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Microorganisms
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Bacterial Pathogens: Biofilm Formation and Eradication
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Bacterial Pathogens: Biofilm Formation and Eradication

A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Biofilm".

Deadline for manuscript submissions: closed (28 February 2026) | Viewed by 2437

Special Issue Editor

Dr. Maria João Vieira

E-Mail Website
Guest Editor
1. Center of Biological Engineering (CEB), Campus de Gualtar, University of Minho, 4710-057 Braga, Portugal
2. LABBELS–Associate Laboratory, Braga/Guimarães, 4710-057 Braga, Portugal
Interests: bacterial biofilm; bacteriocin; bactearial detection

Special Issue Information

Dear Colleagues,

This special issue aims to gain deeper knowledge on the impact of biofilm formation and detachment on bacterial pathogenicity

Biofilms structures of microorganisms and the polymeric matrix they produce, attached to biotic or abiotic surfaces, are known to be a safe place for bacterial pathogens. Bacteria in sessile form can adhere to the surface and grow protected in the biofilm or bacteria can develop new phenotypes in the biofilm.

Chemical and antibiotic treatments are not effective, most of the times, to remove or to kill the microbial  threat. Moreover, they can promote the survival and the development   of new phenotypes such as pathogenicity, resistance to antibiotics, persister and viable but not c formation of viarable cells, production of metabolites that allow cells to resist to very harsh conditions. More, it is known, that biofilms release continually microorganisms to the surrounding media in addiction to esporadic  detachment of large portions of the pellicle, being a source of contamination.

Biofilms, as such, were named in the early 80s by William Costerton and are still a challenge in clinical, industrial and natural settings. In the last 50 years, a large amount of work was carried out in biofilms and the associated bacterial pathogenicity. New insights are being  achieved new methodologies including advanced microscopy  and ohmic approaches.

As such, state of the art original research articles and systematic reviews foccused on biofilm formation and  detachment on bacterial pathogens are welcome.

Dr. Maria João Vieira
Guest Editor

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 submissions that pass pre-check are 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 250 words) can be sent to the Editorial Office for assessment.

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. Microorganisms 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 2700 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

  • bacterial pathogens
  • biofilm
  • biofilm formation
  • biofilm removal
  • pathogenicity in biofilms
  • methods to detect pathogens in biofilms

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Published Papers (4 papers)

