Special Issue "New Insights on Biofilm Antimicrobial Strategies"

A special issue of Antibiotics (ISSN 2079-6382).

Deadline for manuscript submissions: closed (30 September 2020).

Special Issue Editors

Dr. Luís Melo
E-Mail Website
Guest Editor
Laboratory of Research in Biofilms Rosário Oliveira, Centre of Biological Engineering, University of Minho, Braga, Portugal
Interests: bacteriophage; phage-host interactions; biofilms; endolysins; phage therapy; phage genomics and evolution
Special Issues and Collections in MDPI journals
Dr. Nuno F. Azevedo
E-Mail Website
Guest Editor
LEPABE, Department of Chemical Engineering, Faculty of Engineering of the University of Porto, Rua Dr Roberto Frias, 4200-465 Porto, Portugal
Interests: multispecies biofilms; nucleic acid mimics for therapy and diagnostics; development of hybridization-based techniques

Special Issue Information

Dear Colleagues,

The study of microbial biofilms has been gaining interest in recent decades. Generally, these communities consist of cells adhering to surfaces and surrounded by a self-produced exopolymeric matrix that confers protection against external stress. Biofilms can pose a threat to almost every sector of activity, namely, agriculture, food industries, and veterinary and human health. Due to their spatial localization and metabolic state, biofilm cells are very tolerant to antibiotics or disinfectants. Recently, several novel promising therapies that can be alternative, complementary or resensitize to antimicrobials are being developed worldwide.

This Special Issue of Antibiotics deals with different strategies to prevent biofilm formation or control development. The issue welcomes various submission types, such as original research papers, short communications, reviews, case reports, and perspectives.

Potential topics for this Special Issue include different antifouling strategies but are not limited to:

  • Phages or phage-derived enzymes;
  • Use of physical approaches (photoporation; sonoporation);
  • Development of antimicrobial peptides or nucleic acid mimics;
  • Use of natural products;
  • Modification of surfaces to prevent biofilm formation;
  • Combinations of antimicrobial agents with a synergystic effect.

Dr. Luís Melo
Dr. Nuno F. Azevedo
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All papers will be peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Antibiotics is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 1800 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • biofilm prevention
  • biofilm control
  • surface modifications
  • new generation antimicrobial compounds
  • synergistic approaches

Published Papers (16 papers)

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Editorial

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Editorial
New Insights on Biofilm Antimicrobial Strategies
Antibiotics 2021, 10(4), 407; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics10040407 - 09 Apr 2021
Viewed by 580
Abstract
Over the last few decades, the study of microbial biofilms has been gaining interest among the scientific community [...] Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)

