Session Type: ePosters
Session Title: ePosters
Authors(s): L. Ponce-Benavente (1, 2), J. Wagemans (3, 2), R. Lavigne (3), A. Trampuz (4, 2), M. Gonzalez-Moreno (1, 2)
Authors Affiliations(s): (1) Center for Musculoskeletal Surgery, Germany, (2) Charité-Universitätsmedizin Berlin, Germany, (3) Laboratory of Gene Technology, KU Leuven, Belgium, (4) Center for Septic Surgery, Germany
Background:
Increased antimicrobial resistance and the occurrence of biofilm-associated infections require alternative therapies, such as the use of lytic bacteriophages. The employment of phage therapy requires the isolation and characterization of bacteriophages highly active against specific strains in planktonic and biofilm growth state. In this study, we aimed to isolate and characterize bacteriophages active against a broad collection of Staphylococcus aureus clinical isolates.
Methods:One rifampicin-resistant (RRSA-5), one vancomycin-resistant (VRSA-1) and 23 methicillin-resistant S. aureus strains were included in this study. Saliva collected from volunteers and sewage from wastewater treatment plants were screened for the presence of lytic bacteriophages. Whole genome sequencing of the isolated bacteriophages was performed on a MiSeg instrument (Illumina). Host range was determined by spot assay on soft agar overlays. Isothermal microcalorimetry and sonication were used to evaluate the antibiofilm activity of a selected bacteriophage as monotherapy and in combination with staggered application of fosfomycin.
Results:Three lytic bacteriophages were successfully isolated from saliva (n=1) and sewage (n=2). The host range revealed activity against 72% of the isolates from the bacterial collection (Fig.1). Bacteriophage ΦM was selected for further testing against the RRSA-5 strain. A time-killing assay showed the highest decrease in bacteria counts after 6 hours of co-incubation of bacteriophage ΦM and a 24-hours-old-biofilm formed by RRSA-5 (Fig.2). The staggered exposure of RRSA-5 biofilm first to bacteriophage ΦM for 6h, followed by addition of increasing concentration of fosfomycin revealed a remarkable reduction of heat production compared to the growth-control or antimicrobial alone (Fig.3).
Conclusions:Saliva and untreated wastewater appear to be rich sources of bacteriophages active against a broad range of S. aureus isolates. Newly isolated bacteriophages were active against antibiotic-resistant strains, demonstrating to be promising adjuvant therapies for biofilm-associated infections. This study provided valuable data on the combined use of bacteriophage and antibiotics against RRSA, bringing new insights into the potential clinical application. Further investigations leading to improvement of bacteriophage properties against biofilms and their efficacy in experimental models and humans should follow.
Keyword(s): bacteriophage, RRSA, biofilmSession Type: ePosters
Session Title: ePosters
Authors(s): L. Ponce-Benavente (1, 2), J. Wagemans (3, 2), R. Lavigne (3), A. Trampuz (4, 2), M. Gonzalez-Moreno (1, 2)
Authors Affiliations(s): (1) Center for Musculoskeletal Surgery, Germany, (2) Charité-Universitätsmedizin Berlin, Germany, (3) Laboratory of Gene Technology, KU Leuven, Belgium, (4) Center for Septic Surgery, Germany
Background:
Increased antimicrobial resistance and the occurrence of biofilm-associated infections require alternative therapies, such as the use of lytic bacteriophages. The employment of phage therapy requires the isolation and characterization of bacteriophages highly active against specific strains in planktonic and biofilm growth state. In this study, we aimed to isolate and characterize bacteriophages active against a broad collection of Staphylococcus aureus clinical isolates.
Methods:One rifampicin-resistant (RRSA-5), one vancomycin-resistant (VRSA-1) and 23 methicillin-resistant S. aureus strains were included in this study. Saliva collected from volunteers and sewage from wastewater treatment plants were screened for the presence of lytic bacteriophages. Whole genome sequencing of the isolated bacteriophages was performed on a MiSeg instrument (Illumina). Host range was determined by spot assay on soft agar overlays. Isothermal microcalorimetry and sonication were used to evaluate the antibiofilm activity of a selected bacteriophage as monotherapy and in combination with staggered application of fosfomycin.
Results:Three lytic bacteriophages were successfully isolated from saliva (n=1) and sewage (n=2). The host range revealed activity against 72% of the isolates from the bacterial collection (Fig.1). Bacteriophage ΦM was selected for further testing against the RRSA-5 strain. A time-killing assay showed the highest decrease in bacteria counts after 6 hours of co-incubation of bacteriophage ΦM and a 24-hours-old-biofilm formed by RRSA-5 (Fig.2). The staggered exposure of RRSA-5 biofilm first to bacteriophage ΦM for 6h, followed by addition of increasing concentration of fosfomycin revealed a remarkable reduction of heat production compared to the growth-control or antimicrobial alone (Fig.3).
Conclusions:Saliva and untreated wastewater appear to be rich sources of bacteriophages active against a broad range of S. aureus isolates. Newly isolated bacteriophages were active against antibiotic-resistant strains, demonstrating to be promising adjuvant therapies for biofilm-associated infections. This study provided valuable data on the combined use of bacteriophage and antibiotics against RRSA, bringing new insights into the potential clinical application. Further investigations leading to improvement of bacteriophage properties against biofilms and their efficacy in experimental models and humans should follow.
