@article {pmid42294728,
year = {2026},
author = {Mao, Z and Jiang, M and Zhao, Z and Xu, S and Wang, H and Chen, K and Duan, J and Chen, Z and He, D and Xing, P and Wu, QL},
title = {Biofilm-forming traits enrich the plasmid diversity and functional potential in particle-attached bacteria in coastal ecosystems.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0046026},
doi = {10.1128/spectrum.00460-26},
pmid = {42294728},
issn = {2165-0497},
abstract = {UNLABELLED: Planktonic microorganisms play a central role in aquatic biogeochemical processes and are commonly divided into particle-attached (PA) and free-living (FL) fractions. Although these two lifestyles differ in ecological strategy, the contribution of plasmids to their niche differentiation remains poorly resolved. Here, we conducted a plasmid-centric metagenomic analysis of two anthropogenically impacted coastal ecosystems in South China, the Pearl River Estuary (PRE), and Daya Bay (DYB), to determine the environmental and biological drivers of plasmid diversity, and their functional potenitial. We found that plasmid diversity was jointly shaped by different fractions and environmental stressors. The PA fraction contained significantly higher plasmid abundance and richness than the FL fraction, and was enriched in multifunctional and conjugative plasmids. These plasmids were associated with genes adapting to the PA lifestyle or microenvironments, suggesting linkage between particle attachment and plasmid maintenance. Structural equation modeling indicated that different fractions shaped plasmid diversity primarily through biofilm-forming genes. Along an anthropogenic gradient from DYB to PRE, increasing pollution levels were accompanied by higher plasmid diversity and greater abundances of antibiotic and metal resistance genes. Plasmid diversity was strongly correlated with resistance gene abundance. The enrichment of transferable plasmids in the PA fraction, where cell densities are high and intercellular distances are close, suggested that particle-associated habitats favor genetic exchange and the persistence of resistance traits. Together, these results demonstrate that particle-associated microbial communities represent key reservoirs of plasmid diversity and resistance potential in coastal ecosystems and highlight the combined influence of lifestyles and anthropogenic stress on plasmid-mediated microbial adaptation.

IMPORTANCE: Plasmids play an important role in microbial adaptation by mediating horizontal gene transfer, yet the ecological contexts that favor their persistence and diversification in natural environments remain poorly understood. This study showed that particle-attached microbial communities in coastal waters harbored substantially higher plasmid diversity and resistance potential than free-living communities, and that this enrichment is strongly linked to biofilm-associated traits. By demonstrating how particulate habitats and pollution gradients jointly shape plasmid diversity and resistance gene abundance, our findings identify particle-associated microenvironments as critical reservoirs for plasmid-mediated functions in coastal ecosystems. These results advance understanding of how microbial lifestyle and human activities influence microbial evolution and the environmental dissemination of resistance traits.},
}

@article {pmid42294941,
year = {2026},
author = {Friebel, L and Knepper, J-P and Becker, NS and Abbaszade, G and Stückrath, K and Soltwisch, J and Müller, S and Dreisewerd, K and Mascher, T},
title = {Cannibalism shapes biofilm structure and composition in Bacillus subtilis.},
journal = {mBio},
volume = {},
number = {},
pages = {e0052526},
doi = {10.1128/mbio.00525-26},
pmid = {42294941},
issn = {2150-7511},
abstract = {UNLABELLED: In Bacillus subtilis colony biofilms, phenotypic diversification confers tissue-like properties and enhanced competitive fitness within a structural framework that allows both colony expansion and long-term survival via endospore formation. Cannibalism is a sporulation delay strategy, in which one subpopulation produces the sporulation delay protein SDP, the sporulation killing factor SKF, and the epipeptide EPE. These toxins are thought to lyse susceptible nonproducers, thereby releasing nutrients to prevent premature sporulation. However, the molecular mechanisms orchestrating this bacterial type of programmed cell death during biofilm development are poorly understood. Here, we comprehensively characterized mutants defective in either toxin production or the corresponding autoimmunity by a multiscale approach, combining luminescence reporters, colony biopsy, multi-parameter flow cytometry, and MALDI-mass spectrometry imaging to resolve cannibalism function and distribution. The toxins are produced in distinct, only partially overlapping areas of the colony, and are interdependent in their spatial distribution. Both EPE and SDP, but not SKF, are crucial for delaying sporulation. Loss of EPE or SDP autoimmunity resulted in severe morphological changes and stress-induced occurrence of suppressor mutants. The absence of all three toxins led to small, hyper-sporulating colonies with excessive wrinkle formation, indicating that cannibalism is essential for maintaining biofilm structure and lateral expansion. Our results provide the first evidence for the complex interactions between the three cannibalism toxins that shape biofilm architecture through bacterial programmed cell death. Localized toxin production and its spatial distribution affect the spatiotemporal organization, morphology, and subpopulation dynamics within B. subtilis biofilms.

IMPORTANCE: Programmed cell death (PCD) is a ubiquitous and crucial mechanism to structure eukaryotic multicellular tissues. PCD-like processes have also been described in bacteria, but their contribution to multicellular development is poorly understood. Cannibalism in Bacillus subtilis has been described as a sporulation delay strategy, in which one subpopulation produces antimicrobial peptides that kill susceptible nonproducing siblings. Their lysis is thought to release nutrients that delay the sporulation in the producing subpopulation. This study comprehensively analyses the role of the three cannibalism toxins in shaping colony biofilms. By combining MALDI-mass spectrometry imaging, colony biopsy, flow cytometry, and luminescence reporters, we demonstrate that cannibalism toxins are crucial for biofilm structure. They show a discrete and interdependent localization within the colonies. While cannibalism inhibits sporulation and causes severe envelope stress within biofilms, our data challenge the established role of cannibalism-dependent killing as the mechanism behind this sporulation delay.},
}

@article {pmid42297250,
year = {2026},
author = {He, QP and Gati, G and Wang, J},
title = {Overcoming mass transfer limitations in biological methane removal: a "dry" biofilm approach.},
journal = {Bioresource technology},
volume = {},
number = {},
pages = {135158},
doi = {10.1016/j.biortech.2026.135158},
pmid = {42297250},
issn = {1873-2976},
abstract = {Biological methane (CH4) removal from dilute air streams (200-5,000 ppm) is fundamentally constrained by slow mass transfer, primarily due to CH4's low solubility and diffusivity in water. Conventional biofilters, the current state of technology, suffer from long start-up periods, progressive pore clogging, and high transport energy requirements. This study investigates a "dry" biofilm concept designed to overcome these bottlenecks by minimizing the aqueous boundary layer while maintaining microbial activity via capillary-mediated nutrient delivery. Using concentrated biomass of the methanotroph Methylomicrobium buryatense 5GB1C, three generations of membrane-supported reactor configurations were evaluated. The third-generation (G3) design utilized a cellulose-bead capillary support to maintain a physical gap between the membrane and the liquid surface, enabling continuous drainage of metabolic water. Experimental results demonstrated that the "dry" G3 configuration achieved CH4 removal rate of 148.9 mg·m[-2]·hr[-1] at 4000 ppm, representing a 397% improvement over the first-generation floating mesh configuration. At 500 ppm, G3 design achieved a CH4 removal rate of 18.6 mg·m[-2]·hr[-1], corresponding to an over six-fold improvement over one of the highest reported values. Furthermore, the system enabled immediate start-up post-inoculation and maintained an optimal microenvironment pH (8.8-9.0) even as the bulk medium acidified. These results establish that replacing liquid-phase diffusion with drastically faster gas-phase transport provides a high-efficiency framework for mitigating low-concentration CH4 emissions. With the added benefits of minimal pressure drop and easy biomass harvesting via scraping, this dry biofilm approach offers a scalable and sustainable alternative for atmospheric methane mitigation.},
}

@article {pmid42297329,
year = {2026},
author = {Thibodeau, AJ and Mouchet, F and Nguyen, VX and Moura, T and Vivant, M and Pinelli, E and Barret, M},
title = {Sub-inhibitory concentrations of antibiotics modulate antimicrobial resistance fate in a Biofilm-Grazer system.},
journal = {Environmental pollution (Barking, Essex : 1987)},
volume = {},
number = {},
pages = {128470},
doi = {10.1016/j.envpol.2026.128470},
pmid = {42297329},
issn = {1873-6424},
abstract = {Antimicrobial resistance (AMR) represents an escalating global health challenge, primarily driven by the extensive use of antibiotics in agriculture and medicine. Environmental compartments like soil, water, and biofilms contribute to the spread and persistence of resistance genes. Biofilms and wildlife microbiota are recognized as reservoirs for antibiotic-resistant bacteria and antibiotic resistance genes (ARGs) in aquatic ecosystems. The ecological interactions between biofilms and wildlife gut microbiota, under antibiotic selective pressure, and their implication regarding the dissemination of antimicrobial resistance in aquatic ecosystems is poorly understood. This study examined the impact of sub-inhibitory concentrations of sulfamethoxazole, ciprofloxacin and trimethoprim (10 μg. L[-1]) on the resistome in a Biofilm-grazer system, using a river biofilm and Xenopus laevis larvae over a 12-day period. The results indicated that antibiotic pressure resulted in notable variations in microbial composition in both biofilms and gut microbiota. Additionally, comparison of bacterial community composition between gut and biofilm compartments indicated increased microbial exchange between these environments, with stronger effects observed under antibiotic exposure. An increase of the resistome was observed only in biofilm especially under antibiotics exposure. Variations in the grazer's bacterial communities appeared to attenuate the increase of ARGs within the gut. However, grazing activity concomitantly enhanced the resistome abundance and diversity within the biofilm. This study presents an original perspective on the understanding of impact of sub-inhibitory antibiotic concentration on ecosystems though a microcosm experiment, highlighting the necessity for environmental monitoring and further studies across diverse and complex ecological settings to mitigate antibiotic resistance dissemination.},
}

@article {pmid42298526,
year = {2026},
author = {Tunç, AK and Gecer, EN and Erenler, R and Marzi, M},
title = {Biofilm inhibition by silver nanoparticles produced from Stachys spectabilis.},
journal = {BMC complementary medicine and therapies},
volume = {},
number = {},
pages = {},
doi = {10.1186/s12906-026-05438-8},
pmid = {42298526},
issn = {2662-7671},
abstract = {BACKGROUND: Green synthesis of silver nanoparticles (AgNPs) has emerged as an environmentally friendly approach for developing novel antimicrobial materials. Although several Stachys species have been investigated for their pharmacological properties, the antibacterial and antibiofilm activities of AgNPs synthesized from Stachys spectabilis have not previously been evaluated. Therefore, this study aimed to investigate the antimicrobial and antibiofilm potential of Stachys spectabilis-derived AgNPs against clinically relevant biofilm-forming bacterial pathogens.

METHODS: AgNPs were fabricated through an aqueous green synthesis approach and characterized using standard analytical techniques. Antibacterial activity was assessed via liquid microdilution to determine minimum inhibitory concentrations (MICs) against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae. Antibiofilm activity was quantified using the crystal violet assay in 96-well microplates.

RESULTS: Disc diffusion results revealed measurable inhibition zones only in E. coli and P. aeruginosa (6 mm), corresponding largely to the disc diameter and indicating limited nanoparticle diffusion in agar. In contrast, MIC and MBC analyses demonstrated a clear concentration-dependent antibacterial effect: S. aureus showed the highest susceptibility with a MIC of 1024 µg/mL, while the remaining strains exhibited MICs of 2048 µg/mL. Consistently higher MBC values confirmed the inhibitory-to-bactericidal threshold. AgNPs also displayed substantial antibiofilm activity, achieving biomass reductions of 59.7% in P. aeruginosa, 57.0% in E. coli, 54.9% in S. aureus, and 40.1% in K. pneumoniae. Overall, the results demonstrate that although AgNPs exhibit limited diffusion in solid media, they possess concentration-dependent antibacterial and antibiofilm activity in liquid media.

