@article {pmid41033401,
year = {2025},
author = {Díaz-Navarro, M and Crespo, A and Muñoz, P and Guembe, M},
title = {Biomass and metabolic activity staining biofilm techniques are not reliable enough to be used in microbiology laboratories.},
journal = {Journal of microbiological methods},
volume = {},
number = {},
pages = {107285},
doi = {10.1016/j.mimet.2025.107285},
pmid = {41033401},
issn = {1872-8359},
abstract = {Crystal violet and XTT staining quantify biofilms but are influenced by external factors. Testing MSSA, E. coli, and C. albicans refrigerated five weeks revealed weekly variability between researchers. These methods need careful interpretation and should be supported by more reproducible techniques for reliable results.},
}

@article {pmid41033369,
year = {2025},
author = {Namazi, P and Behbahani, BA and Noshad, M and Vasiee, A and Taki, M and Joyandeh, H},
title = {Anti-Listeria Mechanisms, Safety, and Predictive Modeling of Lacticaseibacillus casei UTMB9: Probiotic Profiling Targeting Virulence Gene Expression and Biofilm Formation.},
journal = {Microbial pathogenesis},
volume = {},
number = {},
pages = {108071},
doi = {10.1016/j.micpath.2025.108071},
pmid = {41033369},
issn = {1096-1208},
abstract = {There is growing interest in probiotics due to their potential to confer health benefits. This study aimed to explore the potential probiotic characteristics, gene expression linked to biofilm formation, and anti-biofilm properties of Lacticaseibacillus casei UTMB9. L. casei UTMB9 tolerates acidic environments, with survival rates of 6.90, 7.59, and 7.96 log CFU/mL at pH 2.5, 3.5, and 4.5, respectively, and exhibits slight growth inhibition under bile concentrations up to 0.7 %. Viability declined modestly from 8.51 to 6.61 log colony forming unit (CFU)/mL under simulated gastrointestinal conditions. Surface hydrophobicity (40.9 %), auto-aggregation (30.9 %), co-aggregation (40.8 %), and adhesion (11.9 %) support its adherence potential. Antimicrobial assays showed most potent inhibition against Listeria monocytogenes (9.70 mm) versus E. coli (4.74 mm). Scanning Electron Microscopy (SEM) imaging revealed pronounced damage to L. monocytogenes cells after exposure to cell-free supernatant (CFS), accompanied by significant downregulation of prfA and flaA, and marked antibiofilm suppression. It also demonstrated antioxidant activity (2,2-diphenyl-1-picrylhydrazyl (DPPH) 43.6 %, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) 47.5 %, linoleic acid oxidation inhibition 30.3 %) and moderate anticancer activity (IC50 ≈ 55-62 mg/mL). The strain reduced cholesterol uptake by 41.9 %, was broadly antibiotic-sensitive (except for partial resistance to ampicillin), and lacked biogenic amine production, DNase, or hemolytic activity. In the second part of this study, Gaussian Process Regression (GPR) was used to predict acidity and bile salt. GPR accurately predicted acidity and bile tolerance (MAPE = 0.22 % and 0.18 %; R[2]≥ 0.99). These findings position L. casei as a promising probiotic agent with robust antimicrobial, antibiofilm, antioxidant, and predictive model-supported features.},
}

@article {pmid41032695,
year = {2025},
author = {Ashrafi, B and Heydari, R and Rezaei, F and Beiranvand, B and Pajouhi, N and Rashidipour, M and Taherikalani, M and Soroush, S},
title = {Investigating the release of active compounds and cytotoxicity of thymol/gallic acid/β-cyclodextrin bio-nanocomposite: a targeted strategy with the approach of disrupting the genes involved in the quorum sensing system and biofilm formation in P. aeruginosa (PAO1).},
journal = {Preparative biochemistry & biotechnology},
volume = {},
number = {},
pages = {1-17},
doi = {10.1080/10826068.2025.2563676},
pmid = {41032695},
issn = {1532-2297},
abstract = {The quorum sensing (QS) system and cell-to-cell communication have had a significant impact on biofilm formation and virulence factor increase in Pseudomonas aeruginosa (P. aeruginosa), making this opportunistic pathogen a global concern and potentially life-threatening agent. The present study aimed to create an innovative pharmaceutical bio-nanocomposite (BNC) comprising thymol (THY) and gallic acid (GA) based on β-cyclodextrin (β-CD), which was used to investigate the release kinetics of active compounds, the level of cytotoxicity, antibacterial and anti-biofilm potential, and measuring the expression of genes effective in QS in the strain PAO1 pays P. aeruginosa. Based on this, physicochemical characteristics of the synthesized BNC were determined using Fourier transform infrared spectroscopy analysis (FTIR), UV-vis measurement, dynamic light scattering (DLS), zeta potential, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The BNC's antibacterial and anti-biofilm capabilities were assessed using the PAO1 reference strain of P. aeruginosa and the expression level of QS-effective genes (rhlI, rhlR, lasI, and lasR) in bacteria was also evaluated in the presence of the synthesized BNC. The results of FTIR spectroscopy show the formation of intramolecular hydrogen bonds between THY, GA and β-CD. Absorption peaks of the UV-vis spectroscopy spectrum of the synthesized BNC at wavelengths of 217 and 272 nm are confirmed successful encapsulation of the THY and GA into the β-CD. The maximum size of the synthesized BNC was recorded as 356.3 nm with a polydispersity index (PDI) of 0.816. SEM and TEM micrographs show the presence of THY/GA active compounds in the pores in β-CD and the formation of a dense polymer network. After 360 minutes of release kinetics, more than 70% of the complex's active chemicals had been released. The biological complex's low toxicity is indicated by average cell survival of more than 65% and the ability to preserve the spindle shape of normal fibroblast cells at high concentrations. The PAO1 strain has minimum inhibitory concentrations (MIC) of 323 and 199.6 μg/mL for minimal biofilm inhibition concentration 50% (MBIC50). The decrease in rhlI and rhlR gene expression relative to the control group (without treatment) suggests that the active chemicals released from the biological complex interact and disrupt the QS pathway. Overall, the synthesized pharmaceutical complex has promise as a clever and effective option for future research and practical advances, as well as the development of complementary therapies.},
}

@article {pmid41030757,
year = {2025},
author = {Kurtzman, GM and Horowitz, RA and Johnston, R and Lanphier, L},
title = {Oral Biofilm and Gender-Specific Health Considerations.},
journal = {Cureus},
volume = {17},
number = {8},
pages = {e91289},
pmid = {41030757},
issn = {2168-8184},
abstract = {Oral biofilm plays a central role in the development of periodontal and systemic diseases, with growing evidence highlighting significant gender-specific differences. Hormonal fluctuations in women, during puberty, menstruation, pregnancy, menopause, and with oral contraceptive use, may alter the composition and behavior of oral biofilm, increasing susceptibility to gingival inflammation and periodontal disease. Conditions such as polycystic ovary syndrome (PCOS), osteoporosis, and pregnancy-associated gingivitis further demonstrate the influence of endocrine factors on oral health. In men, higher rates of severe periodontitis are observed, potentially linked to testosterone-related immune responses and behavioral factors with associations to lower sperm counts, increased incidence of prostate cancer, and erectile dysfunction. These distinctions underscore the importance of considering sex-specific biology in both the prevention and management of oral and systemic diseases influenced by biofilm. This study reviews the connections between gender-specific health and oral biofilm.},
}

@article {pmid41029314,
year = {2025},
author = {Jana, D and Manna, T and Guchhait, KC and Panja, S and Karmakar, A and Ballav, S and Hazra, S and Dey, S and Panda, AK and Ghosh, C},
title = {An investigation on anti-biofilm potential of Aegle marmelos fruit extract against multi-drug-resistant Staphylococcus aureus.},
journal = {BMC complementary medicine and therapies},
volume = {25},
number = {1},
pages = {334},
pmid = {41029314},
issn = {2662-7671},
mesh = {*Biofilms/drug effects ; *Plant Extracts/pharmacology ; *Aegle/chemistry ; *Anti-Bacterial Agents/pharmacology ; Fruit/chemistry ; *Staphylococcus aureus/drug effects ; *Drug Resistance, Multiple, Bacterial/drug effects ; Microbial Sensitivity Tests ; Humans ; },
abstract = {BACKGROUND: Staphylococcus aureus, member of ESKAPEE pathogens is a noteworthy contributor to the global crisis rising due to antimicrobial resistance. Biofilms are the primary reason behind the increased antibiotic resistance and tolerance of pathogens. Hence targeting bacterial biofilms has been prioritized as an alternative strategy to counter antibiotic resistance. Aegle marmelos has gained prominence in Indian traditional medicine as seeds, fruits, leaves, bark and roots of this plant are being in use extensively in treating several kinds of ailments by the inhabitants of this subcontinent due to its ethno-pharmacological relevance. The fruit of this plant has been found with remarkable anti-bacterial properties along with other therapeutic efficacies. The present study aimed to identify the anti-biofilm potential of methanolic fruit extract of Aegle marmelos (AMFE) against multi-drug-resistant (MDR) S. aureus strains as a resort to counter the global crisis of antimicrobial resistance for alternative approaches.

RESULTS: MBIC and MBEC of AMFE ranged between 100 and 200 µg.mL[-1] and 300-500 µg.mL[-1], respectively. AMFE could substantially reduce the carbohydrate and protein content of the exo-polymeric substance (EPS), crucial for biofilm production. Expressions of major biofilm promoting genes icaAD and its accessory sarA were down-regulated upon AMFE treatment as revealed from qRT-PCR analysis whereas the quorum sensing gene agr that promotes biofilm detachment was up-regulated. Fluorescence, scanning electron and atomic force microscopic studies confirm the reduction of biofilm biomass upon AMFE treatment. Up to 10 mg.mL[-1] AMFE was non-toxic to human lymphocytes with cell viability of 75.35%. GC-MS and FT-IR studies could detect the bioactive components where 9-octadecenoic acid, n-hexadecanoic acid, 9,12-octadecadienoic acid, methyl 4,7,10- hexadecatrienoate were the major components.

CONCLUSION: Anti-biofilm activity of AMFE towards MDR S. aureus have been established through in vitro biochemical and gene expression studies that were further substantiated by microscopic studies which reveal that AMFE could be explored in the management of S. aureus-associated infections.},
}

*Biofilms/drug effects
*Plant Extracts/pharmacology
*Aegle/chemistry
*Anti-Bacterial Agents/pharmacology
Fruit/chemistry
*Staphylococcus aureus/drug effects
*Drug Resistance, Multiple, Bacterial/drug effects
Microbial Sensitivity Tests
Humans

@article {pmid41029014,
year = {2025},
author = {Soltane, R},
title = {Pubescine as a Novel Antibacterial Agent Against Vancomycin-Resistant Enterococcus: Growth Inhibition, Antibiotic Synergy, and Anti-Biofilm Activity.},
journal = {Current pharmaceutical biotechnology},
volume = {},
number = {},
pages = {},
doi = {10.2174/0113892010399006250923063945},
pmid = {41029014},
issn = {1873-4316},
abstract = {INTRODUCTION: The rise of Vancomycin-Resistant Enterococcus (VRE) has become a major public health concern due to its resistance to conventional antibiotics and ability to form biofilms. The urgent need for novel therapeutic strategies has led to increased interest in natural compounds with antimicrobial potential. Pubescine (PBN), a steroidal alkaloid isolated from Holarrhena pubescens, has demonstrated antimicrobial properties, but its efficacy against VRE remains unexplored.

METHODS: PBN was isolated and purified from Holarrhena pubescens using chromatographic techniques and identified through spectroscopic analysis. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined via broth microdilution assays. Time-kill assays assessed the bacteriostatic or bactericidal nature of PBN. Resistance development was evaluated through prolonged bacterial exposure to subinhibitory concentrations. Synergistic interactions with vancomycin and cefoxitin were analyzed using checkerboard microdilution assays. Biofilm formation and eradication were assessed via crystal violet staining and fluorescence imaging. Metabolic activity and oxidative stress induction were measured using the Alamar Blue assay and Reactive Oxygen Species (ROS) quantification, respectively.

RESULTS: PBN exhibited concentration-dependent inhibition of VRE growth, primarily exerting a bacteriostatic effect without promoting the development of resistance. Checkerboard assays revealed strong synergy between PBN and vancomycin (FICI = 0.1875) and cefoxitin (FICI = 0.3125), suggesting that PBN enhances the efficacy of these antibiotics.

DISCUSSION: PBN significantly reduced biofilm formation and facilitated biofilm disruption at concentrations as low as 4 μg/mL. Metabolic assays demonstrated that PBN suppresses bacterial metabolic activity, while ROS quantification indicated a substantial increase in oxidative stress, suggesting a multi-targeted mechanism of action.

CONCLUSION: These findings establish PBN as a promising antimicrobial agent with potent activity against vancomycin-resistant Enterococcus faecalis. Its ability to enhance antibiotic efficacy, inhibit biofilm formation, and induce oxidative stress underscores its potential as a novel therapeutic strategy against multidrug-resistant infections. Further in vivo studies and pharmacokinetic evaluations are warranted to assess its clinical applicability.},
}

@article {pmid41028885,
year = {2025},
author = {Veerabathiran, A and Subramania, AK and Saikia, M and Manikandamaharaj, TS and Rajendra, SP and Duraisamy, R and Angaiah, S},
title = {Development of 3D-printed Ti-MXene incorporated chitosan/HAP nano-composite soft-bone scaffold and its mechanical, anti-biofilm and cell-viability studies.},
journal = {Scientific reports},
volume = {15},
number = {1},
pages = {33762},
pmid = {41028885},
issn = {2045-2322},
support = {RSPD2025R723//Researchers Supporting Project/ ; },
mesh = {*Chitosan/chemistry ; *Printing, Three-Dimensional ; *Biofilms/drug effects/growth & development ; *Titanium/chemistry ; *Nanocomposites/chemistry ; *Tissue Scaffolds/chemistry ; Cell Survival/drug effects ; *Durapatite/chemistry ; Humans ; Staphylococcus aureus/drug effects/physiology ; Tissue Engineering ; Biocompatible Materials/chemistry ; Bone and Bones ; Materials Testing ; Nitrites ; Transition Elements ; },
abstract = {The prevailing scientific literature suggests that implantable plates demand resurgery, and it will result in corrosion behaviour and the formation of biofilm on the scaffold by Staphylococcus aureus, which is capable of causing a bone surgery-related detrimental effect in two-thirds of the people suffering from osteomyelitis diseases. The development of a nanocomposite scaffold by 3D-bioprinting to improve potent mechanical features with substantial biological characteristics. By incorporating hydroxyapatite (10% w/v) into chitosan (10% w/v) at 1:1 ratio mimicking the natural structure of soft bone tissue. Furthermore, a better structural hydrogel was synthesized for 3D bio-printing through the incorporation of Ti-MXene into the Chitosan/Hydroxyapatite nanocomposite at two distinct ratios. Apart from this, 0.3 mg/mL of Ti-MXene containing 3D-printed nanocomposite scaffold revealed better structural morphology with very less biofilm formation when compared to other 3D-printed scaffolds. Furthermore, mechanical testing such as tensile revealed 23.3 MPa for 0.3 mg/mL of Ti-MXene incorporated Chitosan/HAP nanocomposite. Additionally, this scaffold exhibits a favorable contact angle (74.70°) with a low swelling ratio (27.6%) and degradation rate (1.1%). Further, an in-vitro cell viability test showed a higher cell attachment without cell death. These results find the absence of toxic effect and suggest an enhancement in cell attachment.},
}

*Chitosan/chemistry
*Printing, Three-Dimensional
*Biofilms/drug effects/growth & development
*Titanium/chemistry
*Nanocomposites/chemistry
*Tissue Scaffolds/chemistry
Cell Survival/drug effects
*Durapatite/chemistry
Humans
Staphylococcus aureus/drug effects/physiology
Tissue Engineering
Biocompatible Materials/chemistry
Bone and Bones
Materials Testing
Nitrites
Transition Elements

@article {pmid41028857,
year = {2025},
author = {Kashyap, S and Rathod, Y and Biswas, S},
title = {Murraya koenigii methanolic extract inhibits bacterial growth and biofilm of Staphylococcus aureus and Enterococcus faecalis.},
journal = {Scientific reports},
volume = {15},
number = {1},
pages = {34056},
pmid = {41028857},
issn = {2045-2322},
support = {BT/INF/22/SP42543/2021//Department of Biotechnology, Ministry of Science and Technology, India/ ; BT/INF/22/SP42543/2021//Department of Biotechnology, Ministry of Science and Technology, India/ ; },
mesh = {*Biofilms/drug effects/growth & development ; *Enterococcus faecalis/drug effects/growth & development/physiology ; *Plant Extracts/pharmacology/chemistry ; *Staphylococcus aureus/drug effects/growth & development/physiology ; *Murraya/chemistry ; *Anti-Bacterial Agents/pharmacology/chemistry ; Microbial Sensitivity Tests ; Molecular Docking Simulation ; Methanol/chemistry ; Plant Leaves/chemistry ; Humans ; Bacterial Lysates ; },
abstract = {Hospital-acquired infections caused by Staphylococcus aureus and Enterococcus faecalis are significant global health challenges due to their biofilm-forming ability, also contributing to the derived antibiotic resistance and environmental persistence. This growing resistance poses serious global health challenges, emphasizing the need for better surveillance and new treatments. Plant-derived bioactives have emerged as possible therapeutics to such opportunistic pathogens and they are potential alternatives to traditional antimicrobials. This study investigates the in vitro activity of Murraya koenigii's methanolic (MKM) leaf extract and its compounds against the growth and biofilm-forming ability of S. aureus and E. faecalis. Results revealed that the MKM extract effectively inhibited the growth of S. aureus and E. faecalis at their respective MIC levels. Furthermore, flow cytometry and confocal imaging demonstrated substantial membrane damage in MKM-treated cells compared to DMSO-treated and untreated controls. Additionally, the MKM extract significantly disrupts biofilm formation and leads to reduced extracellular polymeric substance (EPS) production. Scanning electron microscopy provided visual evidence of disrupted biofilm architecture following MKM extract treatment. HR-LC/MS analysis identified bioactive compounds within the extract, which were further evaluated for drug-likeness properties through ADME analysis. In silico molecular docking studies confirmed strong binding affinities of MKM-derived compounds with key biofilm-related receptor proteins, SpA in S. aureus and Esp in E. faecalis. These findings highlight the significant potential of MKM extract as a novel and effective phytotherapeutic resource for developing strategies to combat biofilm-associated infections.},
}

*Biofilms/drug effects/growth & development
*Enterococcus faecalis/drug effects/growth & development/physiology
*Plant Extracts/pharmacology/chemistry
*Staphylococcus aureus/drug effects/growth & development/physiology
*Murraya/chemistry
*Anti-Bacterial Agents/pharmacology/chemistry
Microbial Sensitivity Tests
Molecular Docking Simulation
Methanol/chemistry
Plant Leaves/chemistry
Humans
Bacterial Lysates

@article {pmid41028113,
year = {2025},
author = {Rafique, A and Baig, N and Naim, A and Rafiq, I},
title = {Utilization of clove and cinnamon essential oils as an alternative to inhibit MDR and biofilm producing E. coli from raw chicken meat.},
journal = {Scientific reports},
volume = {15},
number = {1},
pages = {34079},
pmid = {41028113},
issn = {2045-2322},
mesh = {*Biofilms/drug effects/growth & development ; Animals ; *Escherichia coli/drug effects/isolation & purification/physiology/genetics ; Chickens/microbiology ; *Oils, Volatile/pharmacology ; *Meat/microbiology ; *Cinnamomum zeylanicum/chemistry ; *Anti-Bacterial Agents/pharmacology ; *Drug Resistance, Multiple, Bacterial/drug effects ; *Clove Oil/pharmacology ; Microbial Sensitivity Tests ; Syzygium/chemistry ; Food Microbiology ; },
abstract = {The microbial contamination and spoilage found in chicken meat is responsible for food-borne illnesses and outbreaks leading to hospitalizations. E. coli is the most commonly reported microorganism. The dissemination of bacterial strain with biofilm-formation ability and resistance to antimicrobials at the end of the food chain is a global concern; as well. Eco-friendly and novel means like essential oils are required to break these vicious patterns and ensure the longevity of quality food products. The study aimed to probe the prevalence, pattern of antimicrobial resistance and the biofilm formation ability in E. coli isolated from chicken meat samples. It also explored the antimicrobial and anti-biofilm formation ability of clove and cinnamon essential oils. 150 chicken meat samples from different localities of Karachi, Pakistan were isolated and identified by selective culturing and conventional microbiological techniques. Following antibiogram analysis, antibacterial activity of clove and cinnamon essential oils was evaluated. Putative biofilm production ability was also explored using the test tube, microplate reader, and scanning electron microscopy. Finally, the molecular characterization of potentially strong biofilm producers was done along with exploration of the pathogenic gene (PapC). 49 chicken meat samples out of 150 were contaminated with E. coli. 90% (44 isolates) of E. coli were multidrug resistant. 59.2% (29 isolates) were biofilm producers (BPs). Out of 29 BPs, nine (31%) were strong biofilm producers (SBPs). No significant correlations were observed between antimicrobial resistance and biofilm producing ability of E. coli isolates (p value ≥ 0.05). 40% of SBPs were inhibited when subjected to both clove (MIC: 250 to 500 µL/mL) and cinnamon (MIC: 62.5 µL/mL) EOs. Activity of both neat CO and CinO had no significant difference (p value ≥ 0.05). The identity of 3 SBPs (Strains: AR11E, AR12E and AR22E) were further confirmed by molecular identification (16SrRNA) and SEM revealed potential degradation of the bacterial cells with a reduction in count when treated with CinO and CO. Only one strain (AR22E) was positive for the papC gene. The prevalence of E. coli and strong-biofilm producers in retail chicken meat was not very high; however, the majority of the isolates were multi-drug resistant. Therefore, it is important to keep a tab on the prevalence of these commensal and pathogenic microorganisms in retail chicken meat since they are an exposure site close to the consumer. The use of alternative means like essential oils in poultry, meat and meat-products is a good strategy since they have proven efficacy against pathogenic E. coli.},
}

*Biofilms/drug effects/growth & development
Animals
*Escherichia coli/drug effects/isolation & purification/physiology/genetics
Chickens/microbiology
*Oils, Volatile/pharmacology
*Meat/microbiology
*Cinnamomum zeylanicum/chemistry
*Anti-Bacterial Agents/pharmacology
*Drug Resistance, Multiple, Bacterial/drug effects
*Clove Oil/pharmacology
Microbial Sensitivity Tests
Syzygium/chemistry
Food Microbiology

@article {pmid41028035,
year = {2025},
author = {Choudhary, J and Shariff, M},
title = {Characterization of carbapenem-resistant biofilm forming Acinetobacter baumannii isolates from clinical and surveillance samples.},
journal = {Scientific reports},
volume = {15},
number = {1},
pages = {33892},
pmid = {41028035},
issn = {2045-2322},
mesh = {*Biofilms/drug effects/growth & development ; *Acinetobacter baumannii/drug effects/isolation & purification/genetics/physiology ; Humans ; *Carbapenems/pharmacology ; *Acinetobacter Infections/microbiology/drug therapy/epidemiology ; Anti-Bacterial Agents/pharmacology ; Microbial Sensitivity Tests ; beta-Lactamases/genetics ; Cross Infection/microbiology ; Drug Resistance, Multiple, Bacterial/genetics ; Virulence Factors/genetics ; },
abstract = {Acinetobacter baumannii (A. baumannii) is an important nosocomial pathogen responsible for a wide range of human infections. The emergence of multidrug resistance (MDR) causes life-threatening nosocomial infections. Also, the formation of biofilm helps it survive on abiotic surfaces and is transferred through healthcare workers, thereby causing nosocomial infections. Hence, we study the current antibiotic resistance patterns and virulence factors in our clinical and colonizing isolates. A total of 92 isolates (44 colonizing and 48 clinical) of A. baumannii were included in the study. Antibiotic susceptibility testing was performed by VITEK 2. Biofilm formation was assessed by the tissue culture plate method. Polymerase chain reaction (PCR) for oxacillinases, MBLs and biofilm-associated genes were performed. Meropenem resistance was found in 42 (87.5%) of the clinical and 44 (97.7%) of the colonizing isolates. A strongly adherent biofilm was produced by 11 (22.91%) of the clinical and 12 (27.27%) of the colonizing isolates. Biofilm-associated genes, ompA, bap and csuE were present in 45 (93.7%), 47 (97.9%) and 44 (91.6%) of the clinical isolates, respectively and in all the colonizing isolates. blaOXA23-like was more prevalent in colonizing than clinical isolates. blaOXA-58-like and blaOXA-24-like were present in very few isolates. The presence of metallo beta-lactamase (MBLs) was observed to be lower than oxacillinases. NDM1 was present in 15.29%, SIM in 27%, GIM in 14.11%, VIM in 32.9%, SPM in 5.8%, and IMP in 1.2% of the meropenem-resistant isolates. Carbapenem resistance (XDR) is increasing in A.baumannii. Biofilm formation is an important virulence factor responsible for its survival in the hospital environment and causes nosocomial infections. Biofilm-producing isolates were also found to be carbapenem-resistant. Strict disinfection procedures are to be followed to prevent its spread in the hospital.},
}

*Biofilms/drug effects/growth & development
*Acinetobacter baumannii/drug effects/isolation & purification/genetics/physiology
Humans
*Carbapenems/pharmacology
*Acinetobacter Infections/microbiology/drug therapy/epidemiology
Anti-Bacterial Agents/pharmacology
Microbial Sensitivity Tests
beta-Lactamases/genetics
Cross Infection/microbiology
Drug Resistance, Multiple, Bacterial/genetics
Virulence Factors/genetics

@article {pmid41026297,
year = {2025},
author = {Nebo, UC and Arotupin, DJ and Olalemi, AO},
title = {Biodegradation of heavy petroleum polycyclic aromatic hydrocarbons (PAHs) in polluted soil by biofilm-forming Bacillus tropicus UCB and Pseudomonas aeruginosa SYLI isolated from crude oil-contaminated sludge.},
journal = {Biodegradation},
volume = {36},
number = {5},
pages = {97},
pmid = {41026297},
issn = {1572-9729},
mesh = {*Pseudomonas aeruginosa/metabolism/isolation & purification/physiology/genetics ; *Biofilms/growth & development ; *Polycyclic Aromatic Hydrocarbons/metabolism ; Biodegradation, Environmental ; *Petroleum/metabolism ; *Soil Pollutants/metabolism ; *Sewage/microbiology ; *Bacillus/metabolism/isolation & purification/physiology ; Soil Microbiology ; },
abstract = {Crude oil pollution poses a threat to soil ecosystems, particularly in oil-producing regions. This study assessed the biodegradation potential of biofilm-forming Bacillus tropicus UCB and Pseudomonas aeruginosa SYLI isolated from crude oil sludge. Sludge samples were seasonally collected and bacterial counts determined using standard methods while microbial enrichment was conducted in mineral salt medium containing 1% crude oil. Biofilm formation was assessed using Congo red agar and microplate assays. Isolates were identified through cultural, biochemical, and 16S rDNA analysis. Dose-response toxicity test examined degradation across 1%, 3%, 7%, and 10% crude oil concentrations, while PAHs degradation in soil microcosm was analysed using GC-MS. Seasonal variations significantly influenced bacterial populations, with highest count (1.53 × 10[8] CFU/mL) in the dry season and the least 3.17 × 10[6] CFU/mL) during wet season. Optical density peaked at 2.86 nm in enrichment III. Results revealed molecular identities of the isolates as B. tropicus UCB and P. aeruginosa SYLI. Both isolates metabolized crude oil from 1 to 10%, with B. tropicus producing 601 mg/L CO2 with 10% at day 12 and P. aeruginosa yielding 616 mg/L with 1% at day 4. In addition, results showed over 99% removal of low molecular weight PAHs and 75% degradation of high molecular weight PAHs, upon biostimulation. These findings highlight complementary strengths of B. tropicus on high-oil loads and P. aeruginosa rapid initial degradation at lower concentrations. This study suggests that biofilm formation coupled with biostimulation may improve bacterial efficiency in bioremediation. It also represents the first in vitro report on PAHs degradation by Bacillus tropicus.},
}