Research

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Article
Bacteriophage Cocktail-Mediated Inhibition of Pseudomonas aeruginosa Biofilm on Endotracheal Tube Surface
Antibiotics 2021, 10(1), 78; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics10010078 - 15 Jan 2021
Cited by 1 | Viewed by 820
Abstract
Although different strategies to control biofilm formation on endotracheal tubes have been proposed, there are scarce scientific data on applying phages for both removing and preventing Pseudomonas aeruginosa biofilms on the device surface. Here, the anti-biofilm capacity of five bacteriophages was evaluated by [...] Read more.
Although different strategies to control biofilm formation on endotracheal tubes have been proposed, there are scarce scientific data on applying phages for both removing and preventing Pseudomonas aeruginosa biofilms on the device surface. Here, the anti-biofilm capacity of five bacteriophages was evaluated by a high content screening assay. We observed that biofilms were significantly reduced after phage treatment, especially in multidrug-resistant strains. Considering the anti-biofilm screens, two phages were selected as cocktail components, and the cocktail’s ability to prevent colonization of the endotracheal tube surface was tested in a dynamic biofilm model. Phage-coated tubes were challenged with different P. aeruginosa strains. The biofilm growth was monitored from 24 to 168 h by colony forming unit counting, metabolic activity assessment, and biofilm morphology observation. The phage cocktail promoted differences of bacterial colonization; nonetheless, the action was strain dependent. Phage cocktail coating did not promote substantial changes in metabolic activity. Scanning electron microscopy revealed a higher concentration of biofilm cells in control, while tower-like structures could be observed on phage cocktail-coated tubes. These results demonstrate that with the development of new coating strategies, phage therapy has potential in controlling the endotracheal tube-associated biofilm. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Carbapenem-Resistant Klebsiella pneumoniae Clinical Isolates: In Vivo Virulence Assessment in Galleria mellonella and Potential Therapeutics by Polycationic Oligoethyleneimine
Antibiotics 2021, 10(1), 56; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics10010056 - 08 Jan 2021
Cited by 3 | Viewed by 870
Abstract
Klebsiella pneumoniae, one of the most common pathogens found in hospital-acquired infections, is often resistant to multiple antibiotics. In fact, multidrug-resistant (MDR) K. pneumoniae producing KPC or OXA-48-like carbapenemases are recognized as a serious global health threat. In this sense, we evaluated [...] Read more.
Klebsiella pneumoniae, one of the most common pathogens found in hospital-acquired infections, is often resistant to multiple antibiotics. In fact, multidrug-resistant (MDR) K. pneumoniae producing KPC or OXA-48-like carbapenemases are recognized as a serious global health threat. In this sense, we evaluated the virulence of K. pneumoniae KPC(+) or OXA-48(+) aiming at potential antimicrobial therapeutics. K. pneumoniae carbapenemase (KPC) and the expanded-spectrum oxacillinase OXA-48 isolates were obtained from patients treated in medical care units in Lisbon, Portugal. The virulence potential of the K. pneumonia clinical isolates was tested using the Galleria mellonella model. For that, G. mellonella larvae were inoculated using patients KPC(+) and OXA-48(+) isolates. Using this in vivo model, the KPC(+) K. pneumoniae isolates showed to be, on average, more virulent than OXA-48(+). Virulence was found attenuated when a low bacterial inoculum (one magnitude lower) was tested. In addition, we also report the use of a synthetic polycationic oligomer (L-OEI-h) as a potential antimicrobial agent to fight infectious diseases caused by MDR bacteria. L-OEI-h has a broad-spectrum antibacterial activity and exerts a significantly bactericidal activity within the first 5-30 min treatment, causing lysis of the cytoplasmic membrane. Importantly, the polycationic oligomer showed low toxicity against in vitro models and no visible cytotoxicity (measured by survival and health index) was noted on the in vivo model (G. mellonella), thus L-OEI-h is foreseen as a promising polymer therapeutic for the treatment of MDR K. pneumoniae infections. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Increased Intraspecies Diversity in Escherichia coli Biofilms Promotes Cellular Growth at the Expense of Matrix Production
Antibiotics 2020, 9(11), 818; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9110818 - 17 Nov 2020
Cited by 2 | Viewed by 553
Abstract
Intraspecies diversity in biofilm communities is associated with enhanced survival and growth of the individual biofilm populations. Studies on the subject are scarce, namely, when more than three strains are present. Hence, in this study, the influence of intraspecies diversity in biofilm populations [...] Read more.
Intraspecies diversity in biofilm communities is associated with enhanced survival and growth of the individual biofilm populations. Studies on the subject are scarce, namely, when more than three strains are present. Hence, in this study, the influence of intraspecies diversity in biofilm populations composed of up to six different Escherichia coli strains isolated from urine was evaluated in conditions mimicking the ones observed in urinary tract infections and catheter-associated urinary tract infections. In general, with the increasing number of strains in a biofilm, an increase in cell cultivability and a decrease in matrix production were observed. For instance, single-strain biofilms produced an average of 73.1 µg·cm−2 of extracellular polymeric substances (EPS), while six strains biofilms produced 19.9 µg·cm−2. Hence, it appears that increased genotypic diversity in a biofilm leads E. coli to direct energy towards the production of its offspring, in detriment of the production of public goods (i.e., matrix components). Apart from ecological implications, these results can be explored as another strategy to reduce the biofilm burden, as a decrease in EPS matrix production may render these intraspecies biofilms more sensitive to antimicrobial agents. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Biofilm-Induced Antibiotic Resistance in Clinical Acinetobacter baumannii Isolates