CONCLUSIONS: The synthesized silver nanoparticles demonstrated substantial antibacterial and antibiofilm efficacy despite their limited diffusion in solid media. While disc diffusion assays produced minimal inhibition zones, MIC and MBC analyses revealed concentration-dependent inhibitory and bactericidal effects, with S. aureus showing the greatest susceptibility. Additionally, significant reductions in biofilm biomass across all tested species indicate that AgNPs not only inhibit planktonic bacterial growth but also effectively disrupt established biofilms. These findings suggest that AgNPs hold considerable potential as alternative antimicrobial agents, particularly in applications where both planktonic and biofilm-associated bacterial forms must be controlled. To the best of our knowledge, this is the first study to demonstrate the antibacterial and antibiofilm activities of Stachys spectabilis-derived AgNPs, highlighting their potential as sustainable antimicrobial agents for the control of biofilm-associated bacterial infections.},
}

@article {pmid42298784,
year = {2026},
author = {Zainal, M and Ibrahim, MJ and Mohd Sarmin, N' and Ismail, S and Cirillo, N and Dashper, SG and Arzmi, MH},
title = {Phenotypic switch Candidozyma auris (Candida auris) modulates biofilm formation and virulence genes SAP5 and ALS5 in mono- and co-culture environments with Staphylococcus aureus.},
journal = {Biofouling},
volume = {},
number = {},
pages = {1-15},
doi = {10.1080/08927014.2026.2688203},
pmid = {42298784},
issn = {1029-2454},
abstract = {Candidozyma auris (formerly Candida auris) (C. auris), an emerging multidrug-resistant fungal pathogen, forms biofilms as a virulence factor. This study aimed to determine the effect of phenotypic switch on C. auris biofilm formation and virulence gene expression in mono- and co-culture with Staphylococcus aureus. Phenotypic switching was induced by prolonged incubation, and biofilms were developed in RPMI-1640, YEPD, SDB, and BHIYE. The biofilm biomass and total cell count were measured. SAP5 and ALS5 gene expression was quantified using qPCR. The 4th switched generation mono-culture biofilm in BHIYE produced the highest biomass (3.34 ± 0.08) and total cell count (5.66 ± 0.03 log10 cells mL[-1]). In addition, SAP5 and ALS5 expression peaked in the 2nd switched generation mono-culture by 10.43 ± 0.44-fold and 4.764 ± 0.01-fold, respectively. Co-culture biofilms exhibited significantly higher ALS5 expression in selected switched generations compared to unswitched C. auris (p < 0.05). In conclusion, phenotypic switching enhanced biofilm formation and modulated the expression of SAP5 and ALS5 in C. auris.},
}

@article {pmid42299093,
year = {2026},
author = {Yüksel Yence, D},
title = {Prevalence, antibiotic resistance, enterotoxin genes, biofilm formation, and agr typing of Staphylococcus aureus from raw milk and cheese.},
journal = {Polish journal of veterinary sciences},
volume = {29},
number = {2},
pages = {191-202},
doi = {10.24425/pjvs.2026.1262},
pmid = {42299093},
issn = {2300-2557},
mesh = {*Staphylococcus aureus/drug effects/genetics/physiology ; Animals ; *Biofilms/growth & development ; *Enterotoxins/genetics/metabolism ; *Cheese/microbiology ; *Milk/microbiology ; *Drug Resistance, Bacterial ; *Bacterial Proteins/genetics/metabolism ; *Anti-Bacterial Agents/pharmacology ; Gene Expression Regulation, Bacterial/physiology ; Trans-Activators ; },
abstract = {In this study, Staphylococcus aureus was detected in 46% of raw milk and 10% of cheese samples collected in Edirne, Türkiye. All isolates carried the sec and seg enterotoxin genes, while 39% harbored sed, 7% seh, and only 4% of isolates carried either sea or sei. A total of 25% of the isolates exhibited multidrug resistance. The highest resistance rate was observed against penicillin (39%), followed by kanamycin (18%), tetracycline (14%), clindamycin (11%), chloramphenicol and rifampin (7%), and trimethoprim-sulfamethoxazole and gentamicin (4%). Methicillin resistance was found in 11%, and mecA was identified in two isolates. All isolates formed biofilms at 22°C and 37°C, and 82% also at 4°C. Agr typing showed that 21% of isolates belonged to group I, 21% to group II, and 11% to group III, while no group IV isolates were detected. These findings demonstrate that enterotoxigenic, antimicrobial-resistant, and biofilm-forming S. aureus isolates from dairy products may persist along the food chain and represent a potential public health risk, underscoring the importance of continuous microbiological monitoring and preventive strategies within a One Health framework.},
}

*Staphylococcus aureus/drug effects/genetics/physiology
Animals
*Biofilms/growth & development
*Enterotoxins/genetics/metabolism
*Cheese/microbiology
*Milk/microbiology
*Drug Resistance, Bacterial
*Bacterial Proteins/genetics/metabolism
*Anti-Bacterial Agents/pharmacology
Gene Expression Regulation, Bacterial/physiology
Trans-Activators

@article {pmid42300347,
year = {2026},
author = {Pongen, YL and Chinnappa, B and Bollapragreate, SK and Punniyakotti, P and Durairaj, T},
title = {Biofilm mediated mineral alteration of limestone by Bacillus cereus and sulphate reducing bacterial consortia: impacts on quality grading, stability, and composition.},
journal = {Environmental science. Processes & impacts},
volume = {},
number = {},
pages = {},
doi = {10.1039/d5em00841g},
pmid = {42300347},
issn = {2050-7895},
abstract = {Microbial mediation of mineral transformation plays a vital role in both geochemical processes and industrial applications. This study investigates native bacterial communities from Upper Cretaceous limestone fossil deposits in Ariyalur, Tamil Nadu, India, focusing on their role in altering the composition of the limestone. A total of eleven morphologically distinct bacterial isolates and a sulphate-reducing bacterial (SRB) consortium were isolated and screened for urease production and sulphate-reducing ability under site-specific conditions. Biofilm formation, evaluated using confocal laser scanning microscopy (CLSM), revealed isolate-specific differences in biofilm architecture. The urease-positive isolate (CS4), Bacillus cereus, produced a thick matrix-rich biofilm (81.34 ± 0.84 µm) associated with mineral precipitation. In contrast, the SRB consortia formed a thinner biofilm (42.18 ± 0.87 µm), which promoted mineral dissolution. SEM/EDX and XRD analysis confirmed significant changes in the limestone texture and mineral composition of CS4-treated samples, including increased calcium content (∼6.39%) and reduced phosphorus (∼11.68%), sulphur (∼2.33%), and chlorine (∼3.07%), which are elements detrimental to cement quality. The study also indicates that sulphate reduction may be an important factor in the genesis of microbial-mediated limestone fossil formation. These findings suggest that microbial biofilm dynamics and urease-mediated processes can selectively enhance limestone quality, providing new insights into selective mineral leaching, biocalcification, and microbial-mineral interactions with potential industrial relevance.},
}

@article {pmid42300540,
year = {2026},
author = {Chen, X and Zhang, J and Rahimi, S and Kozjek, K and Larsson, L and Mijakovic, I and Pandit, S},
title = {Mechanistic insights into graphene coatings for oral biofilm inhibition and osteoblast compatibility.},
journal = {Journal of materials chemistry. B},
volume = {},
number = {},
pages = {},
doi = {10.1039/d6tb00324a},
pmid = {42300540},
issn = {2050-7518},
abstract = {The initial adhesion of bacterial cells to implant surfaces is a critical step in biofilm formation. Biofilms are complex microbial communities that are much more tolerant to conventional antimicrobial treatments than planktonic cells, often requiring mechanical disruption in addition to antimicrobial treatment. Once established, these biofilms and their self-produced extracellular matrix are difficult to eradicate. As a result, there is growing interest in engineering implant surfaces that can effectively disrupt bacterial adhesion and subsequent biofilm formation. Various surface-modification strategies, including antimicrobial agents and nanomaterial-based coatings, have been investigated. Among these, graphene-based coatings have shown promising antimicrobial properties. However, the mechanisms of their bactericidal activity remain insufficiently understood. We evaluated the antimicrobial efficacy of vertically aligned graphene (VG) coatings against Streptococcus mutans, employing electron microscopy and transcriptomics analysis to elucidate the mode of action. These coatings inhibited biofilm formation through a multifaceted mechanism: (i) reducing bacterial colonization; (ii) mechanical disruption of bacterial membranes by nanoscale protrusions; (iii) modulating expression of the genes associated with membrane integrity, transport, oxidative stress, and cell division. Importantly, the coatings inhibited bacterial adhesion and biofilm formation without affecting osteoblast growth or proliferation. These results indicate that VG coatings could offer a dual benefit by enhancing antimicrobial activity while being compatible for osseointegration, indicating their potential as candidates for next-generation biomedical implants.},
}

@article {pmid42300809,
year = {2026},
author = {Pan, D and Liu, R and Qiu, J and Wu, Z and Su, N and Shi, J and Cheng, N},
title = {Biofilm-mediated antibiotic cross-protection: Acinetobacter baumannii-driven enhancement of Elizabethkingia anopheles.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0334925},
doi = {10.1128/spectrum.03349-25},
pmid = {42300809},
issn = {2165-0497},
abstract = {UNLABELLED: Acinetobacter baumannii is a clinically prevalent multidrug-resistant bacterium, while Elizabethkingia, an emerging pathogen, has shown increasing infection and mortality rates in recent years. Although co-infections involving both pathogens have been reported, their interactions remain poorly understood. In this study, clinical data from 15 co-infected patients revealed that such infections were more frequent in elderly individuals with underlying diseases requiring intensive care and prolonged hospitalization. The two isolates exhibited similar antibiotic resistance profiles. In vitro co-culture experiments showed that Elizabethkingia anophelis alone was sensitive to doxycycline, but after co-culturing with A. baumannii, it survived at concentrations over 16 times its minimum inhibitory concentration (MIC), suggesting that A. baumannii may mediate antibiotic cross-protection. The maximum biomass (OD570) of the co-culture exceeded that of the monoculture under multiple antibiotic conditions. Transcriptomic analysis revealed no significant changes in drug resistance genes in either strain, but A. baumannii showed strong upregulation of the biofilm-associated gene pgaB. GO and KEGG analyses indicated enrichment in biofilm formation, adhesion, and exopolysaccharide synthesis. In contrast, E. anophelis showed reduced expression of its biofilm-related genes. Crystal violet staining, confocal microscopy, and SEM confirmed that co-culturing enhanced and compacted biofilm structure. Moreover, the Galleria mellonella model demonstrated higher virulence in co-infections. These findings suggest that biofilm enhancement mediated by A. baumannii promotes antibiotic tolerance in Elizabethkingia. This study provides new insights into polymicrobial co-infections and supports the development of therapeutic strategies targeting interspecies interactions.