*Pseudomonas aeruginosa/metabolism/isolation & purification/physiology/genetics
*Biofilms/growth & development
*Polycyclic Aromatic Hydrocarbons/metabolism
Biodegradation, Environmental
*Petroleum/metabolism
*Soil Pollutants/metabolism
*Sewage/microbiology
*Bacillus/metabolism/isolation & purification/physiology
Soil Microbiology

@article {pmid41025661,
year = {2025},
author = {Suyama, H and Luu, LDW and Zhong, L and Raftery, MJ and Lan, R},
title = {Proteomic comparison of epidemic Australian Bordetella pertussis biofilm cells.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0171525},
doi = {10.1128/spectrum.01715-25},
pmid = {41025661},
issn = {2165-0497},
abstract = {Bordetella pertussis causes whooping cough, a severe respiratory infectious disease. Studies have compared the currently dominant single nucleotide polymorphism (SNP) cluster I (pertussis toxin promoter allele, ptxP3) and previously dominant SNP cluster II (ptxP1) strains as planktonic cells. Since biofilm formation is linked with B. pertussis pathogenesis in vivo, this study compared the biofilm formation capabilities of representative strains of cluster I and cluster II. Confocal laser scanning microscopy found that the cluster I strain had a denser biofilm structure compared to the cluster II strain. Differences in protein abundance of the biofilm cells were then compared using tandem mass tagging and high-resolution multiple reaction monitoring. In total, 1,453 proteins were identified, of which 40 proteins had significant differential abundance between the two strains in biofilm conditions. Of particular interest was a large increase in the abundance of energy metabolism proteins (cytochrome proteins PetABC and BP3650) in the cluster I strain. When the abundance of these proteins was compared between six additional strains from each cluster, it was found that the protein abundance varied between all strains. These findings suggest that there are large levels of individual proteomic diversity between B. pertussis strains in biofilm conditions despite the highly conserved genome of the species. Overall, this study revealed visual differences in biofilm structure between B. pertussis strains and highlighted strain-specific variation in protein abundance that dominates potential cluster-specific changes that may be linked with the dominance of cluster I strains.IMPORTANCEBordetella pertussis causes whooping cough. The currently circulating cluster I strains have taken over previously dominant cluster II strains. It is important to understand the reasons behind this evolution to develop new strategies against the pathogen. Recent studies have shown that B. pertussis can form biofilms during infection. This study compared the biofilm formation capabilities of a cluster I and a cluster II strain and identified visual differences in the biofilms. The protein abundance between these strains grown in biofilms was compared, and proteins identified with varied abundance were measured with additional strains from each cluster. It was found that despite the highly conserved genetics of the species, there was varied protein abundance between the additional strains. This study highlights that strain-specific variation in protein abundance during biofilm conditions may dominate the cluster-specific changes that may be linked to the dominance of cluster I strains.},
}

@article {pmid41023809,
year = {2025},
author = {Yang, SB and Ku, D and Moon, JH and Lee, JH and Kang, SW and Kim, HK and Kwack, KH},
title = {Complete genome sequence of Streptococcus hominis isolated from subgingival biofilm.},
journal = {BMC genomic data},
volume = {26},
number = {1},
pages = {69},
pmid = {41023809},
issn = {2730-6844},
support = {RS-2024-00358942//National Research Foundation of Korea (NRF)/ ; RS-2023-00280791//National Research Foundation of Korea (NRF)/ ; RS-2024-00404660//Korea Basic Science Institute/ ; },
mesh = {*Biofilms ; *Genome, Bacterial ; *Streptococcus/genetics/isolation & purification/classification ; Humans ; Whole Genome Sequencing ; *Gingiva/microbiology ; Phylogeny ; Adult ; Base Composition ; Molecular Sequence Annotation ; RNA, Ribosomal, 16S/genetics ; },
abstract = {OBJECTIVE: Streptococcus hominis is a recently described species within the genus Streptococcus, yet its genomic characteristics remain poorly understood, particularly in the context of the oral microbiome. Previously, only two complete genomes from non-oral sources were available. To address this gap, we sequenced and analyzed S. hominis strain KHUD_010, isolated from the subgingival biofilm of a healthy Korean adult.

DATA DESCRIPTION: Genomic DNA from KHUD_010 was extracted and confirmed as S. hominis by 16 S rRNA gene sequencing. Whole-genome sequencing using the PacBio Sequel II platform generated 135,974 HiFi reads (N50: 10,345 bp). De novo assembly with SMRT Link v11.0 produced a single circular chromosome of 1,883,665 bp with 39.04% GC content. Annotation via the NCBI Prokaryotic Genome Annotation Pipeline predicted 1,793 protein-coding genes, four rRNA operons (5 S, 16 S, 23 S), and 120 tRNAs. BUSCO analysis showed 99.1% completeness. Comparative genomics with NSJ-17 and UMB6992B revealed 1,416 core, 223 dispensable, and 398 strain-specific gene clusters. KHUD_010 harbored 18 unique gene clusters comprising 20 genes, mostly assigned to COG category L (replication, recombination, repair). This high-quality genome expands the genomic landscape of S. hominis and provides a valuable reference for future studies on oral microbiome diversity and host adaptation.},
}

*Biofilms
*Genome, Bacterial
*Streptococcus/genetics/isolation & purification/classification
Humans
Whole Genome Sequencing
*Gingiva/microbiology
Phylogeny
Adult
Base Composition
Molecular Sequence Annotation
RNA, Ribosomal, 16S/genetics

@article {pmid41020670,
year = {2025},
author = {Lordejani, AS and Tajbakhsh, E and Khamesipour, F and Momtaz, H and Momeni Shahraki, M},
title = {Prevalence, Antibiotic Resistance, and Biofilm Formation of Proteus mirabilis in Dairy Products: Implications for Veterinary and Public Health.},
journal = {Veterinary medicine and science},
volume = {11},
number = {6},
pages = {e70600},
pmid = {41020670},
issn = {2053-1095},
mesh = {*Biofilms/growth & development ; *Proteus mirabilis/physiology/drug effects ; Animals ; *Dairy Products/microbiology ; Iran/epidemiology ; *Drug Resistance, Bacterial ; *Anti-Bacterial Agents/pharmacology ; Cattle ; Prevalence ; Public Health ; Milk/microbiology ; Sheep ; Goats ; *Food Microbiology ; Drug Resistance, Multiple, Bacterial ; },
abstract = {Dairy products are essential components of human and animal nutrition, providing vital nutrients such as proteins, vitamins and minerals. However, their susceptibility to microbial contamination, particularly by pathogens like Proteus mirabilis, poses significant risks to both animal and public health. This study investigated the prevalence, antibiotic resistance patterns and biofilm-forming ability of P. mirabilis in various dairy products, with a focus on raw milk, in Shahrekord, Iran. A total of 480 samples, including raw cow, goat and sheep milk, as well as cheese, yogurt, cream, curd and a traditional Iranian fermented yogurt-based beverage (doogh), were analysed under controlled laboratory conditions. The findings revealed a 12.29% prevalence rate of P. mirabilis, with raw cow's milk showing the highest contamination rate (21.66%). Among the isolates, 88.12% were capable of forming biofilms, and 71.15% exhibited strong biofilm production. Antibiotic susceptibility testing identified a high prevalence of multidrug-resistant strains, with the highest resistance rates observed for cotrimoxazole (59.32%) and gentamicin (50.84%). In addition, 25.42% of the isolates were identified as extended-spectrum beta-lactamase (ESBL) producers, with blaCTX-M being the most prevalent resistance gene. A significant correlation was found between biofilm-forming ability and the presence of antibiotic resistance genes, highlighting the dual challenges of microbial persistence and antimicrobial resistance. These findings emphasize the need for improved hygiene practices in dairy production, targeted biofilm-disrupting strategies and enhanced surveillance programs to mitigate risks to both animal and public health. This study provides critical insights for veterinary professionals and policymakers to develop effective interventions aimed at reducing contamination and combating antimicrobial resistance in dairy products.},
}

*Biofilms/growth & development
*Proteus mirabilis/physiology/drug effects
Animals
*Dairy Products/microbiology
Iran/epidemiology
*Drug Resistance, Bacterial
*Anti-Bacterial Agents/pharmacology
Cattle
Prevalence
Public Health
Milk/microbiology
Sheep
Goats
*Food Microbiology
Drug Resistance, Multiple, Bacterial

@article {pmid41020610,
year = {2025},
author = {Shan, W and Du, F and Zhang, H and Zhang, J and Hu, X and Fan, X and Li, W},
title = {D-histidine exhibited anti-biofilm activity against Aggregatibacter actinomycetemcomitans.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0121625},
doi = {10.1128/spectrum.01216-25},
pmid = {41020610},
issn = {2165-0497},
abstract = {Aggregatibacter actinomycetemcomitans is a key pathogen implicated in periodontitis. The bacterium in biofilms exhibits significant resistance to antimicrobial agents and host immune responses compared to its planktonic form, posing a major challenge for periodontal therapy. Recently, D-histidine has emerged as a promising anti-biofilm agent against Pseudomonas aeruginosa infections. However, its potential application in the oral field remains unexplored. This study investigated the anti-biofilm effect of D-histidine on A. actinomycetemcomitans and examined its influence on the expression of virulence factor genes to elucidate possible underlying mechanisms. Our results demonstrated that D-histidine inhibited biofilm formation and disrupted established biofilms in a concentration-dependent manner, without affecting bacterial growth. Furthermore, D-histidine downregulated the expression of virulence factors by inhibiting quorum sensing (QS)-related genes. Notably, combining D-histidine with antibiotics, such as amoxicillin, minocycline, and metronidazole, synergistically enhanced biofilm eradication and enabled the use of lower antibiotic dosages. These findings support the further evaluation of D-histidine as a potential anti-biofilm agent in the treatment of periodontitis.IMPORTANCEThe increasing prevalence of antibiotic-resistant A. actinomycetemcomitans biofilms posed a significant challenge in periodontitis management. This study demonstrated that D-histidine effectively targeted A. actinomycetemcomitans biofilms by disrupting structural integrity and suppressing virulence gene expression, without exerting bactericidal effects that could promote resistance development. Notably, D-histidine showed potent synergy with minocycline, significantly enhancing biofilm eradication while potentially enabling reduced antibiotic dosages. These findings established D-histidine as a promising adjunctive therapeutic agent, addressing the urgent need for novel approaches to overcome biofilm-associated antibiotic tolerance in periodontal treatment.},
}

@article {pmid41018127,
year = {2025},
author = {Adhikari, S and Khanal, S and Marasini, A and Panthi, P and Adhikari, A and Luitel, H and Pandeya, YR},
title = {Antimicrobial resistance and biofilm-forming ability in Staphylococcus aureus causing clinical bovine mastitis in Chitwan, Nepal.},
journal = {Veterinary and animal science},
volume = {30},
number = {},
pages = {100508},
pmid = {41018127},
issn = {2451-943X},
abstract = {This cross-sectional study, conducted from August to December 2024, investigated the antimicrobial resistance and biofilm-forming abilities of Staphylococcus aureus from clinical bovine mastitis in Chitwan, Nepal. Out of 134 California Mastitis Test-positive milk samples, 32 (23.9%) were confirmed as S. aureus by biochemical tests and species-specific nuc gene PCR. Antimicrobial susceptibility testing of the 32 isolates against 12 antibiotics revealed high resistance rates, particularly to ampicillin (78.1%), nalidixic acid (75.0%), and enrofloxacin (62.5%). The prevalence of multidrug resistance (MDR), defined as resistance to ≥3 antibiotic classes, was alarmingly high, with 27 (84.4%) isolates classified as MDR. Presumptive methicillin-resistant S. aureus (MRSA), detected via cefoxitin resistance, was identified in 14 (43.8%) isolates, all of which were also MDR. Biofilm-forming abilities were assessed qualitatively and quantitatively, with 5 (15.6%) isolates classified as strong biofilm producers. Fisher's exact test revealed no significant association between biofilm formation and overall MDR status (p > 0.05). However, a statistically significant correlation was found between strong biofilm formation and high-level MDR (resistance to ≥ 4 classes) (p < 0.05), as well as between strong biofilm formation and presumptive MRSA status (p < 0.01). These findings highlight the co-existence of high-level resistance and strong virulence phenotypes in S. aureus from bovine mastitis in Nepal, underscoring the urgent need for robust antimicrobial stewardship, enhanced surveillance, and the development of strategies to mitigate biofilm-associated treatment failures.},
}

@article {pmid41016948,
year = {2025},
author = {Kim, MJ and Kim, E and Mangal, U and Seo, JY and Cha, JY and Lee, KJ and Choi, YJ and Kwon, JS and Yu, HS and Choi, SH},
title = {Biofilm-resistant zwitterionic resin-based adhesive for orthodontic bracket-tooth interfaces: an in vitro evaluation.},
journal = {Clinical oral investigations},
volume = {29},
number = {10},
pages = {478},
pmid = {41016948},
issn = {1436-3771},
support = {RS-2023-00249723//Basic Science Research Program through the National Research Foundation (NRF) funded by the Ministry of Education/ ; S3366101//the Ministry of SMEs and Startups (MSS, Korea)/ ; RS-2023-00255335//the Ministry of Science and ICT, the Ministry of Trade, Industry and Energy, the Ministry of Health & Welfare, the Ministry of Food and Drug Safety/ ; 2021R1A2C2091260//National Research Foundation of Korea (NRF)/ ; },
mesh = {*Biofilms/drug effects ; *Orthodontic Brackets/microbiology ; Methacrylates/chemistry ; Microscopy, Electron, Scanning ; In Vitro Techniques ; Phosphorylcholine/analogs & derivatives/chemistry ; Shear Strength ; Materials Testing ; *Dental Bonding ; Microscopy, Confocal ; Surface Properties ; *Resin Cements/chemistry ; Betaine/chemistry/analogs & derivatives ; Humans ; *Dental Cements/chemistry ; },
abstract = {OBJECTIVES: An in vitro study to verify the potential effectiveness of an orthodontic adhesive incorporating a polybetaine zwitterionic mixture in preventing biofilm formation.

MATERIALS AND METHODS: 2-methacryloyloxyethyl phosphorylcholine (MPC) and sulfobetaine methacrylate (SBMA), were added to An adhesive at 1 wt% (MS1) And 3 wt% (MS3). The MS0 group had no zwitterions. Flowability measurement, shear bond strength (SBS), the adhesive remnant index (ARI), and the contact angle were assessed. Bracket bonding was performed, and cross-sections were examined using scanning electron microscopy (SEM). Biofilm formation was analyzed using confocal laser scanning microscopy. Statistical analyses were conducted using R software. Wilcoxon tests with the Holm correction were used for non-parametric data, and pairwise t-tests with the Bonferroni correction were used for parametric data. Significance was set at P < 0.0001.

RESULTS: Flow analysis showed no significant differences in the experimental groups compared to the MS0 group (P > 0.05). The SEM images confirmed uniform bonding in all groups. SBS decreased significantly with higher MS content (P < 0.0001), And ARI scores shifted with the addition of zwitterionic mixtures, increasing scores to 1 And 2. MS3 showed a significantly lower contact angle compared to MS0 (P < 0.05). MS3 exhibited reduced biofilm formation and lower fluorescence intensity (P < 0.05).

CONCLUSIONS: Despite reductions in SBS, all adhesives remained at minimum acceptable levels. The 3 wt% zwitterionic adhesive effectively suppressed biofilm formation and may help prevent demineralization.

CLINICAL RELEVANCE: An orthodontic adhesive containing a zwitterionic mixture can satisfy clinical properties and prevent side effects due to biofilm formation.},
}

*Biofilms/drug effects
*Orthodontic Brackets/microbiology
Methacrylates/chemistry
Microscopy, Electron, Scanning
In Vitro Techniques
Phosphorylcholine/analogs & derivatives/chemistry
Shear Strength
Materials Testing
*Dental Bonding
Microscopy, Confocal
Surface Properties
*Resin Cements/chemistry
Betaine/chemistry/analogs & derivatives
Humans
*Dental Cements/chemistry

@article {pmid41016822,
year = {2025},
author = {Shin, J and Park, DH and Jun, W and Park, OJ and Yun, CH and Im, J and Han, SH},
title = {Optimization of the Purification Process for Lactiplantibacillus plantarum Lipoteichoic Acid with Anti-Biofilm Properties against Dental Pathogens.},
journal = {Journal of microbiology and biotechnology},
volume = {35},
number = {},
pages = {e2506045},
doi = {10.4014/jmb.2506.06045},
pmid = {41016822},
issn = {1738-8872},
mesh = {*Biofilms/drug effects ; *Teichoic Acids/pharmacology/isolation & purification/chemistry ; *Lipopolysaccharides/pharmacology/isolation & purification/chemistry ; Streptococcus mutans/drug effects/physiology ; *Anti-Bacterial Agents/pharmacology/isolation & purification ; Porphyromonas gingivalis/drug effects/physiology ; },
abstract = {Biofilms formed by oral pathogens play a critical role in the initiation and development of various dental diseases by enhancing resistance to dental medicaments. Previously, we reported the potent anti-biofilm activity of Lactobacillus lipoteichoic acid (LTA), a major cell wall component of Gram-positive bacteria, against various dental pathogens. Nevertheless, the practical application of LTA as an anti-biofilm agent is limited due to its complex, cost- and labor-intensive purification processes. Therefore, to minimize the purification processes required to obtain LTA with anti-biofilm activity, we isolated LTA from Lactiplantibacillus plantarum (Lp.LTA) and its intermediates following each purification step, and compared their anti-biofilm activity against dental pathogens. The Lp.LTA intermediates underwent sequential purification after butanol extraction and hydrophobic-interaction chromatography, and these were designated as LTA-Butanol and LTA-HIC, respectively. The results of Western blot analysis demonstrated that LTA-HIC has a higher concentration of LTA than LTA-Butanol. Although both LTA-Butanol and LTA-HIC dose-dependently inhibited Streptococcus mutans and Porphyromonas gingivalis biofilm formation, the anti-biofilm activity of LTA-HIC was superior to that of LTA-Butanol and comparable to Lp.LTA. Furthermore, sodium hydroxide treatment of LTA-HIC diminished its anti-biofilm activity, suggesting that LTA is a key component responsible for the anti-biofilm capacity of LTA-HIC. Collectively, these results demonstrated that LTA-HIC serves as an intermediate in the LTA purification, retaining its anti-biofilm properties and offering a viable solution to the challenges associated with the LTA purification process.},
}

*Biofilms/drug effects
*Teichoic Acids/pharmacology/isolation & purification/chemistry
*Lipopolysaccharides/pharmacology/isolation & purification/chemistry
Streptococcus mutans/drug effects/physiology
*Anti-Bacterial Agents/pharmacology/isolation & purification
Porphyromonas gingivalis/drug effects/physiology

@article {pmid41016310,
year = {2025},
author = {Song, Y and Zhang, X and Kong, Z and Zhang, J and Fukushima, T and Hu, S and Virdis, B},
title = {Enhancing the productivity of caproic acid in open culture chain elongation: A comparative study of biofilm systems.},
journal = {Journal of environmental management},
volume = {394},
number = {},
pages = {127395},
doi = {10.1016/j.jenvman.2025.127395},
pmid = {41016310},
issn = {1095-8630},
abstract = {Growing global energy consumption and climate challenges have emphasized the need for biotechnology-based methods to synthesize valuable chemicals. Biological chain elongation (CE) shows great potential for decarbonization by producing valuable biochemicals - specifically medium-chain fatty acids (MCFAs) - from waste streams containing simple short-chain chemical building blocks like acetic acid and ethanol. However, one of the key parameters that impacts the commercial viability of CE, hence its integration in sustainable chemical manufacturing, is the volumetric productivity. In this study, we compared two biofilm systems using commercially available carriers (respectively AnoxK™ Z-200 and K5) with a planktonic system to examine how biofilms enhance the conversion of acetate and ethanol to caproic acid (a medium chain carboxylic acid). The results show that the Z-200 and K5 systems achieved productivity up to 3.46 ± 0.08 g caproate/L/d and 8.1 ± 0.8 g caproate/L/d, respectively, outperforming the planktonic system at 3.02 ± 0.12 g caproate/L/d. Cycle studies further proved the superior performance of the biofilm systems, as shown by short lag-time and fast reaction kinetics. We validated biofilm formation in the CE process through microscopic visualization using scanning electron microscopy (SEM), confocal laser scan microscopy (CLSM), biomass quantification, and analysis of extracellular polymeric substances (EPS). Analysis of the microbial community through 16S rRNA gene sequencing revealed that the biofilm systems were enriched by functional microbes (including Clostridium sensu stricto 12, Bacteroides, Lachnoclostridium, Caproiciproducens, and Proteiniphilum) previously associated with chain elongation microbiomes. The superior performance in the biofilm systems likely stems from improved biomass concentration, enriched functional microbes, and increased EPS production favouring retention of functional taxa. Overall, this work demonstrates how microbial biofilms can improve productivity of MCFA in CE systems, potentially expanding CE applications and improve decarbonization potential.},
}

@article {pmid41016202,
year = {2025},
author = {Zhang, Y and Huang, C and Qiu, Y and Li, R and Liu, J and Du, Y and Li, D},
title = {Synergistic elimination of bacillus Calmette-Guérin biofilm and tissue restoration facilitated by ultrasound-mediated nanoparticles and antioxidants.},
journal = {International immunopharmacology},
volume = {166},
number = {},
pages = {115582},
doi = {10.1016/j.intimp.2025.115582},
pmid = {41016202},
issn = {1878-1705},
abstract = {Biofilm formation in Mycobacterium tuberculosis (MTB) enhances antibiotic resistance by impeding drug penetration and evading host immunity. This poses a significant challenge to conventional drug therapies, highlighting the urgent need for novel treatment strategies to overcome MTB's biofilm-mediated resistance. This study introduces the development of low-intensity ultrasound-mediated levofloxacin (LEV) and catalase (CAT) -loaded PEG-PLGA nanoparticles (LEV@CAT-NPs) for antimicrobial sonodynamic therapy (aSDT), offering an innovative strategy to combat BCG biofilm infection, by utilizing BCG as a model for MTB. N-acetylcysteine (NAC) was supplemented during the latter stages of the treatment process of anti-infection therapy to facilitate the transformation of macrophages to the M2 phenotype and to promote tissue repair. Ultrasound-mediated LEV@CAT-NPs, along with the subsequent addition of NAC not only enhanced repair at the infection site but also led to a progressive resolution of the inflammatory response in tissues. The treatment regimen induced a shift in macrophage polarization towards the M2 phenotype and modulated cytokine expression, decreasing pro-inflammatory while increasing anti-inflammatory cytokines, which contributed to the restoration of redox balance in the infected tissues. This study proposes a novel therapeutic strategy that not only targets drug-resistant MTB but also promotes tissue repair, highlighting its dual role in infection management.},
}

@article {pmid41015993,
year = {2025},
author = {Setiawan, M and Gaffar, M and Wartati, S and Qanitha, A and Sjahril, R},
title = {Correlation of bacterial biofilm profile based on optical density cut-off with clinical severity in patients with chronic suppurative otitis media tubotympanic type.},
journal = {The Medical journal of Malaysia},
volume = {80},
number = {5},
pages = {537-543},
pmid = {41015993},
issn = {0300-5283},
mesh = {Humans ; *Biofilms/growth & development ; *Otitis Media, Suppurative/microbiology ; Cross-Sectional Studies ; Adult ; Male ; Female ; Chronic Disease ; Middle Aged ; Young Adult ; Adolescent ; Severity of Illness Index ; },
abstract = {INTRODUCTION: Chronic suppurative otitis media (CSOM) is a middle ear infection with a high incidence in ear cases, is often recurrent, and causes hearing impairment. Bacteria in the CSOM frequently form biofilms, which enhance antibiotic resistance and contribute to disease progression. The aim of this study was to determine the correlation of bacterial biofilm profiles based on optical density cut-off with the clinical picture of patients with tubotympanic type CSOM.

MATERIALS AND METHODS: This was a cross-sectional study using a descriptive analytical design. The study was conducted at the tertiary teaching hospital of Hasanuddin University and the network hospital in Makassar, Indonesia, from July 2023 to July 2024. The study population consisted of patients with the CSOM tubotympanic type who met the inclusion criteria. Bacterial cultures and biofilm examinations were performed using the tissue culture plate method. Data were analyzed using SPSS® version 28.

RESULTS: A total of 53 patients with the CSOM tubotympanic type were included in this study. The mean age of the patients was 30±14 years. Pseudomonas aeruginosa was the most dominant bacterium (32.1%), with 20 other bacteria, and all these bacteria formed biofilms with either weak or moderate strength. There was a significant association between biofilm formation and nature of secretion (r=0.395, p=0.003). The chronicity of the disease (r=0.407, p=0.002) and the degree of hearing impairment (r=0.294, p=0.032) were also significant. A significant positive association was found between total clinical score and biofilm formation (r=0.429, p=0.001).