Antibiotics 2020, 9(11), 817; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9110817 - 17 Nov 2020
Cited by 4 | Viewed by 910
Abstract
In order to understand the role of biofilm in the emergence of antibiotic resistance, a total of 104 clinical Acinetobacter baumannii strains were investigated for their biofilm-forming capacities and genes associated with biofilm formation. Selected biofilm-formers were tested for antibiotic susceptibilities when grown [...] Read more.
In order to understand the role of biofilm in the emergence of antibiotic resistance, a total of 104 clinical Acinetobacter baumannii strains were investigated for their biofilm-forming capacities and genes associated with biofilm formation. Selected biofilm-formers were tested for antibiotic susceptibilities when grown in biofilm phase. Reversibility of antibiotic susceptibility in planktonic cells regrown from biofilm were investigated. We found 59.6% of the strains were biofilm-formers, among which, 66.1% were non-multidrug resistant (MDR) strains. Presence of virulence genes bap, csuE, and abaI was significantly associated with biofilm-forming capacities. When strains were grown in biofilm state, the minimum biofilm eradication concentrations were 44, 407, and 364 times higher than the minimum bactericidal concentrations (MBC) for colistin, ciprofloxacin, and imipenem, respectively. Persisters were detected after treating the biofilm at 32–256 times the MBC of planktonic cells. Reversibility test for antibiotic susceptibility showed that biofilm formation induced reversible antibiotic tolerance in the non-MDR strains but a higher level of irreversible resistance in the extensively drug-resistant (XDR) strain. In summary, we showed that the non-MDR strains were strong biofilm-formers. Presence of persisters in biofilm contributed to the reduced antibiotic susceptibilities. Biofilm-grown Acinetobacter baumannii has induced antibiotic tolerance in non-MDR strains and increased resistance levels in XDR strains. To address the regulatory mechanisms of biofilm-specific resistance, thorough investigations at genome and transcription levels are warranted. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Pitfalls Associated with Discriminating Mixed-Species Biofilms by Flow Cytometry
Antibiotics 2020, 9(11), 741; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9110741 - 27 Oct 2020
Cited by 1 | Viewed by 685
Abstract
Since biofilms are ubiquitous in different settings and act as sources of disease for humans, reliable methods to characterize and quantify these microbial communities are required. Numerous techniques have been employed, but most of them are unidirectional, labor intensive and time consuming. Although [...] Read more.
Since biofilms are ubiquitous in different settings and act as sources of disease for humans, reliable methods to characterize and quantify these microbial communities are required. Numerous techniques have been employed, but most of them are unidirectional, labor intensive and time consuming. Although flow cytometry (FCM) can be a reliable choice to quickly provide a multiparametric analysis, there are still few applications on biofilms, and even less on the study of inter-kingdom communities. This work aimed to give insights into the application of FCM in order to more comprehensively analyze mixed-species biofilms, formed by different Pseudomonas aeruginosa and Candida albicans strains, before and after exposure to antimicrobials. For comparison purposes, biofilm culturability was also assessed determining colony-forming units. The results showed that some aspects, namely the microbial strain used, the morphological state of the cells and the biofilm matrix, make the accurate analysis of FCM data difficult. These aspects were even more challenging when double-species biofilms were being inspected, as they could engender data misinterpretations. The outcomes draw our attention towards the need to always take into consideration the characteristics of the biofilm samples to be analyzed through FCM, and undoubtedly link to the need for optimization of the processes tailored for each particular case study. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Synthesis of Electrospun TiO2 Nanofibers and Characterization of Their Antibacterial and Antibiofilm Potential against Gram-Positive and Gram-Negative Bacteria
Antibiotics 2020, 9(9), 572; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9090572 - 03 Sep 2020
Cited by 16 | Viewed by 2506
Abstract
Recently, titanium dioxide (TiO2) nanomaterials have gained increased attention because of their cost-effective, safe, stable, non-toxic, non-carcinogenic, photocatalytic, bactericidal, biomedical, industrial and waste-water treatment applications. The aim of the present work is the synthesis of electrospun TiO2 nanofibers (NFs) in [...] Read more.
Recently, titanium dioxide (TiO2) nanomaterials have gained increased attention because of their cost-effective, safe, stable, non-toxic, non-carcinogenic, photocatalytic, bactericidal, biomedical, industrial and waste-water treatment applications. The aim of the present work is the synthesis of electrospun TiO2 nanofibers (NFs) in the presence of different amounts of air–argon mixtures using sol-gel and electrospinning approaches. The physicochemical properties of the synthesized NFs were examined by scanning and transmission electron microscopies (SEM and TEM) coupled with energy-dispersive X-ray spectroscopy (EDX), ultraviolet-visible spectroscopy and thermogravimetric analyzer (TGA). The antibacterial and antibiofilm activity of synthesized NFs against Gram-negative Pseudomonas aeruginosa and Gram-positive methicillin-resistant Staphylococcusaureus (MRSA) was investigated by determining their minimum bacteriostatic and bactericidal values. The