IMPORTANCE: Hospital infections often involve more than one kind of bacterium, making them harder to treat. In this study, we focused on Acinetobacter baumannii, a well-known drug-resistant hospital pathogen, and Elizabethkingia anophelis, an emerging bacterium linked to severe infections. By analyzing patients and conducting laboratory experiments, we found that when these two bacteria grow together, E. anophelis becomes more tolerant to antibiotics and the mixed community forms stronger biofilms-protective layers that help bacteria survive. This cooperation also increased infection severity in an insect model. Our findings suggest that A. baumannii can enhance the persistence and drug resistance of E. anophelis through biofilm-related interactions. Understanding such bacterial partnerships may help develop better ways to control hospital infections and guide new antimicrobial strategies.},
}

@article {pmid42301295,
year = {2026},
author = {Sarkar, R and Agrawal, RK and Bankoti, K and Singh, J and Kumar, K and Saini, M and Roy, D and Sharma, GK and Singh, V and Chandra, M and Singh, BR},
title = {Characterization of Virulence Factors, Biofilm Production, and Antimicrobial Resistance in Mastitis-Associated Staphylococcus aureus Strains by Phenotypic and Genotypic Methods.},
journal = {Foodborne pathogens and disease},
volume = {},
number = {},
pages = {15353141261460684},
doi = {10.1177/15353141261460684},
pmid = {42301295},
issn = {1556-7125},
abstract = {Staphylococcus aureus, a common cause of bovine mastitis, relies on several virulence factors, with biofilm formation being a key contributor to its pathogenicity. The present study investigated the occurrence of S. aureus as etiological agents in bovine mastitis with a focus on the existence of various virulence factors and antibiotic resistance status. Among 120 milk samples collected from West Bengal and Uttar Pradesh, 36 (30%) S. aureus strains were confirmed by conventional methods and PCR. Phenotypic analysis revealed hemolysin (55.55%) and coagulase production (36.11%), while molecular analysis revealed the presence of leukotoxin (luksF, 19.44%), hemolysin (hlb, 58.33%), coagulase (coa, 63.88%), and toxic shock syndrome toxin (tsst-1, 30.55%) genes. Biofilm production ability was detected in 97.22% (crystal violet assay) and 86.11% (Congo red agar assay) strains. Biofilm-associated genes, namely, icaA (80.55%, 29/36), icaB (75%, 27/36), icaC (69.44%, 25/36), icaD (86.11%, 31/36), and MSCRAMMs genes, namely, clfA (58.33%, 21/36), clfB (75%, 27/36), fnbA (75%, 27/36), fnbB (55.55%, 20/36), bap (38.88%, 14/36), bbp (83.33%, 30/36), ebps (69.44%, 25/36), eno (66.66%, 24/36), fib (41.66%, 15/36), and cna (8.33%, 3/36), were also detected. Antimicrobial resistance was observed in 88.88% isolates, with 72.22% exhibiting multidrug resistance (MDR). Among the isolates, 83.33% were methicillin-resistant S. aureus (MRSA), and mecA, femA, and femB genes were present either singly or in combination in 76.66% of the isolates. Efflux pump protein genes, namely, norA, norB, norC, mdeA, mepA, and sepA, were detected either singly or in combination in S. aureus isolates. 61.53% of MDR-MRSA isolates harbored all six efflux pump genes. According to this study, S. aureus of mastitis origin harbors various virulence, antibiotic resistance, biofilm-forming, and efflux pump genes. Bovine mastitis-derived MDR S. aureus isolates can pose a significant public health risk and need urgent attention to formulate strategies for their control and preventing transfer to the human food chain.},
}

@article {pmid42301356,
year = {2026},
author = {Yu, L and Hu, M and Cao, Y and Zhang, J and Gao, Y and Zhu, F},
title = {Inhibition of biofilm formation by Limosilactobacillus fermentum supernatant against Porphyromonas gingivalis and Fusobacterium nucleatum: an in-vitro study.},
journal = {Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]},
volume = {57},
number = {1},
pages = {},
pmid = {42301356},
issn = {1678-4405},
support = {(HB2020045)//Wuxi Double Hundred Top Talents Project from Health Committee of Wuxi/ ; (M202240)//General Project of Wuxi Municipal Commission of Health and Family Planning/ ; },
mesh = {*Biofilms/drug effects/growth & development ; *Fusobacterium nucleatum/drug effects/physiology/growth & development/genetics ; *Porphyromonas gingivalis/drug effects/physiology/genetics/growth & development ; Carbon-Sulfur Lyases/genetics/metabolism ; Quorum Sensing/drug effects ; Bacterial Proteins/genetics/metabolism ; *Limosilactobacillus fermentum/metabolism/chemistry ; Homoserine/analogs & derivatives/metabolism ; *Anti-Bacterial Agents/pharmacology/metabolism ; Lactones/metabolism ; },
abstract = {BACKGROUND: This study aims to investigate the inhibitory effect of the supernatant of Limosilactobacillus fermentum CCFM1139 on the LuxS/AI-2 quorum sensing system in dual-species biofilms of Porphyromonas gingivalis and Fusobacterium nucleatum, and its anti-biofilm formation efficacy.

METHODS: Construct an in vitro dual-species biofilm model of P. gingivalis and F. nucleatum; after adding the supernatant of L. fermentum CCFM1139 at four stages of biofilm formation (0 h, 12 h, 24 h, 36 h), the following experiments were conducted: (1) Biofilm mass changes were assessed using crystal violet staining; (2) Morphological changes were observed via scanning electron microscopy (SEM); (3) Quantitative Real-Time Reverse Transcription Polymerase Chain Reaction(qRT-PCR) was employed to detect luxS gene expression levels in the biofilm; (4) AI-2 signaling molecule activity was quantified using Vibrio harveyi BB170 bioluminescence assay.

RESULTS: The supernatant of L. fermentum CCFM1139 significantly inhibited the formation of the dual-bacterial biofilm in a time-dependent manner: the inhibition rate peaked at 0 h (65.54%), followed by 60.20% at 12 h, decreasing to 47.09% at 24 h and 26.67% at 36 h. The inhibitory effect during the early phase (0-12 h) was significantly superior to that in the late phase (24-36 h) (p < 0.05). SEM observation revealed that the experimental group biofilm exhibited a loose monolayer structure with increased bacterial spacing, whereas the control group displayed a dense multilayer structure with tightly connected bacteria. qRT-PCR results indicated that the supernatant downregulated luxS gene expression in both P. gingivalis and F. nucleatum: inhibition of P. gingivalis was primarily observed between 0 and 24 h, while inhibition of F. nucleatum was stronger and more persistent. The Vibrio harveyi BB170 bioluminescence assay revealed that the supernatant from L. fermentum CCFM1139 significantly reduced the activity of the AI-2 signaling molecule in the dual-bacterial biofilm (p < 0.05).

CONCLUSION: The supernatant of L. fermentum CCFM1139 effectively reduced biofilm formation by suppressing the LuxS/AI-2 quorum sensing system in P. gingivalis and F. nucleatum, demonstrating its potential as an antimicrobial agent.},
}

*Biofilms/drug effects/growth & development
*Fusobacterium nucleatum/drug effects/physiology/growth & development/genetics
*Porphyromonas gingivalis/drug effects/physiology/genetics/growth & development
Carbon-Sulfur Lyases/genetics/metabolism
Quorum Sensing/drug effects
Bacterial Proteins/genetics/metabolism
*Limosilactobacillus fermentum/metabolism/chemistry
Homoserine/analogs & derivatives/metabolism
*Anti-Bacterial Agents/pharmacology/metabolism
Lactones/metabolism

@article {pmid42301505,
year = {2026},
author = {Xu, X and Du, M and Jiang, Z and Lyu, J and Jiang, W and Zhang, J and Sun, C and Zhang, Y and Wang, J},
title = {Integrated Environmental and Molecular Mechanisms of Navicula sp. Biofilm Induced Settlement and Metamorphosis in Mizuhopecten yessoensis Larvae.},
journal = {Marine biotechnology (New York, N.Y.)},
volume = {28},
number = {4},
pages = {},
pmid = {42301505},
issn = {1436-2236},
mesh = {Animals ; *Metamorphosis, Biological ; Larva/growth & development/physiology ; *Biofilms/growth & development ; Cyclic GMP/metabolism ; *Bivalvia/growth & development/physiology ; },
abstract = {Many marine invertebrate larvae are influenced by environmental factors during development, with diatom biofilms playing a crucial role in the settlement and metamorphosis of bivalve larvae. This study found that Navicula sp. biofilms alter the larval microenvironment by increasing dissolved oxygen levels through photosynthesis and decreasing nitrate, ammonium, and phosphate levels in the surrounding water. Exposure to Navicula sp. biofilms induced directional swimming in larvae and significantly shortened the time required for settlement and metamorphosis. Biofilms formed by Navicula sp. contained effective substances and key infochemicals that promoted the settlement and metamorphosis of Mizuhopecten yessoensis larvae. Soluble polysaccharides containing β-1,4-glycosidic bonds secreted by the biofilm were recognized by the larvae, triggering settlement and metamorphosis signaling. Untargeted and targeted metabolomic analyses revealed increased levels of cGMP (Cyclic guanosine monophosphate), and GMP (Guanosine monophosphate), suggesting that larval settlement and metamorphosis may be associated with cGMP regulation. Based on these results, cGMP was selected for subsequent functional analyses. Treatments with the NO donor SNAP, the cGMP analog 8-Br-cGMP, and the sGC activator BAY 41-2272 significantly promoted metamorphosis, whereas the sGC inhibitor ODQ suppressed metamorphosis in a dose-dependent manner. These findings demonstrate that the NO-sGC-cGMP pathway positively regulates the settlement and metamorphosis of M. yessoensis larvae, with cGMP serving as a key effector. This study provides new insights into the mechanisms underlying larval settlement and metamorphosis in bivalves.},
}

Animals
*Metamorphosis, Biological
Larva/growth & development/physiology
*Biofilms/growth & development
Cyclic GMP/metabolism
*Bivalvia/growth & development/physiology

@article {pmid42282684,
year = {2026},
author = {Lee, BS and Godejohann, M and Mishra, R and Gürtler, F and Deloria, AJ and Liu, M and Leitgeb, R and Drexler, W and Thacker, VV and Berney, M and Haindl, R},
title = {A functional amyloid matrix underpins the PDIM-architected corded superstructure of the Mycobacterium tuberculosis biofilm.},
journal = {bioRxiv : the preprint server for biology},
volume = {},
number = {},
pages = {},
doi = {10.1101/2025.11.07.687260},
pmid = {42282684},
issn = {2692-8205},
abstract = {Mycobacterium tuberculosis (Mtb) biofilm formation is associated with antibiotic tolerance, but its architecture remains poorly understood. Here, we reveal that these biofilms form highly-organized superstructures of cords, and through their deconstruction, provide a new molecular insight into Mtb biofilms. Using multimodal imaging, we demonstrate that the lipid Phthiocerol Dimycocerosate (PDIM) is required for organizing bacilli into foundational cords and contributes specifically to biofilm-associated antibiotic tolerance. In contrast, the ESX-1 secretion system enhances the biochemical complexity of the extracellular matrix. Notably, we identified a functional amyloid matrix that encases bacterial cords or aggregates within the biofilm, likely conferring structural integrity. Together, these findings support a three-component model that distinguishes structural integrity, physical organization, and biochemical maturation, establishing a new architectural framework for Mtb biofilms. Finally, we show that the natural compound epigallocatechin gallate (EGCG) disrupts biofilm formation, highlighting the therapeutic potential of targeting this architecture to overcome drug tolerance in tuberculosis.},
}

@article {pmid42284502,
year = {2026},
author = {Guo, X and Yang, J and An, M and Lin, B and Liu, T and Zhang, L},
title = {Inside-Outside ROS Therapeutic Strategy Based on Piezoelectric Nano-Urchin for Drug-Resistant Bacteria Biofilm Infections.},
journal = {Advanced science (Weinheim, Baden-Wurttemberg, Germany)},
volume = {},
number = {},
pages = {e76086},
doi = {10.1002/advs.76086},
pmid = {42284502},
issn = {2198-3844},
support = {22022402//Natural Science Foundation of China/ ; 21974051//Natural Science Foundation of China/ ; ECNU-SPDH CCTM-202510//East China Normal University-Shanghai Putuo District Central Hospital Collaborative Research Center for Translational Medicine/ ; 2025M771003//China Postdoctoral Science Foundation/ ; GZC20240477//Postdoctoral Fellowship Program of CPSF/ ; },
abstract = {Biofilm-associated infections pose a critical clinical challenge due to their inherent antibiotic resistance and limited therapeutic penetrability. Herein, we engineered a mechano-piezoelectric nano-urchin system, NiCo2S4@UiO-66, which utilizes ultrasound to achieve mechanical biofilm disruption and spatially hierarchical reactive oxygen species (ROS) generation for synergistic antimicrobial therapy. The spiky architecture of NiCo2S4 nano-urchins acts as physical penetrators, mechanically compromising biofilm integrity. Under ultrasound activation, a graded ROS generation mechanism is greatly enhanced via two distinct pathways. Externally, the NiCo2S4 nanozyme activated by piezoelectric UiO-66 successfully catalyzes pathogenic H2O2 at the biofilm periphery into highly destructive ·OH radicals, which not only degrade the extracellular polymeric matrix, but avoids additional oxidative stress. Internally, the mechanically driven piezoelectric UiO-66 component generates long-diffusing singlet oxygen ([1]O2), capable of targeting and eliminating bacteria embedded deep within the biofilm. Driven by the nano-urchin mechanical action, this hierarchical ROS mechanism integrates intra-biofilm [1]O2 production with peripheral ·OH-mediated decomposition, ensuring robust and comprehensive biofilm eradication. In a murine model of methicillin-resistant Staphylococcus aureus (MRSA) infected wounds, the system achieved rapid biofilm clearance and accelerated tissue repair through immunomodulation and angiogenesis promotion. This strategy addresses key limitations of conventional antimicrobial therapies and offers an effective approach for treating multidrug-resistant biofilm infections.},
}

