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Microorganisms
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Feature Papers in Biofilm
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Feature Papers in Biofilm

A topical collection in Microorganisms (ISSN 2076-2607). This collection belongs to the section "Biofilm".

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Editors

Prof. Dr. Ute Römling

E-Mail Website
Collection Editor
Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, Stockholm, Sweden
American Academy of Microbiology (elected fellow)
European Academy of Microbiology (elected fellow)
Distinguished Scholar of COMSTECH
Policy board and member of USERN
Elected Fellow of AcademiaNet
Interests: microbial biofilm formation; cyclic di-nucleotide signaling; pathogen-host interaction; protein quality control; Salmonella typhimurium; Pseudomonas aeruginosa; Candida parapsilosis
Special Issues, Collections and Topics in MDPI journals
Dr. María Guembe

E-Mail Website
Collection Editor
Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, 28007 Madrid, Spain
Interests: biofilms; prostheses; infection; devices; catheter
Special Issues, Collections and Topics in MDPI journals

Topical Collection Information

Dear Colleagues,

This Topical Collection, “Feature Paper in Biofilm”, intends to collect high-quality research articles, short communications, and review articles from all fields of biofilm research. Since the aim of this Topical Collection is to illustrate, through selected works, pioneering research in the field of biofilm, we encourage Editorial Board Members of the Biofilm Section of Microorganisms and beyond to contribute state-of-the-art articles reflecting the latest progress in their research field, or to invite relevant experts and colleagues to do so.

Topics of interest include, but are not limited to:

  • Pathogenesis of biofilms;
  • Biofilm formation of the microbiota;
  • Device-related infections;
  • Chronic tissue-related infections;
  • Innovative strategies for accurate diagnosis of biofilms;
  • Antimicrobial tolerance;
  • New antibacterial approaches;
  • Impact of biofilm formation in biotechnology;
  • Biofilms in nanomedicine.

Prof. Dr. Ute Römling
Dr. María Guembe
Collection 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. 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 collection 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-anonymized 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

  • biofilm
  • infections
  • antibiofilm
  • biofilm formation

Published Papers (6 papers)

