I International Symposium on Quorum Sensing Inhibition

Research Advances in Quorum Sensing Inhibition

Supported by:

Book of abstracts

I International Symposium on Quorum Sensing Inhibition

and Satellite Meeting on Novel Anti-Fouling Strategies

Research Advances in Quorum Sensing Inhibition

I International Symposium on Quorum Sensing Inhibition

Satellite Meeting on Novel Anti-Fouling and Strategies

Book of abstracts

supported by:

EDITED BY ANA OTERO MANUEL ROMERO MARIA ISABEL REYERO

2015

Research Advances in Quorum Sensing Inhibition

I International Symposium on Quorum Sensing Inhibition

Satellite Meeting on Novel Anti-Fouling and Strategies

Book of abstracts

supported by:

EDITED BY

ANA OTERO

MANUEL ROMERO

MARIA ISABEL REYERO

Research Advances in Quorum Sensing Inhibition: I International Symposium on Quorum Sensing Inhibition and Satellite Meeting on Novel Anti-Fouling Strategies, Book of Abstracts/ Edited by Ana Otero, Manuel Romero and Maria Isabel Reyero.

Front cover illustration: The marine alga Delisea pulchra avoids bacterial fouling by secreting furanones that mimic the quorum sensing signals N-acyl homoserine lactones (AHLs), blocking signal reception and avoiding the formation of bacterial biofilm. The picture was published in 1999 and shows the “halo of silence” generated when D. pulchra is embedded in an agar plate with the quorum sensing, pigment producing bacterium Chromobacterium violaceum and N-oxohexanoyl-L-homoserine lactone (Manefiel et al., 1999. Microbiology 145(2):283-291). The picture has been reprinted with the permission of the authors.

© Ana Otero, 2015

Edited by Ana Otero Printed by Campus na nube Servicio de reprografía, edición e impresión dixital da USC

Dep.Legal: C 852‐2015 

  PREFACE

In the last few decades, there have been a number of key scientific milestones which have drastically changed our understanding of the microbial world. These include the discovery, in the 1960s and 1970s, of the existence of bacterial intercellular signals responsible for driving pneumococcal competence, the expression of luminescence in marine vibrios, and fruiting body formation in myxobacteria. Unknown to scientists then, this constituted the tip of the iceberg of a plethora of intricate cell-cell signalling mechanisms developed by bacteria to coordinate group behaviour, later named as Quorum Sensing (QS). These were the first steps which changed the concept of bacteria working as single entities to a behaviour closer to that of multicellular organisms. The burst on the understanding of the different mechanisms governing QS-mediated bacterial responses, during the early 1990’s, revealed a central role for QS in the control of bacterial pathogenicity. This raised the possibility of exploiting QS systems as a novel target in antimicrobial warfare. In the late 1990’s and early 2000’s two important discoveries supported this idea. The first one was when the red seaweed Delisea pulchra was shown to have developed a strategy to prevent bacteria from colonising its surfaces through the production of furanones which mimics the QS N-acylhomoserine lactone signals. These furanones effectively interfered with the QS systems of colonizing bacteria blocking signal reception and hence algal colonisation (Givskov et al., 1996). We have chosen a picture of this phenomenon as the cover for this book. The second discovery was the identification of the AiiA lactonase from Bacillus as the first bacterial enzyme inhibiting QS systems through the degradation of QS signal molecules (Dong et al., 2000). More than 15 years have passed since these discoveries and, during this time, the amount of knowledge on the intricate mechanisms driving QS-mediated signalling in bacteria and the possible strategies to interfere with them has been steadily increasing. As evidenced by the contributions included in this book of abstracts, most efforts have been so far focused on the use of quorum sensing inhibition (QSI) to fight pathogenic bacteria. Still, other applications of QSI such as antifouling, are gaining momentum and will have a special focus on this symposium through a satellite meeting on ‘Novel antifouling strategies’. This is the reason behind the sponsoring of this meeting by the EU-OCEAN funded project Byefouling.

This international symposium was conceived with the purpose of gathering our current knowledge on QSI and evaluate the possibilities of seeing this field applied to sectors ranging from the clinic to the environment in the coming years.

Ana Otero Miguel Cámara

References

Givskov M, de Nys R, Manefield M, Gram L, Maximilien R, Eberl L, Molin S, Steinberg PD, Kjelleberg S. 1996. Eukaryotic interference with homorserine lactone-mediated prokaryotic signalling. J Bacteriol 178:6618-6622.

Dong YH, Xu JL, Li XZ, Zhang LH. 2000. AiiA, an enzyme that inactivates the acylhomoserine lactone quorum-sensing signal and attenuates the virulence of Erwinia carotovora. PNAS 97:3526-3531.

Research Advances in Quorum Sensing Inhibition: I International Symposium on Quorum Sensing Inhibition and Satellite Meeting on Novel Anti-Fouling Strategies, Book of Abstracts/ Edited by Ana Otero, Manuel Romero and Maria Isabel Reyero.

Front cover illustration: The marine alga Delisea pulchra avoids bacterial fouling by secreting furanones that mimic the quorum sensing signals N-acyl homoserine lactones (AHLs), blocking signal reception and avoiding the formation of bacterial biofilm. The picture was published in 1999 and shows the “halo of silence” generated when D. pulchra is embedded in an agar plate with the quorum sensing, pigment producing bacterium Chromobacterium violaceum and N-oxohexanoyl-L-homoserine lactone (Manefiel et al., 1999. Microbiology 145(2):283-291). The picture has been reprinted with the permission of the authors.

© Ana Otero, 2015

Edited by Ana Otero Printed by Campus na nube Servicio de reprografía, edición e impresión dixital da USC

Dep.Legal: C 852‐2015 

  PREFACE

In the last few decades, there have been a number of key scientific milestones which have drastically changed our understanding of the microbial world. These include the discovery, in the 1960s and 1970s, of the existence of bacterial intercellular signals responsible for driving pneumococcal competence, the expression of luminescence in marine vibrios, and fruiting body formation in myxobacteria. Unknown to scientists then, this constituted the tip of the iceberg of a plethora of intricate cell-cell signalling mechanisms developed by bacteria to coordinate group behaviour, later named as Quorum Sensing (QS). These were the first steps which changed the concept of bacteria working as single entities to a behaviour closer to that of multicellular organisms. The burst on the understanding of the different mechanisms governing QS-mediated bacterial responses, during the early 1990’s, revealed a central role for QS in the control of bacterial pathogenicity. This raised the possibility of exploiting QS systems as a novel target in antimicrobial warfare. In the late 1990’s and early 2000’s two important discoveries supported this idea. The first one was when the red seaweed Delisea pulchra was shown to have developed a strategy to prevent bacteria from colonising its surfaces through the production of furanones which mimics the QS N-acylhomoserine lactone signals. These furanones effectively interfered with the QS systems of colonizing bacteria blocking signal reception and hence algal colonisation (Givskov et al., 1996). We have chosen a picture of this phenomenon as the cover for this book. The second discovery was the identification of the AiiA lactonase from Bacillus as the first bacterial enzyme inhibiting QS systems through the degradation of QS signal molecules (Dong et al., 2000). More than 15 years have passed since these discoveries and, during this time, the amount of knowledge on the intricate mechanisms driving QS-mediated signalling in bacteria and the possible strategies to interfere with them has been steadily increasing. As evidenced by the contributions included in this book of abstracts, most efforts have been so far focused on the use of quorum sensing inhibition (QSI) to fight pathogenic bacteria. Still, other applications of QSI such as antifouling, are gaining momentum and will have a special focus on this symposium through a satellite meeting on ‘Novel antifouling strategies’. This is the reason behind the sponsoring of this meeting by the EU-OCEAN funded project Byefouling.

This international symposium was conceived with the purpose of gathering our current knowledge on QSI and evaluate the possibilities of seeing this field applied to sectors ranging from the clinic to the environment in the coming years.