@article {pmid42284781,
year = {2026},
author = {Wang, X and Huang, X and Ge, Z and Zhao, G and Xie, B and Zhan, M and Zhou, S and Su, Y},
title = {Exogenous quorum sensing signal enhances central energy metabolism to fuel biofilm formation and denitrification on microplastics.},
journal = {Journal of hazardous materials},
volume = {514},
number = {},
pages = {142627},
doi = {10.1016/j.jhazmat.2026.142627},
pmid = {42284781},
issn = {1873-3336},
abstract = {Microplastics (MPs) are known to host dense microbial biofilms and form the plastisphere, which serve as significant sites for various biogeochemical processes, including nitrogen transformation. The communication within these complex microbial communities is facilitated by quorum sensing (QS) signals. However, how this inter-bacteria signal crosstalk impacts the colonization and function of key microbes, such as denitrifiers, remains inadequately elucidated. This research delves into the impact of the external signaling molecule N-3-oxododecanoyl-L-homoserine lactone (C12-oxo-HSL) on biofilm development and denitrification processes by the model bacterium Paracoccus denitrificans (P. denitrificans) on microplastic surfaces. Treatment with 10 μM C12-oxo-HSL increased biofilm biomass 2.67-fold and nitrate removal rates 2.61-fold relative to controls, while planktonic biomass remained comparable to or lower than untreated samples, refuting the hypothesis that increased biofilm mass merely reflects accelerated planktonic growth. Transcriptomic analysis unveiled a sophisticated regulatory network. C12-oxo-HSL not only stimulated the expression of genes involved in initial adhesion and motility but also orchestrated a substantial upregulation of key energy metabolism pathways, including glycolysis, the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation. Metabolic upregulation likely increased ATP availability for the augmented production of extracellular polymeric substances, ultimately leading to the formation of a more resilient and efficient biofilm structure. Our findings suggest a potential energy-centric mechanism where exogenous AHLs prime the cellular bioenergetic status to support the structural and functional demands of plastisphere colonization. This highlights the pivotal role of signal-mediated resource allocation in shaping the biogeochemical impact of microplastic pollution.},
}

@article {pmid42286775,
year = {2026},
author = {Uruén, C and Marín, CM and González-Vázquez, LD and Gottschalk, M and Arenas, M and Arenas, J},
title = {Clinically relevant genomic and phenotypic differences in virulence, antimicrobial resistance, and biofilm-associated tolerance between Streptococcus suis lineages ST1 and ST123.},
journal = {Veterinary research},
volume = {57},
number = {1},
pages = {},
pmid = {42286775},
issn = {1297-9716},
support = {PID2023-151032NB-C22//Ministerio de Ciencia e Innovación/Agencia Española de Investigación/ ; PID2020-114617RB-100//Ministerio de Ciencia e Innovación/Agencia Española de Investigación/ ; PID2023-151032NB-C22//MICIU/AEI/10.13039/501100011033/ ; LMP58_21//Gobierno de Aragón/ ; LMP58_21//Gobierno de Aragón, Department of I+D+I project in priority lines/ ; },
mesh = {*Streptococcus suis/genetics/physiology/drug effects/pathogenicity ; *Biofilms ; *Streptococcal Infections/microbiology/veterinary ; Animals ; Virulence/genetics ; *Drug Resistance, Bacterial/genetics ; *Anti-Bacterial Agents/pharmacology ; Spain ; *Swine Diseases/microbiology ; Mice ; Swine ; Phenotype ; Genome, Bacterial ; },
abstract = {Streptococcus suis is a Gram-positive bacterium and an important pathogen in pigs and humans. It can be classified into more than 3000 sequence types (STs), among which the ST1 and ST123 lineages are highly prevalent in Spain. ST1 is a globally distributed lineage, while ST123 has emerged within the last decade only in Spain. In this study, we compared the genotypic and phenotypic characteristics of representative isolates from both lineages to better understand the factors driving the emergence of ST123. Comparative genomic analysis revealed higher genetic variability in ST123 than in ST1. The genomes of both lineages share approximately 1429 genes, representing about 61% of the total genome. Among the lineage-specific genes, we identified 131-143 genes encoding proteins involved in diverse biological functions, including metabolism, regulation, transport, and virulence. Some of these genes were located on genetic islands, encoding for proteins involved in nutrition and catabolism of specific carbohydrates. In mouse infection models, both STs showed a strong capacity to cause systemic infection, although they differed in tissue persistence patterns. In macrophage cultures, ST123 isolates showed reduced adherence and intracellular survival compared with ST1. In contrast, ST123 isolates demonstrated the capacity to acquire ampicillin resistance under laboratory conditions. Moreover, ST123 isolates exhibited increased biofilm formation and enhanced tolerance to β-lactam antibiotics within biofilms compared with ST1. In conclusion, the combination of virulence, increased biofilm-associated antibiotic tolerance, and increased propensity to acquire antimicrobial resistance may explain the rising prevalence of ST123 in Spain. Its recent detection in Italy further supports its potential for expansion and establishment across Europe in the coming years.},
}

*Streptococcus suis/genetics/physiology/drug effects/pathogenicity
*Biofilms
*Streptococcal Infections/microbiology/veterinary
Animals
Virulence/genetics
*Drug Resistance, Bacterial/genetics
*Anti-Bacterial Agents/pharmacology
Spain
*Swine Diseases/microbiology
Mice
Swine
Phenotype
Genome, Bacterial

@article {pmid42287196,
year = {2026},
author = {Wang, Y and Zhang, R and Gao, Z and Liu, X and Pian, Y},
title = {Antimicrobial Peptide SAAP-148 Inhibits Helicobacter pylori and Is Associated with Membrane Disruption, Biofilm Suppression, and Reduced Cell-Associated Urease Activity.},
journal = {Journal of applied microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1093/jambio/lxag145},
pmid = {42287196},
issn = {1365-2672},
abstract = {AIMS: To investigate the in vitro antibacterial activity and related phenotypic effects of the novel antimicrobial peptide SAAP-148 against Helicobacter pylori (H. pylori).

MATERIALS AND METHODS: The minimum inhibitory concentration (MIC) of SAAP-148 against H. pylori strains, including multidrug-resistant isolates, was determined using the broth microdilution method. Its activity after short-term exposure to pH-adjusted conditions and its cytotoxicity toward gastric epithelial cells were evaluated. The effects of SAAP-148 on cell morphology, membrane permeability, established biofilms, bacterial viability, cell-associated urease activity, and ureA/ureB gene expression were assessed. Antibacterial activity against one four-drug-resistant H. pylori clinical isolate was further evaluated by colony counting and OD600 measurement.

RESULTS: SAAP-148 exhibited potent antibacterial activity against H. pylori strains with different resistance profiles, with MIC values ranging from 16-32 µg/mL. SAAP-148 retained antibacterial activity after short-term exposure to pH-adjusted conditions and showed low cytotoxicity toward gastric epithelial cells. SAAP-148 treatment was associated with bacterial morphological damage, increased inner and outer membrane permeability, reduced biofilm biomass and biofilm-associated protein content, decreased cell-associated urease activity, and reduced ureA/ureB mRNA expression. In the dedicated MDR model, SAAP-148 also reduced viable counts of one four-drug-resistant H. pylori clinical isolate.

CONCLUSION: SAAP-148 demonstrated promising in vitro activity against H. pylori and was associated with membrane disruption, antibiofilm effects, and reduced urease-associated readouts. These findings support further preclinical evaluation of SAAP-148 as a potential antimicrobial strategy against difficult-to-eradicate H. pylori.},
}

@article {pmid42287247,
year = {2026},
author = {Xu, D and Luo, L and Zhang, L and Zhang, L and Lu, S and Li, D and Bi, F and Chen, J and Peng, X},
title = {Iron-Hijacking Trojan Horse Nanoplatform Combats Implant-Associated Biofilm Infections Through Immuno-Fibrotic Remodeling.},
journal = {Advanced materials (Deerfield Beach, Fla.)},
volume = {},
number = {},
pages = {e73711},
doi = {10.1002/adma.73711},
pmid = {42287247},
issn = {1521-4095},
support = {82402838//National Natural Science Foundation of China/ ; 82472461//National Natural Science Foundation of China/ ; LHGJ20240264//Henan Provincial Medical Science and Technology Research Joint Venture Project/ ; },
abstract = {Implant-associated biofilm infections persist due to a vicious cycle of biofilm resilience and fibrosis-driven immune exclusion. To break this cycle, we engineered a photothermal nanoplatform (CHPB@H) consisting of curcumin-loaded hollow Prussian blue (CHPB) nanoparticles embedded in an injectable thermosensitive chitosan hydrogel for near-infrared (NIR)-triggered local delivery. Upon NIR irradiation, the platform releases iron ions and curcumin that self-assemble into Fe-curcumin (Fe-Cur) complexes. These complexes act as a molecular "Trojan horse" that hijacks bacterial iron-acquisition pathways to induce ferroptosis. Concurrently, CHPB scavenges reactive oxygen species (ROS) via its intrinsic nanozyme activity and suppresses oxidative stress-driven fibroblast activation, thereby dismantling the fibrotic barrier that excludes immune effector cells. This dual action eliminates biofilms and restores immune cell infiltration into the infection site. In preclinical animal models, the therapy clears established implant infections, remodels the immune-fibrotic microenvironment, and promotes tissue regeneration. This work establishes a "clear-and-remodel" paradigm that integrates active targeted ferroptosis (via iron hijacking) with immuno-fibrotic remodeling, offering a translatable strategy for complex implant-associated biofilm infections.},
}

@article {pmid42287983,
year = {2026},
author = {Chen, P and Zhao, J and Yang, JC and Su, X and Zhang, P and Feng, HT and Tang, BZ},
title = {A membrane-to-nucleus targeting photosensitizer featuring aggregation-induced emission for dual-color imaging-guided antifungal therapy and biofilm disruption.},
journal = {Biomaterials},
volume = {335},
number = {},
pages = {124367},
doi = {10.1016/j.biomaterials.2026.124367},
pmid = {42287983},
issn = {1878-5905},
abstract = {The management of superficial fungal infections is a major global public health burden, driving the need for precise and resistance-free therapies. While photodynamic therapy (PDT) mediated by photosensitizers (PSs) offers a promising alternative, the lack of self-reporting capability in antifungal PSs hampers the accurate control of treatment and increases collateral risk from excessive reactive oxygen species (ROS). In this work, we designed and synthesized a series of aggregation-induced emission (AIE) PSs consisting of triphenylamine and 1H-indene-1,3(2H)-dione units, and found a versatile PS (named as ITTPM) that not only generates ROS efficiently but also simultaneously discriminates between live and dead fungi via a distinct fluorescence signal switch. In live fungi, ITTPM targets the cell membranes emitting red fluorescence. Upon fungal cell death, it relocates to the nuclei, staining them green. Mechanistic investigations via molecule imaging, lipid membrane mimicking experiment, DNA-responsive experiments and molecular docking revealed that ITTPM interacts with fungal membrane phospholipids via its aryl-substituted indanedione fragment and one pyridinium group in live fungi, versus binding to DNA bases via two cationic pyridinium fragments in dead cells. Furthermore, ITTPM shows potent photodynamic antifungal efficacy against planktonic fungi, effectively inhibits biofilm formation, and eradicates mature biofilms. In vivo studies on fungi-infected mouse wounds demonstrated that ITTPM achieved complete fungal eradication, facilitated wound healing and exhibited excellent biocompatibility. This study provides a promising candidate PS for precise clinical treatment of superficial fungal infections and novel insights into developing multifunctional antifungal PSs.},
}