CONCLUSION: All bacteria found in the tubotympanic CSOM formed biofilms. The correlation analysis revealed a significant positive relationship between several clinical variables and biofilm formation. The substantial formation of biofilms may account for the fact that patients with elevated scores frequently experience infections that are challenging to manage with conventional antibiotic treatments.},
}

Humans
*Biofilms/growth & development
*Otitis Media, Suppurative/microbiology
Cross-Sectional Studies
Adult
Male
Female
Chronic Disease
Middle Aged
Young Adult
Adolescent
Severity of Illness Index

@article {pmid41015312,
year = {2025},
author = {Wang, J and Zhu, W and Zhang, X and Liu, S and Ma, J and Xing, D},
title = {Charge-specific impacts of polystyrene nanoplastics on acidogenesis and biofilm adaptation in Ethanoligenens harbinense.},
journal = {Bioresource technology},
volume = {},
number = {},
pages = {133389},
doi = {10.1016/j.biortech.2025.133389},
pmid = {41015312},
issn = {1873-2976},
abstract = {Despite increasing awareness of the risks posed by nanoplastics (NPs) to environmental microbes, the charge-specific effects of functionalized NPs on anaerobic acidogenic bacteria remain poorly understood. This study investigated the impact of functionalized polystyrene (PS) NPs on Ethanoligenens harbinense, a model hydrogen-producing anaerobe. The growth, metabolic, and transcriptomic responses of this bacterium to non-functionalized (PS-NPs), amino-modified (PS-NH2), and carboxyl-modified (PS-COOH) variants were examined. Compared with the control group without NPs addition, PS-NH2 exerted the strongest inhibition, reducing hydrogen and ethanol production by 16% and 20%, respectively, while elevating reactive oxygen species (ROS) level by 148%. It also decreased biomass and down-regulated the expression of ribosome- and translation-related genes. In parallel, biofilm adaptation resulted in an 12% increase in polysaccharide. PS-COOH enhanced biofilm reinforcement with a 21% increase in polysaccharides and up-regulation of bapA and membrane transporter-related genes. Overall, PS-NH2 induced broad transcriptional changes, particularly in pathways related to the phosphotransferase system (PTS), ATP-binding cassette (ABC) transporters, genetic information processing, and signaling/regulatory systems in E. harbinense. These findings provide new insights into how surface charge modifications of NPs affect anaerobic bacterial metabolism and underscore their potential environmental risks.},
}

@article {pmid41015013,
year = {2025},
author = {Sun, H and Li, W and Zhang, S and Yuan, L and Wang, D and Sun, J and Wang, H},
title = {Differential profiles of antibiotic resistance genes in activated sludge and biofilm in wastewater treatment plants.},
journal = {Journal of hazardous materials},
volume = {498},
number = {},
pages = {139955},
doi = {10.1016/j.jhazmat.2025.139955},
pmid = {41015013},
issn = {1873-3336},
abstract = {Wastewater treatment plants (WWTPs) serve as significant sources of antibiotic resistance genes (ARGs) in natural water bodies, with activated sludge and biofilm being the two most critical biological treatment processes in WWTPs. A systematic comparison of ARG composition in these two processes is essential for optimizing the design and operation of wastewater treatment systems. This study collected samples from 16 WWTPs, including one year of longitudinal monitoring data from a full-scale facility and encompassing five biofilm types. The high-throughput sequencing results revealed that the relative abundance of ARGs in activated sludge was significantly higher than in biofilms, with average relative abundances of 2075.05 ppm and 1288.78 ppm, respectively. We also identified plasmids and microbial community structure as key factors contributing to the differences in ARG composition between activated sludge and biofilm. Plasmids primarily influenced the ARGs associated with enzymatic modification mechanisms, while the microbial community structure mainly impacted the abundance of ARGs, particularly through its effect on Bacteroidia. This structural influence was particularly pronounced on ARGs related to enzymatic inactivation, enzymatic modification, efflux pumps, target modification, and target protection mechanisms. These findings provide valuable insights for improving the management of ARGs in WWTPs and contribute to the development of strategies for mitigating ARG proliferation in wastewater treatment systems.},
}

@article {pmid41014987,
year = {2025},
author = {Zhu, B and Pan, X and Guo, H and Chen, Y and Wen, Q},
title = {'Inner membrane - outer gel' PEDOT:PSS/MXene composite material enhances the extracellular electron transfer process by promoting biofilm growth.},
journal = {Biosensors & bioelectronics},
volume = {291},
number = {},
pages = {118021},
doi = {10.1016/j.bios.2025.118021},
pmid = {41014987},
issn = {1873-4235},
abstract = {Against the backdrop of surging global energy demand, microbial fuel cells (MFCs) have garnered significant attention for their potential in green energy conversion. To solve the problem of low efficiency of long-distance electron transfer by microorganisms at the anode interface of MFCs, we propose a novel 'inner membrane - outer gel' PEDOT:PSS/MXene dual-phase hydrogel electrode. The electrode is fabricated via electrochemical polymerization, which further enables it to use a flexible heterogeneous interface to promote three-dimensional (3D) biofilm generation. Experimental results indicate that the composite anode exhibits a charge transfer resistance (Rct) as low as 4.71 Ω, with a maximum power density of 4.55 ± 0.17 W m[-2], which is 1.8 times the maximum power density of a pure hydrogel (only PEDOT:PSS hydrogel). Furthermore, 16S rRNA sequencing revealed that the relative abundance of Geobacter increased to 62.22 %, indicating a significant enrichment of electrogenic microorganisms. Molecular docking simulations further elucidated the electrostatic complementarity and hydrogen bonding interactions between PEDOT and the OmcZ protein, providing theoretical support for efficient electron transfer between conductive nanowires and electrodes within biofilms. Overall, this study provides both experimental and theoretical evidence for the feasibility of the 'inner membrane - outer gel' electrode in enhancing MFC performance, offering new insights into the design of high-performance conductive bioelectrodes in microbial energy conversion devices.},
}

@article {pmid41012569,
year = {2025},
author = {do Nascimento, JL and da Costa, MCV and de Macêdo, LF and de Macêdo, LHC and de Moura, RO and de Mélo, TJA and da Rocha, WRV and de Melo Costa, ACF and Soares-Sobrinho, JL and Silva, DTCD},
title = {Laponite[®]-Based Smart Hydrogels for Sustained Topical Delivery of Silver Sulfadiazine: A Strategy for the Treatment of Contaminated or Biofilm-Forming Wounds.},
journal = {Pharmaceutics},
volume = {17},
number = {9},
pages = {},
doi = {10.3390/pharmaceutics17091234},
pmid = {41012569},
issn = {1999-4923},
support = {382074/2025-4.//National Council for Scientific and Technological Development/ ; grant #01/2025//Universidade Estadual da Paraíba/ ; },
abstract = {Background/Objectives: Silver sulfadiazine (AgSD) is widely used in the topical treatment of burns and infected wounds, but its conventional formulations present drawbacks such as poor water solubility, the need for multiple daily applications, and patient discomfort. To overcome these limitations, this study aimed to develop and evaluate Laponite[®] (LAP)-based hydrogels loaded with AgSD for controlled release and enhanced antimicrobial and antibiofilm efficacy, offering a promising alternative for the treatment of contaminated or biofilm-forming wounds. Methods: Laponite[®]-based hydrogels containing 1% and 1.2% AgSD (LAP@AgSD) were prepared using a one-pot method. The formulations were characterized rheologically, thermally, and structurally. In vitro drug release was assessed using Franz diffusion cells, and mathematical modeling was applied to determine release kinetics. Antibacterial and antibiofilm activities were evaluated against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa using standardized microbiological methods. Results: LAP@AgSD hydrogels exhibited pseudoplastic behavior, high structural integrity, and enhanced thermal stability. In vitro release assays revealed a sustained release profile, best fitted by the Weibull model, indicating diffusion-controlled mechanisms. Antibacterial assays demonstrated concentration-dependent activity, with LAP@AgSD 1.2% showing superior efficacy over LAP@AgSD 1% and comparable performance to the commercial silver sulfadiazine cream (CC-AgSD). Biofilm inhibition was significant for all formulations, with CC-AgSD 1% exhibiting the highest immediate activity, while LAP@AgSD 1.2% provided sustained antibiofilm potential. Conclusions: LAP-based hydrogels are promising smart delivery systems for AgSD, combining mechanical robustness, controlled drug release, and effective antibacterial and antibiofilm activities. These findings support their potential use in topical therapies for infected and chronic wounds, particularly where biofilm formation is a challenge.},
}

@article {pmid41011755,
year = {2025},
author = {Liang, TT and Wen, JQ and Chen, GP and Wang, R and Xu, J and Chen, WY},
title = {Identification and Validation of Promising Targets and Inhibitors of Biofilm Formation in Pseudomonas aeruginosa: Bioinformatics, Virtual Screening, and Biological Evaluation.},
journal = {Pathogens (Basel, Switzerland)},
volume = {14},
number = {9},
pages = {},
doi = {10.3390/pathogens14090855},
pmid = {41011755},
issn = {2076-0817},
support = {CPA:2023ZYS09//Guangdong Pharmaceutical Association/ ; },
mesh = {*Biofilms/drug effects/growth & development ; *Pseudomonas aeruginosa/drug effects/physiology/genetics ; Computational Biology/methods ; *Anti-Bacterial Agents/pharmacology ; Molecular Docking Simulation ; Drug Evaluation, Preclinical ; Bacterial Proteins/metabolism/antagonists & inhibitors/genetics ; Humans ; Protein Interaction Maps ; Clarithromycin/pharmacology ; Azithromycin/pharmacology ; },
abstract = {Pseudomonas aeruginosa, a member of the "ESKAPE" group of bacterial pathogens, exhibits biofilm-forming capacity, a key factor contributing to its resistance to conventional antibiotics and posing significant challenges in clinical treatment. To develop more effective therapeutics against such infections, identifying potential drug targets through bioinformatics analysis is essential. Consequently, we utilized data from the GEO database to investigate differentially expressed genes between planktonic and biofilm groups, and identified drug targets through the construction of a protein-protein interaction (PPI) network and the cytoHubba algorithm. Inhibitors targeting this protein were identified through molecular docking screening of the FDA-approved drug library, and their anti-biofilm activity was validated in vitro. Through bioinformatics analysis, we identified GacS as the drug target in this study for treating biofilm-related infections. Virtual screening revealed that oxidized glutathione (GSSG) and arformoterol tartrate (ARF) are both capable of tightly binding to GacS and demonstrating good stability. In vitro experiments further confirmed that both GSSG and ARF demonstrated anti-biofilm activity, particularly when combined with azithromycin (AZM) or clarithromycin (CAM), significantly enhancing the biofilm inhibition effects of these antibiotics. This combination therapy offers a new and innovative strategy to combat biofilm-associated infections, showcasing the potential of GacS inhibitors in clinical applications. In conclusion, GSSG and ARF may serve as effective GacS inhibitors, and their combination with AZM or CAM could provide a novel approach for treating biofilm-related infections, paving the way for more effective treatment options.},
}

*Biofilms/drug effects/growth & development
*Pseudomonas aeruginosa/drug effects/physiology/genetics
Computational Biology/methods
*Anti-Bacterial Agents/pharmacology
Molecular Docking Simulation
Drug Evaluation, Preclinical
Bacterial Proteins/metabolism/antagonists & inhibitors/genetics
Humans
Protein Interaction Maps
Clarithromycin/pharmacology
Azithromycin/pharmacology

@article {pmid41011744,
year = {2025},
author = {Bridi, V and do Prado, DPG and Ferreira, SRR and Pedretti, CP and Filho, EGP and Santos, WGD and Rezende, HHA},
title = {Antimicrobial Resistance and Biofilm Formation in Bacterial Species Isolated from a Veterinary Hospital.},
journal = {Pathogens (Basel, Switzerland)},
volume = {14},
number = {9},
pages = {},
doi = {10.3390/pathogens14090845},
pmid = {41011744},
issn = {2076-0817},
mesh = {*Biofilms/growth & development ; Microbial Sensitivity Tests ; Hospitals, Animal ; *Anti-Bacterial Agents/pharmacology ; Animals ; *Drug Resistance, Bacterial ; Drug Resistance, Multiple, Bacterial ; Staphylococcus/drug effects/isolation & purification/genetics ; *Bacteria/drug effects/isolation & purification/genetics ; },
abstract = {Micro-organisms are abundant in nature and can also be found in hospital settings, causing high rates of infections. This study aimed to identify bacteria isolated from a veterinary hospital, as well as to perform antimicrobial susceptibility testing using the disk diffusion method (Kirby-Bauer), biofilm production tests using 96-well polystyrene microtiter plates and crystal violet dye, and genetic analysis of the ica operon of Staphylococcus isolates. Three collections were made from eleven surfaces and objects in the hospital's non-critical areas (general areas) and critical areas (surgical center), totaling thirty-three samples. A total of 66 different bacterial isolates were obtained, with 77% (51/66) Gram-positive and 23% (29/66) Gram-negative. Resistance profiles were found for multidrug-resistance (MDR), methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE), and other unidentified species of methicillin-resistant coagulase-negative (MRCNS) and extended-spectrum beta-lactamase (ESBL), as well as biofilm production rates of 57% (38/66) of the isolates. Analysis of the operon genes for Staphylococcus sp. showed divergence in some samples when compared to the phenotypic test performed. In summary, there is a high presence of micro-organisms with resistance and virulence factors spread throughout the various areas of the veterinary hospital.},
}

*Biofilms/growth & development
Microbial Sensitivity Tests
Hospitals, Animal
*Anti-Bacterial Agents/pharmacology
Animals
*Drug Resistance, Bacterial
Drug Resistance, Multiple, Bacterial
Staphylococcus/drug effects/isolation & purification/genetics
*Bacteria/drug effects/isolation & purification/genetics

@article {pmid41011529,
year = {2025},
author = {De Plano, LM and Iaconis, A and Papasergi, S and Mediati, F and Caruso, D and Guglielmino, SPP and Franco, D},
title = {Role of NaCl and Glutamine on Biofilm Production from Pseudomonas aeruginosa.},
journal = {Microorganisms},
volume = {13},
number = {9},
pages = {},
doi = {10.3390/microorganisms13092198},
pmid = {41011529},
issn = {2076-2607},
abstract = {Pseudomonas aeruginosa is an opportunistic pathogen capable of forming antibiotic-resistant biofilms, contributing to persistent infections and treatment failure. Environmental factors such as osmolarity and nutrient availability are known to influence biofilm formation and virulence. In this study, we investigated the effects of NaCl depletion and glutamine supplementation on biofilm production in three P. aeruginosa strains: the laboratory strain ATCC 27853 and two clinical isolates with distinct antibiotic resistance profiles and phenazine production patterns (P. aeruginosa Pr, pyorubrin-producing, and P. aeruginosa Pc, pyocyanin-producing). Bacteria were cultured in standard Luria-Bertani (LB) medium, LB without NaCl, and LB in which yeast extract was replaced by glutamine. For each strain and condition, we assessed growth kinetics, phenazine production, and biofilm formation. Biofilm development was quantified via XTT assays and compared to secondary metabolite profiles. NaCl removal did not substantially affect growth, whereas glutamine supplementation reduced growth, especially in the laboratory strain. Both conditions modulated secondary metabolite production and biofilm formation in a strain-specific manner. In P. aeruginosa ATCC 27853, NaCl depletion significantly increased pyoverdine, pyocyanin, and QS gene expression, while biofilm formation showed significant differences only at 72 h; in contrast, glutamine supplementation affected only pyoverdine. A similar trend was observed in the clinical strain P. aeruginosa Pc, although NaCl depletion did not significantly impact pyoverdine production but already enhanced biofilm formation at 48 h. In P. aeruginosa Pr, only glutamine appeared to alter the considered parameters, increasing pyoverdine production while reducing pyocyanin and biofilm levels, although the absence of NaCl also negatively impacted biofilm formation. These findings highlight the impact of osmotic and nutritional signals on P. aeruginosa virulence traits.},
}

@article {pmid41011426,
year = {2025},
author = {Shen, J and Li, Y and Miao, H},
title = {The PP2A Catalytic Subunit PPH21 Regulates Biofilm Formation and Drug Resistance of Candida albicans.},
journal = {Microorganisms},
volume = {13},
number = {9},
pages = {},
doi = {10.3390/microorganisms13092093},
pmid = {41011426},
issn = {2076-2607},
abstract = {Candida albicans (C. albicans) biofilms exhibit enhanced resistance to conventional antifungal agents; however, the underlying pathogenic mechanisms warrant deeper exploration. Protein phosphatase 2A (PP2A), especially its catalytic activity, is crucial for maintaining physiological balance. This study focused on the role of the PP2A catalytic subunit coding gene PPH21 in biofilm formation and drug resistance of C. albicans. The mutant strain (pph21Δ/Δ) was generated and identified. The oxidative stress was detected by the reactive oxygen species (ROS) and mitochondrial membrane potential (MMP). The autophagic activity was evaluated, and the autophagosomes were observed by transmission electron microscopy (TEM). The biofilm formation was measured by XTT reduction assay, crystal violet (CV) staining, and scanning electron microscopy (SEM). The susceptibility to antifungal agents was examined by XTT reduction assay and spot assay. Additionally, the antioxidant N-acetylcysteine (NAC) was applied to clarify the regulatory effect of C. albicans autophagy on oxidative stress. The pathogenicity of PPH21 in oral C. albicans infection was evaluated through in vivo experiments. We found that PPH21 deletion led to increased oxidative stress and autophagic activities, but it can be reversed by the application of NAC. Moreover, PPH21 deletion also impaired the biofilm formation ability and reduced resistance to antifungal agents. Our findings revealed that PPH21 is involved in both virulence and stress adaptation of C. albicans.},
}

@article {pmid41011382,
year = {2025},
author = {Jotic, A and Cirkovic, I and Bozic, D and Savic Vujovic, K and Milovanovic, J and Folic, M and Trivic, A and Cvorovic, L and Radivojevic, N},
title = {Antibiofilm Effects of N-Acetyl Cysteine on Staphylococcal Biofilm in Patients with Chronic Rhinosinusitis.},
journal = {Microorganisms},
volume = {13},
number = {9},
pages = {},
doi = {10.3390/microorganisms13092050},
pmid = {41011382},
issn = {2076-2607},
support = {451-03-65/2024-03/200110//Ministry of Science, Technological Development and Innovation, the Republic of Serbia/ ; },
abstract = {Staphylococcal bacterial biofilm plays an important role in the pathogenesis and bacterial persistence of chronic rhinosinusitis. N-acetyl cysteine (NAC) has an inhibitory role in biofilm formation, suppressing adhesion and matrix production or favoring dispersal of preformed biofilm. The aim of this study was to examine the in vitro effect of NAC on Staphylococcal biofilm formation by bacterial strains isolated from tissue samples of patients with chronic rhinosinusitis with or without nasal polyps (CRSwNP and CRSsNP). Prospective study included 75 patients with CRS. The biofilm-forming capacity of isolated strains was detected by microtiter-plate method and the effects of sub-inhibitory (1/2x, 1/4x, and 1/8x minimal inhibitory concentration, MIC) and supra-inhibitory minimal concentrations (2x, 4x, and 8xMIC) of NAC on biofilm production were investigated. Staphylococcal bacterial strains were isolated in 54 (72%) patients, and the most frequently isolated species were Staphylococcus aureus (40.7%). Coagulase-negative Staphylococci species were weak producers of biofilm, while S. aureus was a strong biofilm producer. Concentration of 3.1 mg/mL (1/2 MIC) was sufficient to completely prevent biofilm formation in 77.8% of the isolates, where 49.6 mg/mL (8xMIC) led to the complete eradication of formed biofilm in 81.5% of the isolates. The subinhibitory and eradication effects were dose- and strain-dependent. There were no significant differences in MIC values between isolates from patients with CRSwNP and CRSsNP isolates. NAC proved to be effective in inhibiting biofilm formation and reducing formed biofilm by Staphylococcal isolates from patients with CRS. A comparable antibiofilm effect was exhibited in both phenotypes of CRS, indicating that NAC's antibiofilm activity was independent of the underlying clinical phenotype, and more targeted on biofilm matrix components.},
}

@article {pmid41011381,
year = {2025},
author = {Hessling, M and Meulebroeck, W and Alsanius, B},
title = {A Collection and Analysis of Simplified Data for a Better Understanding of the Complex Process of Biofilm Inactivation by Ultraviolet and Visible Irradiation.},
journal = {Microorganisms},
volume = {13},
number = {9},
pages = {},
doi = {10.3390/microorganisms13092048},
pmid = {41011381},
issn = {2076-2607},
support = {BW8-1108//Ministerium für Wirtschaft, Arbeit und Tourismus Baden-Württemberg/ ; },
abstract = {Biofilms are communities of microorganisms that pose a problem in many areas, including the food industry, drinking water treatment, and medicine, because they can contain pathogens and are difficult to eliminate. For this reason, the possibility of biofilm reduction by ultraviolet (UV) or visible light was investigated using data from published reports. Results for different applications, spectral ranges, and microorganisms were compared by performing MANOVA tests. Approximately 140 publications were found that dealt with the irradiation of water or surfaces for biofilm reduction or reduction in biofilm formation. Irradiation of surfaces with UV or visible light in the spectral range 200-525 nm had a positive effect on biofilm reduction and reduction in biofilm formation, although the results for irradiation of water were conflicting. Most investigations were carried out on P. aeruginosa biofilms, but other Gram-positive and Gram-negative bacteria, as well as some fungi and their biofilm sensitivities to irradiation, were also analyzed. Limited data were available for the UVB (280-315 nm) and UVA (315-400 nm) range. Most experiments to date have been carried out in the UVC (100-280 nm) or in the visible violet/blue spectral (400-500 nm) range, with the UVC range being 2-3 orders of magnitude more efficient in terms of applied irradiation dose. Other quantitative statements were difficult to make as the results from the different working groups were highly scattered. Irradiation can reduce the microorganisms in biofilms but does not completely remove biofilms. New biofilm formation can at least be delayed by surface irradiation. Whether it is also possible to prevent the formation of new biofilms in the long term is open to question. Which irradiation wavelengths are optimal for anti-biofilm measures is also still unclear.},
}

@article {pmid41011372,
year = {2025},
author = {Furnica, DT and Falkenstein, J and Dittmer, S and Steinmann, J and Rath, PM and Kirchhoff, L},
title = {Human-like Biofilm Models to Study the Activity of Antifungals Against Aspergillus fumigatus.},
journal = {Microorganisms},
volume = {13},
number = {9},
pages = {},
doi = {10.3390/microorganisms13092040},
pmid = {41011372},
issn = {2076-2607},
abstract = {Aspergillus fumigatus is an opportunistic filamentous fungus that primarily affects the respiratory tract of the human body. Depending on its host's immune response, the pathogen can cause invasive pulmonary aspergillosis (IPA). Biofilm formation by A. fumigatus increases virulence and resistance against antifungals and immune response and is one important factor in IPA development. Here, two human-like models, precision cut lung slices (PCLS) and a biofilm co-culture model, have been developed to test the anti-biofilm activity of voriconazole, amphotericin B, as well as luliconazole against A. fumigatus. In both assays, metabolically active A. fumigatus biofilms were examined at different biofilm developmental stages using an XTT assay. A decrease in the metabolic activity of the fungal biofilms was detected for each of the tested agents in both assays. Significant anti-biofilm effects exist against early-stage biofilm in the co-culture model. In the PCLS assay, amphotericin B showed the strongest inhibition after 24 h. In conclusion, the applied PCLS ex vivo model can be used to study the property and activity of certain antifungal compounds against Aspergillus biofilm. With its close resemblance to human conditions, the PCLS model has the potential for improving the current understanding of biofilm treatments in laboratory settings.},
}

@article {pmid41011371,
year = {2025},
author = {Risheq, S and Sancho, A and Friedman, M and Gati, I and Eliashar, R and Steinberg, D and Gross, M},
title = {Ciprofloxacin-Coated Tympanostomy Tubes with Sustained-Release Varnish: A Novel Strategy to Combat Biofilm Formation by Pseudomonas aeruginosa.},
journal = {Microorganisms},
volume = {13},
number = {9},
pages = {},
doi = {10.3390/microorganisms13092039},
pmid = {41011371},
issn = {2076-2607},
abstract = {OBJECTIVE: The aim of this study is to develop and evaluate the antibacterial and anti-biofilm efficacy of ciprofloxacin-coated tympanostomy tubes (TTs) using a sustained-release varnish (SRV-CIPRO) and introduce a novel tympanic membrane model for preclinical evaluation.

STUDY DESIGN: This was an in vitro experimental study.

SETTING: This study was conducted in a biofilm research laboratory in an academic medical center.

METHODS: Sterile fluoroplastic TTs were coated with SRV-CIPRO or placebo varnish. A novel tympanic membrane (TM) model was developed using a layered agar-plastic system. Antibacterial activity, biofilm inhibition, and bacterial viability were assessed through agar diffusion, MTT, ATP quantification, HR-SEM, and SD-CLSM.

RESULTS: SRV-CIPRO-coated TTs exhibited sustained antibacterial activity for up to 10 days. Compared to the placebo, SRV-CIPRO significantly inhibited biofilm formation, reduced metabolic activity, and decreased bacterial viability (p < 0.05). Imaging confirmed fewer bacterial colonies on SRV-CIPRO TTs. The TM model allowed realistic testing of tube insertion and infection simulation.

CONCLUSION: SRV-CIPRO-coated TTs offer sustained antibiotic delivery, potentially reducing postoperative otorrhea and biofilm-related complications. The TM model provides a platform for preclinical evaluation of middle ear devices.},
}

@article {pmid41011142,
year = {2025},
author = {Maravić-Vlahoviček, G and Kindl, M and Andričević, K and Obranić, S and Vladimir-Knežević, S},
title = {Modulatory Effects of Satureja montana L. Essential Oil on Biofilm Formation and Virulence Factors of Pseudomonas aeruginosa.},
journal = {Pharmaceuticals (Basel, Switzerland)},
volume = {18},
number = {9},
pages = {},
doi = {10.3390/ph18091269},
pmid = {41011142},
issn = {1424-8247},
support = {KK.01.1.1.02.0021 (FarmInova)//European Regional Development Fund/ ; },
abstract = {Background: Antimicrobial resistance is a major global health threat, particularly from pathogens such as Pseudomonas aeruginosa, known for forming biofilms and producing virulence factors that cause persistent infections. Essential oils (EOs) offer promising alternatives to conventional antimicrobial therapy due to their antimicrobial and antibiofilm properties. This study aimed to investigate the modulatory effects of a thymol-rich EO from Satureja montana L. on planktonic growth, biofilm formation, swarming motility, proteolytic activity and pyocyanin production of P. aeruginosa PAO1. Methods: The essential oil, isolated by hydrodistillation from S. montana aerial parts, was analysed by GC-MS. The minimum inhibitory concentration (MIC) of the EO and thymol was determined using the broth microdilution method. Sub-MICs were tested for planktonic growth and biofilm formation. Virulence was assessed by testing swarming motility, proteolytic activity and pyocyanin production. Results: The EO was characterised by a very high content of monoterpenes, with thymol dominating (56.47%). MIC for both EO and thymol was 4 mg/mL. They showed a biphasic effect: higher concentrations significantly inhibited planktonic growth (36-58% reduction; p < 0.05), while lower concentrations promoted it (10-17% increase; p < 0.05). Biofilm biomass varied, but the biofilm index indicated promotion at higher concentrations (0.125-0.5 mg/mL; p < 0.05). Both inhibited swarming at 0.5 mg/mL (thymol was more effective). Thymol decreased proteolytic activity, while EO increased pyocyanin production. Conclusions: S. montana essential oil and thymol show concentration-dependent modulation of P. aeruginosa growth, biofilms and virulence, suggesting their potential as anti-virulence agents, although the biphasic responses require careful dosing.},
}

@article {pmid41011138,
year = {2025},
author = {Elhawary, EA and Soltane, R and Moustafa, MH and Abdelaziz, AM and Said, MA and Zahran, EM},
title = {Sustainable MnO2/MgO Bimetallic Nanoparticles Capped with Sword Fern Methanol Extract Attain Antioxidant/Anti-Biofilm Potential: A UPLC-ESI/LC/MS and Network Pharmacology-Supported Study.},
journal = {Pharmaceuticals (Basel, Switzerland)},
volume = {18},
number = {9},
pages = {},
doi = {10.3390/ph18091262},
pmid = {41011138},
issn = {1424-8247},
support = {25UQU4331312GSSR01//Funder Grant Number Umm Al-Qura University, Saudi Arabia/ ; },
abstract = {Background: Nephrolepis exaltata (sword fern) possesses a considerable amount of phytochemicals and different biological activities. The current study investigates the anti-biofilm potential of greenly synthesized bimetallic nanoparticles of Nephrolepis exaltata leaf methanol extract (NEME-MnO2-MgO BNPs). Methods: The NEME was subjected to UPLC/MS analysis, followed by characterization of its NPs by size, zeta potential, FTIR, entrapment efficiency, and release. Then, antioxidant, antimicrobial and antibiofilm assays were employed, followed by in silico studies. Results: The UPLC/MS analysis of NEME led to the tentative identification of 27 metabolites, mostly phenolics. The MnO2-MgO BNPs presented a uniform size and distribution and exhibited IC50 values of 350 and 215.6 μg/mL, in the DPPH and ABTS assays, respectively. Moreover, the NPs exhibited antimicrobial and anti-biofilm efficacies against Pseudomonas aeruginosa, Klebsiella pneumonia (ATCC-9633), Staphylococcus aureus (ATCC-6538), Escherichia coli, Bacillus cereus, and C. albicans, with MIC values of 250-500 μg/mL. The MnO2-MgO BNPs inhibited Candida albicans biofilms with a % inhibition of 66.83 ± 2.45% at 1/2 MIC. The network pharmacology highlighted epigallocatechin and hyperoside to be the major compounds responsible for the anti-biofilm potential. The ASKCOS facilitated the prediction of the redox transformations that occurred in the green synthesis, while the docking analysis revealed enhanced binding affinities of the oxidized forms of both compounds towards the outer membrane porin OprD of P. aeruginosa, with binding scores of -4.6547 and -5.7701 kcal/mol., respectively. Conclusions: The greenly synthesized Nephrolepis exaltata bimetallic nanoparticles may provide a promising, eco-friendly, and sustainable source for antimicrobial agents of natural origin with potential biofilm inhibition.},
}

@article {pmid41010524,
year = {2025},
author = {Jung, EH and Hwang, G and Kim, KR},
title = {Effect of 6-Shogaol Derived from Ginger (Zingiber officinale) on Dual-Species Biofilm Formation by Streptococcus mutans and Candida albicans.},
journal = {Nutrients},
volume = {17},
number = {18},
pages = {},
doi = {10.3390/nu17182999},
pmid = {41010524},
issn = {2072-6643},
support = {This research was supported by Kyungpook National University Research Fund, 2022.//Kyungpook National University/ ; },
mesh = {*Biofilms/drug effects/growth & development ; *Candida albicans/drug effects/physiology/growth & development ; *Streptococcus mutans/drug effects/physiology/growth & development ; *Catechols/pharmacology/isolation & purification ; *Zingiber officinale/chemistry ; Hydrogen-Ion Concentration ; Plant Extracts/pharmacology ; },
abstract = {BACKGROUND/OBJECTIVES: Dental plaque, a biofilm composed of accumulated oral microorganisms, is a key contributor to various oral diseases. 6-shogaol, a bioactive compound of ginger, is known to have pharmacological activities, including anticancer, anti-inflammatory, and antimicrobial activities. Therefore, we aimed to determine the effects of 6-shogaol on dual-species biofilms of Streptococcus mutans (S. mutans) and Candida albicans (C. albicans).