Ana Otero Miguel Cámara

References

Givskov M, de Nys R, Manefield M, Gram L, Maximilien R, Eberl L, Molin S, Steinberg PD, Kjelleberg S. 1996. Eukaryotic interference with homorserine lactone-mediated prokaryotic signalling. J Bacteriol 178:6618-6622.

Dong YH, Xu JL, Li XZ, Zhang LH. 2000. AiiA, an enzyme that inactivates the acylhomoserine lactone quorum-sensing signal and attenuates the virulence of Erwinia carotovora. PNAS 97:3526-3531.

The Symposium is supported by the EU funded project BYEFOULING and the Universidade de Santiago de Compostela (USC, Spain), with the collaboration of the European Society for Marine Biotechnology.

SCIENTIFIC COMMITTEE:

Dr. Ana Otero, Universidade de Santiago de Compostela (USC), Spain.

Prof. Miguel Cámara, University of Nottingham, UK.

Dr. Yves Dessaux, Institute for integrative biology of the cell. CEA-CNRS-Université Paris- Sud,France.

Dr. Tom Defoirdt, Ghent University, Belgium.

Dr. Karen Tait, Plymouth Marine Laboratory, UK.

Dr. Inmaculada Llamas, Universidad de Granada, Spain.

SATELLITE MEETING SCIENTIFIC AND ORGANISING COMMITTE Dr. Ana Otero, Universidade de Santiago de Compostela (USC), Spain.

Prof. Yehuda Benayhu, Tel Aviv University, Israel Dr. Joao Tedim, Smallmatek, Portugal

Peter Van Aken, Lonza Group Ltd., Switzerland

Dr. Claire Hellio. BioDIMar(UBO/IUEM/LEMAR Bioprospection platform) Université de Bretagne Occidentale, France

LOCAL ORGANIZING COMMITTEE:

Dr. Manuel Romero, Universidade de Santiago de Compostela (USC), Spain.

Celia Mayer, Universidade de Santiago de Compostela (USC), Spain.

Isabel Freire, Universidade de Santiago de Compostela (USC), Spain.

Andrea Muras, Universidade de Santiago de Compostela (USC), Spain.

Hugo Milhazes-Cunha, Universidade de Santiago de Compostela (USC), Spain.

Lorena Silva, Universidade de Santiago de Compostela (USC), Spain.

Isabel Reyero, Universidade de Santiago de Compostela (USC), Spain.

QSI, Santiago de Compostela 2015 

INDEX KEYNOTE LECTURES

Williams P. The art of antibacterial aarfare – Deception through interference with quorum sensing………..……….. 13 Rodríguez Iglesias P. Regulatory aspects of chemical substances under REACH and Biocidal Products Regulation………... 14

INVITED LECTURES

Cámara M. Silencing quorum sensing in Pseudomonas aeruginosa…... 17 Defoirdt T. Quorum sensing inhibition to control bacterial disease: aquaculture as an example... 18 Dessaux Y., Grandclément, C., Faure D. Quorum quenching: actors, biological roles and applications……….…… 19 Tait K. Havenhand, J., Cámara, M., Williams, P. AHLs and macrofouling wihin the marine environment……….…………... 20 Federle MJ. Inhibiting Rgg pheromone receptors to disrupt quorum sensing in Gram- positive bacteria………..…….. 21

ORAL COMUNICATIONS

Streit WR. Metagenomes as potent resources for the identification of novel QQ genes with anti-biofilm activities………..……….. 25 Torres M., Uroz, S., Quesada, E., Llamas, I. Screening of the quorum quenching activity in a metagenomic library from a hypersaline-soil sample taken in Rambla Salada (Murcia, Spain)... 26 Bertini EV, Leguina CV., Castellanos de Figueroa LI, Nieto-Peñalver CG. Inactivation of QS molecules by endophytic yeasts from sugarcane………... 27 Mayer C, Romero M, Muras A, Otero A. Aii20J, a novel thermostable lactonase from Tenacibaculum sp.strain 20J can quench AHL-mediated acid resistance in E.coli……….. 28 Garge S, Nerurkar A. Quorum quenching N-acyl homoserine lactonase from soil isolate Lysinibacillus sp. Gs50………... 29 Rueda NJ, Suarez, MO, Romero N, Correa E, Ordúz S, Flórez A, Flórez AM. Genetic improvement of aiiA gene encoding N-acyl homoserine lactonase from Bacillus thuringiensis by Error-Prone PCR………... 30

The Symposium is supported by the EU funded project BYEFOULING and the Universidade de Santiago de Compostela (USC, Spain), with the collaboration of the European Society for Marine Biotechnology.

SCIENTIFIC COMMITTEE:

Dr. Ana Otero, Universidade de Santiago de Compostela (USC), Spain.

Prof. Miguel Cámara, University of Nottingham, UK.

Dr. Yves Dessaux, Institute for integrative biology of the cell. CEA-CNRS-Université Paris- Sud,France.

Dr. Tom Defoirdt, Ghent University, Belgium.

Dr. Karen Tait, Plymouth Marine Laboratory, UK.

Dr. Inmaculada Llamas, Universidad de Granada, Spain.

SATELLITE MEETING SCIENTIFIC AND ORGANISING COMMITTE Dr. Ana Otero, Universidade de Santiago de Compostela (USC), Spain.

Prof. Yehuda Benayhu, Tel Aviv University, Israel Dr. Joao Tedim, Smallmatek, Portugal

Peter Van Aken, Lonza Group Ltd., Switzerland

Dr. Claire Hellio. BioDIMar(UBO/IUEM/LEMAR Bioprospection platform) Université de Bretagne Occidentale, France

LOCAL ORGANIZING COMMITTEE:

Dr. Manuel Romero, Universidade de Santiago de Compostela (USC), Spain.

Celia Mayer, Universidade de Santiago de Compostela (USC), Spain.

Isabel Freire, Universidade de Santiago de Compostela (USC), Spain.

Andrea Muras, Universidade de Santiago de Compostela (USC), Spain.

Hugo Milhazes-Cunha, Universidade de Santiago de Compostela (USC), Spain.

Lorena Silva, Universidade de Santiago de Compostela (USC), Spain.

Isabel Reyero, Universidade de Santiago de Compostela (USC), Spain.

QSI, Santiago de Compostela 2015 

INDEX KEYNOTE LECTURES

Williams P. The art of antibacterial aarfare – Deception through interference with quorum sensing………..……….. 13 Rodríguez Iglesias P. Regulatory aspects of chemical substances under REACH and Biocidal Products Regulation………... 14

INVITED LECTURES

Cámara M. Silencing quorum sensing in Pseudomonas aeruginosa…... 17 Defoirdt T. Quorum sensing inhibition to control bacterial disease: aquaculture as an example... 18 Dessaux Y., Grandclément, C., Faure D. Quorum quenching: actors, biological roles and applications……….…… 19 Tait K. Havenhand, J., Cámara, M., Williams, P. AHLs and macrofouling wihin the marine environment……….…………... 20 Federle MJ. Inhibiting Rgg pheromone receptors to disrupt quorum sensing in Gram- positive bacteria………..…….. 21

ORAL COMUNICATIONS

Streit WR. Metagenomes as potent resources for the identification of novel QQ genes with anti-biofilm activities………..……….. 25 Torres M., Uroz, S., Quesada, E., Llamas, I. Screening of the quorum quenching activity in a metagenomic library from a hypersaline-soil sample taken in Rambla Salada (Murcia, Spain)... 26 Bertini EV, Leguina CV., Castellanos de Figueroa LI, Nieto-Peñalver CG. Inactivation of QS molecules by endophytic yeasts from sugarcane………... 27 Mayer C, Romero M, Muras A, Otero A. Aii20J, a novel thermostable lactonase from Tenacibaculum sp.strain 20J can quench AHL-mediated acid resistance in E.coli……….. 28 Garge S, Nerurkar A. Quorum quenching N-acyl homoserine lactonase from soil isolate Lysinibacillus sp. Gs50………... 29 Rueda NJ, Suarez, MO, Romero N, Correa E, Ordúz S, Flórez A, Flórez AM. Genetic improvement of aiiA gene encoding N-acyl homoserine lactonase from Bacillus thuringiensis by Error-Prone PCR………... 30