@article {pmid42289118,
year = {2026},
author = {Rashova, M and Kabduova, A and Sailau, Z and Serikberli, G and Nurmukhamed, K and Munaidarova, A},
title = {COMPREHENSIVE ASSESSMENT OF BIOFILM FORMATION AND ANTIMICROBIAL RESISTANCE OF STAPHYLOCOCCUS IN PURULENT-INFLAMMATORY DISEASES.},
journal = {Georgian medical news},
volume = {},
number = {373},
pages = {98-108},
pmid = {42289118},
issn = {1512-0112},
mesh = {*Biofilms/drug effects/growth & development ; Humans ; *Staphylococcal Infections/microbiology/drug therapy ; *Staphylococcus aureus/drug effects/isolation & purification/pathogenicity ; *Anti-Bacterial Agents/pharmacology ; *Drug Resistance, Bacterial ; Microbial Sensitivity Tests ; *Staphylococcus/drug effects ; Staphylococcus epidermidis/drug effects/isolation & purification ; Suppuration/microbiology ; },
abstract = {INTRODUCTION: Treatment of biofilms is a priority in purulent surgery. A biofilm consists of 75-85% extracellular polymeric matrix and 15-20% microbial cells. Polysaccharides, proteins, and extracellular DNA within the matrix protect bacteria from adverse environmental factors (pH, antibiotics, phagocytosis). Communication between bacteria occurs through the "quorum sensing" system. Bacteria within biofilms are 100-1000 times more resistant to antibiotics compared to planktonic forms. This is explained by the limited penetration of antibiotics into the matrix and the reduced metabolic activity of the cells. Bacteria of the genus Staphylococcus, particularly Staphylococcus aureus, are currently recognized as the main causative agents of purulent-inflammatory diseases.

OBJECTIVE OF THE STUDY: to comprehensively assess the biofilm-forming activity of staphylococcal strains isolated from patients with purulent-inflammatory diseases of soft tissues (PIDs), as well as to determine the characteristics of their sensitivity to the main antimicrobial agents.

MATERIALS AND METHODS: To achieve the objectives of the study, we conducted a research project and examined 80 strains of the genus Staphylococcus isolated from purulent-inflammatory diseases. Of these, 50 belonged to the main group (MG) and 30 to the control group (CG). The identification of species characteristics of the 80 strains from the main and control groups was carried out based on their morphological, cultural, and biochemical properties. In addition, these strains were identified using MALDI-TOF mass spectrometry.

RESULTS: Analysis of the obtained results showed that S. aureus was predominant in both groups. During the study, 50 staphylococcal strains and two species-S. aureus and S. epidermidis-were identified among samples collected from patients with purulent-inflammatory diseases. In the control group, out of 30 strains, only one species-S. aureus-was identified. Biofilm-forming activity was also assessed based on microcolony size, and the morphological and tinctorial properties of isolated biofilm samples were studied. The field of view, number of objects, and their proportion within the field of view were calculated using digital images of the samples. The following microcolony sizes were taken into account: up to 10 µm², from 10 to 100 µm², from 100 to 1000 µm², from 1000 to 10,000 µm², and over 10,000 µm².},
}

*Biofilms/drug effects/growth & development
Humans
*Staphylococcal Infections/microbiology/drug therapy
*Staphylococcus aureus/drug effects/isolation & purification/pathogenicity
*Anti-Bacterial Agents/pharmacology
*Drug Resistance, Bacterial
Microbial Sensitivity Tests
*Staphylococcus/drug effects
Staphylococcus epidermidis/drug effects/isolation & purification
Suppuration/microbiology

@article {pmid42289247,
year = {2026},
author = {Lin, Y and Nie, B and Liu, X and Zhang, Q},
title = {Mechanistic insights into superior biofilm formation with heterotrophic nitrification-aerobic denitrification bacteria under polypropylene microplastic stress.},
journal = {Bioresource technology},
volume = {},
number = {},
pages = {135159},
doi = {10.1016/j.biortech.2026.135159},
pmid = {42289247},
issn = {1873-2976},
abstract = {Microplastics may disturb microbial activity and biofilm development in biological wastewater treatment systems, yet the response of three-dimensional rotating biological contactor start-up biofilms to polypropylene microplastic stress remains unclear. This study evaluated a biofilm initiation strategy using heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria (H-3D-RBCs) and compared it with activated sludge-inoculated systems (A-3D-RBCs) under polypropylene microplastic (PP-MP) exposure. H-3D-RBCs showed superior resistance to PP-MP disturbance, with total nitrogen removal decreasing by only 14 %, compared with an approximately 60 % decline in A-3D-RBCs. Respiratory activity inhibition remained below 15 % in H-3D-RBCs but exceeded 90 % in A-3D-RBCs. 16S rRNA gene sequencing showed that PP-MP reduced species richness and diversity in A-3D-RBCs and was associated with a > 90 % loss of core denitrifying genera, including Corynebacterium and Pseudoxanthomonas, whereas H-3D-RBCs maintained community stability and enriched Pseudoxanthomonas to 13.8 %. Metagenomic analysis indicated that PP-MP impaired nitrification and denitrification potential in A-3D-RBCs, as reflected by decreased genes encoding AMO and HAO, a 51.78 % decrease in nosZ abundance, and enhanced dissimilatory nitrate reduction to ammonium (DNRA), which likely intensified competition with denitrification and promoted nitrogen conversion to ammonia. In contrast, H-3D-RBCs suppressed DNRA and maintained high nosZ abundance. Untargeted metabolomics further showed that PP-MP was associated with metabolic disorders in A-3D-RBCs, especially disruptions in alanine, aspartate, and glutamate metabolism and arginine biosynthesis, whereas H-3D-RBCs preserved these key nitrogen metabolic processes. Overall, this study identifies key vulnerabilities of nitrogen-removal biofilms under PP-MP disturbance and provides multi-omics evidence to support the development of microplastic-resistant biofilm wastewater treatment systems.},
}

@article {pmid42290085,
year = {2026},
author = {Cruz, AF and Hamann, PRV and Guimaraes, FEG and Dabul, ANG and Mamani, RCC and Pileggi, M and Neto, MO and Sauda, MR and Valente, GT and Matsui, T and Weiss, TM and Araújo, EA and Polikarpov, I},
title = {Biochemical and Structural Characterization of Two-domain Glycoside Hydrolase PgaB from Serratia marcescens and Its Application for S. aureus Biofilm Degradation.},
journal = {ACS infectious diseases},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsinfecdis.6c00086},
pmid = {42290085},
issn = {2373-8227},
abstract = {Antimicrobial resistance (AMR) is a critical global health threat, with projections estimating up to 10 million deaths annually by 2050. One of the strategies for developing bacterial AMR is the formation of microbial biofilms (BFs). Thus, enzymes capable of degrading BF exopolysaccharides represent potential tools for BF disruption. In this work, we characterize β-1,6-N-acetylglucosaminidase from Serratia marcescens (SmPgaB), a two-domain enzyme with covalently attached GH153 and CE4 modules. Small-angle scattering data demonstrate that SmPgaB is monomeric in solution. We also demonstrate that SmPgaB degrades Staphylococcus aureus biofilms with up to 92% efficiency and inhibits biofilm formation by over 95%. Furthermore, SmPgaB enhances the effectiveness of gentamicin, tetracycline, and chloramphenicol, reducing the viability of planktonic cells by approximately 50% when used in combination with these antibiotics. Confocal laser scanning microscopy confirmed considerable morphological changes in the biofilm post-treatment. These results showcase the potential of β-1,6-N-acetylglucosaminidases as adjunct therapies for BF-related infections, particularly when combined with conventional antibiotics.},
}

@article {pmid42291458,
year = {2026},
author = {Dekkerová, J and Walentín, T and Jozefíková, A and Liščáková, J and Bujdáková, H},
title = {Incidence of carbapenemase-resistant isolates of Klebsiella pneumoniae in a regional hospital in Slovakia; contribution of virulence genes and biofilm production to overall pathogenicity.},
journal = {New microbes and new infections},
volume = {72},
number = {},
pages = {101778},
pmid = {42291458},
issn = {2052-2975},
abstract = {BACKGROUND: The prevalence of carbapenemase (CPE)-producing K. pneumoniae isolates has increased in hospitals worldwide over the past decade. Resistance is often associated with the ability of clinical isolates to form biofilm. The main objective was to investigate biofilm development in 30 hospital isolates of K. pneumoniae with confirmed CPE production as well as to analyze the distribution of resistance genes, the frequency of virulence genes, and their role in the regulation of biofilm.

METHODS: Selected isolates of K. pneumoniae with CPE production were examined for their ability to form biofilms using crystal violet staining. The presence of resistance genes (blaSHV1, blaTEM1, blaCTXM1) and genes (fimH, mrkD, entB) associated with biofilm formation was confirmed by PCR. Quantitative real-time PCR was then performed to compare the expression of biofilm-associated genes in weak, moderate and strong biofilm formers among K. pneumoniae.

RESULTS: More than 50% of all K. pneumoniae isolates were confirmed to be strong biofilm producers. PCR confirmed the presence of resistance-associated genes and genes important for biofilm development in all clinical isolates tested. Additionally, qPCR showed increased regulation of the adhesion-related genes fimH and mrkD in moderate or strong biofilm formers of K. pneumoniae compared to weak biofilm formers.

CONCLUSIONS: The present results provide valuable information on the relationship between increased CPE resistance, biofilm production, and its genetic regulation in K. pneumoniae hospital isolates.},
}

@article {pmid42291756,
year = {2026},
author = {Reichardt, E and Assar, S and Kristensen, MF and Lund, MB and Schlafer, S},
title = {The effect of probiotic bacteria on community composition, microscale pH and matrix architecture in a saliva-derived model of oral biofilm.},
journal = {Journal of oral microbiology},
volume = {18},
number = {1},
pages = {2682457},
pmid = {42291756},
issn = {2000-2297},
abstract = {BACKGROUND: Streptococcus salivarius K12 and M18 have recently gained interest as probiotic organisms for caries control. This study investigated the impact of S. salivarius K12 and M18 supplementation on biofilm virulence in a complex saliva-derived in vitro model compared to treatment with Limosilactobacillus reuteri DSM17938 and no treatment.

METHODS: Biofilms were grown under low (0.05%) and high (1%) sucrose conditions and supplemented with S. salivarius K12, S. salivarius M18 or L. reuteri. Microscale pH dynamics were assessed using confocal microscopy-based pH ratiometry. The biofilm matrix composition was analyzed by glucan and extracellular DNA mapping. Microbial community composition was determined by 16S rRNA gene amplicon sequencing.

RESULTS: At 0.05% sucrose, pH was significantly higher in S. salivarius M18 biofilms, but not in S. salivarius K12 biofilms, compared to control. pH in L. reuteri biofilms was significantly lower. At 1% sucrose, no significant effects of probiotic supplementation on biofilm pH were observed. Shifts in the bacterial composition were minor, most notably an increased abundance of lactobacilli in L. reuteri-treated biofilms at 1% sucrose. No significant effects on the biofilm matrix composition were observed.

CONCLUSIONS: In conclusion, probiotic supplementation exerted only minor effects on biofilm composition and virulence.},
}

@article {pmid42291757,
year = {2026},
author = {Oh, H and Kim, J and Bai, J},
title = {Antibacterial and anti-biofilm effects of postbiotic mediator derived from Jeotgal isolate Lactiplantibacillus plantarum KMC12 against Streptococcus mutans.},
journal = {Journal of oral microbiology},
volume = {18},
number = {1},
pages = {2684130},
pmid = {42291757},
issn = {2000-2297},
abstract = {BACKGROUND: Dental caries, commonly known as tooth decay, is a progressive breakdown of tooth tissues caused by the biofilm-producing bacteria, Streptococcus mutans. S. mutans plays a primary role in the initiation and progression of dental caries by producing a biofilm called dental plaque.