METHODS: Dual-species oral biofilms were formed on hydroxyapatite (HA) disks for 42 h and exposed to 6-shogaol. The pH was measured in the experimental medium, and the biomass, colony-forming unit (CFU) of microbial cells, and insoluble extracellular polysaccharides (EPS) were quantified in the biofilm formed on the HA disk. Confocal laser scanning microscopy (CLSM) was used to assess biofilm morphology, and quantitative polymerase chain reaction was performed to analyze gtf gene expression.

RESULTS: 6-shogaol dose-dependently reduced insoluble EPS, CFU counts, and dry weight of biofilms. The pH was maintained above 5.5 in the 6-shogaol-treated group. CLSM images showed that S. mutans proliferation, C. albicans hyphal development, and EPS production were markedly inhibited in biofilms treated with 6-shogaol. The expression of gtfB and gtfC was significantly downregulated by 6-shogaol.

CONCLUSIONS: These findings suggest that 6-shogaol has the potential to be a promising natural product for the prevention and management of oral biofilm-related oral diseases.},
}

*Biofilms/drug effects/growth & development
*Candida albicans/drug effects/physiology/growth & development
*Streptococcus mutans/drug effects/physiology/growth & development
*Catechols/pharmacology/isolation & purification
*Zingiber officinale/chemistry
Hydrogen-Ion Concentration
Plant Extracts/pharmacology

@article {pmid41009919,
year = {2025},
author = {Murray, RK and Martin, AE and Zipkowitz, S and Jahan, N and Davis, TD and Redman, WK},
title = {α-Amylase-Mediated Antibiotic Degradation and Sequestration in Pseudomonas aeruginosa Biofilm Therapy.},
journal = {Antibiotics (Basel, Switzerland)},
volume = {14},
number = {9},
pages = {},
doi = {10.3390/antibiotics14090941},
pmid = {41009919},
issn = {2079-6382},
abstract = {BACKGROUND: As of 2022, 80% of all documented microbial infections are biofilm-associated: communities of microorganisms adhered to a surface and enclosed in a complex extracellular polymeric substance (EPS). The EPS acts as a physical barrier protecting the bacteria from antimicrobial agents and host immune responses. To combat this hurdle, the application of glycoside hydrolases (GH) has been investigated due to their ability to cleave particular structural polysaccharides within the EPS, thus breaking down the protective barrier and improving antibiotic clearance. While various studies demonstrate the capacity of GHs to improve antibiotic efficacy against biofilms in combination, there is clear differential success between these treatments depending on the GH and antibiotic chosen. Due to the overlap of GH targets and antibiotic structures, it is imperative to ensure that the antibiotics in combinatorial treatments are not degraded by the GH.

METHODS: This study aimed to screen the GH α-amylase produced from Aspergillus oryzae (AO) and Bacillus subtilis (BS), combined with various antibiotics from different classes, charges, and mode of actions by determining MICs. against the bacterium Pseudomonas aeruginosa (PA) of 6 antibiotics with or without α-amylase and treat 2-day PA biofilms with antibiotics with or without GHs. Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) stability assays and Differential Scanning Fluorimetry (DSF) were conducted to determine antibiotic and GH degradation as well as antibiotic sequestration.

RESULTS: Increased MICs in the presence of GHs as well as decreased antibiotic clearance against 2-day biofilms were suggestive of antibiotic degradation. LC-MS/MS stability assays of tetracycline and ciprofloxacin in the presence and absence of α-amylase further demonstrated the α-amylase-mediated antibiotic sequestration. Differential scanning fluorimetry (DSF) assays confirmed α-amylase-antibiotic interactions.

CONCLUSIONS: This study suggests that α-amylase is capable of degrading and sequestering a variety of antibiotics, and the degree to which these phenomena occur varies depending upon the source of the GH. As a potential treatment for biofilm-associated infections, it is imperative that the GH + antibiotic combinations are determined compatible prior to clinical use.},
}

@article {pmid41009848,
year = {2025},
author = {Oliveira, MCF and Canellas, ALB and Berbert, LC and Cardoso, AM and Silva, VA and Garutti, SST and Rangel, DHF and Dias, RCS and Perini, JA and Souza, CRVM and Chagas, TPG and Laport, MS and Pellegrino, FLPC},
title = {Assessment of Antimicrobial Resistance and Virulence of Biofilm-Forming Uropathogenic Escherichia coli from Rio de Janeiro.},
journal = {Antibiotics (Basel, Switzerland)},
volume = {14},
number = {9},
pages = {},
doi = {10.3390/antibiotics14090869},
pmid = {41009848},
issn = {2079-6382},
support = {88887.820714/2023-00//CAPES/ ; E-26/211.519/2019//FAPERJ/ ; E-26/211.209/2021//FAPERJ/ ; E-26/204.045/2024//FAPERJ/ ; E-26/211.284/2021//FAPERJ/ ; 405020/2023-6//CNPq/ ; 309158/2023-0//CNPq/ ; },
abstract = {Background/Objectives: Uropathogenic Escherichia coli (UPEC) is the leading cause of urinary tract infections in both community and hospital settings worldwide. Antimicrobial-resistant UPEC strains pose a significant challenge for effective antibiotic therapy. In this study, 50 bacterial isolates recovered from urine samples of patients attended in different sectors of a public hospital in Rio de Janeiro over five months were analyzed to assess antimicrobial resistance and virulence profiles through broad gene screening. Methods: Biofilm production was assessed using a semi-quantitative adherence assay. PCR was employed to investigate 27 resistance genes, 6 virulence genes, sequence types (STs), and phylogroups. Susceptibility to 25 antimicrobial agents was determined by disk diffusion testing. Furthermore, the pathogenic potential was evaluated in vivo using the Tenebrio molitor larvae infection model. Results: Most UPEC isolates were moderate or strong biofilm producers (41/50; 82%). The sul1 and sul2 resistance genes were the most frequently detected (58%). Two virulence gene patterns were identified: fyuA, iutA, fimH, cnf1 and fyuA, iutA, fimH (13 isolates; 26%). ST131 and ST73 were the most common sequence types (16% each), and phylogroup B2 was the most prevalent (50%). Thirty isolates (60%) were multidrug-resistant, most of which belonged to phylogroup B2. UPEC exhibited dose-dependent lethality, causing 100% mortality at 2.6 × 10[8] CFU/mL within 24 h. Conclusions: These findings reinforce the urgent need for surveillance strategies and effective antimicrobial stewardship in clinical practice.},
}

@article {pmid41009836,
year = {2025},
author = {Oh, Y and Kim, TJ},
title = {Temperature Adaptive Biofilm Formation in Yersinia enterocolitica in Response to pYV Plasmid and Calcium.},
journal = {Antibiotics (Basel, Switzerland)},
volume = {14},
number = {9},
pages = {},
doi = {10.3390/antibiotics14090857},
pmid = {41009836},
issn = {2079-6382},
support = {2021R1F1A1061888//National Research Foundation of Korea/ ; },
abstract = {Background/Objectives: Yersinia enterocolitica is a pathogenic bacterium that forms biofilms, enhancing its persistence and resistance to antimicrobial agents. Biofilm formation in Y. enterocolitica is influenced by environmental factors such as temperature, calcium, and the presence of the virulence plasmid pYV. This study aims to explore how temperature, calcium, and pYV modulate biofilm formation in Y. enterocolitica, with a focus on motility and extracellular polymeric substance (EPS) production as key factors. Methods: Y. enterocolitica strains with and without the pYV plasmid were cultured at two different temperatures (26 °C and 37 °C). The effect of calcium (5 mM) on biofilm formation was tested at both temperatures. Biofilm formation was measured using crystal violet staining, motility was assessed using soft agar plates, and EPS production was quantified to determine its role in biofilm stabilization. Results: At 26 °C, biofilm formation increased in pYV-negative strains, driven primarily by motility and flagellar expression. In contrast, at 37 °C, pYV-positive strains showed strong biofilm formation despite reduced growth, with EPS production as the key stabilizing factor. Calcium modulated biofilm formation in a temperature-dependent manner: at 26 °C, 5 mM calcium modestly reduced biofilm formation in pYV-negative strains, while at 37 °C, it significantly suppressed both EPS production and biofilm formation by approximately 50% in pYV-positive strains. Conclusions: This study reveals a novel regulatory switch where temperature, calcium, and pYV modulate biofilm formation in Y. enterocolitica. These findings suggest that Y. enterocolitica can adapt between motility- and EPS-dominated biofilm strategies depending on environmental conditions. Understanding these mechanisms offers potential targets for controlling biofilm-related persistence in clinical and food safety contexts.},
}

@article {pmid41009754,
year = {2025},
author = {Tegegne, DT and Abbott, IJ and Poźniak, B},
title = {Catheter-Associated Urinary Tract Infections: Understanding the Interplay Between Bacterial Biofilm and Antimicrobial Resistance.},
journal = {International journal of molecular sciences},
volume = {26},
number = {18},
pages = {},
doi = {10.3390/ijms26189193},
pmid = {41009754},
issn = {1422-0067},
support = {N070/0011/24//Wroclaw University of Environmental and Life Sciences/ ; },
mesh = {*Biofilms/drug effects/growth & development ; Humans ; *Urinary Tract Infections/microbiology/drug therapy/etiology ; *Catheter-Related Infections/microbiology/drug therapy ; *Drug Resistance, Bacterial ; Urinary Catheters/microbiology/adverse effects ; Anti-Bacterial Agents/therapeutic use/pharmacology ; Bacteria/drug effects ; },
abstract = {The increasing use of urinary catheters in healthcare, driven by an aging population and escalating antimicrobial resistance, presents both benefits and challenges. While they are essential to managing urinary retention and enabling precise urine output monitoring, their use significantly increases the risk of catheter-associated urinary tract infections (CAUTIs), the most common type of healthcare-associated infection. CAUTI risk is closely linked to the duration of catheterization and the formation of bacterial biofilms on catheter surfaces. These biofilms, often composed of polymicrobial communities encased in an extracellular matrix, promote persistent infections that are highly resistant to conventional antimicrobial therapies. Common CAUTI uropathogens include E. coli, E. faecalis, P. aeruginosa, P. mirabilis, K. pneumoniae, S. aureus, and Candida spp. The complexity and resilience of these biofilm-associated infections underscore the urgent need for innovative treatment strategies. Therefore, dynamic in vitro bladder infection models, which replicate physiological conditions such as urine flow and bladder voiding, have become valuable tools for studying microbial behavior, biofilm development, and therapeutic interventions under real clinical conditions. This review provides an overview of CAUTIs, explores the role of biofilms in sub-optimal responses to antimicrobial treatment and advances in model systems, and presents promising new approaches to combating these infections.},
}

*Biofilms/drug effects/growth & development
Humans
*Urinary Tract Infections/microbiology/drug therapy/etiology
*Catheter-Related Infections/microbiology/drug therapy
*Drug Resistance, Bacterial
Urinary Catheters/microbiology/adverse effects
Anti-Bacterial Agents/therapeutic use/pharmacology
Bacteria/drug effects

@article {pmid41009506,
year = {2025},
author = {Liberatore, A and Bertoldi, A and Balboni, A and Gabrielli, L and Cantiani, A and Lanna, F and Sartori, M and Brogini, S and Giavaresi, G and Lazzarotto, T},
title = {Biofilm Formation, c-di-GMP Production, and Antimicrobial Resistance in Staphylococcal Strains Isolated from Prosthetic Joint Infections: A Pilot Study in Total Hip and Knee Arthroplasty Patients.},
journal = {International journal of molecular sciences},
volume = {26},
number = {18},
pages = {},
doi = {10.3390/ijms26188929},
pmid = {41009506},
issn = {1422-0067},
support = {RF-2019-12370058.//Ministero della Salute/ ; },
mesh = {*Biofilms/growth & development/drug effects ; Humans ; *Cyclic GMP/analogs & derivatives/metabolism ; *Prosthesis-Related Infections/microbiology/drug therapy ; *Staphylococcal Infections/microbiology/drug therapy ; Pilot Projects ; *Arthroplasty, Replacement, Knee/adverse effects ; *Drug Resistance, Bacterial ; Anti-Bacterial Agents/pharmacology ; Female ; *Staphylococcus aureus/drug effects/isolation & purification/genetics/physiology ; Male ; *Arthroplasty, Replacement, Hip/adverse effects ; *Staphylococcus/drug effects/isolation & purification/genetics/physiology ; Aged ; Middle Aged ; Microbial Sensitivity Tests ; },
abstract = {Total joint arthroplasty (TJA) and total joint replacement (TJR) are effective treatments for end-stage osteoarthritis, but prosthetic joint infections (PJIs) remain a significant complication. These infections are often associated with bacteria that form biofilms, which contribute to their persistence and resistance to treatment. The aim of this study was to investigate the biofilm-forming ability, cyclic diguanylic acid (c-di-GMP) production, and the presence of biofilm-associated genes in Staphylococcus aureus and coagulase-negative Staphylococci (CoNS) isolates obtained from synovial fluid samples of patients with PJIs following TJA and TJR. A total of 198 samples were analyzed, with bacterial growth detected in 33 samples (16.7%). Among these, 10 strains of S. aureus and 22 strains of CoNS were identified. Biofilm formation was evaluated using the crystal violet assay, and c-di-GMP levels were measured. A statistically significant linear regression was found between biofilm formation and c-di-GMP production (p = 0.016, R[2] = 0.18). Genetic analysis revealed the presence of biofilm-associated genes, including icaA, clfA, fnbA in S. aureus, and atlE, fbe in CoNS. Furthermore, there was a statistically significant difference in c-di-GMP production between strains harboring the icaA gene and strains without icaA (p = 0.016), while oxacillin resistance was detected more frequently in strains carrying fbe gene (p = 0.031). The study emphasizes the variability in antibiotic resistance profiles among staphylococcal isolates, underscoring the complexity of managing these infections.},
}

*Biofilms/growth & development/drug effects
Humans
*Cyclic GMP/analogs & derivatives/metabolism
*Prosthesis-Related Infections/microbiology/drug therapy
*Staphylococcal Infections/microbiology/drug therapy
Pilot Projects
*Arthroplasty, Replacement, Knee/adverse effects
*Drug Resistance, Bacterial
Anti-Bacterial Agents/pharmacology
Female
*Staphylococcus aureus/drug effects/isolation & purification/genetics/physiology
Male
*Arthroplasty, Replacement, Hip/adverse effects
*Staphylococcus/drug effects/isolation & purification/genetics/physiology
Aged
Middle Aged
Microbial Sensitivity Tests

@article {pmid41008208,
year = {2025},
author = {Huang, X and Yang, H and Wang, C and Yan, S and Ren, X and Sun, Z},
title = {Pterostilbene Eliminates MRSA Independent of Metabolic State and Effectively Prevents Biofilm Formation in Milk Matrices.},
journal = {Foods (Basel, Switzerland)},
volume = {14},
number = {18},
pages = {},
doi = {10.3390/foods14183236},
pmid = {41008208},
issn = {2304-8158},
support = {202203021212430//Basic Research Program of Shanxi Province for the Youth Science and Technology Research/ ; SXBYKY2022007//Project of Scientific Research for Excellent Doctors of Shanxi Province/ ; 2022BQ01//Doctorate Research Starting Foundation of Shanxi Agricultural University/ ; 2025CYJSTX12-10//The earmarked fund for Modern Agro-industry Technology Research System/ ; },
abstract = {The relentless spread of antimicrobial resistance poses a severe threat to global public health, food safety, and environmental security. Natural products with potent antimicrobial activity offer prospective substitutes for traditional antibiotics and chemical preservatives. Here, we demonstrate that pterostilbene (PT), a natural dietary compound, exhibits rapid lytic activity against methicillin-resistant Staphylococcus aureus (MRSA). PT displayed metabolism-independent bactericidal effects, effectively eradicating dormant persister cells within one hour, though its activity was partially attenuated under anaerobic conditions. Mechanistically, PT disrupted membrane integrity by increasing permeability, dissipating membrane potential, depleting cellular ATP, and suppressing reactive oxygen species (ROS) generation. Its efficacy was modulated by membrane phospholipid composition, with phosphatidylglycerol (PG) and cardiolipin (CL) critically influencing antimicrobial potency. Crucially, PT robustly inhibited MRSA biofilm formation in milk. These findings highlight PT's potential as a structurally stable, natural antimicrobial for controlling resilient MRSA, particularly against biofilm-associated and persister subpopulations in food systems.},
}

@article {pmid41007765,
year = {2025},
author = {Agboola, OE and Agboola, O and Ayinla, ZA and Agboola, SS and Olaiya, OE and Oyinloye, OM and Fajana, OM and Idowu, OT and Omotuyi, OI and Ilesanmi, OS and Oyinloye, BE},
title = {Integrated Virtual Screening for Anti-Caries Compounds from Neem: Dual-Target Inhibition of Biofilm Formation and Bacterial DNA Replication.},
journal = {Biomedicines},
volume = {13},
number = {9},
pages = {},
doi = {10.3390/biomedicines13092202},
pmid = {41007765},
issn = {2227-9059},
abstract = {Background: Dental caries arise from polymicrobial biofilms and require interventions that address both local virulence and systemic burden. Methods: A curated set of 124 neem-derived phytochemicals was screened against Streptococcus mutans glucansucrase (3AIC) and Staphylococcus aureus DNA gyrase B (3U2D) using harmonized AutoDock Vina parameters. Ligand standardization and receptor preparation followed conventional protocols. Results: The most favorable docking scores reached -10.7 kcal·mol[-1] for 3AIC and -8.9 kcal·mol[-1] for 3U2D. Redocking produced pose RMSD values of 1.52 Å (3AIC) and 0.96 Å (3U2D). Per-receptor ADMET profiles for the six top-ranked compounds indicated median logP values of 4.93 (3AIC) and 4.52 (3U2D), median TPSA values of 80.3 and 62.9 Å[2], median rotatable bonds of 2.5 and 1.0, and median QED values of 0.41 and 0.76, respectively. Conclusions: An integrated, dual-target screen prioritized neem constituents with plausible local anti-cariogenic activity and physicochemical features compatible with systemic disposition. These in silico findings motivate targeted experimental validation.},
}

@article {pmid41007401,
year = {2025},
author = {Ding, X and An, Q and Kang, H and Li, S and Zhang, S and Yang, H and Dou, X and Ji, Y and Zhao, Y and Fan, H},
title = {Whole-Genome Analysis of a Novel Multidrug-Resistant Escherichia coli Strain from Dairy Calves in Northeast China: Mechanisms of Antibiotic Resistance and Biofilm Formation.},
journal = {Biology},
volume = {14},
number = {9},
pages = {},
doi = {10.3390/biology14091257},
pmid = {41007401},
issn = {2079-7737},
support = {32373085//National Natural Science Foundation of China/ ; ZL2024C020//Natural Science Foundation of Heilongjiang Province/ ; 2024M750390//China postdoctoral science fund/ ; },
abstract = {The use of antibiotics is so widespread in animal husbandry, but negligent management and lack of policies often lead to the massive use of antibiotics on farms. In this study, we collected cases of epidemic calf diarrhea in northeastern China and isolated a new strain of multidrug-resistant Escherichia coli (MDR-E. coli). In order to explore the information of this pathogen in detail, we used whole-genome sequencing to determine the genome sequence, and explored in detail the resistance, pathogenicity, genetic evolution and other biological processes of the strain through bioinformatics analysis. The results showed that the E. coli isolated in this study was a new multidrug-resistant strain with a large number of drug resistance genes (77) and virulence genes (84), including a circular chromosome and five circular plasmids, which are basically impossible to treat by currently commonly used antibiotics. The findings of this study suggest that the prolonged misuse of antibiotics in agricultural settings may contribute to the development of antibiotic-resistant strains of E. coli. This, in turn, has the potential to trigger outbreaks of antibiotic-resistant bacterial diarrhea, leading to substantial economic losses and posing significant public health risks. These results underscore the necessity for the judicious use of antibiotics and will inform the development of pertinent policies and regulations.},
}

@article {pmid41007393,
year = {2025},
author = {Oh, JH and Kim, E and Cho, M},
title = {Biofilm Formation by Rice Rhizosphere Nitrogen-Fixing Microorganisms and Its Effect on Rice Growth Promotion.},
journal = {Biology},
volume = {14},
number = {9},
pages = {},
doi = {10.3390/biology14091249},
pmid = {41007393},
issn = {2079-7737},
support = {PJ017406012025//National Institute of Agricultural Sciences Program/ ; },
abstract = {Excessive nitrogen fertilizer use contributes to environmental pollution and undermines agricultural sustainability. Enhancing symbiotic interactions between rice and nitrogen-fixing microorganisms offers a promising strategy to potentially improve nitrogen use efficiency (NUE). This study investigates the role of rice root exudates in promoting biofilm formation by nitrogen-fixing microbes to enhance nitrogen fixation. Nine nitrogen-fixing microbial strains were evaluated for biofilm formation in response to flavone and apigenin treatments, with Gluconacetobacter diazotrophicus KACC 12358 serving as the reference strain. The most responsive strain was selected, and a library of 1597 natural compounds was screened to identify those that promote biofilm formation in both the selected and reference strains. A. indigens KACC 11682 exhibited the highest biofilm-forming capacity, with apigenin treatment showing an OD595 value approximately 1.4 times higher than the DMSO control. Screening identified 68 compounds that enhanced biofilm formation by more than 500% compared to the control. Among them, eight compounds induced strong biofilm formation (O.D. > 2.0) in A. indigens. Cardamomin, a chalconoid flavonoid, emerged as one of the most effective compounds, showing a 245% increase in biofilm formation. Growth promotion assays showed that A. indigens increased rice fresh weight by approximately 128% compared to untreated controls. This study demonstrates the potential of rice root exudate-derived compounds to promote beneficial symbiosis with nitrogen-fixing microbes. These findings offer a novel approach that may contribute to enhancing rice NUE. Future research will focus on evaluating the long-term effects of these compounds and microorganisms, assessing their applicability in real agricultural settings, and conducting further validation across various rice cultivars.},
}

@article {pmid41005094,
year = {2025},
author = {Russo, S and Muscetta, M and Liotta, I and Rizzo, M and Marra, D and Gentile, G and Förster, B and Caserta, S and Marotta, R and Cocca, M and Vitiello, G},
title = {Sunlight driven photo-treatment of polyhydroxybutyrate microplastics mediated by carbon nanodots-doped ZnO mesocrystals: Induced surface changes boost degradation in soil and biofilm formation.},
journal = {Journal of hazardous materials},
volume = {498},
number = {},
pages = {139933},
doi = {10.1016/j.jhazmat.2025.139933},
pmid = {41005094},
issn = {1873-3336},
abstract = {Plastics are widely used due to their versatility. However, their accumulation as microplastics and nanoplastics in the environment poses a significant threat for the ecosystems. Although biodegradable polymers, like polyhydroxybutyrate (PHB), offer a sustainable alternative, their degradation could still lead to fragmentation and MP accumulation, hence it is essential to enhance biodegradation rates in order to mitigate environmental impacts. This study investigates the integration of photocatalytic pretreatment with natural biodegradation to accelerate PHB biodegradation. Photoactive ZnO mesocrystals doped with biowaste-derived carbon nanodots, prepared via a wet-chemical approach, are proposed to improve the photocatalytic efficiency under sunlight. ZnO mesocrystals doped with 30 wt% of carbon nanodots exhibited superior photocatalytic properties, thus photodegrading PHB microplastics more efficiently than bare ZnO. Phototreated PHB microplastics exhibit an improved biodegradation in soil when exposed for a controlled irradiation time. Additionally, PHB microplastics photo-treated in the presence of doped mesocrystals showed an enhanced microbial colonization indicating improved biofilm formation. These findings highlight the potential of photocatalytic pretreatment in modifying surface properties to boost degradation in soil, thus offering a promising strategy for reducing microplastic accumulation.},
}

@article {pmid41003166,
year = {2025},
author = {Cedeño-Pinargote, AC and Jara-Medina, NR and Pineda-Cabrera, CC and Cueva, DF and Erazo-Garcia, MP and Tejera, E and Machado, A},
title = {Impact of Biofilm Formation by Vaginal Candida albicans and Candida glabrata Isolates and Their Antifungal Resistance: A Comprehensive Study in Ecuadorian Women.},
journal = {Journal of fungi (Basel, Switzerland)},
volume = {11},
number = {9},
pages = {},
doi = {10.3390/jof11090620},
pmid = {41003166},
issn = {2309-608X},
support = {Project ID: 12260 entitled "Adhesión inicial y resistencia antimicrobiana de Candida sp. aisladas de la microbiota humana''//Universidad San Francisco de Quito USFQ/ ; Project ID: 17357 entitled "Alternative approaches for eliminating Biofilms"//Universidad San Francisco de Quito USFQ/ ; Project ID: 16801 entitled ''Characterization of single and mixed Biofilms''//Universidad San Francisco de Quito USFQ/ ; },
abstract = {Candida albicans and Candida glabrata are key fungal pathogens linked to candidiasis, with rising concerns due to antifungal resistance and biofilm abilities. However, data from Latin America remains limited. This study assessed biofilm formation and antifungal susceptibility of vaginal Candida isolates from Ecuadorian women. Biofilm formation at 24 and 48 h was evaluated using biomass and CFU assays and the biofilm formation index. Antifungal resistance in planktonic cells and patient microbiota profiles were also analyzed. Biofilm assessment showed 57.14% of isolates were high biofilm formers, 33.33% intermediate, 4.76% low, and 4.76% non-formers. Planktonic susceptibility testing included fluconazole, voriconazole, posaconazole, caspofungin, anidulafungin, micafungin, flucytosine, and amphotericin B. Micafungin showed the lowest MBEC90 value among tested antifungals, with an average MIC of 0.15 µg/mL, MBIC90 of 1.26 µg/mL, and MBEC90 of 1.86 µg/mL. Fluconazole followed with MIC, MBIC90, and MBEC90 values of 4.19, 63.33, and 66.59 µg/mL. Flucytosine had the highest values (MIC = 11.36 µg/mL; MBIC90 = 244.71 µg/mL; MBEC90 = 245.33 µg/mL). Both micafungin and flucytosine produced similar reductions in viable biofilm cells (1.44 log CFU), while fluconazole induced a slightly lower reduction of 1.39 log CFU. Findings suggest echinocandins may be effective against biofilm-forming Candida in this Ecuadorian population subset.},
}

