Bucharest, Faculty of biology. Dept. of Tel:+40-213118077


275

Tatiana Elisabeta

Genetics, Bd. Miahil Kogalniceanu 36-46, Sector 5, Bucharest 050107, Romania

Fax:+40-213118077

[email protected]

Vejdovszky, Philipp

University of Natural Resources and Life Sciences, Vienna, Muthgasse 18, 1190 Wien, Austria

Tel: 06503196190

[email protected]

Venkateswaran, Kasthuri (keynote)

California Institute of Technology, Jet Propulsion Laboratory Biotechnology and Planetary Protection Group; M/S 89-2 4800 Oak Grove Dr., Pasadena, CA 91109

Tel: (818) 393-1481

Fax: (818) 3934176

[email protected]

Venzmer, Helmuth

Hochschue Wismar, PF 1210 23952 Wismar Tel:0049 13841753231

[email protected]

Verran, Joanna School of Health Care Science; Manchester Metropolitan University; Chester St.; Manchester M1 5GD

Tel: 00-44-161 247 1206

[email protected]

Vessoni-Penna, Thereza Christina

School of Pharmaceutical Science – Universidade de Sao Paulo, Department of Technology, RVA Professor Miguel Mauricio DA Rocha, n°663, Sao Paulo Capital 04622-032. Brazil

Tel: +55 1199020580

Fax: +55 1121264601

[email protected]

Vieira, Denise Universidade de Sao Paulo Faculdade de Ciencias Farmaceutica – Departamento de tecnologia Bioquimico Farmaceutica,Dr. Francisco Da Silva Prado Street,35, 09661-010, Sao Paulo, Brazil

Tel:(55 11)4178-5374

Fax:(5511)4178-5374 [email protected]

Vilhelmsson, Oddur

The University of Akureyri Department of Natural Resource Sciences.Borgir vid Nordurslod, Akureyri,IS-600 Iceland.

Tel: +354 697 4252

Fax: +354 460 8999

[email protected]

Voitl, Christian Institute of Applied Microbiology, Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 11, 1190 Vienna, Austria

Tel: + 43 1 47654 6935

[email protected]

von Werder, Julia

Hochschule Wismar, University of Applied Sciences Technology, Business and Design, Philipp Müller str. 14, PF 1210, 23966Wismar, Germany

Tel: 03841-753486

Fax:03841 753266

[email protected]

Waghmode, Tatoba R.

Department of Biochemistry, Shivaji University Kolhapur-416004, India

[email protected]

Walczak, Maciej

Nicolaus Copernicus University, Department of Envrionmental Microbiology and Biotechnology, P79-017-72-91, Fafanina 9, 87-100 Torun, Poland

Tel.: +48 566114433

Fax.:+48 566114443

[email protected]

Walentowsk, Judyta

Institute of Natural Fibres & Medicinal Plants, UL Wojska Polskiego 71B, PL 60 630 Poznan, Poland

Tel:+48 618455813


276

Fax:+48 618417830

[email protected]

Walker, James T.

HPA, Microbiological Services Division, Porton Down Salisbury. European Space Agency, UK

[email protected]

Wei, Dong-Sheng

Center for Wood Science, Department of Biology, University of Hamburg,Leuschnerstrasse 91d, 21031 Hamburg, Germany

Tel:+49 4073962462

Fax.+49 4073962499

[email protected]

Welgama, Aakash

Institute for Innovation, Design, and Sustainability Research, Robert Gordon University,St. Andrew Street, Aberdeen, AB25 1HG, UK

Tel:00441224262847

[email protected]

Wetzenkircher, Martina

Technisches Museum Wien, Mariahilferstraße 212, 1140 Wien, Austria

Tel: 06648372022

Fax: +01899984444

[email protected]

Wiegner, Katharina

BAM Federal Institute for Materials Research and Testing,Unter den Eichen 44-46, 12203 Berlin, Germany

Tel: +49 3081044872

Fax: +493081044427

[email protected]

Winquist, Erika Department of Biotechnology and Chemical Technology, Aalto University,P>O Box 96081 FI-01051 Laskut, Finland

Tel:+358 505731529 [email protected]

Wongwongsee, Wanwasan

Microbiology, Science, Chulalongkorn University, Phayathai Rd., Pathumwan, Bangkok, 10330, Thailand

Tel: (6684)7025346

Fax:(662)2527576

[email protected]

Yelle, Daniel J. USDA Forest Products Laboratory, Performance Enhanced Biopolymers, One Gifford Pinchot Drive, Madison, Wisconsin, 53726, USA

Tel: 1 608 231 9359

Fax:+1 608 231 9262

[email protected]

Yusof, Mohd. Nizam

Faculty of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam, Selanor, MALAYSIA

Tel:+6012 933 6720

[email protected]

Zafar, Urooj

Faculty of Life Sciences, Manchester University,24-Eston Street,MI3OFF Manchester, UK

Tel: +44 7789058075

[email protected]

Zakharova, Kristina


Tel: + 43 1 47654 6935

[email protected].


277

Zalar, Polona University of Ljubljana, Biotechnical Faculty, Department of Biology, Jamnikarjeva 101, 1000 Ljubljana, Slovenia

Tel:+38613203392

Fax:+3681 2573300

[email protected]

Zanardini, Elisabetta

School of Life Sciences, University of Warwick,Gibber Hill Coventry , CV47AL(UK)

Tel:+442476522557

Fax: +442476523701

[email protected]

Zhang, Jing State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008, & Joint Open Laboratory of Soil and Environment, Institute of Soil Science and Hong Kong Baptist University, Nanjing 210008, China

[email protected]

Zidkova, Ljuba DEKONTA, a.s.,Remediation and Ecological Projects division, Dretovice 109, 273 42 Stehelceves, Czech Republic.

Tel:+420 602126166

Fax:+420312292966

[email protected]

Zucconi, Laura Department of Ecology and Sustainable Economic Development, Università della Tuscia, Largo dell’Università snc, 01100 Viterbo, Italy

Tel:+390761357033

Fax:+390761357751

[email protected]

Sponsors of IBBS-15 Vienna:

15th Intern

ational Biodeterioration& Biodegradation Symposium

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IBBS-15 Abstract book. 15th International Biodeterioration and Biodegradation Symposium

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