QSI, Santiago de Compostela 2015 

Müller C, Birmes FS, Rückert C, Kalinowski J, Fetzner S. Bacterial degradation of the Pseudomonas quinolone signal PQS and identification of new quorum quenching enzymes……….. 31 De la Fuente-Nuñez C, Reffuveille F, Brackman G, Coenye T, Hancock REW. Peptides that eradicate multidrug-resistant biofilms and protect against lethal Pseudomonas aeruginosa infections……… 32 Palliyil S, Downham C, Broadbent I, Charlton K, Porter A. Protective effect of quorum quenching monoclonal antibodies in lethal Pseudomonas infection……….. 33 Ivanova K, Fernandes MM, Tzanov T. Sonochemical coatings of acylase nanoparticles inhibit Pseudomonas aeruginosa biofilm formation and virulence factors production………..…. 34 Utari PD, van Merkerk R, Quax WJ. Utilizing AHL acylase to disrupt biofilm formation of hospital-acquired pathogen Acinetobacter nosocomialis M2………... 35 Bhargava N, Sharma P, Capalash N. Unchartering quorum quenching potential in G.glabra for attenuation of quorum sensing mediated virulence of A.

baumannii……… 36 Slachmuylders L, Brackman G, Coenye T. Insight into the effect of the quorum sensing inhibitor baicalin hydrate on Burkholderia cepacia complex biofilm susceptibility……… 37 Cavaleiro E., Duarte AS, Esteves AC, Correia A, Whitcombe MJ, Piletska EV, Piletsky SA, Chianella I. Methacrylate-based polymers to inhibiotic bacterial quorum sensing……… 38 Pande GSJ, Natrah FMI, Kumar U, Bossier P, Defoirdt T. AHL-degrading bacteria isolated from microalgae protect prawn larvae from Vibrio campbellii infection………... 39 Yang Q, Aamdal-Scheie A, Benneche T, Defoirdt T. Novel quorum sensing-disrupting thiophenones with a promising potential to treat vibriosis in aquaculture………. 40 Ascenso OS, Rui F, Miguel AS, Marques JC, Xavier KB, Ventura MR. Study and modulation of inter-species quorum sensing by AI-2 analogues……….……….. 41 Park H, Lee K, Shin H-K, Holzapfel W. Inhibition of autoinducer-2 leads to gut microbiota modulation……….…. 42 Lee K, Park H, Shin H-K, Holzapfel W. Autoinducer-2 quorum sensing influences on bacterial growth under specific stress conditions………. 43 Talagas A, Perchat S, Lereclus D, Nessler, S. Structural analysis of NprR, a bifunctional quorum sensor of the Bacillus cereus group………..………... 44 Brackman G, Van den Driessche F, Coenye T. How does the quorum sensing inhibitor hamamelitanning increase Staphylococcus aureus biofilm susceptibility?... 45

QSI, Santiago de Compostela 2015 

Murugan K, Sekar K, Al-Sohaibani S. Anticaries, antibiofilm and quorum sensing inhibitory activity of stigmasterol-3-D-glucoside isolated from Mimusops elegi L………... 46 Jaiyen Y, Rui F, Steele V, Murray E, Chan W, Williams P. Discovery of inhibitors of the Staphylococcus……….. 47 García-Contreras R, Castañeda P, Maeda T, Wood TK. Is spreading of resistance against quorum quenchers possible?... 48

POSTER COMUNICATIONS

Zhang Y, Brackman G, Coenye T. Interfering with biofilm formation of bacteria involved in chronic wound infections by enzymatic quorum quenching………... 51 Birmes FS, Fetzner S. Mycobacterium abscessus, an emerging pathogen in cystic fibrosis patients, degrades the Pseudomonas quinolone signal………... 52 Müller C, Birmes FS, Fetzner S. Rhodococcus erythropolis BG43, an isolate able to degrade HHQ, PQS and related alkylhydroxyquinolines……….……… 53 Cocotl-Yañez M, Dafhnis-Calas F, Krasnogor N, Cámara M, Heeb S. Engineered quorum quenching bacterial coatings and skins against Pseudomonas aeruginosa infections………. 54 Porzio E, Andrenacci D, Manco G. Quorum quenching activity of archaeal lactonases against Pseudomonas aeruginosa in a Drosophila infection model………... 55 Torres M, Quesada E, Llamas I. Potential biotechnological applications of two strains of Alteromonas with high quorum-quenching activity………... 56 Freire I, Nascimento P, Reyero I, Muras A, Mayer C, Milhazes-Cunha H, Romero M, Otero A. Use of the quorum quenching strain Tenacibaculum sp. 20J to improve survival in mollusc larvae cultures………. 57 Mayer C, Romero M, Muras A, Rumbo S, Gato E, Tomas M, Otero A. Effect of the quorum quenching lactonase Aii20J on biofilm formation by different strains and mutants of Acinetobacter baumannii……….….… 58 Rey D, Mayer C, Muras A., Romero M, Otero A. Identification and cloning of enzymes responsible for quorum quenching activity in the marine bacterium Maribacter ulvicola…..……….. 59 Singh BN. Disabling Pseudomonas aeruginosa quorum sensing by bio-silver nanoparticles……….. 60 Bodelón G, Montes-García V, López-Puente V, Rodal-Cedeira S, Costas C, Pérez-Juste J, Pastoriza-Santos I, Liz-Marzán LM. Non-invasive plasmonic detection and imaging of quorum sensing in biofilms of Pseudomonas aeruginosa……….…….. 61

QSI, Santiago de Compostela 2015 

Müller C, Birmes FS, Rückert C, Kalinowski J, Fetzner S. Bacterial degradation of the Pseudomonas quinolone signal PQS and identification of new quorum quenching enzymes……….. 31 De la Fuente-Nuñez C, Reffuveille F, Brackman G, Coenye T, Hancock REW. Peptides that eradicate multidrug-resistant biofilms and protect against lethal Pseudomonas aeruginosa infections……… 32 Palliyil S, Downham C, Broadbent I, Charlton K, Porter A. Protective effect of quorum quenching monoclonal antibodies in lethal Pseudomonas infection……….. 33 Ivanova K, Fernandes MM, Tzanov T. Sonochemical coatings of acylase nanoparticles inhibit Pseudomonas aeruginosa biofilm formation and virulence factors production………..…. 34 Utari PD, van Merkerk R, Quax WJ. Utilizing AHL acylase to disrupt biofilm formation of hospital-acquired pathogen Acinetobacter nosocomialis M2………... 35 Bhargava N, Sharma P, Capalash N. Unchartering quorum quenching potential in G.glabra for attenuation of quorum sensing mediated virulence of A.

baumannii……… 36 Slachmuylders L, Brackman G, Coenye T. Insight into the effect of the quorum sensing inhibitor baicalin hydrate on Burkholderia cepacia complex biofilm susceptibility……… 37 Cavaleiro E., Duarte AS, Esteves AC, Correia A, Whitcombe MJ, Piletska EV, Piletsky SA, Chianella I. Methacrylate-based polymers to inhibiotic bacterial quorum sensing……… 38 Pande GSJ, Natrah FMI, Kumar U, Bossier P, Defoirdt T. AHL-degrading bacteria isolated from microalgae protect prawn larvae from Vibrio campbellii infection………... 39 Yang Q, Aamdal-Scheie A, Benneche T, Defoirdt T. Novel quorum sensing-disrupting thiophenones with a promising potential to treat vibriosis in aquaculture………. 40 Ascenso OS, Rui F, Miguel AS, Marques JC, Xavier KB, Ventura MR. Study and modulation of inter-species quorum sensing by AI-2 analogues……….……….. 41 Park H, Lee K, Shin H-K, Holzapfel W. Inhibition of autoinducer-2 leads to gut microbiota modulation……….…. 42 Lee K, Park H, Shin H-K, Holzapfel W. Autoinducer-2 quorum sensing influences on bacterial growth under specific stress conditions………. 43 Talagas A, Perchat S, Lereclus D, Nessler, S. Structural analysis of NprR, a bifunctional quorum sensor of the Bacillus cereus group………..………... 44 Brackman G, Van den Driessche F, Coenye T. How does the quorum sensing inhibitor hamamelitanning increase Staphylococcus aureus biofilm susceptibility?... 45