@article {pmid41002904,
year = {2025},
author = {Liu, M and Liao, B},
title = {Modelling of Diffusion and Reaction of Carbon Dioxide and Nutrients in Biofilm for Optimal Design and Operation of Emerging Membrane Carbonated Microalgal Biofilm Photobioreactors.},
journal = {Membranes},
volume = {15},
number = {9},
pages = {},
doi = {10.3390/membranes15090269},
pmid = {41002904},
issn = {2077-0375},
support = {RGPIN-2019--05087//Natural Sciences and Engineering Research Council of Canada/ ; },
abstract = {The biological performance and carbon dioxide (CO2) flux of the novel and emerging concept of a membrane carbonated microalgal biofilm photobioreactor (MC-MBPBR) for wastewater treatment were investigated using mathematical modelling in conjunction with the finite-difference method. A set of differential equations was established to model the performance of an MC-MBPBR. The impacts of CO2 partial pressure, wastewater characteristics, and biofilm thickness on the concentration profiles and fluxes of CO2 and nutrients (N and P) to the biofilm of the MC-MBPBR were systematically studied. The modelling results showed profound impacts of these parameters on process efficiency (CO2 transfer and N and P removals) and the existence of an optimal biofilm thickness for maximum CO2, N, and P fluxes into the biofilm. Penetration of CO2 through the biofilm into the bulk water phase might occur under certain conditions. An increase in gaseous CO2 and increased influent N and P concentrations led to higher CO2, N, and P fluxes. The optimal biofilm thickness varied with the change in wastewater characteristics and gaseous CO2 concentration. The modelling results were in relatively good agreement with experimental results from the literature. The proposed mathematical models can be used as a powerful tool to optimize the design and operation of the novel MC-MBPBR for wastewater treatment and microalgae cultivation.},
}

@article {pmid41002690,
year = {2025},
author = {de Andrade, JG and Natali, AFF and Loureiro, C and Rodrigues, GWL and Ribeiro, APF and de Freitas, RN and Barzotti, RJ and Oliveira, LC and de Moraes, YGC and Gomes, NA and Chaves-Neto, AH and Martinho, FC and Jacinto, RC},
title = {Synergistic Effect of Sodium Hypochlorite and Carbon Dioxide Against Enterococcus faecalis Biofilm.},
journal = {Dentistry journal},
volume = {13},
number = {9},
pages = {},
doi = {10.3390/dj13090417},
pmid = {41002690},
issn = {2304-6767},
support = {001//the Coordination for the Improvement of Higher Education Personnel, Brazil (CAPES/ ; 2021/02260-6//São Paulo Research Foundation (FAPESP)/ ; },
abstract = {Objectives: This study aimed to evaluate whether the addition of pressurized carbon dioxide (PCD) influences the antimicrobial efficacy of 2.5% sodium hypochlorite (NaOCl) against Enterococcus faecalis biofilm in root canals and dentinal tubules. Methods: Forty extracted human mandibular premolars with single canals were contaminated with E. faecalis for 10 days and randomly assigned to four groups (n = 10): 2.5% NaOCl, 2.5% NaOCl + CO2, sterile saline, and sterile saline + CO2. The pH and temperature of the NaOCl solution were measured before and after CO2 incorporation. Microbial load was assessed by CFU counts before and after irrigation, and in dentin samples from the cervical, middle, and apical thirds. Oxidative stress was evaluated via lipid peroxidation (TBARS), protein carbonyl content, and total protein quantification. Biofilm metabolic activity was analyzed using the XTT reduction assay. Data were analyzed using one-way ANOVA on ranks and two-way repeated measures ANOVA (α = 0.05), a very large effect size (Cohen's d) ≈ 1.756 was assumed. Results: All irrigation protocols significantly reduced bacterial load (p < 0.05). Both NaOCl groups outperformed the saline controls (p = 0.009). The addition of CO2 to NaOCl slightly enhanced disinfection in the main canal but did not improve antimicrobial action in dentinal tubules. CO2 incorporation reduced the pH of NaOCl from ~13.4 to 7.4 and slightly increased the temperature, making the solution more chemically reactive. However, both oxidative stress markers and the XTT assay showed that the combination with CO2 impaired the antimicrobial effectiveness of NaOCl. Conclusions: Despite the improvement in bacterial reduction in the root canal lumen, the combination of PCD with NaOCl failed to enhance intratubular disinfection and reduced the oxidative damage and metabolic inactivation of the biofilm. CO2 pressurization appears to limit the antimicrobial action of NaOCl.},
}

@article {pmid41002344,
year = {2025},
author = {Bao, H and Yu, F and Dai, P and Guo, B and Xu, Y},
title = {Construction of an Electrochemical Impedance Spectroscopy Matching Method Based on Adaptive Multi-Error Driving and Application Testing for Biofilm Impedance Verification.},
journal = {Biosensors},
volume = {15},
number = {9},
pages = {},
doi = {10.3390/bios15090604},
pmid = {41002344},
issn = {2079-6374},
support = {JGCG2024210//Zhejiang Provincial Department of Education/ ; JGCG2024210//Zhejiang Provincial Department of Education；/ ; },
mesh = {*Biofilms ; *Dielectric Spectroscopy/methods ; Algorithms ; *Biosensing Techniques ; Serum Albumin, Bovine/analysis ; Cattle ; Animals ; },
abstract = {Electrochemical impedance spectroscopy (EIS) is a technique used to analyze the kinetics and interfacial processes of electrochemical systems. The selection of an appropriate equivalent circuit model for EIS interpretation was traditionally reliant on expert experience, rendering the process subjective and prone to error. To address these limitations, an automated framework for both model selection and parameter estimation was proposed. The methodology was structured such that initial model screening was performed by a global heuristic search algorithm, adaptive optimization was guided by an integrated XGBoost-based error feedback mechanism, and precise parameter estimation was achieved using a Differential Evolution-Levenberg-Marquardt (DE-LM) algorithm. When evaluated on a purpose-built dataset comprising 4.8 × 10[5] spectra across diverse circuit and biofilm scenarios, a model classification accuracy of 96.32% was achieved, and a 72.3% reduction in parameter estimation error was recorded. The practical utility of the method was validated through the quantitative analysis of bovine serum albumin-Clenbuterol hydrochloride (BSA-CLB), wherein an accuracy of 95.2% was demonstrated and a strong linear correlation with target concentration (R[2] = 0.999) was found. Through this approach, the limitations of traditional black-box models were mitigated by resolving the physical meaning of parameters. Consequently, the automated and quantitative monitoring of processes such as biofilm formation was facilitated, enabling the efficient evaluation of antimicrobial drugs or anti-fouling coatings.},
}

*Biofilms
*Dielectric Spectroscopy/methods
Algorithms
*Biosensing Techniques
Serum Albumin, Bovine/analysis
Cattle
Animals

@article {pmid41002336,
year = {2025},
author = {Yöney, B and Obořilová, R and Lacina, K and Farka, Z and Skládal, P},
title = {Pathogen-on-a-Chip: Impedance-Based Detection of Biofilm Formation of Staphylococcus aureus and Staphylococcus epidermidis.},
journal = {Biosensors},
volume = {15},
number = {9},
pages = {},
doi = {10.3390/bios15090596},
pmid = {41002336},
issn = {2079-6374},
support = {CZ.02.01.01/00/22_008/0004596//Ministry of Education of Czech Republic/ ; },
mesh = {*Biofilms/growth & development ; *Staphylococcus epidermidis/physiology ; *Staphylococcus aureus/physiology ; Dielectric Spectroscopy ; *Biosensing Techniques ; Electrodes ; Microscopy, Atomic Force ; *Lab-On-A-Chip Devices ; Electric Impedance ; },
abstract = {Bacterial biofilms are complex microbial communities that contribute to the pathogenesis of chronic infections. Therefore, it is crucial to detect biofilm-associated infections in early stages as their delayed treatment becomes more complicated. Herein, we describe a label-free electrochemical impedance spectroscopy (EIS) method for detecting biofilm formation by Staphylococcus aureus and Staphylococcus epidermidis. Printed circuit board-based biamperometric gold electrodes were modified with poly-L-lysine to enhance bacterial attachment to the sensor surface. Formation and inhibition of biofilms were evaluated based on changes in charge transfer resistance (Rct). The control Rct value increased by ~90 kΩ for S. epidermidis biofilm and by ~60 kΩ for S. aureus biofilms. Antibiotic-treated samples exhibited similar values to those using the control. In addition, biofilm formation was evaluated through optical microscopy using safranin staining, and the micrographs suggest significant biomass on the electrodes, whereas the control appeared clear. Atomic force microscopy was used to visualize the biofilm on the electrode surface, obtain cross-sectional profiles, and evaluate its roughness. The roughness parameters indicate that S. aureus forms a rougher biofilm than S. epidermidis, while S. epidermidis forms a more compact biofilm. These findings suggest that the optimized EIS-based method effectively monitors changes related to biofilms and serves as a promising tool for evaluation of new anti-biofilm agents, such as antibiotics, phages or antibodies.},
}

*Biofilms/growth & development
*Staphylococcus epidermidis/physiology
*Staphylococcus aureus/physiology
Dielectric Spectroscopy
*Biosensing Techniques
Electrodes
Microscopy, Atomic Force
*Lab-On-A-Chip Devices
Electric Impedance

@article {pmid41001999,
year = {2025},
author = {Giovanni, A and Shi, YZ and Wang, PC and Tsai, MA and Chen, SC},
title = {Comparative Evaluation of Oral Biofilm and Killed Cell Vaccines Against Streptococcus iniae in Four-Finger Threadfin Fish (Eleutheronema tetradactylum): Immune Response and Protection Efficacy.},
journal = {Journal of fish diseases},
volume = {},
number = {},
pages = {e70062},
doi = {10.1111/jfd.70062},
pmid = {41001999},
issn = {1365-2761},
support = {113-2313-B-020-001//National Science and Technology Council/ ; },
abstract = {Mariculture, a significant component of the maritime industry that focuses on marine food production, faces challenges in maintaining productivity during bacterial disease outbreaks, particularly in high-value aquaculture such as the four-finger threadfin fish in Taiwan. Streptococcosis, caused by Streptococcus iniae, is a major contributor to the mortality of the four-finger threadfin (Eleutheronema tetradactylum). Recurrent streptococcosis outbreaks have highlighted the pressing need for highly effective vaccination strategies. Given its safety, environmental friendliness, and protective effects, vaccination is widely acknowledged as an effective means of preventing aquatic diseases. An innovative approach involves using biofilm-forming S. iniae as vaccine candidates for aquaculture. This study presents an effective approach for developing a biofilm-based vaccine by cultivating S. iniae on chitosan particles, facilitating robust biofilm formation and enhancing immune responses in four-finger threadfin fish. For comparison, a formalin-killed cell (FKC) vaccine, prepared from whole-cell S. iniae, was evaluated. Immune responses were examined in the blood, mucus, and gut lavage from both the vaccinated and control groups. These responses include immune-related gene expression, antibody titers, and lysozyme activity. At 30 days post-vaccination, the biofilm vaccine group exhibited elevated antibody titers, with values of 0.23 ± 0.02 in serum, 0.09 ± 0.01 in mucus, and 0.16 ± 0.01 in gut lavage. Following vaccination, both the FKC and biofilm vaccines significantly upregulated the expression of key proinflammatory cytokines (tumour necrosis factor-α, interleukin [IL]-10, IL-12) in the spleen and kidney, indicating robust activation of the innate immune response. However, the biofilm vaccine induced markedly higher expression of these cytokines, highlighting its stronger stimulation of innate immune responses. These results suggest that the biofilm-based formulation stimulates early immune signalling pathways that are critical for protection against S. iniae infection. In the challenge experiments, the relative percent survival was of 22.85% for the biofilm and 42.8% for the FKC vaccine groups. This study demonstrates that while both FKC and biofilm vaccines activated innate and adaptive immunity, the FKC vaccine provided higher protection (RPS 42.8% vs. 22.85%), indicating that strong immunogenicity does not always translate into effective protection and that oral vaccine strategies require further refinement. Further optimisation of oral vaccine formulations is required to improve the protective efficacy of biofilm-based vaccines in aquaculture.},
}

@article {pmid41001849,
year = {2025},
author = {Li, L and Zhao, Z and Gao, Y and Jiang, X and Liu, H and Guo, X and Huang, X and Zhou, L and Liu, C and Shen, XC},
title = {Dynamic Metal-Phenolic Coordinated Hydrogel for Synergistic Photothermal/Chemodynamic Therapy against Biofilm-Infected Wounds and Real-Time Monitoring.},
journal = {ACS applied materials & interfaces},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsami.5c17481},
pmid = {41001849},
issn = {1944-8252},
abstract = {The development of multifunctional hydrogel dressings integrating injectability, self-healing capability, tissue adhesion, multimodal antibacterial mechanisms, and real-time wound status monitoring remains a critical challenge for combating bacterial biofilms and accelerating wound healing. Herein, we present a dynamically cross-linked nanocomposite hydrogel (QCS-TA/LDH-panis) via Fe[3+]/Mn[2+]-mediated coordination between tannic acid (TA)-modified quaternized chitosan (QCS-TA) and polysulfonatoaniline-intercalated FeMn-layered double hydroxide (LDH-panis). The LDH-panis nanohybrids, synthesized through in situ polymerization of 3-sulfonatoaniline within FeMn-LDH interlayers, exhibit a near-infrared (NIR)-responsive photothermal effect (η = 64.3%) and pH/H2O2-activated peroxidase-like activity for biofilm-disrupting hydroxyl radical ([•]OH) generation. Concurrently, the QCS-TA matrix enables a "capture-and-kill" mechanism via electrostatic interactions (quaternary ammonium groups) and bacterial affinity adhesion (catechol/pyrogallol moieties). Under near-infrared (NIR) irradiation, synergistic mild photothermal/chemodynamic therapy (mPTT/CDT) combined with contact-killing achieved >95% eradication of Staphylococcus aureus and Escherichia coli biofilms. Notably, the hydrogel's conductivity enabled real-time monitoring of wound exudate and temperature fluctuation during the healing progression. In vivo evaluations confirmed accelerated infected wound regeneration (98.2% closure in 12 days) through biofilm elimination, inflammatory suppression, reepithelialization, and collagen deposition. This multifunctional hydrogel unifies dynamic adaptability, multimodal antibacterial therapy, and sensing intelligence, offering a promising strategy for the clinical management of biofilm-associated infection.},
}

@article {pmid41000992,
year = {2025},
author = {Li, J and Squyres, GR and Duong, K and Reichhardt, C and Parsek, MR and Newman, DK},
title = {Extracellular matrix chemistry tunes bacterial biofilm metabolism and optimizes fitness.},
journal = {bioRxiv : the preprint server for biology},
volume = {},
number = {},
pages = {},
doi = {10.1101/2025.09.18.677115},
pmid = {41000992},
issn = {2692-8205},
abstract = {Chemically complex extracellular matrices define cellular microenvironments and shape cell behavior. We hypothesized a composition-properties-function relationship in these natural living materials, where interactions among matrix components govern material properties and cellular physiology. Using Pseudomonas aeruginosa biofilms as a model system, we show that electrostatic interactions between the cationic polysaccharide Pel and extracellular DNA (eDNA) regulate retention of pyocyanin (PYO), a redox-active metabolite that supports anaerobic metabolism via extracellular electron transfer (EET). Biofilm-mimetic hydrogels and natural biofilms revealed that altering Pel's charge via pH adjustment or chemical acetylation, or tuning the Pel:eDNA ratio, predictably modulates PYO retention and EET efficiency. Functionally, a lower Pel:eDNA ratio enhances metabolism under oxygen limitation, whereas a higher ratio promotes survival under antibiotic stress. These findings highlight how matrix chemistry encodes tunable material properties that confer biofilm fitness advantages and establish a materials-based framework for understanding extracellular matrices in multicellular communities.},
}

@article {pmid41000730,
year = {2025},
author = {Gloag, E and Marshall, CW and Kubota, N and Deaver, SE and Deshote, B and Cooper, VS and Wozniak, DJ},
title = {Pseudomonas aeruginosa biofilm-deficient mutants undergo parallel adaptation during chronic infection.},
journal = {bioRxiv : the preprint server for biology},
volume = {},
number = {},
pages = {},
doi = {10.1101/2025.09.20.677542},
pmid = {41000730},
issn = {2692-8205},
abstract = {Pseudomonas aeruginosa readily adapts to infection by acquiring stable and heritable mutations. Previously, we discovered that the first adaptations in a porcine wound model were rugose small-colony variants (RSCVs) caused by mutations in the wsp operon. These mutants overproduce Pel and Psl biofilm exopolysaccharides that improve defense against host responses. To identify other mechanisms of host adaptation that lead to hyperbiofilm phenotypes, we created a mutant with an activated wsp pathway but unable to produce these exopolysaccharides. Porcine wounds were infected with this mutant and biopsies were sampled at days 7, 14 and 35. Small colony variants were isolated from the wound, and whole genome sequencing revealed these variants had acquired mutations in genes in lipopolysaccharide and type IV pili biosynthesis, with wzy and pilU genes being most commonly targeted. pilU mutants were associated with a hyperbiofilm phenotype that outcompeted the parental strain, and wzy mutants were associated with a hyperbiofilm phenotype and increased tolerance to host antimicrobial products. We further identified that several variants had acquired large genome deletions that spanned up to 320 consecutive genes and other variants with high copy numbers of Pf6 filamentous phage. Together our results suggest that the hyperbiofilm phenotype is adaptive in chronic infections and that P. aeruginosa has redundant and diverse pathways to generate this phenotype.},
}

@article {pmid40999847,
year = {2025},
author = {Piovesan, LF and de Lima Fontes, M and Gimenes, MDS and Ozelin, SD and Claro Monteiro, G and Miguel Silva, J and Barbosa, ML and Lourenção Brighenti, F and Barud, HDS},
title = {Orodispersible film based on hyaluronic acid and morin for dental biofilm control.},
journal = {Biofouling},
volume = {},
number = {},
pages = {1-12},
doi = {10.1080/08927014.2025.2560090},
pmid = {40999847},
issn = {1029-2454},
abstract = {Orodispersible films (ODFs) are an innovative oral drug delivery method benefiting pediatric, geriatric, and non-compliant patients. They are portable, easy to swallow, and enhance bioavailability. Hyaluronic acid (HA) stands out among hydrophilic polymers for oral delivery of antimicrobial agents. This study evaluated the physicochemical properties, in vitro release profile, and antimicrobial/antibiofilm activity of HA-based ODFs combined with the flavonoid morin, known for its antimicrobial properties. Antimicrobial activity and microbial viability were assessed via biomass quantification. The films were thin (12-27 µm), flexible, homogeneous, and mechanically resistant. A burst release of morin was observed, reaching complete release at 210 min. Cytotoxicity analysis confirmed the non-toxic profile, showing cell viability. HA-morin films significantly reduced Streptococcus mutans biofilm mass, viability, and acidogenicity compared to the controls. Findings confirmed the non-toxic, and their significant antibiofilm activity against S. mutans. This innovative mucoadhesive system has potential for managing dental diseases and oral drug delivery.},
}

@article {pmid40999251,
year = {2025},
author = {Khan, MS and Jahan, N and Khatoon, R and Ansari, FM and Ahmad, S},
title = {Risk factors and clinical outcomes of multidrug-resistant and biofilm-producing infections in diabetic foot ulcers: a two-year cohort study.},
journal = {World journal of microbiology & biotechnology},
volume = {41},
number = {10},
pages = {346},
pmid = {40999251},
issn = {1573-0972},
mesh = {Humans ; Male ; *Biofilms/growth & development/drug effects ; *Diabetic Foot/microbiology/drug therapy ; Female ; Middle Aged ; Risk Factors ; *Drug Resistance, Multiple, Bacterial ; Prospective Studies ; Aged ; Anti-Bacterial Agents/therapeutic use/pharmacology ; Adult ; *Bacteria/drug effects/isolation & purification/classification ; Treatment Outcome ; Cohort Studies ; },
abstract = {This two-year prospective study analysed 124 diabetic foot ulcer (DFU) patients admitted to the Department of Surgery to identify clinical and microbiological predictors of multidrug-resistant (MDR) and biofilm-producing infections. The cohort was predominantly male (82.3%), with a mean age of 54.3 ± 11.6 years and a high prevalence of tobacco use (63%) and illiteracy (48.4%). Most patients (91.1%) exhibited poor glycaemic control (mean HbA1c 8.3 ± 1.3%), with neuropathy (79%) and retinopathy (72.6%) as the most common comorbidities. Ulcer severity ranged from Wagner grade 1 to 4, with 60.5% persisting for over one month. Microbiological analysis revealed 59.7% monomicrobial and 33.9% polymicrobial infections, with biofilm formation detected in 60.5% of isolates. Multivariable logistic regression showed significant associations between MDR isolates and tobacco use (OR: 4.06, P = 0.049), polymicrobial infections (OR: 20.75, P < 0.001), and biofilm formation (OR: 3.84, P = 0.033). Predictors of biofilm production included male sex (OR: 7.89, P = 0.023), neuropathy (OR: 4.44, P = 0.040), prolonged ulcer duration (OR: 0.09, P = 0.001), non-necrotic ulcers (OR: 0.16, P = 0.014), and MDR organisms (OR: 3.64, P = 0.034). Clinical outcomes included a 30.6% amputation rate (22.6% minor, 8.1% major) and an 8.9% mortality rate, with a mean hospital stay of 22.9 ± 9.2 days. These findings highlight the multifactorial nature of MDR development and biofilm-associated infections, emphasizing the critical roles of biofilm formation, polymicrobial infections, and tobacco use in DFU management. Early intervention and tailored therapies are essential to mitigate these risks and improve patient outcomes.},
}

Humans
Male
*Biofilms/growth & development/drug effects
*Diabetic Foot/microbiology/drug therapy
Female
Middle Aged
Risk Factors
*Drug Resistance, Multiple, Bacterial
Prospective Studies
Aged
Anti-Bacterial Agents/therapeutic use/pharmacology
Adult
*Bacteria/drug effects/isolation & purification/classification
Treatment Outcome
Cohort Studies

@article {pmid40998038,
year = {2025},
author = {Pourhajibagher, M and Moghaddam, EK and Moeininejad, M and Esboei, BR and Bahador, A},
title = {Ex vivo assessment of blue laser-activated berberine-loaded nanoniosomes in enhancing photodynamic therapy against Streptococcus mutans biofilm on tooth enamel.},
journal = {Photodiagnosis and photodynamic therapy},
volume = {},
number = {},
pages = {105229},
doi = {10.1016/j.pdpdt.2025.105229},
pmid = {40998038},
issn = {1873-1597},
abstract = {BACKGROUND: Dental caries is a biofilm-mediated disease primarily caused by Streptococcus mutans, which produces extracellular polysaccharides via glucosyltransferases such as GtfB, promoting biofilm formation and cariogenicity. Photodynamic therapy (PDT) has gained attention as a promising alternative to conventional antimicrobials. This study investigates the efficacy of blue laser-activated berberine-loaded nanoniosomes (nNios@Ber) in enhancing PDT against S. mutans biofilms formed on tooth enamel.

MATERIALS AND METHODS: An ex vivo model using human tooth enamel slabs was employed to cultivate S. mutans biofilms. Berberine was encapsulated in nanoniosomes (nNios@Ber) to improve stability and delivery. Following determination of minimum inhibitory concentration (MIC) of nNios@Ber, biofilms were treated with various concentrations of nNios@Ber combined with blue laser irradiation (405 ± 10 nm nm) to activate PDT. Additionally, quantitative real-time PCR was used to quantify gtfB gene expression.

RESULTS: The MIC of nNios@Ber against S. mutans was found to be 15.6 μg/mL. All PDT-treated groups showed significant reductions in biofilm viability and a dose-dependent downregulation of gtfB expression. Specifically, gtfB expression decreased by 4.8-, 5.7-, and 7.4-fold in the 2 × MIC, 4 × MIC, and 8 × MIC nNios@Ber plus blue laser groups, respectively (P < 0.05). Neither nNios@Ber nor laser alone produced statistically significant effects (P > 0.05). These findings confirm that nNios@Ber's anti-biofilm efficacy is enhanced when photoactivated by blue laser.

CONCLUSION: Blue laser-activated nNios@Ber significantly inhibit S. mutans biofilm formation and virulence gene expression, offering an effective and targeted approach for caries management. This photoactivated nNios@Ber enhances the photodynamic activity, providing a promising adjunct or alternative to conventional antimicrobial agents such as CHX in oral healthcare.},
}

@article {pmid40997643,
year = {2025},
author = {Lim, TW and Lee, J and Ab Ghani, SM and Burrow, MF and McGrath, C},
title = {Ultrasonic Cleaning and its Effects on Denture Biofilm: A Systematic Review.},
journal = {International dental journal},
volume = {75},
number = {6},
pages = {103921},
doi = {10.1016/j.identj.2025.103921},
pmid = {40997643},
issn = {1875-595X},
abstract = {INTRODUCTION AND AIMS: Denture biofilm is associated with various oral and systemic diseases. Therefore, an effective denture-cleaning method should be adopted by denture wearers. This systematic review aimed to evaluate the efficacy of ultrasonic denture cleaning in reducing plaque quantity and microbial load and changing the microbial composition.

METHODS: Clinical studies published in English with no restriction on the year of publication that compared ultrasonic denture cleaning with other cleaning methods were included. Literature was searched across 5 electronic databases (PubMed, Embase, Web of Science, Scopus, and Cochrane Central Register for Controlled Trials). An additional search via other methods was performed until June 2025. The Revised Cochrane Handbook risk-of-bias tools (RoB 2) for individually randomised and parallel-group trials, cluster-randomised trials, and crossover trials were employed for the risk of bias assessments. A narrative synthesis of the included studies was generated.

RESULTS: A total of 2,811 articles were identified. Fifteen studies underwent full-text article screening, and an additional 4 articles were added through manual search. Ultimately, 10 randomised controlled clinical trials reporting the efficacy of ultrasonic denture cleaning contributed to this review. All included studies, assessed using the RoB 2, were considered to be of 'some concerns' to a 'high risk' of bias. The present review identified a diverse range of ultrasonic cleaning protocols. From the synthesis of the 10 included studies, positive outcomes were observed in reducing denture plaque quantity and microbial load, particularly when a chemical cleansing method was combined with ultrasonic cleaning.

CONCLUSION: There was consistent evidence from randomised clinical trials that supported ultrasonic cleaning reduced denture plaque quantity and modified microbial load, particularly when supplemented with an effervescent denture cleanser.