AIM: This study aimed to evaluate the antibacterial and anti-biofilm effects of the neutralized postbiotic mediator (nPM) derived from Lactiplantibacillus plantarum KMC12 and to assess its potential as a therapeutic agent against S. mutans.

METHODS: KMC12 nPM was prepared through centrifugation, filtration and lyophilization. Its antibacterial and anti-biofilm effects were investigated through MIC/MBC assays, membrane permeability tests, biofilm formation quantification, EPS production measurement, RT-qPCR analysis of biofilm-related genes, and cytotoxicity assays using Caco-2 cells.

RESULTS: KMC12 nPM exhibited antibacterial activity with MIC and MBC values of 128 and 256  mg/mL, respectively, and caused approximately 87% membrane damage at 512 mg/mL. Biofilm formation was inhibited by 36% at 256 mg/mL and 83% at 512 mg/mL. Furthermore, KMC12 nPM significantly downregulated gtfB, gtfD, ftf, aguD and atpD gene expression and reduced EPS production without showing cytotoxicity.

CONCLUSION: These results suggest KMC12 nPM as a promising, novel therapeutic agent for inhibiting S. mutans biofilm formation and caries prevention.},
}

@article {pmid42291886,
year = {2026},
author = {Bhujugade, SR and Patil, HV and Patil, SR},
title = {Biofilm-Forming Methicillin-Resistant Staphylococcus aureus: A Comprehensive Phenotypic and Genotypic Review.},
journal = {Cureus},
volume = {18},
number = {5},
pages = {e108813},
pmid = {42291886},
issn = {2168-8184},
abstract = {Methicillin-resistant Staphylococcus aureus (MRSA) is a major global pathogen, capable of forming biofilms that confer enhanced antimicrobial tolerance, virulence, and persistence in both hospital and community settings. Biofilms are structured microbial communities encased in extracellular polymeric substances, composed of polysaccharides, proteins, and extracellular DNA. Complex genetic networks, including the ica operon, the agr quorum-sensing system, the sarA regulatory system, and stress-response genes such as sigB, regulate MRSA biofilm formation. Phenotypic methods (Congo red agar, microtiter plate, and tube test) and genotypic assays (PCR-based detection of ica, fnbA/B, clfA/B, sarA, and agr) provide insights into biofilm capacity and virulence potential. Clinically, MRSA biofilms contribute to device-related infections, chronic wounds, and recurrent infections, challenging conventional therapies such as vancomycin, daptomycin, and rifampicin. Emerging strategies, including enzymatic matrix degradation, bacteriophage therapy, quorum-sensing inhibitors, and nanoparticle-based drug delivery, offer potential alternatives. This review synthesizes current knowledge on MRSA biofilm phenotypes and genotypes, highlighting molecular mechanisms, clinical significance, and therapeutic approaches.},
}

@article {pmid42293286,
year = {2026},
author = {Hillman, EBM and Walters, JRF and Carson, D and Rijpkema, S},
title = {Response to Kårhus et al. Regarding "Ruminococcus gnavus and Biofilm Markers in Feces From Primary Bile Acid Diarrhea Patients Indicate New Disease Mechanisms and Potential for Diagnostic Testing".},
journal = {Gastro hep advances},
volume = {5},
number = {7},
pages = {100951},
pmid = {42293286},
issn = {2772-5723},
}

@article {pmid42293296,
year = {2026},
author = {Bodle, KB and Vahidi, G and Johnson, E and Walker, D and Parker, A and Jones, C and Goeres, D and Peyton, BM and Smith, HJ},
title = {Assessing a method for single- and multidomain biofilm growth in a novel biofilm reactor: shear stress, ruggedness, repeatability, and reproducibility.},
journal = {Biofilm},
volume = {11},
number = {},
pages = {100372},
pmid = {42293296},
issn = {2590-2075},
abstract = {Numerous lab-scale bioreactor systems exist to cultivate biofilms under desired growth conditions. However, existing systems have not been able to simultaneously deliver low shear stress, high gas transfer, and intermittent wetting. The industrial surfaces biofilm reactor (ISBR) was specifically developed to overcome these limitations by simultaneously providing these conditions. This study presents the first standard operating procedure (SOP) for the growth of a model biofilm-forming organism, Pseudomonas aeruginosa, within the ISBR, providing a reproducible framework for investigating biofilm development under conditions that closely mimic industrial environments. Statistical analyses confirmed the SOP's repeatability, ruggedness, and reproducibility using biofilm density and biovolume data, supported by an analytical shear stress model for ISBR coupons. Biofilm densities were rugged to recycle and rotation rate deviations but increased with influent feed rate. Conversely, biovolumes were rugged to influent feed and rotation rate changes but declined with higher recycle rates. Both metrics demonstrated excellent repeatability and reproducibility, underscoring the need to assess these characteristics for consistent biofilm formation. Additionally, as a proof-of-concept, two multidomain biofilms (a Legionella pneumophila-Vermamoeba vermiformis model and a Rhodotorula mucilaginosa yeast-P. aeruginosa bacterial coculture) were cultivated and analyzed. Both exhibited high repeatability, and the coculture remained stable over a month-long growth period.},
}

@article {pmid42293568,
year = {2026},
author = {Tareau, AS and Parente, AMES and Furtado, AA and Barreau, M and Le, H and Amorim-Carmo, B and de Sousa, LHN and Maillot, O and Gonzalez, M and Forge, A and Tahrioui, A and Resende, JM and Lesouhaitier, O and Mendonça Araújo, R and Fernandes-Pedrosa, MF and Chevalier, S},
title = {Stigmurin derivatives as potent-biofilm eradicating agents against the major human opportunistic pathogen Pseudomonas aeruginosa.},
journal = {Frontiers in microbiology},
volume = {17},
number = {},
pages = {1827386},
pmid = {42293568},
issn = {1664-302X},
abstract = {Pseudomonas aeruginosa is a versatile opportunistic pathogen whose capacity to form biofilms contributes to persistent and difficult-to-treat infections, particularly in clinical environments. Conventional antibiotics often fail to eliminate biofilm-embedded bacteria, highlighting the need for alternative therapeutic strategies. Here, we investigated the anti-biofilm activity of Stigmurin, an antimicrobial peptide identified in the venom of Tityus stigmurus, together with six lysine-enriched analogs. The peptides were synthesized and characterized at a physicochemical level, and their effects on bacterial growth, biofilm development, and disruption of pre-established biofilms were evaluated under both static and dynamic conditions. Their influence on pyocyanin production, membrane fluidity, and the tridimensional structure of the most active analog, StigA31, was also analyzed. The peptides revealed no marked impact on bacterial growth or biofilm formation. However, several of them were able to disrupt mature biofilms on polystyrene pegs. Stigmurin (24 h exposition) reduced biofilm biomass by nearly 47%, while StigA31 achieved dispersal levels of up to 35%, even at nanomolar concentrations. Under dynamic flow conditions, both peptides strongly reduced biofilm biovolume and thickness within 2 h and altered matrix exopolysaccharide detection. None of the compounds stimulated pyocyanin production. Stigmurin altered membrane fluidity, whereas StigA31 displayed a distinct mechanism that may be associated with its helical conformation. These findings indicate that Stigmurin analogs, particularly StigA31, promote P. aeruginosa biofilm dispersal through non-bactericidal mechanisms. Their activity at sub-inhibitory concentrations highlights their potential as promising candidates for use in strategies aimed at controlling biofilm-associated infections.},
}

@article {pmid42294700,
year = {2026},
author = {Milani, G and Cortimiglia, C and Bellotti, G and Lucchini, F and Cocconcelli, PS},
title = {Locus of Adhesion and Autoaggregation (LAA) pathogenicity island genes hes and sisA are involved in virulence and biofilm formation in LEE-negative Shiga toxin-producing Escherichia coli (STEC).},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0335225},
doi = {10.1128/spectrum.03352-25},
pmid = {42294700},
issn = {2165-0497},
abstract = {Shiga toxin-producing Escherichia coli (STEC) is a major foodborne pathogen capable of causing severe diseases such as hemolytic uremic syndrome (HUS), particularly in children. While the locus of enterocyte effacement (LEE) has long been associated with STEC virulence, an increasing number of LEE-negative strains are being linked to human infections. Some strains harbor the locus of adhesion and autoaggregation (LAA), a pathogenicity island comprising genes implicated in adhesion and biofilm formation. In this study, we investigated the role of two LAA-associated genes, hes and sisA, in the virulence of the LEE-negative STEC strain UC4224 isolated from raw milk cheese. Using lambda Red recombineering, we generated mutant strains with deletions in stx1, stx2, and/or the hes-sisA region. Galleria mellonella larvae were employed as an in vivo infection model to evaluate pathogenicity. Mutants were also assessed for their ability to produce biofilm and adhere to epithelial cell lines. Our results showed that deletion of hes-sisA significantly decreased larval mortality, biofilm formation, and adhesion to Caco-2 intestinal cells. These findings suggest that the hes and sisA genes may have a role in STEC colonization and persistence, representing potential determinants of human infection. This work highlights the importance of LAA-associated hes and sisA in the pathogenicity of LEE-negative STEC strains and suggests that alternative virulence factors beyond Shiga toxins are critical to disease progression. Further studies are warranted to explore the functional roles of these and other genes within the LAA region across different STEC backgrounds and to improve their contribution to human infection.IMPORTANCEShiga toxin-producing Escherichia coli (STEC) is the third most common cause of gastrointestinal illness in Europe. Cattle act as asymptomatic reservoirs, transmitting infection to humans mainly through contaminated raw milk and meat. Several outbreaks linked to raw milk products have been reported in recent years, prompting EU Rapid Alert System notifications, including in 2024. These infections represent a major public health and economic concern. In 2019, the European Food Safety Authority recommended enhanced surveillance and a broader assessment of STEC pathogenicity, emphasizing virulence factors beyond Shiga toxins and intimin. This study investigates a specific genomic region within a pathogenicity island to elucidate its role in virulence and biofilm formation. The findings highlight additional molecular markers that may contribute to STEC pathogenic potential. Incorporating these markers into molecular screening and surveillance can improve the identification of high-risk STEC clones, supporting more accurate risk assessment and targeted public health interventions.},
}

@article {pmid42282683,
year = {2026},
author = {Taddei, SM and Deka, N and Marin, A and Hunt, BC and Guterman, LB and Ma, M and Qu, J and Armbruster, CE},
title = {L-Fucose-Dependent Biofilm Formation by Escherichia coli Enhances Polymicrobial Interactions and Antibiotic Tolerance on Urinary Catheters.},
journal = {bioRxiv : the preprint server for biology},
volume = {},
number = {},
pages = {},
doi = {10.64898/2026.06.01.729324},
pmid = {42282683},
issn = {2692-8205},
abstract = {Urinary tract infections are common healthcare associated infections, a large subset of which are caused by indwelling catheters. Long term catheterization causes persistent, asymptomatic, polymicrobial colonization despite catheters changes and antibiotic usage. In these polymicrobial populations, P. mirabilis , E. faecalis , and E. coli were found as the most common co-colonizing species. We investigated how interactions between P. mirabilis , E. coli , and E. faecalis contribute to biofilm formation and colonization of urinary catheters. Our results show that the interaction between these three species leads to enhanced biofilm biomass driven by an increase in total protein content of the biofilm. Biofilm enhancement required all three species and was also media-dependent, especially for dual-species combinations. Importantly, triple species biofilms also demonstrate biofilm enhancement when established under flow conditions in a biofilm reactor model using silicone urinary catheters. Additionally, triple species biofilm enhancement occurred in co-colonizing isolates from catheterized patients and was found to be specific to interactions between these three species. Triple species biofilms also demonstrated a species-dependent resistance to two commonly used antibiotics, ciprofloxacin and nitrofurantoin. By examining priority effects, E. coli was found to be the main facilitator of biofilm enhancement in a flow model. Finally, proteomics revealed that an L-fucose utilization pathway in E. coli was a key contributor to triple species biofilm enhancement. Overall, our results demonstrate the significant impact of polymicrobial interactions on biofilm formation in the catheterized environment and highlight ways in which complex microbial interplay and priority effects can shape the establishment of persistent colonization.},
}