QSI, Santiago de Compostela 2015 

Murugan K, Sekar K, Al-Sohaibani S. Anticaries, antibiofilm and quorum sensing inhibitory activity of stigmasterol-3-D-glucoside isolated from Mimusops elegi L………... 46 Jaiyen Y, Rui F, Steele V, Murray E, Chan W, Williams P. Discovery of inhibitors of the Staphylococcus……….. 47 García-Contreras R, Castañeda P, Maeda T, Wood TK. Is spreading of resistance against quorum quenchers possible?... 48

POSTER COMUNICATIONS

Zhang Y, Brackman G, Coenye T. Interfering with biofilm formation of bacteria involved in chronic wound infections by enzymatic quorum quenching………... 51 Birmes FS, Fetzner S. Mycobacterium abscessus, an emerging pathogen in cystic fibrosis patients, degrades the Pseudomonas quinolone signal………... 52 Müller C, Birmes FS, Fetzner S. Rhodococcus erythropolis BG43, an isolate able to degrade HHQ, PQS and related alkylhydroxyquinolines……….……… 53 Cocotl-Yañez M, Dafhnis-Calas F, Krasnogor N, Cámara M, Heeb S. Engineered quorum quenching bacterial coatings and skins against Pseudomonas aeruginosa infections………. 54 Porzio E, Andrenacci D, Manco G. Quorum quenching activity of archaeal lactonases against Pseudomonas aeruginosa in a Drosophila infection model………... 55 Torres M, Quesada E, Llamas I. Potential biotechnological applications of two strains of Alteromonas with high quorum-quenching activity………... 56 Freire I, Nascimento P, Reyero I, Muras A, Mayer C, Milhazes-Cunha H, Romero M, Otero A. Use of the quorum quenching strain Tenacibaculum sp. 20J to improve survival in mollusc larvae cultures………. 57 Mayer C, Romero M, Muras A, Rumbo S, Gato E, Tomas M, Otero A. Effect of the quorum quenching lactonase Aii20J on biofilm formation by different strains and mutants of Acinetobacter baumannii……….….… 58 Rey D, Mayer C, Muras A., Romero M, Otero A. Identification and cloning of enzymes responsible for quorum quenching activity in the marine bacterium Maribacter ulvicola…..……….. 59 Singh BN. Disabling Pseudomonas aeruginosa quorum sensing by bio-silver nanoparticles……….. 60 Bodelón G, Montes-García V, López-Puente V, Rodal-Cedeira S, Costas C, Pérez-Juste J, Pastoriza-Santos I, Liz-Marzán LM. Non-invasive plasmonic detection and imaging of quorum sensing in biofilms of Pseudomonas aeruginosa……….…….. 61

QSI, Santiago de Compostela 2015 

Huedo P, Yero D, Martínez-Servat S, Daura X, Gibert I. New insights into the QS regulatory network of Stenotrophomonas maltophilia………... 63 Morohoshi T, Shiogai S, Ochiai S, Azuma T, Ishizuka M, Ikeda T. Inhibition of biofilm formation of Gram-negative bacteria using quorum-quenching compounds………... 64 Lu C, Maurer C, Kirsch B, Steinbach A, Weiβ S, Pawar S, Hartmann RW. Quinolone- based PqsR antagonists provide in vivo proof-of-concept for PQS-targeting anti-virulence strategy………... 65 Thomann A, Martins AG, Brengel C, Weidel E, Plaza A, Börger C, Empting M, Hartmann RW. Biological evaluation of an in vivo-potent dual target PQS-quorum sensing inhibitor that hinders biofilm formation………..…….... 66 Lafayette I, Dubern J-F,Do QT, Lazenby J, Halliday N, Heeb S, Williams P, Cámara M.

Discovery of novel small-molecules for the inhibition of Pseudomonas quinolone signalling... 67 Zender M, Kirsch B, Maurer CK, Empting M, Hartmann RW. Development of novel antagonists of PqsR, an important regulator of Pseudomonas aeruginosa virulence……... 68 Kalia M, Kumar V, Kumar Singh P, Dohare S, Narvi SS, Agarwal, V. Inhibition of quorum sensing controlled virulence phenotypes in Pseudomonas aeruginosa PAO1 by using

eucalyptus oil……….………. 69 Pérez-López M, Soto-Hernández M, García-Contreras R, Martínez-Vázquez M, San Miguel-Chávez R, Castillo-Juárez I. Regulatory effect of fatty acids from seeds (Amaranthus hypochondriacus L., Salvia hispánica L.and Helianthus annuus L.) in the quorum sensing systems of Chromobacterium violaceum……….…… 70 Muñoz-Casares N, Soto-Hernández M, García-Contreras R, Martínez-Vázquez M, Aguilar-Rodríguez S, Montes de Oca Mejía M, Castillo-Juárez I. Phytochemical study and evaluation of quorum sensing system inhibition of barks Ceiba spp………... 71 Busetti A, Flynn P, Graham, WG, Gilmore B.F. Quorum sensing inhibition and attenuation of virulence by atmospheric pressure non thermal plasma……... 72 Tait K, Ransome E, Munn C. Bacterial communication and climate change:

consequences for the coral ecosystem………..…………..….… 73 Sato R, Yamaguchi T, Someya N, Ikeda T, Morohoshi T. Analysis of N-acylhomoserine lactone-degrading mechanisms in the coagulase-negative staphylococci……….….. 74 Pérez-Pascual D, Metton C, Besset C, Monnet V, Gardan R. SHP-RovS cell-cell communication mechanism, a target for quorum quenching?... 75 Endo EH, Nakamura TU, Nakamura CV, Días Filho BP. Anti-biofilm effect of Rosmarinus officinalis against MRSA Staphylococcus aureus……….… 76 Ryu Eun-Ju, Sim J, Ko Y-K, Jung K-S, Choi B-K. Application of quorum sensing

  QSI, Santiago de Compostela 2015   

Muras A, Mayer C, Romero M, Ferrer M, Mira A., Otero, A. Interference with quorum sensing signals to prevent biofilm formation by Streptococcus mutans………..…... 78

SATELLITE METING ON NOVEL ANTI-FOULING STRATEGIES

INVITED LECTURES

Burguess JG. Towards commercialization of a novel marine nuclease for biofilm removal and prevention……… 81 Hellio C. Biomimetics approaches for the development of new antifouling strategies………. 82

ORAL COMUNICATIONS

Tedim J, Simon C. The FP7-Ocean EU project BYEFOULING: an interdisciplinary approach to novel anti-fouling strategies……….. 85 Stübing D, van Haare J. Synergistic fouling control technologies – project overview and first results……….. 86 Blas E, Barros R, Guedella E. New biocoating for corrosion inhibition in metal surfaces………... 87 Piller C, Hoch-Gunter E, Weis M, Larroze S,Teixeira T,Goldenberg L, Antipov A, Benayahu Y. Effects of novel anti-fouling paints on aquatic species of different trophic levels……… 88 Rodríguez-Ezpeleta N, Menchaca I, Zorita I, Alonso L, Franco J. Monitoring biofouling on hard substrata through DNA based approaches……….………..……. 89 Wittig L, Grunwald I, Wunder A, Isaksson D. Bio-based and bio-active biofouling control strategies……….…… 90 Martín-Rodríguez AJ, Álvarez-Méndez SJ, Martín VS, Fernández JJ. Novel quorum sensing disruptors and their antifouling implications………..…. 91 Louzao I, Sui C, Winzer K, Fernandez-Trillo F, Alexander C. Polymer mediated bacterial clustering: cell-adhesive properties of cationic homo- and copolymers……….….. 92 Matteucci G, Esposito M, Fiesoletti F, James V, Napolano L, Pascale C, Rossini P, Testoni A. Test on the environmental impacts of two commercial antifouling coating systems……… 93

QSI, Santiago de Compostela 2015 

Huedo P, Yero D, Martínez-Servat S, Daura X, Gibert I. New insights into the QS regulatory network of Stenotrophomonas maltophilia………... 63 Morohoshi T, Shiogai S, Ochiai S, Azuma T, Ishizuka M, Ikeda T. Inhibition of biofilm formation of Gram-negative bacteria using quorum-quenching compounds………... 64 Lu C, Maurer C, Kirsch B, Steinbach A, Weiβ S, Pawar S, Hartmann RW. Quinolone- based PqsR antagonists provide in vivo proof-of-concept for PQS-targeting anti-virulence strategy………... 65 Thomann A, Martins AG, Brengel C, Weidel E, Plaza A, Börger C, Empting M, Hartmann RW. Biological evaluation of an in vivo-potent dual target PQS-quorum sensing inhibitor that hinders biofilm formation………..…….... 66 Lafayette I, Dubern J-F,Do QT, Lazenby J, Halliday N, Heeb S, Williams P, Cámara M.