topological and morphological alteration caused by TiO2 NFs in bacterial cells was further analyzed by SEM. TiO2 NFs that were calcined in a 25% air-75% argon mixture showed maximum antibacterial and antibiofilm activities. The minimum inhibitory concentration (MIC)/minimum bactericidal concentration (MBC) value of TiO2 NFs against P. aeruginosa was 3 and 6 mg/mL and that for MRSA was 6 and 12 mg/mL, respectively. The MIC/MBC and SEM results show that TiO2 NFs were more active against Gram-negative P. aeruginosa cells than Gram-positive S. aureus. The inhibition of biofilm formation by TiO2 NFs was investigated quantitatively by tissue culture plate method using crystal violet assay and it was found that TiO2 NFs inhibited biofilm formation by MRSA and P. aeruginosa in a dose-dependent manner. TiO2 NFs calcined in a 25% air-75% argon mixture exhibited maximum biofilm formation inhibition of 75.2% for MRSA and 72.3% for P. aeruginosa at 2 mg/mL, respectively. The antibacterial and antibiofilm results suggest that TiO2 NFs can be used to coat various inanimate objects, in food packaging and in waste-water treatment and purification to prevent bacterial growth and biofilm formation. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Communication
Biofilm Control Strategies: Engaging with the Public
Antibiotics 2020, 9(8), 465; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9080465 - 30 Jul 2020
Cited by 1 | Viewed by 1193
Abstract
There are few peer-reviewed publications about public engagement with science that are written by microbiologists; those that exist tend to be a narrative of an event rather than a hypothesis-driven investigation. However, it is relatively easy for experienced scientists to use a scientific [...] Read more.
There are few peer-reviewed publications about public engagement with science that are written by microbiologists; those that exist tend to be a narrative of an event rather than a hypothesis-driven investigation. However, it is relatively easy for experienced scientists to use a scientific method in their approach to public engagement. This short communication describes three public engagement activities hosted by the authors, focused on biofilm control: hand hygiene, plaque control and an externally applied antimicrobial coating. In each case, audience engagement was assessed using quantitative and/or qualitative methods. A critical evaluation of the findings enabled the construction of a public engagement ‘tick list’ for future events that would enable a hypothesis-driven approach with more effective communication activities and more robust evaluation. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Carbon Nanotube/Poly(dimethylsiloxane) Composite Materials to Reduce Bacterial Adhesion
Antibiotics 2020, 9(8), 434; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9080434 - 22 Jul 2020
Cited by 6 | Viewed by 756
Abstract
Different studies have shown that the incorporation of carbon nanotubes (CNTs) into poly(dimethylsiloxane) (PDMS) enables the production of composite materials with enhanced properties, which can find important applications in the biomedical field. In the present work, CNT/PDMS composite materials have been prepared to [...] Read more.
Different studies have shown that the incorporation of carbon nanotubes (CNTs) into poly(dimethylsiloxane) (PDMS) enables the production of composite materials with enhanced properties, which can find important applications in the biomedical field. In the present work, CNT/PDMS composite materials have been prepared to evaluate the effects of pristine and chemically functionalized CNT incorporation into PDMS on the composite’s thermal, electrical, and surface properties on bacterial adhesion in dynamic conditions. Initial bacterial adhesion was studied using a parallel-plate flow chamber assay performed in conditions prevailing in urinary tract devices (catheters and stents) using Escherichia coli as a model organism and PDMS as a control due to its relevance in these applications. The results indicated that the introduction of the CNTs in the PDMS matrix yielded, in general, less bacterial adhesion than the PDMS alone and that the reduction could be dependent on the surface chemistry of CNTs, with less adhesion obtained on the composites with pristine rather than functionalized CNTs. It was also shown CNT pre-treatment and incorporation by different methods affected the electrical properties of the composites when compared to PDMS. Composites enabling a 60% reduction in cell adhesion were obtained by CNT treatment by ball-milling, whereas an increase in electrical conductivity of seven orders of magnitude was obtained after solvent-mediated incorporation. The results suggest even at low CNT loading values (1%), these treatments may be beneficial for the production of CNT composites with application in biomedical devices for the urinary tract and for other applications where electrical conductance is required. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Analysing the Initial Bacterial Adhesion to Evaluate the Performance of Antifouling Surfaces
Antibiotics 2020, 9(7), 421; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9070421 - 17 Jul 2020
Cited by 1 | Viewed by 761
Abstract
The aim of this work was to study the initial events of Escherichia coli adhesion to polydimethylsiloxane, which is critical for the development of antifouling surfaces. A parallel plate flow cell was used to perform the initial adhesion experiments under controlled hydrodynamic conditions [...] Read more.