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2026

Jump to: 2025

12 pages, 611 KB  
Article
From Broadband to Single-Tone Stimulation: Frequency-Selective Response of Staphylococcus aureus Biofilms to Electric Fields
by Marco Balato, Emanuela Roscetto, Maria Rosaria Catania, Martina Aversa, Carlo Petrarca, Massimo Vitelli, Umberto Galdiero, Luigi Costanzo, Valeria Nocerino and Giovanni Balato
Microorganisms 2026, 14(7), 1516; https://doi.org/10.3390/microorganisms14071516 - 11 Jul 2026
Viewed by 330
Abstract
Biofilm-associated infections caused by Staphylococcus aureus are highly resistant to antimicrobial treatments. Low-intensity electric fields have shown promise as an antibiofilm strategy; however, the role of frequency remains poorly understood. In this study, mature S. aureus biofilms were exposed to low-intensity alternating electric [...] Read more.
Biofilm-associated infections caused by Staphylococcus aureus are highly resistant to antimicrobial treatments. Low-intensity electric fields have shown promise as an antibiofilm strategy; however, the role of frequency remains poorly understood. In this study, mature S. aureus biofilms were exposed to low-intensity alternating electric fields (12.5 mV/cm) across broadband (10 Hz–10 MHz) frequency range and band-limited sub-ranges, as well as to selected single-tone frequencies within the interval 1–100 kHz. Antibiofilm activity was assessed in terms of cell culturability (CFU/mL) and biofilm biomass. A significant frequency-dependent effect was observed (p < 0.001), with maximal activity observed only at specific frequencies within the range 1–100 kHz, whereas other frequencies were ineffective. These findings demonstrate that the electrical antibiofilm effect is characterized by a frequency selective response, which is similar to a band-pass filter with a nearly flat shape inside the band-pass range and strongly attenuated effects outside such a band. This frequency-selective response supports the hypothesis of a biophysical mechanism involving membrane and matrix interactions and highlights the potential for developing targeted, frequency-optimized electroceutical strategies. Full article
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19 pages, 1194 KB  
Article
Enhanced Biofilm Formation by ICU-Associated Stenotrophomonas maltophilia Isolates: A Potential Contributor to Persistence and Clonal Dissemination
by Giovanni Di Bonaventura, Giovanni Gherardi, Martina Barchitta, Antonella Agodi and Arianna Pompilio
Microorganisms 2026, 14(7), 1471; https://doi.org/10.3390/microorganisms14071471 - 3 Jul 2026
Viewed by 297
Abstract
Stenotrophomonas maltophilia is an emerging multidrug-resistant opportunistic pathogen in intensive care units (ICUs) and cystic fibrosis (CF), where biofilm formation may favor persistence, device-associated colonization/infection, and clonal dissemination. This study compared biofilm formation, clonal relatedness, biofilm phenotypes, and motility in 37 ICU-associated and [...] Read more.
Stenotrophomonas maltophilia is an emerging multidrug-resistant opportunistic pathogen in intensive care units (ICUs) and cystic fibrosis (CF), where biofilm formation may favor persistence, device-associated colonization/infection, and clonal dissemination. This study compared biofilm formation, clonal relatedness, biofilm phenotypes, and motility in 37 ICU-associated and 42 CF-associated S. maltophilia isolates. Biofilm formation on polystyrene was quantified by crystal violet assay and expressed both as absolute biomass and as a growth-normalized Biofilm Index, calculated to account for differences in planktonic growth. Genetic diversity was assessed by pulsed-field gel electrophoresis, while swimming and twitching motility were evaluated using agar-based assays. ICU isolates showed a higher prevalence of biofilm formation, greater biofilm biomass, and higher growth-normalized Biofilm Index values than CF isolates. They also displayed lower genetic diversity and more frequent cross-transmission, supporting the circulation of selected hospital-associated lineages. Conversely, CF isolates showed greater heterogeneity and a more complex biofilm pattern, consistent with adaptation to a distinct chronic airway environment. Motility was not associated with biofilm formation, suggesting that the enhanced biofilm phenotype of ICU isolates is not explained by swimming or twitching alone. Overall, these findings support a setting-specific model in which enhanced biofilm-forming capacity may contribute to S. maltophilia ICU persistence and clonal dissemination, highlighting the need for targeted surveillance and careful device management. Full article
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13 pages, 703 KB  
Communication
Differential Responses of Salmonella enterica Typhimurium, S. enteritidis, and S. infantis to Chlorine Dioxide In Vitro: Impacts on Growth and Biofilm Development
by Inkar Castellanos-Huerta, Jacob Lum, Guillermo Romero, Aaron Forga, Billy M. Hargis and Danielle Graham
Microorganisms 2026, 14(5), 1058; https://doi.org/10.3390/microorganisms14051058 - 8 May 2026
Viewed by 462
Abstract
Salmonella enterica is a significant Gram-negative bacterium possessing over 2500 serovars capable of affecting both animals and humans and disseminating widely due to its adaptability, genetic diversity, and ability to form biofilms. Different serovars, such as S. enterica Typhimurium (ST), Enteritidis (SE), and [...] Read more.