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Research

23 pages, 2120 KB  
Article
Epidemiological Study of the Relationship Between Antimicrobial Resistance Genes and Biofilm-Forming Capacity in Pathogens Causing Chronic Wound Infections
by Silvia Ioana Musuroi, Adela Voinescu, Corina Musuroi, Delia Muntean, Florin George Horhat, Luminita Mirela Baditoiu, Oana Izmendi, Andrei Cosnita, Valentin Ordodi, Zorin Crainiceanu, Edward Seclaman and Monica Licker
Microorganisms 2026, 14(5), 1117; https://doi.org/10.3390/microorganisms14051117 - 14 May 2026
Viewed by 208
Abstract
Chronic wounds represent a major complication of underlying conditions such as diabetes mellitus, arterial ischemia, surgical wound and burns. This study aimed at the phenotypic and molecular characterization of antimicrobial resistance for a selection of bacterial isolates, originating from wounds harvested from patients [...] Read more.
Chronic wounds represent a major complication of underlying conditions such as diabetes mellitus, arterial ischemia, surgical wound and burns. This study aimed at the phenotypic and molecular characterization of antimicrobial resistance for a selection of bacterial isolates, originating from wounds harvested from patients hospitalized in the Vascular Surgery and Plastic Surgery wards. The microbiological diagnosis of wound infections was established according to the laboratory’s working protocol. PCR screening of antibiotic resistance genes was performed using a real-time PCR, while the microtiter plate assay was used to determine the biofilm-forming capacity. Testing of biofilm susceptibility to meropenem and amikacin was performed on Calgary biofilm device. Of the 88 bacterial isolates studied, 78.40% were Gram-negative bacilli (GNB)—Klebsiella pneumoniae (K.P), Pseudomonas aeruginosa (P.A), Proteus mirabilis (P.M), Acinetobacter baumannii (A.B), while the remaining 21.60% were Gram-positive cocci (GPC)—Staphylococcus aureus (S.A). All A.B isolates and 92.59% of K.P were carriers of β-lactamase- and carbapenemase-encoding genes, while 57.89% of S. aureus isolates were carriers of mecA (methicillin-resistant). Strong biofilm-forming isolates (B+++) were more frequent in P.A than in K.P (p = 0.002) and P.M (p = 0.02), with a frequency comparable to that of A.B strains (p = 0.212). When analyzing the biofilm reaction to meropenem, a significantly lower susceptibility was detected in the biofilm for K.P isolates, compared to the planktonic ones. Most GNB have been extensively multidrug-resistant, particularly K.P and A.B. Isolates from chronic wounds are major biofilm-formers. A strong and statistically significant association has been identified in the case of K.P and P.M between the presence of resistance genes and the biofilm-forming capacity. These findings highlight the need for a customized therapeutic approach for each chronic wound, considering the mechanisms underlying treatment resistance. These include bacterial virulence factors and the wound microenvironment colonized by the biofilm and the relative contribution of each to the overall resistance profile. Full article
(This article belongs to the Special Issue Bacterial Pathogens: Biofilm Formation and Eradication)
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15 pages, 1574 KB  
Article
Engineered Phage Modulates Quorum Sensing and Biofilm Formation in Pseudomonas aeruginosa
by Domenico Franco, Salvatore Papasergi, Francesco Mediati, Salvatore P. P. Guglielmino and Laura Maria De Plano
Microorganisms 2026, 14(5), 1028; https://doi.org/10.3390/microorganisms14051028 - 30 Apr 2026
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Abstract
Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen frequently associated with chronic and biofilm-related infections, largely driven by quorum sensing (QS)-related genes/phenotypes. In this study, we investigated the antivirulence activity of an engineered M13-derived phage-display particle (P9b), selected for specific binding to P. aeruginosa [...] Read more.
Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen frequently associated with chronic and biofilm-related infections, largely driven by quorum sensing (QS)-related genes/phenotypes. In this study, we investigated the antivirulence activity of an engineered M13-derived phage-display particle (P9b), selected for specific binding to P. aeruginosa, which acts as a non-lytic modulator of QS through specific binding to a bacterial surface target. P9b induced a transient delay in early planktonic growth, without affecting long-term proliferation. In contrast, P9b significantly reduced biofilm-associated metabolic activity and pyocyanin production, consistent with an effect on QS-regulated pathways. Transcriptional analysis revealed significant downregulation of key QS regulators (lasI, lasR, rhlI, and rhlR) and modulation of phenazine biosynthesis genes (phzM downregulation and phzS upregulation), suggesting interference with QS-dependent regulatory circuits. Notably, P9b retained binding capacity and antibiofilm activity across clinically relevant P. aeruginosa isolates. Overall, these findings indicate that P9b acts as a selective, non-lytic modulator of virulence-associated traits, attenuating QS-regulated phenotypes without bactericidal effects. This study supports the potential of engineered filamentous phages as targeted antivirulence platforms for the development of innovative strategies against persistent and biofilm-associated infections. Full article
(This article belongs to the Special Issue Bacterial Pathogens: Biofilm Formation and Eradication)
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21 pages, 1342 KB  