The aim of this work was to study the initial events of Escherichia coli adhesion to polydimethylsiloxane, which is critical for the development of antifouling surfaces. A parallel plate flow cell was used to perform the initial adhesion experiments under controlled hydrodynamic conditions (shear rates ranging between 8 and 100/s), mimicking biomedical scenarios. Initial adhesion studies capture more accurately the cell-surface interactions as in later stages, incoming cells may interact with the surface but also with already adhered cells. Adhesion rates were calculated and results shown that after some time (between 5 and 9 min), these rates decreased (by 55% on average), from the initial values for all tested conditions. The common explanation for this decrease is the occurrence of hydrodynamic blocking, where the area behind each adhered cell is screened from incoming cells. This was investigated using a pair correlation map from which two-dimensional histograms showing the density probability function were constructed. The results highlighted a lower density probability (below 4.0 × 10−4) of the presence of cells around a given cell under different shear rates irrespectively of the radial direction. A shadowing area behind the already adhered cells was not observed, indicating that hydrodynamic blocking was not occurring and therefore it could not be the cause for the decreases in cell adhesion rates. Afterward, cell transport rates from the bulk solution to the surface were estimated using the Smoluchowski-Levich approximation and values in the range of 80–170 cells/cm2.s were obtained. The drag forces that adhered cells have to withstand were also estimated and values in the range of 3–50 × 10−14 N were determined. Although mass transport increases with the flow rate, drag forces also increase and the relative importance of these factors may change in different conditions. This work demonstrates that adjustment of operational parameters in initial adhesion experiments may be required to avoid hydrodynamic blocking, in order to obtain reliable data about cell-surface interactions that can be used in the development of more efficient antifouling surfaces. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Phytochemical Composition and In Vitro Biological Activity of Iris spp. (Iridaceae): A New Source of Bioactive Constituents for the Inhibition of Oral Bacterial Biofilms
Antibiotics 2020, 9(7), 403; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9070403 - 11 Jul 2020
Cited by 7 | Viewed by 1108
Abstract
The inhibition and eradication of oral biofilms is increasingly focused on the use of plant extracts as mouthwashes and toothpastes adjuvants. Here, we report on the chemical composition and the antibiofilm activity of 15 methanolic extracts of Iris species against both mono-(Pseudomonas [...] Read more.
The inhibition and eradication of oral biofilms is increasingly focused on the use of plant extracts as mouthwashes and toothpastes adjuvants. Here, we report on the chemical composition and the antibiofilm activity of 15 methanolic extracts of Iris species against both mono-(Pseudomonas aeruginosa, Staphylococcus aureus) and multi-species oral biofilms (Streptococcus gordonii, Veillonella parvula, Fusobacterium nucleatum subsp. nucleatum, and Actinomyces naeslundii). The phytochemical profiles of Iris pallida s.l., Iris versicolor L., Iris lactea Pall., Iris carthaliniae Fomin, and Iris germanica were determined by ultra-high performance liquid chromatography-high-resolution tandem mass spectroscopy (UHPLC-HRMS/MS) analysis, and a total of 180 compounds were identified among Iris species with (iso)flavonoid dominancy. I. pallida, I. versicolor, and I. germanica inhibited both the quorum sensing and adhesion during biofilm formation in a concentration-dependent manner. However, the extracts were less active against maturated biofilms. Of the five tested species, Iris pallida s.l. was the most effective at both inhibiting biofilm formation and disrupting existing biofilms, and the leaf extract exhibited the strongest inhibitory effect compared to the root and rhizome extracts. The cytotoxicity of the extracts was excluded in human fibroblasts. The inhibition of bacterial adhesion significantly correlated with myristic acid content, and quorum sensing inhibition correlated with the 7-β-hydroxystigmast-4-en-3-one content. These findings could be useful for establishing an effective tool for the control of oral biofilms and thus dental diseases. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Article
Nanovectorized Microalgal Extracts to Fight Candida albicans and Cutibacterium acnes Biofilms: Impact of Dual-Species Conditions
Antibiotics 2020, 9(6), 279; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9060279 - 26 May 2020
Cited by 4 | Viewed by 1032
Abstract
Biofilm-related infections are a matter of concern especially because of the poor susceptibility of microorganisms to conventional antimicrobial agents. Innovative approaches are needed. The antibiofilm activity of extracts of cyanobacteria Arthrospira platensis, rich in free fatty acids, as well as of extract-loaded [...] Read more.
Biofilm-related infections are a matter of concern especially because of the poor susceptibility of microorganisms to conventional antimicrobial agents. Innovative approaches are needed. The antibiofilm activity of extracts of cyanobacteria Arthrospira platensis, rich in free fatty acids, as well as of extract-loaded copper alginate-based nanocarriers, were studied on single- and dual-species biofilms of Candida albicans and Cutibacterium acnes. Their ability to inhibit the biofilm formation and to eradicate 24 h old biofilms was investigated. Concentrations of each species were evaluated using flow cytometry. Extracts prevented the growth of C. acnes single-species biofilms (inhibition > 75% at 0.2 mg/mL) but failed to inhibit preformed biofilms. Nanovectorised extracts reduced the growth of single-species C. albicans biofilms (inhibition > 43% at 0.2 mg/mL) while free extracts were weakly or not active. Nanovectorised extracts also inhibited preformed C. albicans biofilms by 55% to 77%, whereas the corresponding free extracts were not active. In conclusion, even if the studied nanocarrier systems displayed promising activity, especially against C. albicans, their efficacy against dual-species biofilms was limited. This study highlighted that working in such polymicrobial conditions can give a more objective view of the relevance of antibiofilm strategies by taking into account interspecies interactions that can offer additional protection to microbes. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Review