Salmonella enterica is a significant Gram-negative bacterium possessing over 2500 serovars capable of affecting both animals and humans and disseminating widely due to its adaptability, genetic diversity, and ability to form biofilms. Different serovars, such as S. enterica Typhimurium (ST), Enteritidis (SE), and Infantis (SI), display varying traits and survival strategies in harsh environments. Biofilms, composed of proteins, lipids, and DNA, enable bacteria to survive stresses such as pH changes, nutrient shortages, temperature fluctuations, and disinfectants. Evaluating disinfectants on inert surfaces is crucial for understanding their effectiveness and impact on poultry. This study assessed the efficacy of chlorine dioxide (ClO2) disinfectant against ST, SE, and SI growth, biofilm formation, and biofilm removal at varying concentrations in vitro. Results showed serotype-dependent and condition-specific responses, with SE and SI being more affected than ST, which may be associated with differences in oxidative stress response mechanisms, highlighting the need for tailored disinfection protocols. Full article
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20 pages, 2387 KB  
Article
Persistence and Risk Assessment of Biofilm-Forming MDR and XDR Bacteria on Non-Poultry Meat Contact Surfaces in Wah Cantt, Pakistan
by Lubna Shakoor, Shumaila Naz, Anas Rashid and Muhammad Idrees
Microorganisms 2026, 14(5), 1051; https://doi.org/10.3390/microorganisms14051051 - 7 May 2026
Viewed by 639
Abstract
Biofilms on meat-contact surfaces pose critical food safety risks. This study investigates the interplay between biofilm architecture, metabolic vigor, and antimicrobial resistance on retail surfaces in Pakistan. Screening 300 isolates from 120 surfaces identified 42 high-risk biofilm formers. Comprehensive phenotypic screening revealed that [...] Read more.
Biofilms on meat-contact surfaces pose critical food safety risks. This study investigates the interplay between biofilm architecture, metabolic vigor, and antimicrobial resistance on retail surfaces in Pakistan. Screening 300 isolates from 120 surfaces identified 42 high-risk biofilm formers. Comprehensive phenotypic screening revealed that standard visual assays severely underestimate the viability of environmental strains. Biofilm biomass and metabolic activity correlated positively (Spearman’s ρ = 0.656, p < 0.001). Crucially, Ordinary Least Squares regression established that metabolic vigor, rather than physical biomass, independently predicts resistance severity. Phenotypic profiling revealed a high-risk landscape with 81.8% multidrug-resistant and 18.2% extensively drug-resistant isolates, including resistance to colistin and Linezolid. Alarmingly, 79.5% of critical resistance phenotypes compromised WHO Reserve category antibiotics, escalating to 100% on mincer machines. Ecological analysis demonstrated surface-driven partitioning; porous wood boards fostered diverse Enterobacteriaceae, while mincers selected for uniformly resistant clades. These findings highlight processing machinery as resilient reservoirs for untreatable pathogens, necessitating targeted anti-biofilm measures, such as matrix-degrading enzymes. Bridging a critical knowledge gap, this study is among the earliest integrated ecological analyses combining phylogenetic, metabolic, and resistance profiling in Pakistan’s non-poultry meat sector. Full article
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20 pages, 4954 KB  
Article
Mechanistic Insights into the Inhibition of Yersinia enterocolitica Biofilm Formation by Lipoic Acid
by Sichen Liao, Siqi Yang, Guoli Gong, Zhenbin Liu, Jiayi Zhang, Hongbo Li, Qing Sun, Haizhen Mo, Liangbin Hu and Lu Tian
Microorganisms 2026, 14(3), 558; https://doi.org/10.3390/microorganisms14030558 - 28 Feb 2026
Cited by 1 | Viewed by 855
Abstract
Yersinia enterocolitica is a foodborne pathogen that forms biofilms on surfaces, enhancing its survivability and increasing bacterial resistance, which poses a significant challenge to public health. Therefore, developing effective strategies to inhibit biofilm formation is crucial. Lipoic acid (LA) is a compound with [...] Read more.
Yersinia enterocolitica is a foodborne pathogen that forms biofilms on surfaces, enhancing its survivability and increasing bacterial resistance, which poses a significant challenge to public health. Therefore, developing effective strategies to inhibit biofilm formation is crucial. Lipoic acid (LA) is a compound with antibiofilm properties. This study investigates the effects of LA on biofilm formation by Y. enterocolitica BNCC 108930 (a standard strain from the BeNa Culture Collection). Biofilm formation, maturation, removal, and cell viability were evaluated by crystal violet staining, extracellular polysaccharide assay, Methylthiazolyldiphenyl-tetrazolium bromide assays, motility, and quorum sensing (QS) assays. The results indicate that LA interferes with the early stages of biofilm formation by compromising cell membrane integrity and reducing cellular adhesion. Furthermore, 2.5 mg/mL of LA reduced biofilm biomass (with a 48 h treatment inhibition rate of 51.46 ± 1.29%) and extracellular polysaccharide production (with a relative inhibition rate of 30.09 ± 1.8%), while significantly reducing the metabolic activity of bacteria within the biofilm (inhibition rate over 85%) compared to the untreated group. Confocal laser scanning microscopy and field emission gun scanning electron microscopy confirm that LA induces a sparse biofilm structure, reduced aggregation, and decreased biofilm thickness to 21.33 ± 2.27 μm. Motility and QS assays demonstrate that LA affects flagellar motility and the secretion of N-acyl homoserine lactones. Transcriptome analysis revealed downregulation of genes involved in the QS system and biofilm formation (e.g., lsrA, lsrC, lsrD, lsrR, and oppA), as well as upregulation of genes related to bacterial chemotaxis and flagellar assembly (e.g., RS19655, RS15590, fliE, fliJ, fliP, fliA, and fliK). These alterations suggest that LA inhibits Y. enterocolitica biofilm formation by affecting intercellular communication and flagellar motility. This study highlights the antibiofilm properties of LA, providing a theoretical basis for potential applications in microbial and biofilm control. Full article
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2025