Discovery of novel small-molecules for the inhibition of Pseudomonas quinolone signalling... 67 Zender M, Kirsch B, Maurer CK, Empting M, Hartmann RW. Development of novel antagonists of PqsR, an important regulator of Pseudomonas aeruginosa virulence……... 68 Kalia M, Kumar V, Kumar Singh P, Dohare S, Narvi SS, Agarwal, V. Inhibition of quorum sensing controlled virulence phenotypes in Pseudomonas aeruginosa PAO1 by using

eucalyptus oil……….………. 69 Pérez-López M, Soto-Hernández M, García-Contreras R, Martínez-Vázquez M, San Miguel-Chávez R, Castillo-Juárez I. Regulatory effect of fatty acids from seeds (Amaranthus hypochondriacus L., Salvia hispánica L.and Helianthus annuus L.) in the quorum sensing systems of Chromobacterium violaceum……….…… 70 Muñoz-Casares N, Soto-Hernández M, García-Contreras R, Martínez-Vázquez M, Aguilar-Rodríguez S, Montes de Oca Mejía M, Castillo-Juárez I. Phytochemical study and evaluation of quorum sensing system inhibition of barks Ceiba spp………... 71 Busetti A, Flynn P, Graham, WG, Gilmore B.F. Quorum sensing inhibition and attenuation of virulence by atmospheric pressure non thermal plasma……... 72 Tait K, Ransome E, Munn C. Bacterial communication and climate change:

consequences for the coral ecosystem………..…………..….… 73 Sato R, Yamaguchi T, Someya N, Ikeda T, Morohoshi T. Analysis of N-acylhomoserine lactone-degrading mechanisms in the coagulase-negative staphylococci……….….. 74 Pérez-Pascual D, Metton C, Besset C, Monnet V, Gardan R. SHP-RovS cell-cell communication mechanism, a target for quorum quenching?... 75 Endo EH, Nakamura TU, Nakamura CV, Días Filho BP. Anti-biofilm effect of Rosmarinus officinalis against MRSA Staphylococcus aureus……….… 76 Ryu Eun-Ju, Sim J, Ko Y-K, Jung K-S, Choi B-K. Application of quorum sensing

  QSI, Santiago de Compostela 2015   

Muras A, Mayer C, Romero M, Ferrer M, Mira A., Otero, A. Interference with quorum sensing signals to prevent biofilm formation by Streptococcus mutans………..…... 78

SATELLITE METING ON NOVEL ANTI-FOULING STRATEGIES

INVITED LECTURES

Burguess JG. Towards commercialization of a novel marine nuclease for biofilm removal and prevention……… 81 Hellio C. Biomimetics approaches for the development of new antifouling strategies………. 82

ORAL COMUNICATIONS

Tedim J, Simon C. The FP7-Ocean EU project BYEFOULING: an interdisciplinary approach to novel anti-fouling strategies……….. 85 Stübing D, van Haare J. Synergistic fouling control technologies – project overview and first results……….. 86 Blas E, Barros R, Guedella E. New biocoating for corrosion inhibition in metal surfaces………... 87 Piller C, Hoch-Gunter E, Weis M, Larroze S,Teixeira T,Goldenberg L, Antipov A, Benayahu Y. Effects of novel anti-fouling paints on aquatic species of different trophic levels……… 88 Rodríguez-Ezpeleta N, Menchaca I, Zorita I, Alonso L, Franco J. Monitoring biofouling on hard substrata through DNA based approaches……….………..……. 89 Wittig L, Grunwald I, Wunder A, Isaksson D. Bio-based and bio-active biofouling control strategies……….…… 90 Martín-Rodríguez AJ, Álvarez-Méndez SJ, Martín VS, Fernández JJ. Novel quorum sensing disruptors and their antifouling implications………..…. 91 Louzao I, Sui C, Winzer K, Fernandez-Trillo F, Alexander C. Polymer mediated bacterial clustering: cell-adhesive properties of cationic homo- and copolymers……….….. 92 Matteucci G, Esposito M, Fiesoletti F, James V, Napolano L, Pascale C, Rossini P, Testoni A. Test on the environmental impacts of two commercial antifouling coating systems……… 93

  QSI, Santiago de Compostela 2015 

POSTER COMUNICATIONS

Arregui L, Soler A, Liébana R, Santos A, Marquina D, Muñagorri F, Serrano S. A tool for controlling biofouling on membrane bioreactor (MBR) wastewater systems based on the selection of quorum quenching bacteria………..……….. 97 Fernández-Hermida X. Hidroboya: The “Sample & Hold” platform to overcome the fouling problem……….…….…. 98 Hoch-Gutner E, Piller C, Avelelas F, Martins R, Malheiro E, Maia F, Tedim J, Benayahu Y. Novel anti-fouling nanomaterials: effects on macrofoulers from the Eastern Mediterranean and the northern Gulf of Aqaba (Red Sea)………... 99 Freire I, Muras A, Reyero I, Otero A. Inhibitory activity in extracts of the marine microalga Isochrysis aff. galbana clon T-ISO against different marine microalgae……….……. 100 Muras A, Freire I, Mayer C, Otero A. Effect of Bacillus licheniformis CECT20T on biofilm formation by marine bacteria………... 101 Muras A, Mayer C, Freire I, Otero A. Monitoring bacterial biofilm development with xCELLigence® technology……….…….…………. 102 Author index...……….………. 103 Notes………... 111

  QSI, Santiago de Compostela 2015   

Keynote lectures

  QSI, Santiago de Compostela 2015 

POSTER COMUNICATIONS

Arregui L, Soler A, Liébana R, Santos A, Marquina D, Muñagorri F, Serrano S. A tool for controlling biofouling on membrane bioreactor (MBR) wastewater systems based on the selection of quorum quenching bacteria………..……….. 97 Fernández-Hermida X. Hidroboya: The “Sample & Hold” platform to overcome the fouling problem……….…….…. 98 Hoch-Gutner E, Piller C, Avelelas F, Martins R, Malheiro E, Maia F, Tedim J, Benayahu Y. Novel anti-fouling nanomaterials: effects on macrofoulers from the Eastern Mediterranean and the northern Gulf of Aqaba (Red Sea)………... 99 Freire I, Muras A, Reyero I, Otero A. Inhibitory activity in extracts of the marine microalga Isochrysis aff. galbana clon T-ISO against different marine microalgae……….……. 100 Muras A, Freire I, Mayer C, Otero A. Effect of Bacillus licheniformis CECT20T on biofilm formation by marine bacteria………... 101 Muras A, Mayer C, Freire I, Otero A. Monitoring bacterial biofilm development with xCELLigence® technology……….…….…………. 102 Author index...……….………. 103 Notes………... 111

  QSI, Santiago de Compostela 2015   

Keynote lectures

ISQSI, Santiago de Compostela 2015  Keynote Lectures 

The art of antibacterial warfare - Deception through interference with quorum sensing