CLINICAL RELEVANCE: Ultrasonic cleaning combined with chemical cleansing should be adopted by older adults to reduce denture plaque coverage and microbial loads.},
}

@article {pmid40997209,
year = {2025},
author = {Ivanov, M and Đorđevski, N and Dabić, J and Stojković, D},
title = {Cutibacterium acnes growth and biofilm formation is inhibited by flavonoids.},
journal = {Natural product research},
volume = {},
number = {},
pages = {1-5},
doi = {10.1080/14786419.2025.2565276},
pmid = {40997209},
issn = {1478-6427},
abstract = {Cutibacterium acnes is the cause of inflammatory acne and implanted devices associated infections. This study investigated flavonoids belonging to the 4 classes: flavanones (hesperidin, neohesperidin, naringenin, naringin, and eriodictyol), flavones (diosmetin, diosmin), isoflavone (genistein), and flavonols (kaempferol, galangin) as inhibitors of C. acnes growth in planktonic and biofilm form and assessed its potential to impact the integrity of cell membrane. Flavonoids could inhibit C. acnes growth, minimal inhibitory concentration (MIC) was in range 50-200 mg/L, with the lowest MIC (50 mg/L) for neohesperidin and eriodictyol. Minimal biofilm inhibitory concentrations were in range 25-200 mg/L, with the lowest values and highest antibiofilm potential recorded for eriodictyol, genistein, and kaempferol. Impact on cell membrane was significant in the presence of almost all examined flavonoids, as determined by crystal violet uptake assay. Flavonoids, especially eriodictyol, represent promising underexplored agents for combating infections and processes linked with the presence of C. acnes.},
}

@article {pmid40996457,
year = {2025},
author = {Jenjitwanich, A and Marx, H and Sauer, M},
title = {Metabolic Shifts Induced by pH Variation in Yarrowia lipolytica Biofilm.},
journal = {FEMS microbiology letters},
volume = {},
number = {},
pages = {},
doi = {10.1093/femsle/fnaf101},
pmid = {40996457},
issn = {1574-6968},
abstract = {The yeast Yarrowia lipolytica adapts its metabolite production based on cultivation conditions, with the pH value playing a critical role. At pH 3, most Y. lipolytica strains produce polyols, while at pH 5, they accumulate predominantly organic acids. Y. lipolytica has demonstrated the ability to transition from a planktonic, free-floating state to an immobilized state as a biofilm. This study aims to clarify the effects of pH level and carbon sources on the physiological state of Y. lipolytica when grown in a biofilm state. These pH variations were applied to the same biofilm culture to assess the capacity of given Y. lipolytica cells to undergo metabolic shifts and recovery under changing environmental conditions. Interestingly, a pH shift from 3 to 5 leads-as expected-to a metabolic shift from polyols to citric acid. However, the shift back to pH 3 does not revert back to polyols as major products. This study not only revealed an unexpected production pattern but also provided benefits for the industrial process in general. Understanding biofilm cultivation methods supports continuous bioprocesses using the immobilized nature of biofilm. pH-alternating experiments reveal how environmental condition fluctuations affect biofilm culture physiology.},
}

@article {pmid40994640,
year = {2025},
author = {Martins, SB and Teixeira, SC and de Souza, G and Alhatlani, BY and Abdallah, EM and Ambrosio, SR and Silva, TS and Bastos, JK and Tanimoto, MH and Barbosa, BF and Ferro, EAV and Martins, CHG},
title = {In vitro assessment of Brazilian red propolis against mycobacteria: antibacterial potency, synergy, inhibition of biofilm formation, and intramacrophage effects.},
journal = {Frontiers in pharmacology},
volume = {16},
number = {},
pages = {1630134},
pmid = {40994640},
issn = {1663-9812},
abstract = {BACKGROUND: Tuberculosis persists as a major global health threat and remains the leading cause of death from infectious disease. Efforts to control the disease are increasingly hampered by the emergence of drug-resistant Mycobacterium tuberculosis strains. At the same time, non-tuberculous mycobacteria are an expanding clinical concern, with few effective therapies available. Brazilian red propolis (BRP) has shown broad-spectrum antibacterial activity, yet its efficacy against mycobacteria is poorly characterized.

METHODS: This study evaluated the in vitro antimycobacterial potential of a crude hydroalcoholic extract of BRP (CHEBRP). Minimum inhibitory concentrations were determined against drug-susceptible and rifampicin-resistant M. tuberculosis strains (M. tuberculosis H37Rv-ATCC 27294, clinical isolate, and rifampicin-resistant clinical isolate; M. kansasii ATCC 12478 and clinical isolate; M. avium ATCC 25291 and clinical isolate). Fractional inhibitory concentration indices were calculated to assess interactions with isoniazid and rifampicin. Biofilm inhibition was measured, and cytotoxicity was assessed in RAW 264.7 macrophages. Intracellular activity was quantified using infected macrophage cultures.

RESULTS: CHEBRP exhibited potent activity against most M. tuberculosis strains tested, including rifampicin-resistant strains. Its combination with isoniazid or rifampicin yielded an indifferent interaction, supporting the feasibility of co-administration. CHEBRP significantly inhibited biofilm formation, showed minimal cytotoxicity toward macrophages, and achieved substantial clearance of intracellular bacilli.

CONCLUSION: These in vitro findings highlight CHEBRP as a promising candidate for adjunctive antimycobacterial therapy. Further studies should investigate its in vivo efficacy, pharmacokinetics, and activity against a broader range of mycobacterial species.},
}

@article {pmid40994194,
year = {2025},
author = {Wang, S and Cai, L and Edwards, AN and Melton, SJ and Retterer, ST and Doktycz, MJ and Morrell-Falvey, JL},
title = {Pseudomonas aeruginosa Adhesion and Biofilm Formation on Poly(l-lysine)-Tethered Hydrogels: Synergistic Effect of Substrate Stiffness and Positive Charge Density.},
journal = {Langmuir : the ACS journal of surfaces and colloids},
volume = {},
number = {},
pages = {},
doi = {10.1021/acs.langmuir.5c02709},
pmid = {40994194},
issn = {1520-5827},
abstract = {Infections associated with antibacterial-resistant Pseudomonas aeruginosa (P. aeruginosa) are the major cause of morbidity and mortality of patients, presenting one of the greatest therapeutic challenges for treatment of community-acquired and nosocomial infections. To develop antimicrobial hydrogel coatings to control the adhesion and subsequent biofilm formation of P. aeruginosa, we have used photo-cross-linked poly(ethylene glycol) diacrylate (PEGDA) hydrogels with varied cross-linking densities and covalently grafted poly(l-lysine) (PLL) at different weight compositions (ϕPLL). Both surface stiffness and positive charge density of the hydrogels were efficiently tuned over a broad range to investigate their effects on two main strains of P. aeruginosa, PA01 and PA14. We found that both number and viability of attached cells were positively correlated with the hydrogel stiffness, leading to thicker and larger coverage of cell colonies at 72 h postseeding on the stiffer substrates. The dependence of both PA01 and PA14 strains on ϕPLL, however, was nonmonotonic. Positive charges from dissociated amine groups in the grafted PLL chains significantly promoted initial adhesion and proliferation of both strains at low ϕPLL and developed into the thickest biofilms on the stiffest hydrogels grafted with ϕPLL of 1-2%. Nevertheless, on the softest hydrogels grafted with PLL at high ϕPLL of 7-10%, the bacteria no longer attached or survived. These results not only improved our fundamental understanding of bacteria-material interactions but also provided a series of PLL-grafted PEGDA hydrogels with controlled stiffness and positive charge density as ideal surface coating materials to prevent bacterial infections.},
}

@article {pmid40992437,
year = {2025},
author = {Kobayashi, F and Kawahara, T and Narai-Kanayama, A and Odake, S},
title = {Nanobubble water for the effective removal of biofilm formed by Escherichia coli.},
journal = {Journal of microbiological methods},
volume = {},
number = {},
pages = {107276},
doi = {10.1016/j.mimet.2025.107276},
pmid = {40992437},
issn = {1872-8359},
abstract = {The removal of biofilm (BF) formed by Escherichia coli using nanobubble (NB) water prepared with different gases and two generation methods was investigated. The E. coli BF removal efficiencies of all NB water samples tested were significantly higher than those of the non-NB water. In particular, nitrogen (N2) NB water exerted the greatest effect on the removal of E. coli BF. The surface tension of N2NB water was the lowest, although that of airNB water was the highest. The dissolved oxygen (O2) concentration in the O2NB water was drastically increased, although that in the N2NB and carbon dioxide (CO2) NB water was decreased. The pH of CO2NB water was significantly decreased, although that of N2NB, airNB, and O2NB water was increased. The E coli BF removal efficiencies of N2NB water were not different in the NB generators between ejector and shearing types. However, the surface tension, pH, and size distribution varied among the NB generator types. Therefore, it was found that NB water removed E. coli BF, and that its efficiencies differed depending on the gas type.},
}

@article {pmid40991694,
year = {2025},
author = {Hansen, KH and Golcuk, M and Byeon, CH and Tunc, A and Plechinger, EB and Dueholm, MKD and Conway, JF and Andreasen, M and Gur, M and Akbey, Ü},
title = {Structural basis of Pseudomonas biofilm-forming functional amyloid FapC formation.},
journal = {Science advances},
volume = {11},
number = {39},
pages = {eadx7829},
doi = {10.1126/sciadv.adx7829},
pmid = {40991694},
issn = {2375-2548},
mesh = {*Biofilms/growth & development ; *Amyloid/chemistry/metabolism/ultrastructure ; Cryoelectron Microscopy ; Molecular Dynamics Simulation ; *Pseudomonas aeruginosa/physiology/metabolism ; *Bacterial Proteins/chemistry/metabolism ; *Pseudomonas/physiology ; Protein Conformation ; },
abstract = {Biofilm-protected Pseudomonas aeruginosa causes chronic infections that are difficult to treat. FapC, the major biofilm-forming functional amyloid in Pseudomonas, is essential for biofilm integrity, yet its structural details remain unresolved. Using an integrative structural biology approach, we combine a solution nuclear magnetic resonance-based structural ensemble of unfolded monomeric FapC, a ~3.3-angstrom-resolution cryo-electron microscopy (cryo-EM) density map of FapC fibril, and all-atom molecular dynamics (MD) simulations to capture the transition from the unfolded to folded monomer to the fibrillar fold, providing a complete structural view of FapC biogenesis. Cryo-EM reveals a unique irregular triple-layer β solenoid cross-β fibril composed of a single protofilament. MD simulations initiated from monomeric and fibrillar FapC mapped structural transitions, offering mechanistic insights into amyloid assembly and disassembly. Understanding FapC reveals how Pseudomonas exploits functional amyloids for biofilm formation, and establishes a structural and mechanistic foundation for developing therapeutics targeting biofilm-related infection and antimicrobial resistance.},
}

*Biofilms/growth & development
*Amyloid/chemistry/metabolism/ultrastructure
Cryoelectron Microscopy
Molecular Dynamics Simulation
*Pseudomonas aeruginosa/physiology/metabolism
*Bacterial Proteins/chemistry/metabolism
*Pseudomonas/physiology
Protein Conformation

@article {pmid40991639,
year = {2025},
author = {Rasheed, N and Ali, X and -Ud-Din, S and Khan, I and Hassan, A and Rasheed, MA},
title = {The function of Anr in the differential effects of oxygen levels on biofilm development and nitrogenase performance in Pseudomonas stutzeri A1501.},
journal = {PloS one},
volume = {20},
number = {9},
pages = {e0333183},
doi = {10.1371/journal.pone.0333183},
pmid = {40991639},
issn = {1932-6203},
mesh = {*Biofilms/growth & development/drug effects ; *Pseudomonas stutzeri/genetics/physiology/enzymology/metabolism ; *Oxygen/metabolism/pharmacology ; *Nitrogenase/metabolism/genetics ; *Bacterial Proteins/genetics/metabolism ; Gene Expression Regulation, Bacterial ; Nitrogen Fixation/genetics ; *Transcription Factors/genetics/metabolism ; Sigma Factor/genetics/metabolism ; },
abstract = {Pseudomonas stutzeri A1501 exhibits a rare and notable trait: nitrogenase activity, which functions under microaerophilic conditions with limited oxygen availability. Optimal biofilm formation occurs in minimal media under nitrogen-depleted conditions. Anr is a transcription regulator with a widespread influence that accelerates the process of biofilm development. The lack of Anr adversely affects nitrogen fixation by regulating the activity of nifA, nifH, and ntrC. The anr insertion mutant significantly reduced the nitrogenase activity. Nitrogenase activity demonstrated considerable variation at different oxygen concentrations. The quantitative reverse transcription polymerase chain reaction (qRT-PCR) demonstrated a reduction in the expression of nif island genes during nitrogen fixation in the absence of anr gene. The discovery revealed that different oxygen levels in the environment significantly influence nitrogenase activity and biofilm formation. The qRT-PCR investigation demonstrated an upregulation of narL gene expression during biofilm formation, suggesting that reduced oxygen levels initiate a signaling cascade that activates the anr gene. This influences both the expression of RpoS (sigma factor) genes and the process of biofilm formation.},
}

*Biofilms/growth & development/drug effects
*Pseudomonas stutzeri/genetics/physiology/enzymology/metabolism
*Oxygen/metabolism/pharmacology
*Nitrogenase/metabolism/genetics
*Bacterial Proteins/genetics/metabolism
Gene Expression Regulation, Bacterial
Nitrogen Fixation/genetics
*Transcription Factors/genetics/metabolism
Sigma Factor/genetics/metabolism

@article {pmid40990949,
year = {2025},
author = {Oster, S and Bollinger, E and Schreiner, VC and Schmitt, T and Filker, S and Bundschuh, M},
title = {Functional stability despite structural changes in freshwater biofilm communities exposed to an antibiotic and an herbicide - the role of nutrient conditions.},
journal = {FEMS microbiology ecology},
volume = {},
number = {},
pages = {},
doi = {10.1093/femsec/fiaf094},
pmid = {40990949},
issn = {1574-6941},
abstract = {Freshwater autotrophic biofilms play a vital role in primary production and nutrient cycling in freshwater ecosystems but are increasingly exposed to chemical stressors such as antibiotics or herbicides. Although nutrient availability may modulate biofilm sensitivity, its impact on biofilm responses to these stressors remains poorly understood. In four independent experiments, we investigated the functional (ash-free dry weight and chlorophyll a, b and c) and structural (16S/18S rRNA metabarcoding) responses of stream-derived biofilms under low- and high-nutrient levels to chronic exposure (14 days) to the antibiotic ciprofloxacin and the herbicide propyzamide in laboratory stream microcosms. High-nutrient levels strongly increased biofilms functional responses and altered the community composition. Chemical exposure led to pronounced shifts in prokaryotic (ciprofloxacin) and eukaryotic (propyzamide) communities, but without significant effects on functional responses, suggesting functional redundancy and ecological buffering capacity of freshwater biofilms. These results highlight the critical role of nutrient supply in biofilm responses and the need for caution when extrapolating laboratory results to field conditions.},
}

@article {pmid40990775,
year = {2025},
author = {Favaro, JC and Geha, O and Canavarros, TN and Nascimento, A and Lopes, MB and Guiraldo, RD and Berger, SB},
title = {Addition of silver nanoparticles to universal adhesive: In vitro effect on biofilm and shear bond strength.},
journal = {Brazilian dental journal},
volume = {36},
number = {},
pages = {e236255},
doi = {10.1590/0103-644020256255},
pmid = {40990775},
issn = {1806-4760},
mesh = {*Silver/chemistry/pharmacology ; *Biofilms/drug effects ; *Shear Strength ; *Metal Nanoparticles/chemistry ; Cattle ; Animals ; Streptococcus mutans/drug effects ; In Vitro Techniques ; Materials Testing ; *Dental Cements/chemistry ; *Dental Bonding ; },
abstract = {Silver nanoparticles (AgNPs) have been incorporated into dental materials at low concentrations to provide antibacterial action without compromising mechanical properties. This in vitro study evaluated the antibiofilm effect and bond strength of an experimental universal adhesive with AgNPs (EAg). Streptococcus mutans biofilm was induced by incubating samples with 0.01% and 0.02% AgNPs, based on a pilot study, compared to a control (experimental without AgNPs, ES) in a 20% sucrose medium. This was followed by sonication and counting of viable cells after 1 and 7 days (n = 9). Enamel and dentine bovine micro shear bond strength test (μ-SBS) was performed (n=10) with EAg0.01% and controls (ES and the commercial OPT (Optbond Universal; Kerr). μ-SBS data of enamel and dentin were evaluated for normality and homogeneity by Shapiro-Wilk and Levene, respectively, resulting in normality. Therefore, they were subjected to ANOVA. The failure type was evaluated using a stereoscopic magnifying glass at '40 and categorized as adhesive, cohesive, and mixed failure. The 0.01% concentration demonstrated the antibiofilm effect at the lowest AgNP concentration and was selected for the μ-SBS test. For μ-SBS ANOVA, there were no statistically significant differences between experimental and commercial adhesives (p<0.05). Evaluation of failure mode showed a predominance of adhesive failure on both substrates for all adhesives. The EAg exhibited antibiofilm activity with adhesive performance statistically similar to that of the commercial adhesive. Experimental universal adhesive containing silver nanoparticles showed antibacterial activity without compromising μ-SBS to enamel and dentin.},
}

*Silver/chemistry/pharmacology
*Biofilms/drug effects
*Shear Strength
*Metal Nanoparticles/chemistry
Cattle
Animals
Streptococcus mutans/drug effects
In Vitro Techniques
Materials Testing
*Dental Cements/chemistry
*Dental Bonding

@article {pmid40990450,
year = {2025},
author = {Wang, H and Li, Y and Chen, H and Yu, P and Shi, B and Alvarez, PJJ},
title = {Perfluorooctanoic Acid Can Increase Biofilm Resilience and Virulence in Drinking Water Distribution Systems.},
journal = {Environmental science & technology},
volume = {},
number = {},
pages = {},
doi = {10.1021/acs.est.5c07943},
pmid = {40990450},
issn = {1520-5851},
abstract = {Despite the prevalence of perfluorooctanoic acid (PFOA) in drinking water distribution systems (DWDSs), its effects on biofilm formation and tolerance to residual disinfectants remain largely unknown. Here, we report that DWDS-relevant levels of PFOA (1-80 ng/L) promoted Pseudomonas aeruginosa biofilm formation (by 1.8- to 4.5-fold) and chlorine tolerance (by 7-22%). PFOA also enhanced biofilm tolerance to common antibiotics, increasing their minimum inhibitory concentrations by 25-67%. In a flow-through system, biofilm evolution toward higher resilience was attributed to PFOA-exerted oxidative stress, which fortuitously upregulated quorum sensing and stimulated extracellular polymeric substances production. PFOA exposure also increased bacterial chemotaxis and activated cyaA and cyaB genes, which enhanced the cAMP/Vfr signaling pathway, contributing to the upregulation of virulence factor genes. PFOA-induced chlorine and antibiotic resistance and increased virulence were also verified with a mesocosm system and a real DWDS pipeline biofilm. Overall, these results underscore the need for proactive microbial risk assessment associated with such indirect effects of emerging pollutants in DWDSs.},
}

@article {pmid40990384,
year = {2025},
author = {Fortes, CV and Ribeiro, AB and Wever, B and Sakly, A and Oliveira, VC and Clemente, LM and Watanabe, E and Silva-Lovato, CH},
title = {Evaluation of biofilm formation, adhesive strength and effectiveness of cleaning protocols on adhesive-containing acrylic resin specimens: An in vitro study.},
journal = {Dental and medical problems},
volume = {},
number = {},
pages = {},
doi = {10.17219/dmp/176231},
pmid = {40990384},
issn = {2300-9020},
abstract = {BACKGROUND: Denture adhesives promote greater stability and retention of dentures. However, they can also facilitate biofilm formation related to oral diseases.

OBJECTIVES: The study aimed to evaluate the influence of 2 adhesives on the microbial load of mixed biofilm and adhesive strength. Additionally, the objective was to assess the effect of 3 hygiene protocols on the microbial load and cell metabolism of this biofilm.

MATERIAL AND METHODS: The study compared Corega Ultra Cream (CCA) and OlivaFix® Gold (OFA) adhesives by evaluating the biofilm formation of Candida albicans, Candida glabrata, Staphylococcus aureus, and Streptococcus mutans by colony-forming unit (CFU), as well as adhesive strength. The implemented hygiene protocols included brushing and immersion in water (BW), 0.15% triclosan (BT0.15%), or 0.25% sodium hypochlorite (BSH0.25%). The control groups were either without adhesive (CG) or without any hygiene protocols (CGwH). The one-way and two-way analyses of variance (ANOVAs) with Tukey's post hoc test and a generalized linear model with Bonferroni adjustment were used for statistical analysis (α = 0.05).

RESULTS: The microbial load of C. albicans was higher when OFA was used (p < 0.001). The microbial loads of C. glabrata and S. mutans were similar between adhesives and higher in the CG (p < 0.001). The influence of the adhesives on the microbial load of S. aureus was not statistically significant (p = 0.287). The adhesive strength promoted by OFA was greater and more stable than when CCA was used (p = 0.007). The immersion in sodium hypochlorite led to a reduction in the microbial load of C. albicans (p < 0.001), C. glabrata (p = 0.002) and S. mutans (p = 0.012), independent of the adhesive. For S. aureus, the microbial load was lower with OFA/BSH0.25% (p = 0.022). All hygiene protocols resulted in a decreased cell metabolism when compared to the CGwH (p < 0.001).

CONCLUSIONS: Brushing with BSH0.25% solution was the most effective hygiene protocol, resulting in a reduction in the microbial load and metabolism. This protocol may be recommended as a first-line option for the disinfection of dentures.},
}

@article {pmid40989607,
year = {2025},
author = {Salman, SAK and Al Muhana, BMM},
title = {Detection of biofilm genes in MDR Staphylococcus aureus isolated from human and cattle urinary tract infections in Babylon Governorate, Iraq.},
journal = {Open veterinary journal},
volume = {15},
number = {6},
pages = {2551-2561},
pmid = {40989607},
issn = {2218-6050},
mesh = {Animals ; Cattle ; *Biofilms/growth & development ; *Staphylococcus aureus/genetics/physiology/drug effects/isolation & purification ; Humans ; *Urinary Tract Infections/microbiology/veterinary/epidemiology ; Iraq/epidemiology ; *Staphylococcal Infections/microbiology/veterinary/epidemiology ; *Drug Resistance, Multiple, Bacterial/genetics ; *Cattle Diseases/microbiology/epidemiology ; Anti-Bacterial Agents/pharmacology ; },
abstract = {BACKGROUND: Staphylococcus aureus (S. aureus) is a major pathogenic bacterium in veterinary medicine and human health. It is one of the most important bacterial agents causing urinary tract infection (UTI). The increasing incidence of multidrug-resistant and biofilm-forming S. aureus is a great problem today. The aim of the study was to investigate the phenotypic and genotypic aspects of biofilm formation in multiple drug-resistant (MDR) S. aureus isolated from UTI infection in human and cattle in Babylon, Iraq.

METHODOLOGY: A total of 168 and 172 urine samples were collected from UTI infection in humans and cattle, respectively, during the period from November 2023 to February 2024. Morphological and biochemical identification was used to diagnose S. aureus. Additional confirmation was performed by the automated Vitek 2 compact system.

RESULTS: Among the 168 human isolates, 24 (14.2%) and 172 cattle isolates, 16 (9.3%) isolates were diagnosed as S. aureus. Genotypically identification of the isolates was performed using the 16s RNA gene. Twenty-two (91.6%) from 24 human source S. aureus isolates and 8 (50%) from 16 animal source S. aureus isolates were found to be MDR according to the Vitek 2 compact system. Results revealed that 19 (86.3%) and 7(87.5%) MDR isolates were phenotypically positive for biofilm production in concern to human and animal source isolates, respectively. Genotypically, three polysaccharide intracellular adhesion genes and one MSCRAMMs (fib) were screened in MDR bacteria using specific primers. The prevalence rate of genes was icaA (100%), icaB (100%), and icaC (100%) in all 30 MDR isolates. fib gene were present in 63.6% of human isolates and 75% of cattle source isolates.

CONCLUSION: The study has shown that the biofilm-forming S. aureus that causes UTI were resistant to multiple antibiotic agents. These findings underscore the necessity of development effective treatment approaches to control UTI infections in humans and animals.},
}

Animals
Cattle
*Biofilms/growth & development
*Staphylococcus aureus/genetics/physiology/drug effects/isolation & purification
Humans
*Urinary Tract Infections/microbiology/veterinary/epidemiology
Iraq/epidemiology
*Staphylococcal Infections/microbiology/veterinary/epidemiology
*Drug Resistance, Multiple, Bacterial/genetics
*Cattle Diseases/microbiology/epidemiology
Anti-Bacterial Agents/pharmacology

@article {pmid40988982,
year = {2025},
author = {Strzelecki, P and Ferté, T and Klimczuk, T and Zielińska-Jurek, A and Szalewska-Pałasz, A and Nowicki, D},
title = {Trans-Cinnamaldehyde-Driven Silver Nanoparticles: Dual Role in Targeting Biofilm Disruption and Control of Biofilm‑Forming Pathogens via Impairing Ferrous Ion Uptake.},
journal = {Nanotechnology, science and applications},
volume = {18},
number = {},
pages = {387-403},
pmid = {40988982},
issn = {1177-8903},
abstract = {PURPOSE: Biofilm-related infections, especially those associated with medical devices like catheters, pose significant clinical challenges due to their resistance to conventional treatments. This study investigates a green chemistry-based approach to synthesize silver nanoparticles (AgNPs) stabilized with trans-cinnamaldehyde (t-CA) and evaluates their potential for combating microbial biofilms and based on novel mechanism of action.

METHODS: Silver nanoparticles (t-CA-AgNPs) were synthesized using t-CA as both a reducing and stabilizing agent. The NPs were then thoroughly characterized using UV-Vis spectroscopy, X-ray diffraction (XRD), electron microscopy (TEM, SEM, STEM), and dynamic light scattering (DLS). We evaluated its antimicrobial potential against the most prevalence biofilm-forming pathogens including Pseudomonas aeruginosa, Escherichia coli and Candida albicans using minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) assays. Moreover, we investigated the mechanism of action of t-CA-AgNPs underlying biofilm inhibition. Biofilm formation and structure were verified by SEM imagining.

RESULTS: DLS analysis confirmed that t-CA-AgNPs had an average particle diameter of 2.5 nm, coupled with a notably negative zeta potential (-45 mV), indicative of good colloidal stability. t-CA-AgNPs displayed potent antimicrobial properties, with MIC values ranging from 26 to 412 µg/mL and MBC values from 103 to 825 µg/mL. Biofilm formation inhibitory properties reached 88.74% of inhibition for P. aeruginosa and 70.60% for E. coli. Moreover, we found potent metal ion-chelating capabilities, importantly, in binding and reducing ferrous ions, the crucial factor of biofilm formation. Furthermore, t-CA-AgNPs substantially impaired biofilm development on catheter surfaces, underscoring their robust antibiofilm potential.