Article
Construction of a Mutant Library of Avibacterium paragallinarum Transposons and Screening and Preliminary Study of Genes Related to Biofilm Formation
by Bingbing Fan, Qishuang Su, Yan Shao, Weidong Sun, Jingming Zhou, Xiangan Han and Wei Jiang
Microorganisms 2026, 14(4), 783; https://doi.org/10.3390/microorganisms14040783 - 30 Mar 2026
Viewed by 554
Abstract
Avibacterium paragallinarum (Av. paragallinarum), the causative agent of infectious coryza, imposes substantial economic burdens on the poultry industry by inducing growth retardation in broilers and reducing egg production in laying hens by up to 40%. Disease control is hindered by the [...] Read more.
Avibacterium paragallinarum (Av. paragallinarum), the causative agent of infectious coryza, imposes substantial economic burdens on the poultry industry by inducing growth retardation in broilers and reducing egg production in laying hens by up to 40%. Disease control is hindered by the limited efficacy of available vaccines and the increasing prevalence of antibiotic resistance—challenges that are exacerbated by the pathogen’s capacity to form biofilms, which facilitate bacterial persistence and enhance drug tolerance. To systematically elucidate the genetic determinants underlying biofilm formation in Av. Paragallinarum, we constructed a high-density random mutant library using mini-Tn5 transposon mutagenesis, comprising 3106 individual mutants. Phenotypic screening via crystal violet staining identified 188 mutants displaying altered biofilm-forming capacity relative to the wild-type strain, including 172 with enhanced and 16 with reduced biofilm formation. Sequencing of transposon insertion sites in these mutants revealed 105 disrupted genes involved in diverse biological pathways, including amino acid metabolism, quorum sensing, and transmembrane transport. A representative subset of eight mutants was selected for detailed phenotypic characterization. Their biofilm phenotypes were consistent with the initial screening results; certain mutants exhibited markedly enhanced biofilm formation (e.g., Tn-2206), whereas others, including Tn-1504, Tn-2428, and Tn-2859, showed significant reductions in biofilm production. Notably, these three biofilm-deficient mutants—harboring disruptions in a TonB-dependent receptor (Tn-1504), a GntP family permease (Tn-2428), and a hypothetical protein (Tn-2859)—displayed drastically attenuated virulence in vitro. Compared with the wild-type strain, these mutants exhibited reductions in cytotoxicity (up to 66.38%), cell adhesion (up to 50.68%), and invasive capacity, while maintaining normal growth kinetics. These findings indicate that the identified genes may play crucial roles in biofilm-associated virulence and highlight Tn-1504, Tn-2428, and Tn-2859 as promising candidates for the development of live attenuated vaccines. Collectively, this study provides a comprehensive genetic foundation for the rational design of novel anti-biofilm strategies against Av. paragallinarum. Full article
(This article belongs to the Special Issue Bacterial Pathogens: Biofilm Formation and Eradication)
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19 pages, 2608 KB  
Article
Regulatory Effects of Lactobacillus crispatus and Lactobacillus rhamnosus on the Formation and Composition of Gardnerella Biofilms
by Hanyu Qin, Yun Liu, Sheng Yin, Zhengyuan Zhai and Bingbing Xiao
Microorganisms 2026, 14(3), 569; https://doi.org/10.3390/microorganisms14030569 - 2 Mar 2026
Viewed by 1054
Abstract
Bacterial vaginosis (BV), the most common lower genital tract infection among women of childbearing age, is characterized by a decline in Lactobacillus populations and the excessive proliferation of anaerobic bacteria. Clinically, metronidazole remains the first-line therapeutic agent. However, the increasing recurrence rate has [...] Read more.
Bacterial vaginosis (BV), the most common lower genital tract infection among women of childbearing age, is characterized by a decline in Lactobacillus populations and the excessive proliferation of anaerobic bacteria. Clinically, metronidazole remains the first-line therapeutic agent. However, the increasing recurrence rate has become an urgent clinical challenge. An important factor of BV recurrence is the persistent presence of Gardnerella biofilms, which enhances pathogenic resistance to antibiotics. In contrast, a healthy vaginal microbiome, predominated by Lactobacillus, exerts protective effects by producing antimicrobial compounds that inhibit BV pathogen colonization and restore microbial homeostasis. Given this, Lactobacillus preparations have gained widespread attention for their adjunctive therapeutic potential in BV management. Accordingly, in this study, we selected two extensively investigated Lactobacillus species, Lactobacillus crispatus and Lactobacillus rhamnosus, to evaluate their inhibitory capacity against Gardnerella biofilms. Our findings suggest that hydrogen peroxide and D-lactic acid are prominent bioactive components involved in the inhibition of Gardnerella biofilm formation by these Lactobacillus species, though the potential contribution of bacteriocins and other uncharacterized factors cannot be excluded. Notably, this inhibitory activity is not accompanied by alterations to the composition of pre-formed biofilms. This study clarifies the anti-biofilm mechanism of specific Lactobacillus, providing a valuable reference for future research on probiotic-based strategies for the treatment of BV. Full article
(This article belongs to the Special Issue Bacterial Pathogens: Biofilm Formation and Eradication)
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