Paul Williams

Centre for Biomolecular Sciences, School of Life Sciences, University of Nottingham, University Park, Nottingham, U.K. E-mail: [email protected]

Keywords: Quorum sensing inhibitors, Staphylococcus aureus, agr system, autoinducing peptides

Abstract

The ability of unicellular micro-organisms to synchronize their behaviour at the population level through quorum sensing (QS) is now recognized as a common adaptive response to environmental challenges. QS enables bacterial populations to mount co-operative responses that improve access to nutrients or particular environmental niches, promote collective defence against other competitor prokaryotes or higher organisms and facilitate survival through differentiation into morphological forms better able to combat environmental threats. Through QS, bacterial populations can modify their nature and dynamics, and act as a community accomplishing tasks that individual bacterial cells would find impossible to achieve. Although we now appreciate that QS signal molecules are chemically diverse, their basic mechanism of action is conserved in that the structure of the QS signal molecule contains information that is directed by a ‘‘sender’’ cell/organism to a ‘‘receiver’’ cell/organism. Since QS often controls virulence and biofilm development, it is an attractive target for anti-infective agents particularly given the global health threat posed by multi-antibiotic resistant bacteria and the lack of new antibiotics entering the clinic. Other potential advantages of targeting QS include expansion of the repertoire of bacterial targets, preserving the host microflora and reducing the pressures that drive selection for resistance.

The common architecture of QS systems provides multiple molecular targets for agents that interfere with QS-mediated cell-to-cell communication, namely (i) the biosynthesis of the signal molecule by the ‘‘sender’’ cell, (ii) the functionality and availability of the signal itself, and (iii) the reception/decoding of the message contained in QS signal molecule by the ‘‘receiver’’ cell. With respect to the discovery of QS inhibitors (QSIs), an appreciation of the need to conserve steric requirements for optimal QS ligand/receptor/synthase interactions, has enabled QSIs to be readily discovered through structural modification of native QS signals. Bacterial strains originally developed as biosensors for QS signal molecules have proved useful as screens for QSIs from chemical libraries and natural products. Recent progress in exploiting our current knowledge of QS systems as targets for anti-infective agents will be explored for bacterial pathogens in general but in particular for Staphylococcus aureus.

ISQSI, Santiago de Compostela 2015  Keynote Lectures 

The art of antibacterial warfare - Deception through interference with quorum sensing

Paul Williams

Centre for Biomolecular Sciences, School of Life Sciences, University of Nottingham, University Park, Nottingham, U.K. E-mail: [email protected]

Keywords: Quorum sensing inhibitors, Staphylococcus aureus, agr system, autoinducing peptides

Abstract

The ability of unicellular micro-organisms to synchronize their behaviour at the population level through quorum sensing (QS) is now recognized as a common adaptive response to environmental challenges. QS enables bacterial populations to mount co-operative responses that improve access to nutrients or particular environmental niches, promote collective defence against other competitor prokaryotes or higher organisms and facilitate survival through differentiation into morphological forms better able to combat environmental threats. Through QS, bacterial populations can modify their nature and dynamics, and act as a community accomplishing tasks that individual bacterial cells would find impossible to achieve. Although we now appreciate that QS signal molecules are chemically diverse, their basic mechanism of action is conserved in that the structure of the QS signal molecule contains information that is directed by a ‘‘sender’’ cell/organism to a ‘‘receiver’’ cell/organism. Since QS often controls virulence and biofilm development, it is an attractive target for anti-infective agents particularly given the global health threat posed by multi-antibiotic resistant bacteria and the lack of new antibiotics entering the clinic. Other potential advantages of targeting QS include expansion of the repertoire of bacterial targets, preserving the host microflora and reducing the pressures that drive selection for resistance.

The common architecture of QS systems provides multiple molecular targets for agents that interfere with QS-mediated cell-to-cell communication, namely (i) the biosynthesis of the signal molecule by the ‘‘sender’’ cell, (ii) the functionality and availability of the signal itself, and (iii) the reception/decoding of the message contained in QS signal molecule by the ‘‘receiver’’ cell. With respect to the discovery of QS inhibitors (QSIs), an appreciation of the need to conserve steric requirements for optimal QS ligand/receptor/synthase interactions, has enabled QSIs to be readily discovered through structural modification of native QS signals. Bacterial strains originally developed as biosensors for QS signal molecules have proved useful as screens for QSIs from chemical libraries and natural products. Recent progress in exploiting our current knowledge of QS systems as targets for anti-infective agents will be explored for bacterial pathogens in general but in particular for Staphylococcus aureus.

ISQSI, Santiago de Compostela 2015  Keynote Lectures 

Regulatory aspects of chemical substances under REACH and Biocidal Products Regulation

Pilar Rodríguez Iglesias

European Chemicals Agency (ECHA). P.O. Box 400, FI-00121, Helsinki, Finland. E- mail: [email protected]

Keywords: REACH, chemicals, biocides, substance, ECHA

Abstract

REACH is a regulation of the European Union (Regulation (EC) 1907/2006), adopted to improve the protection of human health and the environment from the risks that can be posed by chemicals, while enhancing the competitiveness of the EU chemicals industry. It also promotes alternative methods for the hazard assessment of substances in order to reduce the number of tests on animals.

In principle, REACH applies to all chemical substances (i.e. those used in industrial processes but also in our day-to-day lives). REACH places the burden of proof on companies. To comply with the regulation, companies must identify and manage the risks linked to the substances they manufacture and market in the EU. They have to demonstrate to ECHA how the substance can be safely used, and they must communicate the risk management measures to the users.

If the risks cannot be managed, authorities can restrict the use of substances in different ways.

In the long run, the most hazardous substances should be substituted with less dangerous ones.

REACH stands for Registration, Evaluation, Authorisation and Restriction of Chemicals. It entered into force on 1 June 2007.

The Biocidal Products Regulation (BPR, Regulation (EU) 528/2012) concerns the placing on the market and use of biocidal products, which are used to protect humans, animals, materials or articles against harmful organisms like pests or bacteria, by the action of the active substances contained in the biocidal product. This regulation aims to improve the functioning of the biocidal products market in the EU, while ensuring a high level of protection for humans and the environment.

All biocidal products require an authorisation before they can be placed on the market, and the active substances contained in that biocidal product must be previously approved. There are, however, certain exceptions to this principle.

The BPR aims to harmonise the market at Union level, simplify the approval of active substances and authorisation of biocidal products, and introduce timelines for Member State evaluations, opinion-forming and decision-making. It also promotes the reduction of animal testing by introducing mandatory data sharing obligations and encouraging the use of alternative testing methods.

The regulation is applicable since 1 September 2013, with a transitional period for certain provisions.

  ISQSI, Santiago de Compostela 2015   

Invited lectures

ISQSI, Santiago de Compostela 2015  Keynote Lectures 

Regulatory aspects of chemical substances under REACH and Biocidal Products Regulation

Pilar Rodríguez Iglesias

European Chemicals Agency (ECHA). P.O. Box 400, FI-00121, Helsinki, Finland. E- mail: [email protected]

Keywords: REACH, chemicals, biocides, substance, ECHA

Abstract

REACH is a regulation of the European Union (Regulation (EC) 1907/2006), adopted to improve the protection of human health and the environment from the risks that can be posed by chemicals, while enhancing the competitiveness of the EU chemicals industry. It also promotes alternative methods for the hazard assessment of substances in order to reduce the number of tests on animals.

In principle, REACH applies to all chemical substances (i.e. those used in industrial processes but also in our day-to-day lives). REACH places the burden of proof on companies. To comply with the regulation, companies must identify and manage the risks linked to the substances they manufacture and market in the EU. They have to demonstrate to ECHA how the substance can be safely used, and they must communicate the risk management measures to the users.

If the risks cannot be managed, authorities can restrict the use of substances in different ways.

In the long run, the most hazardous substances should be substituted with less dangerous ones.

REACH stands for Registration, Evaluation, Authorisation and Restriction of Chemicals. It entered into force on 1 June 2007.