CONCLUSION: Presented here t-CA-AgNPs exhibit significant antimicrobial and antibiofilm activity. By effectively targeting critical elements in biofilm formation, such as ferrous ions, coupled with antimicrobial potential of both active compounds, these green-synthesized NPs have potential applications in significantly improving the safety and effectiveness of medical devices. However, further studies are needed to ensure their efficacy in clinical use.},
}

@article {pmid40987911,
year = {2025},
author = {Rajalakshmi, E and J L, ES and Balakrishnan, A and Leela, KV and Ramya, M},
title = {Exploring the impact of quercetin on the growth and biofilm formation of Leptospira interrogans.},
journal = {Folia microbiologica},
volume = {},
number = {},
pages = {},
pmid = {40987911},
issn = {1874-9356},
abstract = {Leptospira interrogans, the causative agent of leptospirosis, presents major challenges due to its ability to form environmental biofilms, persistent virulence, and emerging antimicrobial resistance. Plant-derived bioactive compounds offer promising alternatives to conventional antibiotics in targeting bacterial biofilms. This study investigates the antibiofilm and antimicrobial potential of quercetin, a plant-derived polyphenol, against L. interrogans. Quercetin significantly inhibited biofilm formation (91%), induced 44% biofilm dispersion at 500 µg/mL, and reduced viable cell counts by 92% at 1000 µg/mL compared to the untreated control. These effects were further validated using fluorescence-activated cell sorting (FACS), which demonstrated reduction in the live cells in the presence of quercetin. The expression levels of key genes associated with virulence and biofilm regulation were assessed using quantitative RT-PCR, revealing downregulation of csrA and upregulation of lipL32 in quercetin-treated cells. Potential molecular interactions between several leptospiral proteins and quercetin were proven through AutoDock Vina and PyMOL. These findings highlight quercetin's potential as an anti-leptospiral agent for managing leptospiral biofilm formation and dispersions.},
}

@article {pmid40985635,
year = {2025},
author = {Zhao, H and Dufour, D and Ghobaei, N and Bozec, L and Lévesque, CM},
title = {DNA adenine methylation influences gene expression and biofilm formation in Streptococcus mutans.},
journal = {Applied and environmental microbiology},
volume = {},
number = {},
pages = {e0109425},
doi = {10.1128/aem.01094-25},
pmid = {40985635},
issn = {1098-5336},
abstract = {UNLABELLED: Streptococcus mutans, a key oral pathogen, utilizes quorum sensing to regulate biofilm formation-a major virulence factor in the development of dental caries (tooth decay). Our recent research uncovered a complex interplay between the CSP-ComDE quorum sensing pathway and the Type II DpnII restriction-modification (R-M) system in S. mutans. The DpnII R-M system methylates adenine at 5'-GATC-3' sites and cleaves unmethylated DNA, significantly influencing foreign DNA acquisition and gene expression. In this study, we investigated the impact of a ΔRM mutant, which lacks adenine methylation, on biofilm formation. The ΔRM mutant formed fragile biofilms that easily detach from surfaces, with significantly reduced exopolysaccharide content and increased extracellular DNA, which appears to be associated with membrane vesicle production rather than cell lysis. RNA-seq analysis revealed only few differentially expressed genes directly involved in biofilm formation, such as gtfC, suggesting that the biofilm defect may result from indirect effects or alternative regulatory mechanisms. Notably, the downregulation of mutanobactin-related genes and upregulation of genes involved in de novo purine nucleotide biosynthesis point to novel pathways influenced by DNA methylation. These findings contribute to a deeper understanding of the multifactorial nature of biofilm formation and the role of epigenetic modifications in microbial behavior.

IMPORTANCE: This study highlights the critical role of DNA methylation in regulating biofilm formation and virulence in Streptococcus mutans. By examining the interplay between adenine methylation, extracellular DNA (eDNA), membrane vesicles (MVs), and glucan production, we provide new insights into the complex biology of biofilm development. Our findings challenge traditional views by emphasizing the importance of MVs and eDNA in maintaining biofilm integrity. Understanding these epigenetics modifications not only advances our knowledge of microbial regulation but also identifies novel targets for antimicrobial therapy. Since adenine methylation is rare or absent in mammalian cells, targeting this modification presents a promising strategy to disrupt biofilm formation and combat bacterial infections. The insights gained from this study may inform the development of innovative approaches to manage biofilm-associated infections and improve oral health outcomes.},
}

@article {pmid40985596,
year = {2025},
author = {Arbulu, S and Oftedal, TF and Kjos, M},
title = {Transcriptomic response in planktonic and biofilm-associated cells of S. mutans treated with sublethal concentrations of chlorhexidine.},
journal = {FEMS microbiology letters},
volume = {},
number = {},
pages = {},
doi = {10.1093/femsle/fnaf100},
pmid = {40985596},
issn = {1574-6968},
abstract = {Chlorhexidine, an antimicrobial with a broad inhibitory spectrum, is commonly used to treat oral infections as an active ingredient in mouthwash. While typically used at high concentrations (1-2 mg/ml), oral bacteria can be exposed to sublethal concentrations due to the bioavailability and protective barrier of biofilms (dental plaques). Sublethal concentrations can cause transcriptional remodelling of bacteria such as Streptococcus mutans, a key player in dental caries. Using an RNA-seq approach, this report provides a compendium on the effect of sublethal concentrations of chlorhexidine on the transcriptome of S. mutans as planktonic cells and in biofilm states. S. mutans showed major transcriptional remodelling between planktonic and biofilm states. The transcriptional response towards chlorhexidine was more pronounced in planktonic cells compared to sessile cells. However, the response observed for biofilm-associated cells was not specific to chlorhexidine, as the transcriptional response in biofilms exposed to the β-lactam amoxicillin was similar to those observed for chlorhexidine. Furthermore, we found that S. mutans modulates transcription of a multitude of ABC transporters both in planktonic and biofilm-associated cells upon exposure to these antimicrobials.},
}

@article {pmid40983296,
year = {2025},
author = {Yang, T and Han, Y and Zhang, M and Li, L and Chen, M and Li, N and Wang, X},
title = {Quorum sensing enhances extracellular electron uptake of electrotrophic biofilm via metabolic cascade for aerobic biochemical oxygen demand sensing.},
journal = {Bioresource technology},
volume = {439},
number = {},
pages = {133363},
doi = {10.1016/j.biortech.2025.133363},
pmid = {40983296},
issn = {1873-2976},
abstract = {Electrotrophic biocathodes enable rapid biochemical oxygen demand (BOD) sensing but face challenges like slow colonization, low current, and poorly understood microbial interactions. This study employed exogenous quorum sensing signals (C6-HSL and 3O-C12-HSL) to enhance biofilm development on oxygen-reducing biocathodes. These two N-acyl-homoserine lactones (AHLs) accelerated formation and boosted current density 5-6 times. Candidatus Tenderia served as key electrotrophs, while non-electroactive Moheibacter promoted early adhesion and extracellular polymeric substance (EPS) production. Metagenomics showed upregulation of genes related to iron metabolism, adhesion, and electron transfer, alongside increased fulvic-like shuttle secretion. A cascade model was proposed: AHLs initiate non-electrotrophic EPS production, facilitating electrotrophs colonization and electron transfer. AHL-enhanced biofilm showed improved BOD sensing sensitivity and linearity, providing a new strategy and mechanistic basis for developing efficient biocathode biosensors.},
}

@article {pmid40982215,
year = {2025},
author = {Chalana, A and Thakur, V and Gupta, V and Sharma, P and Capalash, N},
title = {Synergistic Combination of Imipenem and 7-Fluoroindole Inhibits β-lactamases and Biofilm in Carbapenem-resistant Acinetobacter baumannii.},
journal = {Journal of applied microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1093/jambio/lxaf234},
pmid = {40982215},
issn = {1365-2672},
abstract = {BACKGROUND: Acinetobacter baumannii is a nosocomial opportunistic pathogen responsible for hospital and community-acquired infections. It has been categorised as a high-priority ESKAPE pathogen due to escalating resistance, underscoring the urgent need for effective treatment options. This study explores the use of 7-Fluoroindole (7-FI) as an adjuvant to imipenem for combating Carbapenem-Resistant Acinetobacter baumannii (CRAB).

METHODS: Clinical strains of A. baumannii were subjected to disc diffusion assay for antibiotic resistance profile, and Fractional Inhibitory Concentration (FIC) of 7-FI was determined with imipenem by checkerboard assay. Expression of β-lactamases, efflux pumps, and outer membrane proteins was determined by qRT-PCR. Biofilm inhibition by 7-FI in combination with imipenem was evaluated by Confocal Laser Scanning Microscopy.

RESULTS: 7-FI reduced the MIC of imipenem by 2 to 8-fold in CRAB strains, and also enhanced susceptibility to meropenem. Mechanistically, 7-FI inhibited the β-lactamase activity by 2.3 to 3.8-fold and downregulated the expression of Class B (blaNDM-1) and D (blaOXA-23 and blaOXA-51) β-lactamases. The efflux pump activity was reduced by 2 to 8.6-fold, and expression of efflux pump genes adeB, adeJ, abeM and emrA was also significantly (p < 0.001) downregulated. 7-FI in combination with imipenem inhibited biofilm formation by 80% even at 1 × FIC (1/4 MIC of imipenem and 1/3 MIC of 7-FI).

CONCLUSION: The combination of 7-FI and imipenem synergistically mitigated carbapenem resistance in A. baumannii by inhibiting the activity of β-lactamases, downregulating the expression of β-lactamases and efflux pumps. The study shows 7-FI as a useful adjuvant to imipenem against CRAB strains.},
}

@article {pmid40981720,
year = {2025},
author = {Sakyi Opoku, NYP and Mishra, A and Fletcher, H and Nizet, V and Abdul-Mutakabbir, JC},
title = {Enhancing Antimicrobial Susceptibility Testing for Acinetobacter baumannii Using Physiologically Relevant Culture Media and Biofilm Formation Assays.},
journal = {Current protocols},
volume = {5},
number = {9},
pages = {e70207},
doi = {10.1002/cpz1.70207},
pmid = {40981720},
issn = {2691-1299},
mesh = {*Acinetobacter baumannii/drug effects/physiology ; *Biofilms/drug effects/growth & development ; Microbial Sensitivity Tests/methods ; *Culture Media/chemistry ; *Anti-Bacterial Agents/pharmacology ; Humans ; },
abstract = {Acinetobacter baumannii is a high-risk pathogen associated with increased patient morbidity and mortality. Host-pathogen interactions amplify its virulence, in part by promoting biofilm formation-a crucial factor in antimicrobial resistance and persistence. Given the bacterium's strong propensity for acquiring resistance, antimicrobial susceptibility testing (AST) is essential for guiding effective therapeutic interventions. However, discrepancies have been observed between in vitro AST results and therapeutic outcomes, with some antimicrobials being deemed to show in vivo efficacy despite appearing ineffective in vitro. This discordance may stem from traditional AST protocols, which rely on bacteriological media such as Mueller Hinton broth (MHB) optimized for bacterial growth but not for mimicking the host environment. Moreover, conventional AST does not account for virulence traits such as biofilm formation, which further contribute to treatment failure. Incorporating physiologically relevant culture media, such as Roswell Park Memorial Institute (RPMI) 1640 medium, alongside assessment of biofilm formation may improve the predictive value of AST. This work outlines two complementary protocols for improving AST interpretation in A. baumannii infections. Basic Protocol 1 compares minimum inhibitory concentration (MIC) values generated using MHB and RPMI. Basic Protocol 2 evaluates biofilm formation in MHB, tryptic soy broth (TSB; control), and RPMI, with and without antimicrobial exposure. Together, these approaches aim to inform alternative AST strategies that better reflect in vivo conditions and optimize therapeutic decision-making. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Comparing A. baumannii minimum inhibitory concentration (MIC) results in bacteriological (MHB) versus physiological (RPMI) media Basic Protocol 2: Comparing A. baumannii isolate(s) biofilm formation following assays completed in bacteriological culture media (MHB and control TSB) and physiological medium (RPMI).},
}

*Acinetobacter baumannii/drug effects/physiology
*Biofilms/drug effects/growth & development
Microbial Sensitivity Tests/methods
*Culture Media/chemistry
*Anti-Bacterial Agents/pharmacology
Humans

@article {pmid40981469,
year = {2025},
author = {Maree, M and Ushijima, Y and Krama, A and Sasaki, M and Miyata, T and Higashide, M and Nguyen, LTT and Morikawa, K},
title = {Mixed-biofilm natural transformation assay reveals the presence of staphylococci in human environments that can transfer SCCmec to Staphylococcus aureus.},
journal = {mSphere},
volume = {},
number = {},
pages = {e0044225},
doi = {10.1128/msphere.00442-25},
pmid = {40981469},
issn = {2379-5042},
abstract = {Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen that causes healthcare-, community-, and livestock-associated infections. The methicillin resistance gene mecA is embedded in the mobile genetic element termed Staphylococcal Cassette Chromosome (SCCmec). SCCmec is shared among staphylococci inhabiting human and animal hosts, which are recognized epidemiologically as the genetic reservoir of SCCmec. However, the ability of diverse methicillin-resistant staphylococci (MRS) to serve as SCCmec donors for S. aureus has not been tested experimentally. Here, we investigated the ability of 157 MRS isolates from pets, meat, livestock, and humans to transfer SCCmec to methicillin-sensitive S. aureus strains using a recently developed natural transformation protocol in mixed biofilms. We found that 25 out of 157 isolates were able to transfer SCCmec to S. aureus. The most effective donor species were S. epidermidis (~33% of the tested isolates), S. felis (40%), and S. capitis (30%). Isolates from meat and livestock (collected in Vietnam and Thailand) had lower transfer rates of SCCmec (5% and 3%, respectively), compared to human and pet isolates from Japan (35% and 25%, respectively). The SCCmec transfer depended on site-specific integration/excision mediated by an intact attB site, which is recognized by the SCC recombinase Ccr. Our study experimentally demonstrates the presence of SCCmec donors in our living environments, highlighting the importance of specific staphylococcal species.IMPORTANCEHow MRSA emerges has long been the pivotal question regarding the ever-increasing burden of antimicrobial resistance (AMR) issues for over half a century. Extensive research efforts in bacteriology, epidemiology, genome biology, and healthcare fields have led to the common understanding that SCCmec is transmitted among distinct staphylococcal species. However, global efforts to provide empirical evidence for intercellular SCCmec transmission have yielded limited results. We recently established the mixed-biofilm transformation assay to evaluate intercellular and interspecies SCCmec transmission. This novel assay system allows us to gain insight into the question "How MRSA emerges," and here, we provide the first experimental results about the potential donor species and habitats. This is the first report to show the ability of staphylococci from distinct sources to transfer SCC to S. aureus. Moreover, the new finding of S. felis as an effective donor that is not commensal to humans reinforces the importance of the One Health concept.},
}

@article {pmid40981351,
year = {2025},
author = {García, MT and Morán, MC and Pons, R and Hafidi, Z and Bautista, E and Vazquez, S and Pérez, L},
title = {Arginine-Derived Cationic Surfactants Containing Phenylalanine and Tryptophan: Evaluation of Antifungal Activity, Biofilm Eradication, Cytotoxicity, and Ecotoxicity.},
journal = {Journal of xenobiotics},
volume = {15},
number = {5},
pages = {},
doi = {10.3390/jox15050140},
pmid = {40981351},
issn = {2039-4713},
support = {Grant PID2022-136354-NBI00//MCIN/AEI/10.13039/501100011033 and by "ERDF A way of making Europe/ ; PTA2020-018511-I//MCIN/AEI/10.13039/501100011033/ ; Grant PRE2022-000837//MCIN/AEI/ 10.13039/501100011033 and "ESF Investing in your future/ ; COOPA23026//CSIC/ ; },
abstract = {Due to the growing emergence of bacterial and fungal resistance, there is an urgent need for novel antimicrobial compounds. Cationic surfactants are effective antimicrobial agents; however, traditional quaternary ammonium compounds (QACs) are increasingly scrutinized due to their cytotoxicity, poor biodegradability, and harmful effects on aquatic ecosystems. While the antimicrobial efficacy of many new biocides, including QACs, has been extensively studied, comprehensive experimental strategies that simultaneously assess antimicrobial activity, mammalian cell toxicity, and ecotoxicity remain limited. Recent studies have reported that amino-acid-based surfactants containing arginine-phenylalanine and arginine-tryptophan exhibit excellent antibacterial activity and are biodegradable. This work extends their biological characterization to evaluate their potential applications. Specifically, we examined how variations in the head group architecture and hydrophobic moiety influence antifungal and antibiofilm activity. We also assessed how these structural parameters impact cytotoxicity and ecotoxicity. These compounds demonstrated strong activity against a wide range of Candida strains. Their hydrophobic character primarily influenced both antifungal efficacy and cytotoxicity. Importantly, these surfactants exhibited potent antimicrobial and antibiofilm effects at non-cytotoxic concentrations. Notably, their aquatic toxicity was significantly lower than that of conventional QACs.},
}

@article {pmid40978156,
year = {2025},
author = {Heine, N and Bittroff, K and Szafrański, SP and Duitscher, M and Behrens, W and Vollmer, C and Mikolai, C and Kommerein, N and Debener, N and Frings, K and Heisterkamp, A and Scheper, T and Torres-Mapa, ML and Bahnemann, J and Stiesch, M and Doll-Nikutta, K},
title = {Influence of species composition and cultivation condition on peri-implant biofilm dysbiosis in vitro.},
journal = {Frontiers in oral health},
volume = {6},
number = {},
pages = {1649419},
pmid = {40978156},
issn = {2673-4842},
abstract = {INTRODUCTION: Changes in bacterial species composition within oral biofilms, known as biofilm dysbiosis, are associated with the development of severe oral diseases. To better understand this process and help establish early detection systems, models are needed which replicate oral biofilm dysbiosis in vitro - ideally by also mimicking natural salivary flow conditions.

METHODS: For this purpose, the present study cultivated two different combinations of oral commensal and pathogenic strains - Streptococcus oralis, Actinomyces naeslundii, Veillonella dispar/parvula, Fusobacterium nucleatum and Porphyromonas gingivalis - comparatively within an established flow chamber model on the implant material titanium, and statically in 6-well plates for 21 days. Biofilm morphology, species distribution, and bacterial metabolism were analyzed by fluorescence microscopy, molecular biological methods, and metabolic interaction prediction.

RESULTS: Biofilm growth and composition were strongly influenced by bacterial species selection, and to a more minor extent, by cultivation conditions. Within the model containing V. dispar and a laboratory P. gingivalis strain, a diversification of commensal species was observed over time along with a significantly reduced pH-value. In contrast, the model containing V. parvula and the clinical isolate P. gingivalis W83, a dysbiotic shift with increased pathogen levels, pH-value, and virulence factors was achieved.

CONCLUSION: Within the present study, different in vitro oral multispecies biofilm models were successfully developed. Depending on bacterial species selection, these models were able to depict the infection-associated dysbiotic shift in species composition under flow conditions solely by intrinsic interactions and without the use of external stimuli.},
}

@article {pmid40977362,
year = {2025},
author = {Kivimaki, SE and Dempsey, S and Camper, C and Tani, JM and Ray, WK and Hicklin, IK and Helm, RF and Blaby-Haas, CE and Brown, AM and Melville, SB},
title = {Type IV Pili-Associated Secretion of a Biofilm Matrix Protein From Clostridium perfringens That Forms Intermolecular Isopeptide Bonds.},
journal = {Molecular microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1111/mmi.70020},
pmid = {40977362},
issn = {1365-2958},
support = {DE-AC02-05CH11231//Office of Science, Office of Basic Energy Sciences, of the U.S. Department of Energy/ ; //U.S. Department of Energy Joint Genome Institute/ ; //Office of Science of the U.S. Department of Energy/ ; },
abstract = {Clostridium perfringens is a gram-positive, anaerobic, spore-forming bacterial pathogen of humans and animals. C. perfringens also produces type IV pili (T4P) and has two complete sets of T4P-associated genes, one of which has been shown to produce surface pili needed for cell adherence. One hypothesis about the second set of T4P genes is that they comprise a type II secretion system (TTSS) like those found in gram-negative bacteria, but for gram-positive bacteria, the TTSS would aid transit across the thick peptidoglycan (PG) layer. The secretome of mutants lacking type IV pilins was examined, and a single protein, BsaC (CPE0517), was identified as being dependent on pilin PilA3 for secretion. The bsaC gene is in an operon with genes encoding a SipW signal peptidase and two putative biofilm matrix proteins, BsaA and BsaB, both of which have remote homology to Bacillus subtilis biofilm protein TasA. Since BsaA forms long oligomers that are secreted, we analyzed BsaA monomer interactions with de novo modeling. These models projected that the monomers formed isopeptide bonds as part of a donor strand exchange process. Mutations in residues predicted to form the isopeptide bonds led to the loss of oligomerization, supporting an exchange and lock mechanism, and isopeptide bonds were detected by mass spectrometry methods. Phylogenetic analysis showed the BsaA family of proteins is widespread among bacteria and archaea, but only a subset is predicted to form isopeptide bonds.},
}

@article {pmid40976822,
year = {2025},
author = {Eladl, A and Abbas, HA and Elbaramawi, SS and El-Hamid, MIA and Nazeih, SI},
title = {The anti-inflammatory drugs diclofenac and ketorolac inhibit urease activity and biofilm formation by uropathogenic Proteus mirabilis.},
journal = {Folia microbiologica},
volume = {},
number = {},
pages = {},
pmid = {40976822},
issn = {1874-9356},
abstract = {Proteus mirabilis is a major bacterium responsible for catheter-associated urinary tract infections (CAUTIs). Urease enzyme has a great role in the pathogenesis of Proteus mirabilis. Using urease, Proteus mirabilis can decompose urea to produce ammonia that increases urine pH and enhances crystal precipitation and crystalline biofilm formation. This leads to catheter blockage and pyelonephritis, respectively. Urease inhibitors are of great value in controlling this problem. Diclofenac sodium and ketorolac tromethamine, with their metal chelating activities, can inactivate urease by chelation of nickel ion in the active site of urease. This study investigated the ability of diclofenac sodium and ketorolac tromethamine to inhibit urease activity in Proteus mirabilis and crystalline biofilm formation. Diclofenac sodium and ketorolac tromethamine showed comparable activities against urease in cell lysates and whole cultures, with subsequent inhibition of pH increase and crystal formation in artificial urine. Diclofenac sodium showed higher biofilm inhibition and downregulation of urease genes ureR and ureC in RT-qPCR. The docking study showed the ability of both drugs to bind to urease enzyme and to chelate nickel ions in the active site of urease, suggesting that nickel chelation is the mode of inhibition of urease enzyme. In conclusion, diclofenac sodium and ketorolac tromethamine are two urease inhibitors that may be useful in treating Proteus mirabilis CAUTI.},
}

@article {pmid40976444,
year = {2025},
author = {Eltawab, R and Li, K and Abdelfattah, A and Elsamahy, T and Jia, H and Cheng, L},
title = {Harnessing methane for improved nitrogen removal in membrane aerated biofilm reactors: towards sustainable wastewater practices.},
journal = {Bioresource technology},
volume = {},
number = {},
pages = {133362},
doi = {10.1016/j.biortech.2025.133362},
pmid = {40976444},
issn = {1873-2976},
abstract = {This review examines the role of methane as an electron donor in Membrane Aerated Biofilm Reactor (MABR) systems, which conventionally use gas-permeable membranes to deliver oxygen directly to stratified biofilms for wastewater treatment. The integration of methane provides a sustainable carbon source for denitrification while also mitigating methane emissions. Methanotrophic denitrification enables methanotrophs to couple methane oxidation with nitrate or nitrite reduction, linking nitrogen removal with greenhouse gas mitigation. This dual function improves treatment efficiency, reduces energy use and operational costs, and addresses drawbacks of conventional MABRs, such as limited denitrification in low-carbon wastewater, by supplying a cost-effective carbon source. Methane-based MABRs, however, introduce challenges including methane-oxygen balance, microbial dynamics, and safety. Recent solutions include Layer-Structured Membranes (LSMs) and artificial intelligence-driven control, which improve biofilm stability, optimize gas transfer, and ensure long-term performance. Collectively, these advances underscore methane-based MABRs as a promising technology for sustainable and low-carbon wastewater treatment.},
}

@article {pmid40976139,
year = {2025},
author = {Maric, T and Tanaka, Y and Li, Z and Wu, J and Li, J and Guan, J and Boisen, A},
title = {Micro/nanomotors for biofilm remediation: Nanoarchitectonic breakthroughs with a focus on regulatory and clinical translation challenges.},
journal = {Biomaterials},
volume = {326},
number = {},
pages = {123715},
doi = {10.1016/j.biomaterials.2025.123715},
pmid = {40976139},
issn = {1878-5905},
abstract = {Biofilm-associated bacterial infections, notorious for their resistance to standard therapies, pose a critical challenge in clinical practice. Micro and nanomotors (MNMs) have emerged as dynamic tools capable of penetrating biofilm matrices and enabling targeted antimicrobial delivery through autonomous motion. Recent advances in nanoarchitectonic design, spanning fuel-free or chemical propulsion, biohybrid systems, and multimodal actuation, significantly enhance their therapeutic precision and biocompatibility. This review critically examines the evolution of MNM materials, geometries, and designs, emphasizing their mechanical disruption of extracellular polymeric substances and synergistic bactericidal effects. Innovations such as cascade-driven MNMs and stimuli-responsive platforms demonstrate >90 % biofilm eradication in vitro and accelerated wound healing in vivo. What distinguishes this review from existing literature is its integrated focus on regulatory and translational barriers to clinical adoption, an aspect seldom addressed in prior MNM reviews. In addition to advances in materials and design, we discuss challenges that must be overcome for clinical translation, including long-term biosafety, degradation, scalable manufacturing under Good Manufacturing Practice (GMP), and regulatory ambiguities surrounding nanoscale medical devices. We outline a path forward for addressing these barriers by emphasizing the need for standardized toxicity testing, stronger interdisciplinary collaboration, and the use of emerging regulatory tools such as Safe(r) Innovation Approaches (SIA), the EU's Safe and Sustainable by Design (SSbD) initiative, and regulatory sandboxes to help accelerate clinical translation. By integrating material and design innovation with regulatory foresight, MNM technology holds transformative potential for combating antibiotic-resistant infections and redefining the eradication of biofilms.},
}

@article {pmid40975913,
year = {2025},
author = {Aydin, D and Akin, ID and Call, DR and Beyenal, H},
title = {Strong alone, weak together: biofilm tensile strength in kangaroo rat burrows.},
journal = {Enzyme and microbial technology},
volume = {192},
number = {},
pages = {110752},
doi = {10.1016/j.enzmictec.2025.110752},
pmid = {40975913},
issn = {1879-0909},
abstract = {Desert kangaroo rats (Dipodomys deserti) construct burrows that create protected micro-niches favorable to increased microbial activity and biofilm formation. Biofilms within these burrows bind sand particles together, increase the tensile strength of the burrow sand and burrow stability. Our previous work 1) demonstrated that kangaroo rat burrow sand exhibits higher tensile strength than surrounding surface sand due to the cementation by biofilms, and 2) characterized the microbial biofilm communities within kangaroo rat burrow sand and isolated abundant microorganisms. In this study, isolated species are used to quantify tensile strength of pure and mixed biofilms using the microcantilever technique. Mono-species biofilms of Aspergillus tamarii (59.30 ± 4.36 kPa) exhibited the highest tensile strength, while Neobacillus niacini (9.45 ± 1.98 kPa) showed the lowest. Dual-species biofilms displayed synergistic or antagonistic effects, depending on species combinations. Mixing N. niacini with Peribacillus frigoritolerans increased tensile strength to 55.11 ± 2.51 kPa, whereas combining A. tamarii with N. niacini reduced the tensile strength to 18.98 ± 2.54 kPa. Mixing up to five species reduced tensile strength to a minimum value of 2.16 kPa. We conclude that biofilms formed by microbial isolates from burrow sand individually had higher tensile strength, but when all were mixed, the tensile strength decreased, making them weaker.},
}

@article {pmid40975416,
year = {2025},
author = {Bi, Z and Wang, X and Fu, H and Huang, Y},
title = {Carbon source shaped microbial ecology, metabolism and performance in biofilm system for simultaneous phosphorus recovery and nitrogen removal.},
journal = {Environmental research},
volume = {},
number = {},
pages = {122800},
doi = {10.1016/j.envres.2025.122800},
pmid = {40975416},
issn = {1096-0953},
abstract = {The application of biofilm-based phosphorus enrichment technology has been hampered by the limited information on the performance, microbial interactions and metabolic patterns of dominant functional bacteria, especially those fed with complex carbon sources conditions. In this study, three representative carbon sources contained in real sewage, i.e., volatile fatty acids (VFAs), glucose, and amino acids were selected as the complex carbon sources. The comparison in phosphorus removal/enrichment performance, carbon utilization, and metabolic characteristics were performed during the biofilm system changed the sole carbon source (acetate sodium) feeding to complex carbon source feeding gradually. The performance reduction and instability were observed in initial stage of complex carbon source feeding, while the phosphorus removal/enrichment efficiency improved significantly after long-term acclimation by extending the anaerobic HRT. The concentration of phosphorus enrichment solution exceeded 50 mg/L, meanwhile the total nitrogen and total phosphorus removal efficiencies over 82% and 97%, respectively. Intriguing, intracellular organic phosphorus (OP) contents fluctuated with phosphorus uptake and release, which may be a hint of the important role of OP in PAOs energy conversion. Complex carbon sources induced the succession of biofilm community, especially the enrichment of hydrolytic fermentation bacteria, and a more intricate microbial interaction network among functional microbiota. The co-occurrence of the EMP and ED pathways during glycolysis implied more extensive carbon utilization pathways, and amino acids was speculated to complement intracellular energy metabolism via the tricarboxylic acid cycle (TCA). This study demonstrated that the biofilm systems have great potential to simultaneously achieve phosphorus removal and enrichment by using complex carbon sources in sewage wastewater.},
}

@article {pmid40975365,
year = {2025},
author = {Preetham, V and Saidulu, D and Tiwary, CS and Gupta, AK},
title = {Role of biofilm activity on novel interwoven lattice-shaped 3D printed biocarrier for enhanced removal of organics and nutrients with reduced footprint: Performance assessment and microbial community dynamics.},
journal = {Bioresource technology},
volume = {},
number = {},
pages = {133356},
doi = {10.1016/j.biortech.2025.133356},
pmid = {40975365},
issn = {1873-2976},
abstract = {Simultaneous nitrification, denitrification, and phosphorus removal (SNDPR) remains challenging in conventional biological systems due to microbial complexity and limitations of traditional carriers like clogging and mass transfer. This study introduces a novel 3D-printed carrier (3D-PC) with high theoretical specific surface area (TSSA) to enhance organics and nutrient removal. A sequencing batch biofilm reactor was operated under optimized anaerobic/oxic/anoxic cycles at chemical oxygen demand (COD) to nitrogen (C/N) ratios of 10, 5, and 2.5 without external carbon addition. The highest removal at C/N 10 achieved 98% total nitrogen (TN) and 85% phosphate phosphorus (PO43--P). Even at low C/N 2.5, 70% TN and 73% PO4[3-]-P were removed, highlighting 3D-PC's efficiency under carbon-limited conditions. The improved performance is attributed to the carrier's high TSSA, porous design, and stable biofilm growth. Achieving good SNDPR at just a 15% carrier fill ratio demonstrates the potential of this compact system for cost-effective and efficient wastewater treatment. This shows the potential of 3D-PC in wastewater treatment.},
}

@article {pmid40975186,
year = {2025},
author = {Song, Y and Li, J and Zhang, Y and Su, L and Qin, S and Wu, C and Song, G},
title = {Biofilm Maturation in Carbapenem-resistant Pseudomonas aeruginosa Is Regulated by the sRNA PA213 and Its Corresponding Encoded Small Protein.},
journal = {International journal of antimicrobial agents},
volume = {},
number = {},
pages = {107625},
doi = {10.1016/j.ijantimicag.2025.107625},
pmid = {40975186},
issn = {1872-7913},
abstract = {OBJECTIVES: Biofilm formation is a key factor contributing to the persistence and resistance of carbapenem-resistant Pseudomonas aeruginosa (CRPA). This study aims to elucidate the regulatory role of PA213, a previously uncharacterized and hypoxia-inducible small RNA, in promoting CRPA biofilm maturation.