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Review
Biofilms as Promoters of Bacterial Antibiotic Resistance and Tolerance
Antibiotics 2021, 10(1), 3; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics10010003 - 23 Dec 2020
Cited by 8 | Viewed by 2089
Abstract
Multidrug resistant bacteria are a global threat for human and animal health. However, they are only part of the problem of antibiotic failure. Another bacterial strategy that contributes to their capacity to withstand antimicrobials is the formation of biofilms. Biofilms are associations of [...] Read more.
Multidrug resistant bacteria are a global threat for human and animal health. However, they are only part of the problem of antibiotic failure. Another bacterial strategy that contributes to their capacity to withstand antimicrobials is the formation of biofilms. Biofilms are associations of microorganisms embedded a self-produced extracellular matrix. They create particular environments that confer bacterial tolerance and resistance to antibiotics by different mechanisms that depend upon factors such as biofilm composition, architecture, the stage of biofilm development, and growth conditions. The biofilm structure hinders the penetration of antibiotics and may prevent the accumulation of bactericidal concentrations throughout the entire biofilm. In addition, gradients of dispersion of nutrients and oxygen within the biofilm generate different metabolic states of individual cells and favor the development of antibiotic tolerance and bacterial persistence. Furthermore, antimicrobial resistance may develop within biofilms through a variety of mechanisms. The expression of efflux pumps may be induced in various parts of the biofilm and the mutation frequency is induced, while the presence of extracellular DNA and the close contact between cells favor horizontal gene transfer. A deep understanding of the mechanisms by which biofilms cause tolerance/resistance to antibiotics helps to develop novel strategies to fight these infections. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Review
Strategies to Reduce Biofilm Formation in PEEK Materials Applied to Implant Dentistry—A Comprehensive Review
Antibiotics 2020, 9(9), 609; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9090609 - 16 Sep 2020
Cited by 3 | Viewed by 1172
Abstract
Polyether-ether-ketone (PEEK) has emerged in Implant Dentistry with a series of short-time applications and as a promising material to substitute definitive dental implants. Several strategies have been investigated to diminish biofilm formation on the PEEK surface aiming to decrease the possibility of related [...] Read more.
Polyether-ether-ketone (PEEK) has emerged in Implant Dentistry with a series of short-time applications and as a promising material to substitute definitive dental implants. Several strategies have been investigated to diminish biofilm formation on the PEEK surface aiming to decrease the possibility of related infections. Therefore, a comprehensive review was carried out in order to compare PEEK with materials widely used nowadays in Implant Dentistry, such as titanium and zirconia, placing emphasis on studies investigating its ability to grant or prevent biofilm formation. Most studies failed to reveal significant antimicrobial activity in pure PEEK, while several studies described new strategies to reduce biofilm formation and bacterial colonization on this material. Those include the PEEK sulfonation process, incorporation of therapeutic and bioactive agents in PEEK matrix or on PEEK surface, PEEK coatings and incorporation of reinforcement agents, in order to produce nanocomposites or blends. The two most analyzed surface properties were contact angle and roughness, while the most studied bacteria were Escherichia coli and Staphylococcus aureus. Despite PEEK’s susceptibility to biofilm formation, a great number of strategies discussed in this study were able to improve its antibiofilm and antimicrobial properties. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Review
Insights into Host–Pathogen Interactions in Biofilm-Infected Wounds Reveal Possibilities for New Treatment Strategies
Antibiotics 2020, 9(7), 396; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9070396 - 10 Jul 2020
Cited by 3 | Viewed by 1222
Abstract
Normal wound healing occurs in three phases—the inflammatory, the proliferative, and the remodeling phase. Chronic wounds are, for unknown reasons, arrested in the inflammatory phase. Bacterial biofilms may cause chronicity by arresting healing in the inflammatory state by mechanisms not fully understood. Pseudomonas [...] Read more.
Normal wound healing occurs in three phases—the inflammatory, the proliferative, and the remodeling phase. Chronic wounds are, for unknown reasons, arrested in the inflammatory phase. Bacterial biofilms may cause chronicity by arresting healing in the inflammatory state by mechanisms not fully understood. Pseudomonas aeruginosa, a common wound pathogen with remarkable abilities in avoiding host defense and developing microbial resistance by biofilm formation, is detrimental to wound healing in clinical studies. The host response towards P. aeruginosa biofilm-infection in chronic wounds and impact on wound healing is discussed and compared to our own results in a chronic murine wound model. The impact of P. aeruginosa biofilms can be described by determining alterations in the inflammatory response, growth factor profile, and count of leukocytes in blood. P. aeruginosa biofilms are capable of reducing the host response to the infection, despite a continuously sustained inflammatory reaction and resulting local tissue damage. A recent observation of in vivo synergism between immunomodulatory and antimicrobial S100A8/A9 and ciprofloxacin suggests its possible future therapeutic potential. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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Other

