Open research questions in Bacterial biofilms and quorum sensing

What the literature leaves open

• The need to optimize and validate reporter systems for each target bacterium. The lack of a suitable gene insertion system for Pta.

  Macroscopic and microscopic visualization of Pseudomonas syringae pv. tabaci infection using complementary bioluminescent and fluorescent reporter systems · 2026 · DOI

• Looking ahead, future research should increasingly focus on integrated bioinformatics workflows that combine deep learning–based screening approaches with molecular docking, molecular dynamics simulations, and high-resolution experimental validation, including proteomic and systems-level analyses.

  Bioinformatics-Driven, Plant-Based Antibiotic Research Against Quorum Sensing and Biofilm Formation in Pseudomonas aeruginosa and Escherichia coli Multiresistant Microbes · 2026 · DOI

• However, under environmental conditions such as freeze–thaw stress, the molecular mechanisms underlying the up- and downregulation of QS system genes remain poorly understood, warranting further investigation.

  Survival and stress responses of Listeria monocytogenes and Pseudomonas aeruginosa in single- and dual-species biofilms on stainless steel under freeze–thaw conditions · 2026 · DOI

• However, whether these effects are mediated through direct modulation of quorum sensing regulatory pathways remains to be demonstrated by molecular studies. Therefore, although the postbiotic exhibited promising in vitro anti-virulence activity, its safety profile and potential effects on host cells remain to be determined before further translational or clinical investigations.

  Fermented sausage–derived Latilactobacillus sakei postbiotic inhibits biofilm formation and quorum sensing-related virulence in clinical antibiotic-resistant Pseudomonas aeruginosa · 2026 · DOI

• However, since we did not measure residual methanol concentrations, the precise relationship between the methanol feeding rate and the product formation rate remains unclear. However, the productivity of the BslA protein in these two prokaryotic cell systems has not been reported to date, given that the scope of those earlier investigations was limited to functional and structural analyses of the protein.

  Expression Profiling of Recombinant Biofilm Surface Layer Protein A in Pichia pastoris Under Constant Dissolved Oxygen and Oxygen-Limited Fermentation · 2026 · DOI

• Whilst our RNA-seq analysis highlights pathways, including the upregulation of TonB-dependent receptors, that suggest metabolite-mediated interactions in these model biolm organisms, the specic metabolites contributing to enhanced biomass remain unknown.

  Microbial Interactions Shape Spatial Organisation and Transcriptional Responses in a Model Mixed-Species Biofilm · 2026 · DOI

• Studies of S. aureus biofilms and sepsis provide context without resolving this P. aeruginosa-specific gap. The epithelial lactate/H3K18la decrease was reported in a full-length study, whereas the proposed macrophage increase rests on a supplement abstract. Different bacterial perturbations, cell types, and experimental systems underlie these reports.

  Lactate dynamics at the Pseudomonas aeruginosa–host interface: bacterial metabolism, host defense, and emerging links to histone lactylation · 2026 · DOI

• It highlights the need for a framework that integrates the dimensions of biofilm resilience mechanisms. The gap in understanding the chemical composition and functional roles of the EPS matrix is also identified.

  The architecture and behavioral phenotype of microbial cells housed in citadel-like biofilms · 2026 · DOI

• Standardized biofilm testing, representative animal models, and regulatory pathways tailored for antibiofilm agents are lacking. Safety, efficacy, and quality standards must address nanomaterials, responsive surfaces, and combination therapies. The clinical validation of daptomycin for prosthetic joint infections illustrates these translational hurdles.

  Emerging Multimodal Strategies for Bacterial Biofilm Eradication: A Comprehensive Review · 2025 · DOI

• In order to effectively deal with this situation, we need to take a multifaceted approach. First, strengthen infection control measures to reduce the spread of MRSA.

  Unmasking MRSA’s Armor: Molecular Mechanisms of Resistance and Pioneering Therapeutic Countermeasures · 2025 · DOI

• Future work will focus on the following: (1) improving efficiency through architectural optimization and advanced pruning strategies; (2) expanding the dataset to include a wider range of damage types and environmental variations to enhance model robustness and generalization ability; and (3) conducting on-site deployment and validation in commercial production systems.