@article {pmid42275129,
year = {2026},
author = {Consuegra-Asprilla, JM and González-Idarraga, M and Montoya-Carrascal, S and Tello-Tobón, DX and Gómez, AA and Martins, FS and González, Á},
title = {Effect of postbiotics on biofilm formation and gene expression in Candida spp. isolates from patients with recurrent vulvovaginal candidiasis.},
journal = {Medical mycology},
volume = {},
number = {},
pages = {},
doi = {10.1093/mmy/myag061},
pmid = {42275129},
issn = {1460-2709},
abstract = {First-line treatment for vulvovaginal candidiasis (VVC) has reduced efficacy in patients with recurrent VVC (RVVC), prompting the search for therapeutic alternatives such as probiotics. Recently, postbiotics (cellular components and metabolites derived from probiotics) have gained relevance because they offer similar benefits to probiotics without the risks associated with live microorganisms. This study aimed to evaluate the anti-biofilm effect of cell-free supernatants (CFSs) derived from probiotic strains against Candida spp. isolates from patients with RVVC, as well as their impact on the expression of genes associated with biofilm formation. Therefore, the anti-biofilm activity of CFSs from Lacticaseibacillus rhamnosus DM-UFMG 63, Bifidobacterium longum 5,1A and Lactobacillus acidophilus NCFM SD 5221 was evaluated in 40 Candida spp. isolates. The CFSs from L. acidophilus NCFM SD 5221 and Lc. rhamnosus 63 were analyzed using the XTT assay, and their effect on gene expression was measured by qPCR. At an estimated protein concentration of ∼200μg/mL, these CFSs significantly inhibited biofilm formation by approximately 50% (p<0.01)). This phenotypic effect correlated with the downregulation of key biofilm-associated genes (ALS3, HWP1, EFG1, TEC1, and UME6), alongside a concomitant upregulation of the transcriptional repressor NRG1. The findings of this study demonstrate that CFSs derived from Lc. rhamnosus DM-UFMG 63 and L. acidophilus NCFM SD 5221 exhibit significant antagonistic activity against biofilm formation in clinical isolates from patients with RVVC. Consequently, these cell-free supernatants represent a promising therapeutic and prophylactic alternative for the management of recurrent vulvovaginal candidiasis.},
}

@article {pmid42276153,
year = {2026},
author = {Diessner, NM and Stones, DH and Read, A},
title = {Analyzing Bacterial Biofilm Formation on Cranioplasty Implants.},
journal = {Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons},
volume = {},
number = {},
pages = {},
doi = {10.1016/j.joms.2026.05.067},
pmid = {42276153},
issn = {1531-5053},
abstract = {BACKGROUND: Cranioplasty (CP) is associated with high surgical site infections, compared to other deep-buried implant surgeries, including total knee and hip replacements.

PURPOSE: This study purpose was to measure and compare in vitro biofilm formation between polyether ether ketone (PEEK) and titanium CP implants.

STUDY DESIGN, AND SETTING: To address the research purpose, the investigator designed and implemented an in vitro cross-sectional study. Samples matching custom-made CP implants design were produced at NHS North Bristol trust. Microbiology testing was carried out at University of Gloucestershire, School of Education Health and Science and Gloucestershire Hospitals NHS Foundation Trust.

PREDICTOR VARIABLES: The predictor variable was between the implant materials unfinished titanium (cp-Ti), pressed titanium (polished cp-Ti), titanium (Ti-6Al-4V), and PEEK.

MAIN OUTCOME VARIABLES: The primary outcome variable was the development of Staphylococcus aureus (S aureus) biofilm on prepared CP samples. Colony-forming unit test were undertaken and images were taken 24 hours post incubation, and Image J was used to count the colonies. Crystal violet biofilm assays were undertaken and absorbance levels of each well (570 um wavelength) and is proportional to the concentration of the crystal violet staining of S aureus biofilm formed on the sample surfaces.

COVARIATES: Not applicable.

ANALYSES: One-way analysis of variance statistical analysis and Tukey's post-hoc test were used to compare parameters between various groups. The level of statistical significance was set at P value .05.

RESULTS: The studies used n = 40 (100%) samples, cp-Ti n = 10 (25%), polished cp-Ti n = 10 (25%), Ti-6Al-4V n = 10 (25%), and PEEK n = 10 (25%). Microbiology tests were used to measure the biofilm formation of S aureus on manufactured samples. PEEK had a statistically significant higher biofilm formation when compared to polished cp-Ti (P < .04) and cp-Ti (P < .05). However, there was not statistically significant result between PEEK and Ti-6Al-4V (P < .12).

CONCLUSION AND RELEVANCE: This study found that commercially pure titanium and Ti-6Al-4V has more resistance to biofilm formation by S aureus than PEEK on CP implant surfaces. Titanium CP implants may help lower S aureus adhesion compared to other materials, possibly limiting the risk of infections related to S aureus and improving patients' safety.},
}

@article {pmid42276819,
year = {2026},
author = {Zhang, Q and Lin, R and Zhao, Y and Zhan, P and Zhao, X and Zou, W},
title = {Biofilm-mediated antibiotic tolerance in bacterial pathogens: Integrated molecular networks and novel therapeutic avenues.},
journal = {Virulence},
volume = {},
number = {},
pages = {2687214},
doi = {10.1080/21505594.2026.2687214},
pmid = {42276819},
issn = {2150-5608},
abstract = {The stable structure of biofilms and the characteristics of the bacteria within them make biofilms an important barrier for bacteria to resist external stress, and a key factor contributing to the difficulty of eradicating clinical infections. This article reviews the multi-stage formation process of biofilms, the various mechanisms of antibiotic tolerance and resistance (such as physical barriers, metabolic adaptations, horizontal gene transfer, etc.), as well as the integrated regulatory roles of molecular networks like quorum sensing (QS) and cyclic diguanosine monophosphate (c-di-GMP). These multiple protective mechanisms in biofilms compose a closed "structure-function" loop system. In the past few years, the emergence of new anti-biofilm intervention approaches (matrix-degrading enzymes, phage therapy, nanomaterials, gene editing, etc.) revealed the possibility to break the limitations of conventional antibiotics by compromising structural integrity or interfering with signaling pathways, providing new ideas for drug-resistance infection control.},
}

@article {pmid42277069,
year = {2026},
author = {Singh, S and Sharma, S and Kumar, A},
title = {Pyruvate carboxylase is critical for biofilm formation in Mycobacterium tuberculosis.},
journal = {NPJ biofilms and microbiomes},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41522-026-01044-1},
pmid = {42277069},
issn = {2055-5008},
support = {IA/S/20/2/505220//Wellcome Trust DBT India Alliance/ ; },
abstract = {Mycobacterium tuberculosis (Mtb) forms biofilms. Biofilm formation is critical in Mtb's virulence. The metabolic pathways governing mycobacterial biofilm formation remain largely unexplored. This study evaluated how different carbon sources influence Mtb biofilm formation. Fermentable substrates-glucose, glycerol, and pyruvate-significantly promoted thick, mature biofilm formation compared to non-fermentable alternatives. In this study, we analyzed the role of pyruvate carboxylase (Pca), an anaplerotic enzyme that converts pyruvate to oxaloacetate, in biofilm formation, as it regulates carbon metabolic flux into the tricarboxylic acid (TCA) cycle and gluconeogenesis. To this end, a transposon mutant of the pca gene was investigated for its ability to form biofilms. We observed that the pca transposon mutant is deficient in pellicle, submerged, and macrocolony biofilm formation. Confocal microscopy indicated that the pca mutant does not accumulate extracellular cellulose in the biofilms and has lower biomass. These defects could be rescued upon complementing the mutant with an episomal copy of the pca gene or adding glucose or pyruvate to the medium. These observations suggest that biofilm formation requires a regulated flow of carbon flux into gluconeogenesis and the TCA cycle to ensure the supply of precursors for EPS biosynthesis and sustained energy production to fuel EPS synthesis.},
}

@article {pmid42280735,
year = {2026},
author = {Wang, J and Li, Y and Zhang, H and Wang, W and Yao, L and Makar, RS and Chen, Z and Han, H},
title = {Biofilm-Forming Enterobacter sp. W5 Mitigates Cadmium and Polystyrene Microplastic Stress in Wheat via Synergistic Immobilization and Proteomic Reprogramming.},
journal = {Plants (Basel, Switzerland)},
volume = {15},
number = {11},
pages = {},
doi = {10.3390/plants15111698},
pmid = {42280735},
issn = {2223-7747},
support = {42377039//National Natural Science Foundation of China/ ; },
abstract = {Cadmium (Cd) and polystyrene (PS) microplastic co-contamination in agricultural soils poses a potential threat to food security. Some functional microorganisms in soil can alleviate the dual stress of Cd and PS on crops. In this study, a biofilm-forming bacterium, Enterobacter sp. W5, was isolated from heavy metal-contaminated rhizosphere soil. Strain W5 exhibited Cd removal efficiency (46.3%) and strong biofilm-forming capacity (OD570 = 5.05), and it effectively colonized PS microplastic surfaces. XPS analysis detected bacterial functional groups (C-O-C, C=O) and PS-associated signals (O-C=O), which may act synergistically in Cd[2+] adsorption. Furthermore, XPS and XRD analyses revealed the presence of Cd-containing precipitates (including CdS, CdO, and Cd3(PO4)2). In hydroponic wheat experiments, W5 inoculation alleviated Cd-PS combined stress, thus significantly promoting plant growth and reducing Cd accumulation by 22.6% in roots and by 34.2% in aboveground tissues. Subcellular distribution analysis revealed that W5 enhanced Cd retention in root cell walls, thereby limiting its translocation to active cellular compartments. Proteomic analysis identified a set of 11 consistently downregulated proteins, including A0A3B6HQ68 and A0A3B6KJV9, which were enriched in secondary metabolite biosynthesis pathways. Bioinformatic analysis suggests that these proteins may be associated with Cd stress responses, though their exact roles remain to be verified. Collectively, this study provides a valuable microbial resource and mechanistic insights into the application of biofilm-forming bacteria for mitigating combined heavy metal-microplastic pollution in agricultural systems.},
}

@article {pmid42282017,
year = {2026},
author = {Chaulagain, D and Paul, B and Leslie, S and Artigues-Lleixà, M and Cros, MP and Toloza, L and Tang, Z and Hoover, A and Güell, M and Karig, D},
title = {Rational assembly of synthetic marine biofilm community with chitinase production.},
journal = {Research square},
volume = {},
number = {},
pages = {},
doi = {10.21203/rs.3.rs-9419003/v1},
pmid = {42282017},
issn = {2693-5015},
abstract = {Highly diverse multispecies biofilms are ubiquitous in microbial ecosystems; however, our current understanding of biofilm dynamics is limited to single species or low richness studies. We aimed to design a multispecies biofilm with a targeted function, chitinase production, using natural marine bacteria. We present a top-down assembly approach to design functional biofilm communities. Using our method, we found that final community membership was established within 24 hours, regardless of nutrient availability. However, cultivation in nutrient-rich media enabled rapid identification of the competitive dominant taxon, Pseudoalteromonas , among the 17 initial isolates used in the assembly. By repeating community assembly in a low-nutrient medium without these highly competitive taxa, we achieved the highest species diversity in the biofilm. The resulting multispecies biofilm exhibited chitinase production and maintained ~ 50% persistence during peak invasion. By comparison, a single species chitinase-producing biofilm formed lower biomass and suffered higher displacement during invasion. Importantly, one member that withstood invasion challenge in the multispecies community was completely undetectable at seven days post-invasion as a single species biofilm, indicating collective invasion resilience in the multispecies community. Further evidence of cooperation for coexistence is supported by increased β-N-acetylglucosaminidase, enzyme that hydrolyzes chitin oligomers, in the 14-member community at later timepoints, while the detected exochitinase activity remained stable. Our findings present a streamlined strategy to assemble diverse and functional biofilm communities for targeted biofilm engineering in marine and applied microbiome contexts, and our achievement of engineered function using natural bacteria offers a powerful complement to synthetic biology.},
}

