Synergistic Antimicrobial Interaction between Honey and Phage against Escherichia coli Biofilms

Chronic wounds afford a hostile environment of damaged tissues that allow bacterial proliferation and further wound colonization. Escherichia coli is among the most common colonizers of infected wounds and it is a prolific biofilm former. Living in biofilm communities, cells are protected, become mo...

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Main Authors: Ana Oliveira, Henrique G. Ribeiro, Ana C. Silva, Maria D. Silva, Jessica C. Sousa, Célia F. Rodrigues, Luís D. R. Melo, Ana F. Henriques, Sanna Sillankorva
Format: Article
Language:English
Published: Frontiers Media S.A. 2017-12-01
Series:Frontiers in Microbiology
Subjects:
Online Access:http://journal.frontiersin.org/article/10.3389/fmicb.2017.02407/full
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author Ana Oliveira
Henrique G. Ribeiro
Ana C. Silva
Maria D. Silva
Jessica C. Sousa
Célia F. Rodrigues
Luís D. R. Melo
Ana F. Henriques
Sanna Sillankorva
author_facet Ana Oliveira
Henrique G. Ribeiro
Ana C. Silva
Maria D. Silva
Jessica C. Sousa
Célia F. Rodrigues
Luís D. R. Melo
Ana F. Henriques
Sanna Sillankorva
author_sort Ana Oliveira
collection DOAJ
description Chronic wounds afford a hostile environment of damaged tissues that allow bacterial proliferation and further wound colonization. Escherichia coli is among the most common colonizers of infected wounds and it is a prolific biofilm former. Living in biofilm communities, cells are protected, become more difficult to control and eradicate, and less susceptible to antibiotic therapy. This work presents insights into the proceedings triggering E. coli biofilm control with phage, honey, and their combination, achieved through standard antimicrobial activity assays, zeta potential and flow cytometry studies and further visual insights sought by scanning electron microscopy and transmission electron microscopy. Two Portuguese honeys (PF2 and U3) with different floral origin and an E. coli-specific phage (EC3a), possessing depolymerase activity, were tested against 24- and 48-h-old biofilms. Synergic and additive effects were perceived in some phage–honey experiments. Combined therapy prompted similar phenomena in biofilm cells, visualized by electron microscopy, as the individual treatments. Honey caused minor membrane perturbations to complete collapse and consequent discharge of cytoplasmic content, and phage completely destroyed cells leaving only vesicle-like structures and debris. Our experiments show that the addition of phage to low honey concentrations is advantageous, and that even fourfold diluted honey combined with phage, presents no loss of antibacterial activity toward E. coli. Portuguese honeys possess excellent antibiofilm activity and may be potential alternative therapeutic agents in biofilm-related wound infection. Furthermore, to our knowledge this is the first study that assessed the impacts of phage–honey combinations in bacterial cells. The synergistic effect obtained was shown to be promising, since the antiviral effect of honey limits the emergence of phage resistant phenotypes.
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spelling doaj.art-a12077a0f8f64954ac9886f70db341962022-12-21T18:50:33ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2017-12-01810.3389/fmicb.2017.02407274474Synergistic Antimicrobial Interaction between Honey and Phage against Escherichia coli BiofilmsAna OliveiraHenrique G. RibeiroAna C. SilvaMaria D. SilvaJessica C. SousaCélia F. RodriguesLuís D. R. MeloAna F. HenriquesSanna SillankorvaChronic wounds afford a hostile environment of damaged tissues that allow bacterial proliferation and further wound colonization. Escherichia coli is among the most common colonizers of infected wounds and it is a prolific biofilm former. Living in biofilm communities, cells are protected, become more difficult to control and eradicate, and less susceptible to antibiotic therapy. This work presents insights into the proceedings triggering E. coli biofilm control with phage, honey, and their combination, achieved through standard antimicrobial activity assays, zeta potential and flow cytometry studies and further visual insights sought by scanning electron microscopy and transmission electron microscopy. Two Portuguese honeys (PF2 and U3) with different floral origin and an E. coli-specific phage (EC3a), possessing depolymerase activity, were tested against 24- and 48-h-old biofilms. Synergic and additive effects were perceived in some phage–honey experiments. Combined therapy prompted similar phenomena in biofilm cells, visualized by electron microscopy, as the individual treatments. Honey caused minor membrane perturbations to complete collapse and consequent discharge of cytoplasmic content, and phage completely destroyed cells leaving only vesicle-like structures and debris. Our experiments show that the addition of phage to low honey concentrations is advantageous, and that even fourfold diluted honey combined with phage, presents no loss of antibacterial activity toward E. coli. Portuguese honeys possess excellent antibiofilm activity and may be potential alternative therapeutic agents in biofilm-related wound infection. Furthermore, to our knowledge this is the first study that assessed the impacts of phage–honey combinations in bacterial cells. The synergistic effect obtained was shown to be promising, since the antiviral effect of honey limits the emergence of phage resistant phenotypes.http://journal.frontiersin.org/article/10.3389/fmicb.2017.02407/fullE. colihoneybacteriophagebiofilmssynergy
spellingShingle Ana Oliveira
Henrique G. Ribeiro
Ana C. Silva
Maria D. Silva
Jessica C. Sousa
Célia F. Rodrigues
Luís D. R. Melo
Ana F. Henriques
Sanna Sillankorva
Synergistic Antimicrobial Interaction between Honey and Phage against Escherichia coli Biofilms
Frontiers in Microbiology
E. coli
honey
bacteriophage
biofilms
synergy
title Synergistic Antimicrobial Interaction between Honey and Phage against Escherichia coli Biofilms
title_full Synergistic Antimicrobial Interaction between Honey and Phage against Escherichia coli Biofilms
title_fullStr Synergistic Antimicrobial Interaction between Honey and Phage against Escherichia coli Biofilms
title_full_unstemmed Synergistic Antimicrobial Interaction between Honey and Phage against Escherichia coli Biofilms
title_short Synergistic Antimicrobial Interaction between Honey and Phage against Escherichia coli Biofilms
title_sort synergistic antimicrobial interaction between honey and phage against escherichia coli biofilms
topic E. coli
honey
bacteriophage
biofilms
synergy
url http://journal.frontiersin.org/article/10.3389/fmicb.2017.02407/full
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