  OMB-YOLO-tiny: A Lightweight Detection Model for Damaged Pleurotus ostreatus Based on Enhanced YOLOv8n · 2025 · DOI

• Future studies should concentrate on the rational design of combinatorial therapies that employ synergistic, multi-target approaches. As most current investigations are limited to in vitro settings, future work can prioritize translational and preclinical in vivo models to assess long-term antifouling and biofilm eradication performance, host tissue compatibility, and immune responses.

  Targeting Bacterial Biofilms on Medical Implants: Current and Emerging Approaches · 2025 · DOI

• Further studies addressing the mechanisms of antibiotic tolerance and resistance in parallel to virulence factor expression are required to be studied across growth phases. Also, future research should be extended to clinical strains to validate the findings of the current study and enhance their clinical applicability.

  Growth-Phase-Dependent Modulation of Quorum Sensing and Virulence Factors in Pseudomonas aeruginosa ATCC 27853 by Sub-MICs of Antibiotics · 2025 · DOI

• There is a scarcity of clinical studies validating the proposed therapeutic alternatives, particularly concerning natural compounds. Despite the wealth of laboratory data and the promise demonstrated by flavonoids, terpenes, and quinones in inducing oxidative stress in S. aureus, the transition of these findings into practical applications remains uncertain and challenging.

  What Do We Know About Staphylococcus aureus and Oxidative Stress? Resistance, Virulence, New Targets, and Therapeutic Alternatives · 2025 · DOI

• Finally, in silico ADMET profiling and cytotoxicity assessments using Caenorhabditis elegans suggest that these aniline derivatives hold promise as therapeutic candidates, warranting further investigation.

  Broad‐Spectrum Antimicrobial and Antibiofilm Activities of Halogenated Anilines Against Uropathogenic Escherichia coli and ESKAPE Pathogens · 2025 · DOI

• The study requires the use of specific strains and techniques to assess biofilm formation. The study needs to investigate the effects of biofilm formation on virulence and drug resistance.

  Differences in Biofilm Formation of Listeria monocytogenes and Their Effects on Virulence and Drug Resistance of Different Strains · 2024 · DOI

• It highlights the need for more effective methods to control biofilms.

  Hidden Places for Foodborne Bacterial Pathogens and Novel Approaches to Control Biofilms in the Meat Industry · 2024 · DOI

• Chemical Modifications for Enhanced Bioavailability and Target Affinity Structural refinement of core pharmacophores is critical to improving drug-like properties.

  Anti-QS Strategies Against Pseudomonas aeruginosa Infections · 2025 · DOI

• Through comprehensive elaboration on the functional mechanisms of these various c-di-GMP effectors, three key aspects have been systematically revealed, as laid out below, establishing the role of effectors as central schedulers in bacterial environmental adaptation mechanisms. The evolution and divergence of effector domains lay the molecular foundation for their functional diversity.

  Dynamic Executors of Bacterial Signals: Functional Versatility and Regulatory Networks of c-di-GMP Effectors · 2025 · DOI

• Investigating the effects of different microchannel geometries on biofilm growth and hydraulic response, - Examining the impact of varying flow rates and pressures on biofilm development, - Studying the role of microbial community composition on biofilm-hydrodynamic…

  Shear redistribution in confined biofilm systems: decoupling structure and function in engineered water environments · 2026 · DOI

• The lack of understanding of how geometric confinement redistributes shear during biofilm growth - The limitation of bulk metrics in evaluating biofilm responses - The need to investigate biofilm-hydrodynamics coupling in confined…

  Shear redistribution in confined biofilm systems: decoupling structure and function in engineered water environments · 2026 · DOI

• This design helps distinguish candidate canonical pyocin regions from genomic regions containing phage-associated features, addressing a key limitation in the annotation of phage-derived elements in bacterial genomes.