Perspective
Bacteriophage Therapy for Clinical Biofilm Infections: Parameters That Influence Treatment Protocols and Current Treatment Approaches
Antibiotics 2020, 9(11), 799; https://0-doi-org.brum.beds.ac.uk/10.3390/antibiotics9110799 - 12 Nov 2020
Cited by 7 | Viewed by 673
Abstract
Biofilm infections are extremely difficult to treat, which is secondary to the inability of conventional antibiotics to eradicate biofilms. Consequently, current definitive treatment of biofilm infections requires complete removal of the infected hardware. This causes significant morbidity and mortality to patients and therefore [...] Read more.
Biofilm infections are extremely difficult to treat, which is secondary to the inability of conventional antibiotics to eradicate biofilms. Consequently, current definitive treatment of biofilm infections requires complete removal of the infected hardware. This causes significant morbidity and mortality to patients and therefore novel therapeutics are needed to cure these infections without removal of the infected hardware. Bacteriophages have intrinsic properties that could be advantageous in the treatment of clinical biofilm infections, but limited knowledge is known about the proper use of bacteriophage therapy in vivo. Currently titers and duration of bacteriophage therapy are the main parameters that are evaluated when devising bacteriophage protocols. Herein, several other important parameters are discussed which if standardized could allow for more effective and reproducible treatment protocols to be formulated. In addition, these parameters are correlated with the current clinical approaches being evaluated in the treatment of clinical biofilm infections. Full article
(This article belongs to the Special Issue New Insights on Biofilm Antimicrobial Strategies)
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