Jump to: 2026

14 pages, 3325 KB  
Article
Formation of Mono-Organismal and Mixed Staphylococcus aureus and Streptococcus mutans Biofilms in the Presence of NaCl
by Yusuke Iwabuchi, Hiroko Yoshida, Shuichiro Kamei, Toshiki Uematsu, Masanori Saito and Hidenobu Senpuku
Microorganisms 2025, 13(5), 1118; https://doi.org/10.3390/microorganisms13051118 - 13 May 2025
Cited by 3 | Viewed by 1896
Abstract
Staphylococcus aureus, an opportunistic bacterium found in the oral cavity, has been reported as a causative agent of infective endocarditis and pneumonia. Salt is an essential mineral for cell maintenance in the human body. This study was conducted to clarify how salt [...] Read more.
Staphylococcus aureus, an opportunistic bacterium found in the oral cavity, has been reported as a causative agent of infective endocarditis and pneumonia. Salt is an essential mineral for cell maintenance in the human body. This study was conducted to clarify how salt affects the formation of biofilms by S. aureus and Streptococcus mutans, pathogens implicated in dental caries. Bacteria were cultivated with various concentrations of NaCl on a 96-well microtiter plate in tryptic soy broth with 0.25% sucrose or 0.25% glucose (TSBs and TSBg, respectively) for 16 h. The effects of glucosyltransferase in S. mutans membrane vesicles (MVs) and extracellular DNA during biofilm formation were also analyzed. S. aureus biofilms were induced by 0.004–0.25 M NaCl but not by NaCl at concentrations greater than 0.25 M in TSBs. The mixed S. aureus and S. mutans biofilms gradually grew and were constructed by dead cells in a NaCl concentration-dependent manner in both TSBs and TSBg. Moreover, biofilms were slightly induced by glucan generation mediated by the glucosyltransferases in MVs under high-salinity conditions. The formation of mixed-species S. aureus and S. mutans biofilms increased in the presence of both extracellular DNA and MVs. Therefore, extracellular DNA, MVs, and dead cells are factors that promote S. aureus biofilm formation under harsh conditions containing NaCl. The sugar (sucrose and glucose) ingestion-induced S. mutans biofilm may be a risk factor for infection by opportunistic pathogens such as S. aureus in individuals who consume food and drinks containing high concentrations of salt. Full article
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