  TattleTail: A Rule-Based Tool for Identifying Canonical Pyocin-Encoding Gene Clusters in Pseudomonas aeruginosa · 2026 · DOI

• Optimization and validation of reporter systems for each target bacterium, - Development of new methods to overcome autofluorescence problems, - Application of the developed system to other plant-pathogenic…

  Macroscopic and microscopic visualization of Pseudomonas syringae pv. tabaci infection using complementary bioluminescent and fluorescent reporter systems · 2026 · DOI

• These metabolites also affect protozoan predators by influencing their physiology, modulating motility, encystation, and virulence (Figure 1). In response, protozoa employ adaptive strategies including metabolic reprogramming, oxidative stress management, and post-translational modifications such as protein S-sulfuration (see Table 1 for summary).

  The Role of Biofilm-Derived Compounds in Microbial and Protozoan Interactions · 2025 · DOI

• Mapping QS dynamics continuously across different growth phases under the stress of sub-MIC antibiotics from different classes fills a critical gap in understanding the pathophysiology of P.

  Growth-Phase-Dependent Modulation of Quorum Sensing and Virulence Factors in Pseudomonas aeruginosa ATCC 27853 by Sub-MICs of Antibiotics · 2025 · DOI

Most-cited papers in Bacterial biofilms and quorum sensing

• Bacterial Biofilms: A Common Cause of Persistent Infections · Science · 1999 · 10,040 citations
• Antibiotic resistance of bacteria in biofilms · The Lancet · 2001 · 3,809 citations
• The Involvement of Cell-to-Cell Signals in the Development of a Bacterial Biofilm · Science · 1998 · 2,774 citations
• Bacterial Persistence as a Phenotypic Switch · Science · 2004 · 2,750 citations
• Tolerance and resistance of microbial biofilms · Nature Reviews Microbiology · 2022 · 1,066 citations
• Quorum sensing triggers the stochastic escape of individual cells from Pseudomonas putida biofilms · Nature Communications · 2015 · 884 citations
• Dental biofilms: difficult therapeutic targets · Periodontology 2000 · 2002 · 882 citations
• Implication of Surface Properties, Bacterial Motility, and Hydrodynamic Conditions on Bacterial Surface Sensing and Their Initial Adhesion · Frontiers in Bioengineering and Biotechnology · 2021 · 830 citations
• Strategies for combating bacterial biofilm infections · International Journal of Oral Science · 2014 · 821 citations
• Dental plaque as a biofilm and a microbial community – implications for health and disease · BMC Oral Health · 2006 · 770 citations

Most recent work

• Next-Generation Strategies for Controlling Foodborne Pathogens: Precision Antimicrobials, Biofilm Disruption, and Emerging Molecular Interventions · Foods · 2026
• Understanding Pseudomonas aeruginosa Biofilms: Quorum Sensing, c-di-GMP Signaling, and Emerging Antibiofilm Approaches · Microorganisms · 2026
• Emerging non-antibiotic strategies for implant-associated biofilm infections by reprogramming the dysregulated immune microenvironment · npj Biofilms and Microbiomes · 2026
• High-throughput design of defined microbial consortia for crop protection · Annals of Botany · 2026
• Chemically modified and inactivated bacteria enable intra-biofilm drug delivery and long-term immunity against implant infections · Nature Biomedical Engineering · 2026
• Detection to Disruption: A Comprehensive Review of Bacterial Biofilms and Therapeutic Advances · Antibiotics · 2026
• Fatty Acids as Prebiotics and Their Role in Antibiofilm Activity · Antibiotics · 2026
• Lactic Acid Bacteria as Natural Antimicrobials: Biofilm Control in Food and Food Industry · Antibiotics · 2026
• Bacterial biofilm conundrum: insight into the frontiers of antibiotic resistance and state-of-the-art anti-biofilm interventions · Frontiers in Cellular and Infection Microbiology · 2026
• Autoinducer 2 as a universal language in microbial consortia: decoding molecular mechanisms, ecological impacts, and application · Gut Microbes · 2026

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