METHODS: We collected 113 clinical isolates for species identification and antimicrobial profiling. Comparative transcriptomic analyses were conducted between carbapenem-susceptible and resistant strains to identify functional sRNA candidates. The expression and structure of PA213 were characterized using qRT-PCR, RACE, and secondary structure prediction. A Flag-tagged PA213 fusion protein was confirmed by Western blot. PA213's phenotypic effects were assessed through assays of biofilm formation, motility, pyocyanin production, and extracellular matrix regulation. Biofilm architecture was visualized by SEM, and viability was analyzed via CFU counts and PI staining. Protein partners were identified via GST-pulldown and LC-MS, and interaction with OprI was confirmed through immunofluorescence and biochemical validation. The impact of PA213 on quorum sensing (QS) gene expression was examined by RT-qPCR based on RNA-seq datasets.

RESULTS: PA213 is a novel, uncharacterized sRNA that is highly expressed in clinical CRPA isolates. This sRNA featured a typical stem-loop structure and a 279-nucleotide open reading frame (ORF), which encoded a low-molecular-weight polypeptide. PA213 was significantly upregulated in clinical CRPA under hypoxia and encoded a small peptide with high conservation. Functionally, PA213 acted as a multifunctional regulator, enhancing biofilm formation, inhibiting pyocyanin synthesis, promoting swimming motility, and reducing virulence and pathogenicity. It enhanced biofilm formation during microcolony (12-48 h) and maturation (48-72 h) phases, without increasing EPS production or relying on the las/rhl QS pathway. It ensured sufficient functional bacterial density and maintained the three-dimensional structure of the biofilm by modulating viable bacterial proliferation. Mechanistically, the protein encoded by PA213 directly interacted with OprI, mediating spatial stabilization of mature biofilms.

CONCLUSION: PA213 functions as a bifunctional sRNA-peptide regulator that promotes CRPA biofilm maturation via non-coding and protein-mediated mechanisms, offering new insights into hypoxia-responsive biofilm regulation and antimicrobial resistance evolution.},
}

@article {pmid40975037,
year = {2025},
author = {Fan, X and Dai, C and Han, J and Lou, S and Zhang, N and He, M and Zhu, Z and Li, P and Chen, X and Xu, X},
title = {Nordihydroguaiaretic acid (NDGA) exhibits potent anti-biofilm and antimicrobial activity against methicillin-resistant Staphylococcus aureus and Candida albicans.},
journal = {Phytomedicine : international journal of phytotherapy and phytopharmacology},
volume = {148},
number = {},
pages = {157265},
doi = {10.1016/j.phymed.2025.157265},
pmid = {40975037},
issn = {1618-095X},
abstract = {INTRODUCTION: The global rise of antimicrobial resistance (AMR), particularly among biofilm-forming bacteria and fungi, has created an urgent need for novel therapeutics. Nordihydroguaiaretic acid (NDGA), a plant-derived polyphenolic lignan, has demonstrated promising biological activities, yet its potential as a dual-action antimicrobial agent remains underexplored.

OBJECTIVES: This study aimed to evaluate the antimicrobial and anti-biofilm activities of NDGA against methicillin-resistant Staphylococcus aureus (MRSA) and Candida albicans, elucidate its mechanism of action, and assess its in vivo efficacy and biosafety.

MATERIALS AND METHODS: NDGA's antimicrobial activities were assessed through MIC, time-kill, and biofilm disruption assays against standard and clinical isolates. Transcriptomic profiling and deep-learning-guided molecular docking were used to identify key microbial targets and regulatory pathways. In vivo efficacy was validated in Galleria mellonella and murine systemic infection models. Cytotoxicity, hemolysis, and acute toxicity assays were conducted to evaluate biosafety.

RESULTS: NDGA exhibited potent antimicrobial activity, with MIC values of 32 μg/ml for S. aureus USA300 and 64 μg/ml for C. albicans SC5314. It effectively eradicated persister cells and disrupting mature biofilms of both MRSA and C. albicans. Transcriptomic analysis revealed that NDGA modulated multiple microbial virulence and biofilm-regulating pathways, including arginine biosynthesis in MRSA and ergosterol metabolism in C. albicans. Docking studies confirmed strong binding affinity of NDGA to critical microbial targets. In vivo, NDGA significantly improved survival rates, reduced pathogen burden, and alleviated tissue damage, showing comparable efficacy to vancomycin and fluconazole. NDGA demonstrated favorable biosafety with low cytotoxicity, minimal hemolysis, and no observable acute toxicity in mammalian models.

CONCLUSION: NDGA is a promising antimicrobial candidate capable of disrupting biofilms and overcoming drug resistance in both bacterial and fungal infections. Its multitargeted mode of action, coupled with in vivo efficacy and biosafety, supports further development as a next-generation anti-infective agent.},
}

@article {pmid40974892,
year = {2025},
author = {Xiu, J and Geng, B and Liu, M and Han, X and Zhang, D and Bai, X},
title = {Biofilm induced microplastics and microbial metabolites release from Polypropylene Random pipes in drinking water distribution systems.},
journal = {Water research},
volume = {288},
number = {Pt A},
pages = {124626},
doi = {10.1016/j.watres.2025.124626},
pmid = {40974892},
issn = {1879-2448},
abstract = {While plastic pipes are extensively employed in water infrastructure owing to their durability and corrosion resistance‌, their long-term deployment in drinking water distribution systems (DWDS) fosters biofilm proliferation through microbial colonization. However, biofilm-induced material alterations and the concurrent release of microplastics (MPs) and microbial secondary metabolites remain undercharacterized. To elucidate biofilm-mediated corrosion in polypropylene random copolymer (PPR) pipes and their implications for water safety, this research employed a multi-methodological framework of high-resolution scanning electron microscopy (SEM), flow cytometric quantification coupled with 16S rRNA sequencing, and gas chromatography-mass spectrometry (GC-MS). The results demonstrate that biofilm colonization induces progressive surface deterioration, with SEM revealing sequential roughness increase (60 days), microcracks formation (90 days), and pore formation (120-150 days). Biofilm-induced corrosion increased microplastics release (2.1-fold versus control) and enhanced leaching of microbial metabolites, including organophosphate flame retardants (TEP 18-fold, TCEP 5.2-fold), phthalates (DEHP 14-fold), and antioxidant derivatives (2,4-DTBP 20-fold, BHB 41-fold). PPR-degrading bacterial communities were dominated by Sphingobium, Bradyrhizobium, Comamonadaceae, and Sediminibacterium. GC-MS analysis detected released compounds with microbial degradation pathways evidenced by dodecanal accumulation (11-fold) and 1-dodecanol depletion (0.04-fold). These findings confirmed that biofilm development accelerate PPR material aging through both biofilm-induced corrosion and biochemical degradation, while introducing multiple water quality risks. These findings revealed that biofilm development accelerates PPR material aging through synergistic biofilm-induced corrosion and biochemical degradation, concurrently introducing multiple water quality risks via significantly elevated contaminant release. This study elucidates the mechanisms and quantifies the impacts of biofilm-induced corrosion in PPR pipes, highlighting the urgent need for biofilm control or material improvements in end DWDS.},
}

@article {pmid40973254,
year = {2025},
author = {Wangpaiboon, K and Maneechan, M and Yenpech, N and Chirachanchai, S and Jiraroj, D and Tungasmita, DN and Panpetch, P and Krusong, K and Charoenwongpaiboon, T and Pichyangkura, R},
title = {Unraveling role of the glucan-binding domain in mutansucrase-mediated glucan synthesis and biofilm-related properties.},
journal = {Carbohydrate polymers},
volume = {369},
number = {},
pages = {124270},
doi = {10.1016/j.carbpol.2025.124270},
pmid = {40973254},
issn = {1879-1344},
mesh = {*Biofilms/growth & development ; *Glucans/chemistry/biosynthesis/metabolism ; *Streptococcus mutans/enzymology/physiology ; Dextrans/metabolism/chemistry ; *Glucosyltransferases/metabolism/chemistry ; *Bacterial Proteins/metabolism/chemistry ; Protein Domains ; },
abstract = {Mutansucrase (MUS) catalyzes the formation of α-1,3-linked glucan (mutan) from sucrose. Most mutansucrase are produced by Mutan Streptococcus species, which are considered primary etiological agents of dental caries. Previous investigations have posited MUS as one of the crucial virulence factors contributing to biofilm formation. This study investigated the role of the glucan-binding domains (GBDs) in MUS activity and the properties of the resulting glucan product. Our findings showed GBDs rarely participate in α-1,3 glucan elongation and have negligible binding affinity for them. Conversely, GBDs exhibited high affinity for α-1,6 glucans (dextran). Notably, using dextran as an acceptor for α-1,3 glucan extension significantly enhanced MUS transglycosylation and processivity, leading to higher molecular weight glucans and gel-like material formation, emphasizing MUS's role in biofilm development. Although GBDs didn't directly determine glycosidic linkage type, their cooperative interaction with the catalytic domain modulated the physicochemical properties and structural diversity of the synthesized glucans. Furthermore, intrinsic glucan conformation influenced chain elongation, offering a new perspective on biofilm synthesis beyond the enzyme's role. This work also characterized glucans synthesized by MUS via both common sucrose reactions and dextran priming process, providing insights for addressing biofilm challenges and developing new biomaterial applications.},
}

*Biofilms/growth & development
*Glucans/chemistry/biosynthesis/metabolism
*Streptococcus mutans/enzymology/physiology
Dextrans/metabolism/chemistry
*Glucosyltransferases/metabolism/chemistry
*Bacterial Proteins/metabolism/chemistry
Protein Domains

@article {pmid40972766,
year = {2025},
author = {Upadhyay, A and Pal, D and Kumar, A},
title = {Development of the combinatorial dye assay for improved detection of the Salmonella Typhi biofilm.},
journal = {Journal of microbiological methods},
volume = {},
number = {},
pages = {107272},
doi = {10.1016/j.mimet.2025.107272},
pmid = {40972766},
issn = {1872-8359},
abstract = {Typhoidal biofilms resist antibiotics and are inadequately stained by traditional dyes. This study presents a 0.5 % combinatorial dye, prepared by mixing crystal violet and safranin in a 1:1 ratio, resulting in a final concentration of 0.25 % crystal violet and 0.25 % safranin (total 0.5 %). The formulation enhances staining intensity, quantification, and detection of typhoidal biofilms.},
}

@article {pmid40969145,
year = {2025},
author = {Khan, A and Aasim, M and Bibi, NS and Saddique, H and Ullah, H and Khan, Z and Aziz, U and Khan, NZ and Ali, W and Muhammad, N},
title = {Utilization of physicochemical approach to propose a strategy for biofilm inhibition.},
journal = {Biofouling},
volume = {},
number = {},
pages = {1-12},
doi = {10.1080/08927014.2025.2559318},
pmid = {40969145},
issn = {1029-2454},
abstract = {Biofilm development, which occurs on numerous surfaces, can reduce the efficiency and increase operating costs in bioprocesses and fermentation. The current study proposes a strategy for biofilm inhibition by investigating the interactions between microorganisms and surfaces using an extended Derjaguin-Landau-Verwey-Overbeek (xDLVO) approach and cell partition index (CPI) technique. Glass slide and Petri dish surfaces were modified with different surfactants. The results show that modification increased CPI values and altered the interaction behavior from attractive to repulsive, between microbial cells and different surfaces. Secondary energy values calculated by xDLVO theory between microbial cells and modified surfaces were repulsive. Meanwhile, the secondary energy values calculated for microbial cells and unmodified glass slide (-31 kT) and Petri dish surfaces (-27 kT) were attractive between cells and surfaces. The current study has opened a window for research in the field of biofilm inhibition through a surface energetics approach.},
}

@article {pmid40966435,
year = {2025},
author = {Skliar, I and Kryvtsova, M and Kostenko, Y and Savenko, M},
title = {Biofilm-forming Ability of Anaerobic Bacterial Strains Isolated from Patients Diagnosed with Periodontitis.},
journal = {Prague medical report},
volume = {126},
number = {3},
pages = {144-150},
doi = {10.14712/23362936.2025.22},
pmid = {40966435},
issn = {1214-6994},
mesh = {*Biofilms/growth & development ; Humans ; *Periodontitis/microbiology ; *Bacteria, Anaerobic/physiology/isolation & purification ; Male ; Adult ; Middle Aged ; Female ; },
abstract = {Bacterial biofilms represent the primary causative agents of pathogenic processes within the oral cavity. Biofilm microorganisms exhibit heightened resistance to adverse environmental factors. The objective of this study was to ascertain the biofilm-forming potential of clinical strains of anaerobic microorganisms isolated from the periodontal pocket of patients diagnosed with periodontitis. The study of biofilm formation was carried out by the method (Djordjevic et al., 2002) using flat-bottomed 96-well polystyrene microtitration plates. Biofilm formation was determined by staining with 0.1% crystal violet. The study utilised clinical strains isolated from the periodontal pocket of 61 patients diagnosed with grade III chronic periodontitis. Of these, 30 strains belonging to three bacterial species were selected for further analysis: Porphyromonas asaccharolytica, Slackia exigua, and Schaalia odontolytica. Among the Porphyromonas asaccharolytica strains, 80% formed biofilm; the figure for Slackia exigua was 90%, and for Schaalia odontolytica - 80%. In total, 25 strains were capable of biofilm formation. The results indicate that the isolated strains of Porphyromonas asaccharolytica, Slackia exigua, and Schaalia odontolytica exhibited a significant capacity for biofilm formation (83.3% of the strains formed biofilm), particularly Slackia exigua strains, which exhibited the highest number of strains with high biofilm formation ability - 5 (16.6%). It is also noteworthy that these microorganisms exhibited a moderate persistence frequency (16.4% each), of the total sample of microbial biocenoses from inflammatory periodontal soft tissue areas.},
}

*Biofilms/growth & development
Humans
*Periodontitis/microbiology
*Bacteria, Anaerobic/physiology/isolation & purification
Male
Adult
Middle Aged
Female

@article {pmid40961771,
year = {2025},
author = {Ogundipe, TT and Beitia, S and Zhang, L and Zhang, X and Obe, T},
title = {Differences in microbial composition of litter and water line biofilm of broiler farms as influenced by water quality history.},
journal = {Poultry science},
volume = {104},
number = {11},
pages = {105832},
doi = {10.1016/j.psj.2025.105832},
pmid = {40961771},
issn = {1525-3171},
abstract = {Microbiome profiling of litter and drinking water lines of poultry environments is essential to understanding their role in the persistence and transmission of opportunistic and pathogenic microorganisms. While the litter environment is well characterized, the drinking water system has received less attention, despite its potential as a control point for Salmonella. In this study, we identified two groups of five farms based on historic water quality issues: normal sulfur-iron (NSIF) and high sulfur-iron (HSIF) content with the aim of assessing the incidence/quantity of Salmonella and characterize the microbiota of the litter and water line biofilms within the two farm groups. Our findings showed no differences in Salmonella incidence and quantity between the litter of both groups, while the biofilm of one of the HSIF farms had Salmonella. Common serotypes, such as Enteritidis, Infantis, Kentucky, and Typhimurium, were identified in the litter of both groups, while Agona, Alachua, and Schwarzengrund were unique to the NSIF group and Ouakam and Worthington to the HSIF group. Generally, the litter exhibited high microbial diversity compared to the biofilm. The litter of the NSIF group had lower alpha diversity than the HSIF group. Conversely, NSIF biofilms had higher alpha diversity than the HSIF biofilms. Distinct differences in microbial community composition were observed in the litter and biofilm of the two farm groups. Aerococcus, Lactobacillus, and Staphylococcus dominated the litter of the two groups, whereas probiotic (NSIF) and pathogenic (HSIF) Bacillus was the most prevalent in the biofilms. These results suggest that water quality issues could influence the microbiota of poultry house environments, particularly in water line biofilms. Effective removal of these biofilms is crucial for controlling Salmonella at pre-harvest poultry production.},
}

@article {pmid40961761,
year = {2025},
author = {Khanal, S and Luitel, H and Adhikari, S and Marasini, A and Bhattarai, RK},
title = {Molecular characterization, antimicrobial resistance profiling, and biofilm analysis of Salmonella isolates from dead-in-shell embryonated eggs.},
journal = {Poultry science},
volume = {104},
number = {11},
pages = {105773},
doi = {10.1016/j.psj.2025.105773},
pmid = {40961761},
issn = {1525-3171},
abstract = {This study investigates the molecular characterization, antimicrobial resistance (AMR) profiling, and biofilm formation of Salmonella isolates from dead-in-shell embryonated eggs collected from 15 hatcheries in Bharatpur, Chitwan. A total of 210 samples were analyzed using cultural, biochemical, and molecular techniques, including PCR for virulence gene detection. The study revealed a prevalence rate of 11.42 %, with isolates demonstrating significant AMR, particularly to cephalexin (100 %), ampicillin (95.83 %), and nalidixic acid (91.67 %). Moreover, 83.33 % (n = 20) of the isolates were classified as multidrug-resistant (MDR), with a mean multiple antibiotic resistance index of 0.51. Biofilm formation analysis indicated that 70.83 % of the isolates could produce biofilms, with 41.67 % classified as weak producers and 16.67 % as strong producers. Molecular analysis identified key virulence genes, including invA (100 %), spvC (50 %), spvB1 (33.33 %), and the biofilm-associated gene agfA (12.5 %), emphasizing their roles in pathogenicity, systemic infections, and biofilm formation. Our findings highlight the critical public health risks posed by MDR Salmonella, which can persist in the food chain and compromise human and animal health. This research underscores the need for enhanced biosecurity in hatcheries, judicious use of antimicrobials, and periodic monitoring of bacterial resistance and virulence factors to mitigate the economic and health impacts of Salmonella infections in poultry production systems.},
}

@article {pmid40961754,
year = {2025},
author = {Chen, H and Bi, M and Li, S and Yang, S and Zhang, Y and Zhao, L and Jiang, X and Yu, T},
title = {Inhibition of Listeria monocytogenes biofilm formation by phenyllactic acid in pasteurized milk is associated with suppression of the Agr system.},
journal = {International journal of food microbiology},
volume = {444},
number = {},
pages = {111440},
doi = {10.1016/j.ijfoodmicro.2025.111440},
pmid = {40961754},
issn = {1879-3460},
abstract = {The biofilm-forming propensity of Listeria monocytogenes on food-contact surfaces presents critical challenges to food safety. Phenyllactic acid (PLA), a microbial-derived organic acid compound, exhibits potent biofilm inhibition against this pathogen, yet its mechanistic basis remains undefined. This study investigated PLA's effects on the Agr system, which promotes L. monocytogenes biofilm formation, to explore its potential as a biofilm control agent. Quantitative biofilm analysis through crystal violet-based microplate assays demonstrated decreased biofilm biomass under PLA treatment. Quantitative real time PCR (RT-qPCR) revealed marked downregulation of the agr system components, paralleled by diminished β-galactosidase activity in agr promoter-reporter constructs. These evidences suggested that PLA can inhibit L. monocytogenes biofilm formation by suppressing the Agr system in medium. Microscale thermophoresis analysis excluded direct molecular interactions between PLA and AgrC/AgrA proteins. Our data demonstrated that signal molecule autoinducing peptide (AIP) produced by AgrBD in L. monocytogenes was a cyclic hexapeptide whose biosynthesis was inhibited by PLA. Notably, PLA maintained its biofilm-inhibitory capacity by suppressing the Agr system in pasteurized milk. This preserved activity across distinct experimental environments highlights the application potential of PLA as a food-safe antimicrobial agent.},
}

@article {pmid40961672,
year = {2025},
author = {Chen, TT and Yan, ZW and Cai, FY and He, QX and You, HH and Rensing, C and Lü, J and Zhou, S},
title = {Carbon quantum dots boost microbial biofilm-based hydrovoltaic electricity generation.},
journal = {Water research},
volume = {288},
number = {Pt A},
pages = {124593},
doi = {10.1016/j.watres.2025.124593},
pmid = {40961672},
issn = {1879-2448},
abstract = {Microbial biofilm-based hydrovoltaic electricity generators (BioHEGs) have recently been developed as promising and readily available platforms for green energy harvesting, despite their unsatisfactory output performances and unspecified mechanisms regarding electric current production. Herein, carbon quantum dots (CQDs) were used to construct a nano-biohybrid system with Shewanella oneidensis MR-1 (S. oneidensis), through which the CQDs/S. oneidensis BioHEG achieved a maximum open-circuit voltage of ca. 0.65 V and short-circuit current density of ca. 5.23 μA·cm[-2]. In addition, both the hydrovoltaic effect and electrical conductivity of CQDs/S. oneidensis nano-biohybrids were noticeably improved due to enhanced secretion of extracellular polymeric substances (EPS) and accelerated electron transfer upon CQDs implantation, thereby leading to a nearly 14-fold increase in output power density compared to the bare S. oneidensis cells. Studies aimed to elucidate the underlying mechanism indicated that the hybridization of CQDs and S. oneidensis greatly promoted the metabolic synthesis of outer membrane c-type cytochromes (OM c-Cyts) and the extracellular secretion of riboflavin (RF), which was demonstrated to be decisive in the current producing process of the CQDs/S. oneidensis BioHEG. This work thus proposes a viable strategy to boost the hydrovoltaic electricity generation capacity of microbial biofilms and provides a new perspective on the mechanism of accelerated electron transfer pathways inside BioHEGs.},
}

@article {pmid40959855,
year = {2025},
author = {Wang, Y and Wu, F and Hua, L and Gao, C and Wang, S and Liu, Y and Ren, Y and Shi, L and van der Mei, HC and Li, Y},
title = {Bimetal-Phenolic Framework to Combat Bacterial Infections via Synergistic Biofilm Dispersal, Bacterial Killing and Immune Modulation.},
journal = {Advanced science (Weinheim, Baden-Wurttemberg, Germany)},
volume = {},
number = {},
pages = {e13863},
doi = {10.1002/advs.202513863},
pmid = {40959855},
issn = {2198-3844},
support = {52293383//National Natural Science Foundation of China/ ; 52422307//National Natural Science Foundation of China/ ; 22475156//National Natural Science Foundation of China/ ; 22275043//National Natural Science Foundation of China/ ; 52203184//National Natural Science Foundation of China/ ; LR24H180001//Zhejiang Provincial Natural Science Foundation Outstanding Youth Project/ ; ZY2023031//Wenzhou Municipal Science and Technology Major Project/ ; CXTD202501043//Zhejiang Clinovation Pride/ ; 2025WMU-X001//First Affiliated Hospital of Wenzhou Medical University/ ; },
abstract = {The presence of drug resistance and extracellular matrix protection in biofilms makes it increasingly difficult to control bacterial infections using antibiotics. Therefore, there is an urgent need to develop new non-antibiotic approaches to eradicate drug-resistant bacterial infections. Here, a bimetal-phenolic framework (Que-Fe-CeMPF) is constructed by direct self-assembly of coordinated Fe[3+] and Ce[4+] ions with the polyphenol quercetin (Que). Que-Fe-CeMPF enhanced hydroxyl radical (·OH) generation, particularly in an acidic environment and presence of H2O2, compared with single metal-phenolic frameworks (Que-FeMPF and Que-CeMPF). · OH damaged bacterial cell walls, resulting in intracellular protein loss and bacterial cell death. Additionally, Que-Fe-CeMPF effectively dispersed biofilms by degrading matrix eDNA, allowing easier ·OH penetration, resulting in higher killing efficiency compared to Que-FeMPF and Que-CeMPF. Que-Fe-CeMPF stimulated macrophages to adopt an M2-like phenotype, suppressing excessive immune activation and promoting tissue repair at the infection site. As a combined effect of bacterial killing, biofilm degradation, and immune-modulation, the infectious pneumonia caused by Pseudomonas aeruginosa in mice is more effectively eradicated by Que-Fe-CeMPF than by free quercetin or the antibiotic ciprofloxacin. Moreover, Que-Fe-CeMPF is less prone to resistance development in pathogens compared to ciprofloxacin. Thus, Que-Fe-CeMPF is a promising non-antibiotic antimicrobial agent with multimodal activity for controlling drug-resistant bacterial infections.},
}