Keyword(s): bacteriophage, RRSA, biofilm
Center for Musculoskeletal Surgery, Charité-Universitätsmedizin Berlin, Germany
Session Type: ePosters
Session Title: ePosters
Authors(s): L. Ponce-Benavente (1, 2), J. Wagemans (3, 2), R. Lavigne (3), A. Trampuz (4, 2), M. Gonzalez-Moreno (1, 2)
Authors Affiliations(s): (1) Center for Musculoskeletal Surgery, Germany, (2) Charité-Universitätsmedizin Berlin, Germany, (3) Laboratory of Gene Technology, KU Leuven, Belgium, (4) Center for Septic Surgery, Germany
Background:
Increased antimicrobial resistance and the occurrence of biofilm-associated infections require alternative therapies, such as the use of lytic bacteriophages. The employment of phage therapy requires the isolation and characterization of bacteriophages highly active against specific strains in planktonic and biofilm growth state. In this study, we aimed to isolate and characterize bacteriophages active against a broad collection of Staphylococcus aureus clinical isolates.
Methods:One rifampicin-resistant (RRSA-5), one vancomycin-resistant (VRSA-1) and 23 methicillin-resistant S. aureus strains were included in this study. Saliva collected from volunteers and sewage from wastewater treatment plants were screened for the presence of lytic bacteriophages. Whole genome sequencing of the isolated bacteriophages was performed on a MiSeg instrument (Illumina). Host range was determined by spot assay on soft agar overlays. Isothermal microcalorimetry and sonication were used to evaluate the antibiofilm activity of a selected bacteriophage as monotherapy and in combination with staggered application of fosfomycin.
Results:Three lytic bacteriophages were successfully isolated from saliva (n=1) and sewage (n=2). The host range revealed activity against 72% of the isolates from the bacterial collection (Fig.1). Bacteriophage ΦM was selected for further testing against the RRSA-5 strain. A time-killing assay showed the highest decrease in bacteria counts after 6 hours of co-incubation of bacteriophage ΦM and a 24-hours-old-biofilm formed by RRSA-5 (Fig.2). The staggered exposure of RRSA-5 biofilm first to bacteriophage ΦM for 6h, followed by addition of increasing concentration of fosfomycin revealed a remarkable reduction of heat production compared to the growth-control or antimicrobial alone (Fig.3).
Conclusions:Saliva and untreated wastewater appear to be rich sources of bacteriophages active against a broad range of S. aureus isolates. Newly isolated bacteriophages were active against antibiotic-resistant strains, demonstrating to be promising adjuvant therapies for biofilm-associated infections. This study provided valuable data on the combined use of bacteriophage and antibiotics against RRSA, bringing new insights into the potential clinical application. Further investigations leading to improvement of bacteriophage properties against biofilms and their efficacy in experimental models and humans should follow.
Keyword(s): bacteriophage, RRSA, biofilmSession Type: ePosters
Session Title: ePosters
Authors(s): L. Ponce-Benavente (1, 2), J. Wagemans (3, 2), R. Lavigne (3), A. Trampuz (4, 2), M. Gonzalez-Moreno (1, 2)
Authors Affiliations(s): (1) Center for Musculoskeletal Surgery, Germany, (2) Charité-Universitätsmedizin Berlin, Germany, (3) Laboratory of Gene Technology, KU Leuven, Belgium, (4) Center for Septic Surgery, Germany
Background:
Increased antimicrobial resistance and the occurrence of biofilm-associated infections require alternative therapies, such as the use of lytic bacteriophages. The employment of phage therapy requires the isolation and characterization of bacteriophages highly active against specific strains in planktonic and biofilm growth state. In this study, we aimed to isolate and characterize bacteriophages active against a broad collection of Staphylococcus aureus clinical isolates.
Methods:One rifampicin-resistant (RRSA-5), one vancomycin-resistant (VRSA-1) and 23 methicillin-resistant S. aureus strains were included in this study. Saliva collected from volunteers and sewage from wastewater treatment plants were screened for the presence of lytic bacteriophages. Whole genome sequencing of the isolated bacteriophages was performed on a MiSeg instrument (Illumina). Host range was determined by spot assay on soft agar overlays. Isothermal microcalorimetry and sonication were used to evaluate the antibiofilm activity of a selected bacteriophage as monotherapy and in combination with staggered application of fosfomycin.
Results:Three lytic bacteriophages were successfully isolated from saliva (n=1) and sewage (n=2). The host range revealed activity against 72% of the isolates from the bacterial collection (Fig.1). Bacteriophage ΦM was selected for further testing against the RRSA-5 strain. A time-killing assay showed the highest decrease in bacteria counts after 6 hours of co-incubation of bacteriophage ΦM and a 24-hours-old-biofilm formed by RRSA-5 (Fig.2). The staggered exposure of RRSA-5 biofilm first to bacteriophage ΦM for 6h, followed by addition of increasing concentration of fosfomycin revealed a remarkable reduction of heat production compared to the growth-control or antimicrobial alone (Fig.3).
Conclusions:Saliva and untreated wastewater appear to be rich sources of bacteriophages active against a broad range of S. aureus isolates. Newly isolated bacteriophages were active against antibiotic-resistant strains, demonstrating to be promising adjuvant therapies for biofilm-associated infections. This study provided valuable data on the combined use of bacteriophage and antibiotics against RRSA, bringing new insights into the potential clinical application. Further investigations leading to improvement of bacteriophage properties against biofilms and their efficacy in experimental models and humans should follow.
Keyword(s): bacteriophage, RRSA, biofilm