The Biocidal Products Regulation (BPR, Regulation (EU) 528/2012) concerns the placing on the market and use of biocidal products, which are used to protect humans, animals, materials or articles against harmful organisms like pests or bacteria, by the action of the active substances contained in the biocidal product. This regulation aims to improve the functioning of the biocidal products market in the EU, while ensuring a high level of protection for humans and the environment.

All biocidal products require an authorisation before they can be placed on the market, and the active substances contained in that biocidal product must be previously approved. There are, however, certain exceptions to this principle.

The BPR aims to harmonise the market at Union level, simplify the approval of active substances and authorisation of biocidal products, and introduce timelines for Member State evaluations, opinion-forming and decision-making. It also promotes the reduction of animal testing by introducing mandatory data sharing obligations and encouraging the use of alternative testing methods.

The regulation is applicable since 1 September 2013, with a transitional period for certain provisions.

  ISQSI, Santiago de Compostela 2015   

Invited lectures

    ISQSI, Santiago de Compostela 2015  Invited lectures 

Silencing quorum sensing in Pseudomonas aeruginosa Miguel Cámara

Centre of Biomolecular Sciences, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, United Kingdom. E-mail: [email protected] Keywords: Pseudomonas aeruginosa, PQS, alkylquinolones, biofilms, antimicrobials, peptide nucleic acids

Abstract

Pseudomonas aeruginosa is a major opportunistic pathogen in cystic fibrosis, wound and nosocomial infections due to its high levels of intrinsic and acquired resistance to antibiotics, presenting a serious problem to the public health systems. This organism can adapt to many different environments using highly intricate regulatory networks. Quorum sensing (QS) plays a central role in this adaptation processes and hence is key for the success of this organism during infection regulating the production of many virulence traits. P. aeruginosa has several quorum sensing systems including the LasR/I and RhlR/I systems and their cognate signal molecules, N-(3-oxododecanoyl)-L-homoserine lactone and N-butanoyl-L-homoserine lactone respectively. P. aeruginosa also produces 2-alkyl-4-quinolones (AQ) signal molecules as QS molecules. The major P. aeruginosa AQ signal molecules are 2-heptyl-4-quinolone (HHQ) and 2-heptyl-3-hydroxy-4-quinolone (PQS). These QS systems play an important role in biofilm formation and their resistance to antibiotics. We have shown they are active in vivo during human infection, especially in the CF lung, and can interfere with host responses to infection.

For the above reasons, QS in P. aeruginosa represents a target for the development of novel antibacterials which can attenuate bacterial virulence so that the pathogen fails to adapt to the host environment. Interference with either the synthesis or the transduction of a QS signal is core to this attenuation. We are currently investigating several ways to inhibit QS in P.

aeruginosa. These include: (i) searching for molecules which can antagonise the interaction of PQS/HHQ with their cognate regulatory protein PqsR and (ii) using antisense PNAs (Peptide Nucleic Acids) which prevent the translation of pqsA, the first gene of the pqsA-E operon involved in the biosynthesis of AQ and their response. Using these two approaches we have identified QS inhibitors which can attenuate the virulence of P. aeruginosa and most importantly sensitise biofilms to antibiotics. Details of these studies and their potential impact in the clinic will be present.

    ISQSI, Santiago de Compostela 2015  Invited lectures 

Silencing quorum sensing in Pseudomonas aeruginosa Miguel Cámara

Centre of Biomolecular Sciences, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, United Kingdom. E-mail: [email protected] Keywords: Pseudomonas aeruginosa, PQS, alkylquinolones, biofilms, antimicrobials, peptide nucleic acids

Abstract

Pseudomonas aeruginosa is a major opportunistic pathogen in cystic fibrosis, wound and nosocomial infections due to its high levels of intrinsic and acquired resistance to antibiotics, presenting a serious problem to the public health systems. This organism can adapt to many different environments using highly intricate regulatory networks. Quorum sensing (QS) plays a central role in this adaptation processes and hence is key for the success of this organism during infection regulating the production of many virulence traits. P. aeruginosa has several quorum sensing systems including the LasR/I and RhlR/I systems and their cognate signal molecules, N-(3-oxododecanoyl)-L-homoserine lactone and N-butanoyl-L-homoserine lactone respectively. P. aeruginosa also produces 2-alkyl-4-quinolones (AQ) signal molecules as QS molecules. The major P. aeruginosa AQ signal molecules are 2-heptyl-4-quinolone (HHQ) and 2-heptyl-3-hydroxy-4-quinolone (PQS). These QS systems play an important role in biofilm formation and their resistance to antibiotics. We have shown they are active in vivo during human infection, especially in the CF lung, and can interfere with host responses to infection.

For the above reasons, QS in P. aeruginosa represents a target for the development of novel antibacterials which can attenuate bacterial virulence so that the pathogen fails to adapt to the host environment. Interference with either the synthesis or the transduction of a QS signal is core to this attenuation. We are currently investigating several ways to inhibit QS in P.

aeruginosa. These include: (i) searching for molecules which can antagonise the interaction of PQS/HHQ with their cognate regulatory protein PqsR and (ii) using antisense PNAs (Peptide Nucleic Acids) which prevent the translation of pqsA, the first gene of the pqsA-E operon involved in the biosynthesis of AQ and their response. Using these two approaches we have identified QS inhibitors which can attenuate the virulence of P. aeruginosa and most importantly sensitise biofilms to antibiotics. Details of these studies and their potential impact in the clinic will be present.

ISQSI, Santiago de Compostela 2015  Invited lectures 

Quorum sensing inhibition to control bacterial disease:

aquaculture as an example

Tom Defoirdt

Laboratory of Aquaculture & Artemia Reference Center, Ghent University. Rozier 44, 9000 Gent, Belgium. E-mail: [email protected]

Keywords: Gnotobiotic models, quorum sensing inhibitors, AHL, AI-2, CAI-1, indole

Abstract

Due to large-scale use of antibiotics, many pathogenic bacteria have acquired (multiple) resistance, and therefore, alternative treatments to control disease are needed. More than ten years ago, we proposed inhibition of quorum sensing, bacterial cell-to-cell communication, as a novel strategy to control infections caused by antibiotic-resistant bacteria in aquaculture.

In this presentation, I will discuss our current knowledge on the impact of quorum sensing and quorum sensing disruption on the virulence of aquaculture pathogens towards different host organisms. We found that the three-channel quorum sensing system of Vibrio harveyi regulates the expression of different virulence factors and the virulence of the bacterium towards gnotibiotic brine shrimp (Artemia franciscana) and giant river prawn (Macrobrachium rosenbergii) larvae. Remarkably, the impact of the three signal molecules on virulence of Vibrio harveyi was different for the two hosts, with acylhomoserine lactones (AHLs) playing no role in virulence towards brine shrimp and cholerae autoinducer-1 (CAI-1) playing no role in virulence towards river prawn. In contrast to what we observed in Vibrio harveyi, the three channel quorum sensing system of Vibrio anguillarum was found to play no role in virulence towards gnotobiotic sea bass (Dicentrarchus labrax) larvae. However, we found that another signalling molecule, indole, played a major role in virulence of this pathogen. Finally, we found that AHL quorum sensing controls the virulence of Aeromonas species towards larvae of the freshwater fish species burbot (Lota lota).

The most important quorum sensing-disrupting agents reported thus far include compounds that interfere with quorum sensing signal detection and signal transduction, and signal molecule- degrading bacteria. Quorum sensing-disrupting compounds studied in our laboratory include antagonistic AHL molecules, brominated furanones, brominated thiophenones and cinnamaldehyde. All of these compounds were found to increase survival of aquaculture animals when challenged to pathogenic bacteria. Furthermore, we found that metabolites produced by some micro-algae that are frequently used in aquaculture are also able to disrupt quorum sensing in Gram-negative bacteria. Finally, we found that signal molecule-degrading bacteria isolated from aquaculture settings have a positive effect on survival of challenged aquaculture animals, e.g. giant river prawn larvae challenged to Vibrio harveyi.

In conclusion, the data we obtained thus far indicate that quorum sensing disruption is a valid alternative biocontrol strategy for aquaculture, that biocontrol agents with quorum sensing- disrupting activity can be obtained from the aquatic environment and that these agents have a beneficial effect on cultured organisms.

ISQSI, Santiago de Compostela 2015  Invited lectures 

Quorum quenching: actors, biological roles and applications Yves Dessaux, Catherine Grandclément, Denis Faure

Department of Microbiology, Institute for integrative biology of the cell, CEA-CNRS- Université Paris-Sud, avenue de la terrasse, 91198 Gif sur Yvette, France.

E-mail: [email protected]

Keywords: quorum sensing inhibitor, quorum quenching enzyme, screening, metagenomics, biological control, soft-rot bacteria

Abstract

Numerous bacterial populations are able to monitor their own population density and regulate the expression of some genes accordingly. This regulatory process is termed quorum sensing (QS). It relies upon the production of QS signal(s) that each individual cell of a given bacterial population produces. The overall concentration of the signal(s) therefore mimics the cell density of this population. Once a threshold concentration of the signal is perceived by a receptor, the QS-regulated genes are expressed or repressed.

Quorum quenching (QQ) refers to all processes involved in the disturbance of the quorum sensing. QQ molecular actors are diverse in nature (enzymes, chemical compounds), mode of action (QS-signal cleavage, site competition…), and targets as all the main steps of the QS pathway that are synthesis, accumulation and perception of the QS signals may be affected.

Usually enzymes inactivating QS signals are named QQ enzymes, while (bio)chemicals disrupting QS-pathways are called QS inhibitors (or QSIs). Rather than listing all known enzymes and QSI molecules, the presentation will focus on a limited number of cases chosen to exemplify the variety of actors and origins. In relation, the occurrence of quenching functions in uncultivable micro-organisms will be discussed.

The biological role of the QQ phenomenon remains an intriguing - and to a certain extent unanswered - question. QSIs could be described as elements of a chemical warfare that involves organisms competing for the same ecological niche. This description most often results, however, from circumstantial observations and arguments. QQ enzymes could contribute to the recycling or the clearing (riddance) of the QS signals. However, in a well- documented case that implicates a QS molecule of the N-acyl homoserine lactone class, experimental results revealed that a microbial QQ enzyme may be involved in a detoxification process rather than in a fine tuning of QS-regulated functions.

Whatever the biological roles of QQ are, QSI molecules and QQ enzymes have been used in numerous exploratory applications in medicine, agriculture, environmental sciences and technology. Several examples of such applications have been or will be presented by others in the meeting. Because food production and preservation will become a critical parameter in the next decades, this presentation will mostly focus on applied outcomes in agriculture, and especially on the development of procedures that aim at countering plant-macerating pathogens of the Pectobacterium genus.

ISQSI, Santiago de Compostela 2015  Invited lectures 

Quorum sensing inhibition to control bacterial disease:

aquaculture as an example

Tom Defoirdt

Laboratory of Aquaculture & Artemia Reference Center, Ghent University. Rozier 44, 9000 Gent, Belgium. E-mail: [email protected]

Keywords: Gnotobiotic models, quorum sensing inhibitors, AHL, AI-2, CAI-1, indole

Abstract

Due to large-scale use of antibiotics, many pathogenic bacteria have acquired (multiple) resistance, and therefore, alternative treatments to control disease are needed. More than ten years ago, we proposed inhibition of quorum sensing, bacterial cell-to-cell communication, as a novel strategy to control infections caused by antibiotic-resistant bacteria in aquaculture.

In this presentation, I will discuss our current knowledge on the impact of quorum sensing and quorum sensing disruption on the virulence of aquaculture pathogens towards different host organisms. We found that the three-channel quorum sensing system of Vibrio harveyi regulates the expression of different virulence factors and the virulence of the bacterium towards gnotibiotic brine shrimp (Artemia franciscana) and giant river prawn (Macrobrachium rosenbergii) larvae. Remarkably, the impact of the three signal molecules on virulence of Vibrio harveyi was different for the two hosts, with acylhomoserine lactones (AHLs) playing no role in virulence towards brine shrimp and cholerae autoinducer-1 (CAI-1) playing no role in virulence towards river prawn. In contrast to what we observed in Vibrio harveyi, the three channel quorum sensing system of Vibrio anguillarum was found to play no role in virulence towards gnotobiotic sea bass (Dicentrarchus labrax) larvae. However, we found that another signalling molecule, indole, played a major role in virulence of this pathogen. Finally, we found that AHL quorum sensing controls the virulence of Aeromonas species towards larvae of the freshwater fish species burbot (Lota lota).

The most important quorum sensing-disrupting agents reported thus far include compounds that interfere with quorum sensing signal detection and signal transduction, and signal molecule- degrading bacteria. Quorum sensing-disrupting compounds studied in our laboratory include antagonistic AHL molecules, brominated furanones, brominated thiophenones and cinnamaldehyde. All of these compounds were found to increase survival of aquaculture animals when challenged to pathogenic bacteria. Furthermore, we found that metabolites produced by some micro-algae that are frequently used in aquaculture are also able to disrupt quorum sensing in Gram-negative bacteria. Finally, we found that signal molecule-degrading bacteria isolated from aquaculture settings have a positive effect on survival of challenged aquaculture animals, e.g. giant river prawn larvae challenged to Vibrio harveyi.

In conclusion, the data we obtained thus far indicate that quorum sensing disruption is a valid alternative biocontrol strategy for aquaculture, that biocontrol agents with quorum sensing- disrupting activity can be obtained from the aquatic environment and that these agents have a beneficial effect on cultured organisms.

ISQSI, Santiago de Compostela 2015  Invited lectures 

Quorum quenching: actors, biological roles and applications Yves Dessaux, Catherine Grandclément, Denis Faure

Department of Microbiology, Institute for integrative biology of the cell, CEA-CNRS- Université Paris-Sud, avenue de la terrasse, 91198 Gif sur Yvette, France.

E-mail: [email protected]

Keywords: quorum sensing inhibitor, quorum quenching enzyme, screening, metagenomics, biological control, soft-rot bacteria

Abstract

Numerous bacterial populations are able to monitor their own population density and regulate the expression of some genes accordingly. This regulatory process is termed quorum sensing (QS). It relies upon the production of QS signal(s) that each individual cell of a given bacterial population produces. The overall concentration of the signal(s) therefore mimics the cell density of this population. Once a threshold concentration of the signal is perceived by a receptor, the QS-regulated genes are expressed or repressed.

Quorum quenching (QQ) refers to all processes involved in the disturbance of the quorum sensing. QQ molecular actors are diverse in nature (enzymes, chemical compounds), mode of action (QS-signal cleavage, site competition…), and targets as all the main steps of the QS pathway that are synthesis, accumulation and perception of the QS signals may be affected.

Usually enzymes inactivating QS signals are named QQ enzymes, while (bio)chemicals disrupting QS-pathways are called QS inhibitors (or QSIs). Rather than listing all known enzymes and QSI molecules, the presentation will focus on a limited number of cases chosen to exemplify the variety of actors and origins. In relation, the occurrence of quenching functions in uncultivable micro-organisms will be discussed.

The biological role of the QQ phenomenon remains an intriguing - and to a certain extent unanswered - question. QSIs could be described as elements of a chemical warfare that involves organisms competing for the same ecological niche. This description most often results, however, from circumstantial observations and arguments. QQ enzymes could contribute to the recycling or the clearing (riddance) of the QS signals. However, in a well- documented case that implicates a QS molecule of the N-acyl homoserine lactone class, experimental results revealed that a microbial QQ enzyme may be involved in a detoxification process rather than in a fine tuning of QS-regulated functions.

Whatever the biological roles of QQ are, QSI molecules and QQ enzymes have been used in numerous exploratory applications in medicine, agriculture, environmental sciences and technology. Several examples of such applications have been or will be presented by others in the meeting. Because food production and preservation will become a critical parameter in the next decades, this presentation will mostly focus on applied outcomes in agriculture, and especially on the development of procedures that aim at countering plant-macerating pathogens of the Pectobacterium genus.