@article {pmid42264144,
year = {2026},
author = {Darrouzet, E and Luche, S and Diemer, H and Cianférani, S and Lafond-Fenonjoie, D and Rabilloud, T and Lelong, C},
title = {Growth mode-dependent proteomic responses of Bacillus subtilis to cerium oxide nanoparticles: Pellicle biofilm versus swarming.},
journal = {Journal of proteomics},
volume = {},
number = {},
pages = {105696},
doi = {10.1016/j.jprot.2026.105696},
pmid = {42264144},
issn = {1876-7737},
abstract = {Proteomic analysis revealed that Bacillus subtilis exhibits markedly different physiological adaptations under pellicle biofilm and swarming growth conditions, and that these lifestyles strongly influence the bacterial response to CeO2 nanoparticles. In pellicle biofilms, proteins involved in respiration, amino acid acquisition, Mn/Fe uptake, and SUF-mediated iron‑sulfur cluster synthesis were upregulated, together with oxidative stress defense systems, indicating adaptation to microaerobic and heterogeneous biofilm conditions. In contrast, swarming cells displayed increased sporulation-associated processes and a stronger stringent response. Exposure to CeO2 nanoparticles induced a pronounced response, particularly under swarming conditions, where central carbon metabolism enzymes were strongly repressed and stringent response pathways were reinforced. In biofilms, CeO2 effects were more moderate, with limited metabolic perturbation and a slight stimulation of biofilm formation. The contrasting responses between lifestyles appear primarily linked to differences in metabolic state, oxidative stress physiology, and nanoparticle accessibility within the biofilm matrix rather than to direct nanoparticle toxicity alone. Overall, these findings demonstrate that nanoceria impacts B. subtilis physiology in a growth mode-dependent manner and highlight the importance of considering bacterial lifestyle when evaluating nanoparticle toxicity. SIGNIFICANCE: This research provides a biologically understanding of how Bacillus subtilis, an important bacterium for soil, plant, and animal health, adapts to environmental stress under more physiologically realistic growth conditions. Using shotgun proteomics, we demonstrated that pellicle biofilm and swarming lifestyles are associated with profoundly distinct physiological states, notably in metabolism, oxidative stress management, metal homeostasis, and developmental regulation. Extending this approach to exposure to cerium oxide nanoparticles (CeO2 NPs), a widely distributed environmental nanomaterial, we showed that bacterial responses are strongly dependent on growth mode. Swarming cells exhibited pronounced metabolic repression and activation of stringent response pathways, whereas biofilms displayed more limited perturbations together with slight stimulation of biofilm formation. These contrasting responses appear to result primarily from lifestyle-dependent differences in metabolic activity, oxidative stress physiology, and nanoparticle accessibility within the biofilm matrix rather than from direct nanoparticle toxicity alone. Since regulatory toxicology frameworks such as REACH often overlook subtle physiological adaptations, this study highlights the importance of considering microbial lifestyle and physiological context when assessing the ecological risks of emerging chemicals and nanomaterials.},
}

@article {pmid42264315,
year = {2026},
author = {Kashi, M and Hariri, Y and Kahbazi, M and Chegini, Z and Shariati, A},
title = {Natural compounds targeting multidrug-resistant Acinetobacter baumannii: Anti-Biofilm, anti-resistance, and synergistic strategies.},
journal = {Microbial pathogenesis},
volume = {217},
number = {},
pages = {108621},
doi = {10.1016/j.micpath.2026.108621},
pmid = {42264315},
issn = {1096-1208},
abstract = {Acinetobacter baumannii has emerged as a major cause of nosocomial infections. The emergence of drug-resistant strains has made most current antibiotics ineffective. The limited number of antibiotics currently in development has prompted the development of innovative strategies. One of these strategies is the use of natural compounds with potent antibacterial activity. Among them, carvacrol, thymol, cinnamaldehyde, curcumin, quercetin, eucalyptol, linalool, catechin, limonene, geraniol, coumarin, and eugenol have demonstrated considerable activity against A. baumannii. Natural compounds primarily act by compromising the integrity of the cell membrane, leading to leakage of intracellular contents and cell lysis and death. These compounds also inhibit ATP synthesis, induce oxidative stress, and interact with OmpA. Inhibition of efflux pumps, interaction with enzymes such as PBP, OXA, IMP, VIM, and NDM, disruption of secretion systems, and induction of DNA damage are among the mechanisms by which these compounds act against antibiotic resistance and may help limit the emergence of further resistance. Natural compounds also showed synergistic effects with conventional antibiotics, including imipenem, ciprofloxacin, gentamicin, and colistin, and could re-sensitize bacteria to these agents. To enhance the antibacterial and antibiofilm activity of natural compounds, nanoformulations have been used as effective vehicles, improving their bioavailability and therapeutic effectiveness. These findings provide valuable insights into the potential of natural compounds as effective agents against A. baumannii and its biofilm, offering promising avenues for developing novel strategies to prevent and manage drug-resistant A. baumannii infections, overcome antibiotic resistance, and provide a foundation for future therapeutic innovations.},
}

@article {pmid42264402,
year = {2026},
author = {Li, C and Tan, Y and Ma, S and Wang, J and Bai, W and Li, Z and Gao, S and Zhao, Q and Qin, J and Ye, Z},
title = {Concentration-dependent roles of hydrazine in immobilized denitrifying biofilm for industrial wastewater treatment.},
journal = {Bioresource technology},
volume = {},
number = {},
pages = {135122},
doi = {10.1016/j.biortech.2026.135122},
pmid = {42264402},
issn = {1873-2976},
abstract = {Hydrazine-bearing industrial wastewater is challenging to treat biologically because hydrazine can simultaneously act as a reducing substrate and a microbial inhibitor. In this study, an immobilized denitrifying biofilm system was used to evaluate the concentration-dependent effects of hydrazine on denitrification performance, electron contribution, and microbial response under anoxic conditions. Under sufficient co-substrate conditions, 5-10 mg/L hydrazine was effectively removed, with a maximum removal efficiency of approximately 94%, while stable denitrification was maintained. Nitrogen-15 isotope tracing showed that approximately 31% of the electrons released from hydrazine oxidation were transferred to denitrification-coupled nitrate reduction, indicating that hydrazine can partially contribute reducing equivalents in the denitrifying biofilm. However, elevated hydrazine concentrations impaired hydrazine oxidation and denitrification, induced nitrite and ammonium accumulation, and reduced carbon utilization. Mechanistic analyses showed that this deterioration was associated with oxidative stress, membrane damage, and inhibition of key enzymes, particularly nitrite reductase and hydroxylamine oxidoreductase. Metagenomic analysis further revealed a stress-induced shift in the microbial community from central carbon metabolism toward compensatory pathways. Overall, this study provides mechanistic and process-level insights into the feasibility and operational limitations of using immobilized denitrifying biofilms for treating hydrazine-bearing industrial wastewater.},
}

@article {pmid42264454,
year = {2026},
author = {Tuan, DA and Masak, J},
title = {Poultry-derived Lactiplantibacillus sp. PCE3 cell-free supernatant inhibits Candida albicans biofilm biomass and hyphal morphogenesis in vitro.},
journal = {Journal of applied microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1093/jambio/lxag142},
pmid = {42264454},
issn = {1365-2672},
abstract = {AIMS: To evaluate whether the cell-free supernatant (CFS) of a poultry-derived Lactiplantibacillus sp. strain PCE3 can inhibit biofilm biomass accumulation and hyphal morphogenesis of Candida albicans ATCC 10231 in vitro.

METHODS AND RESULTS: Biofilm biomass was quantified by crystal violet staining after 48 h growth in RPMI 1640 plus 2% glucose with 10%, 20%, or 40% (v/v) CFS. Filamentation was induced in RPMI 1640 plus 10% fetal bovine serum at 37°C and assessed qualitatively by light microscopy, scanning electron microscopy, and representative 24 h microscopy observations. Planktonic growth was summarized by 24 h colony-forming unit counts. CFS inhibited biofilm biomass in a concentration-dependent manner, with mean inhibition of 29.6 ± 8.3%, 48.1 ± 7.2%, and 79.8 ± 5.8% at 10%, 20%, and 40% (v/v), respectively (all p < 0.05 vs control). Under hypha-inducing conditions, 40% CFS shifted morphology from extensive filamentation in untreated controls to predominantly moderate or suppressed filamentation, and representative microscopy supported persistence of this non-filamentous phenotype over the observation period. Planktonic viable counts decreased by up to approximately 1.1 log10 at 40% CFS, indicating partial growth inhibition without eradication.

CONCLUSIONS: Poultry-derived Lactiplantibacillus sp. strain PCE3 CFS showed in vitro anti-virulence activity against C. albicans, reducing biofilm biomass and restraining filamentation more strongly than planktonic growth. These findings support avian-derived postbiotic preparations as an underexplored source of anti-Candida activity for further characterization and validation.},
}

@article {pmid42269295,
year = {2026},
author = {Zhao, J and Gong, J and Li, J},
title = {Synergistic nZVI-biochar and biofilm remediation of thiamethoxam at soil-water interfaces.},
journal = {Journal of hazardous materials},
volume = {514},
number = {},
pages = {142650},
doi = {10.1016/j.jhazmat.2026.142650},
pmid = {42269295},
issn = {1873-3336},
abstract = {The widespread use of neonicotinoid insecticides such as thiamethoxam poses significant ecological risks due to their persistence and mobility, particularly at the soil-water interface (SWI). This study investigated a nanoscale zero-valent iron-loaded biochar composite (nZVI-BC) synergized with indigenous phototrophic biofilms to enhance in situ degradation. While biochar alone immobilized thiamethoxam, it also prolonged its persistence in soil. The nZVI-BC composites significantly reduced pesticide leaching (49.7-62.6%) and accelerated its soil degradation, achieving 85.3-96.6% removal within 40 days, which was 2.1-2.4 times greater than biochar alone, while reducing the pesticide's half-life to 6.73-14.6 days. Phototrophic biofilms were the primary driver at the SWI, mediating aqueous-phase degradation (half-life of 0.67 days). Integrated analysis revealed that nZVI provided electrons for reductive transformation and enriched functional microbes, while biochar acted as a stable carrier and electron shuttle, facilitating microbial co-metabolism. This work demonstrates a synergistic "adsorption-reduction-biodegradation" strategy, effectively overcoming the limitation of biochar's contaminant retention and offering a sustainable approach to pesticide remediation at critical environmental interfaces.},
}

@article {pmid42269853,
year = {2026},
author = {Karim, ME and Eubank, T and Begum, K and Alam, MJ and Garey, KW},
title = {Reproducibility Assessment of the Crystal Violet Biofilm Assay in Clostridioides difficile.},
journal = {Anaerobe},
volume = {},
number = {},
pages = {103056},
doi = {10.1016/j.anaerobe.2026.103056},
pmid = {42269853},
issn = {1095-8274},
abstract = {OBJECTIVES: Crystal violet staining is widely used assay to quantitate bacterial biofilms. However, reproducibility remains a persistent challenge, particularly for anaerobic pathogens such as Clostridioides difficile. The purpose of this study was to assess the reproducibility of the crystal violet biofilm assay in C. difficile, systematically assessing the impact of microplate surface chemistry, plate format, and isolate-level heterogeneity on assay reproducibility.

METHODS: Reference strains (CD630 and R20291) and fifteen clinical isolates were grown anaerobically in tissue-culture treated and untreated 24-, 48-, and 96-well microplates. Biofilm biomass was quantified spectrophotometrically after 48h using crystal violet staining. All experiments were performed in three independent biological replicates. Reproducibility was assessed using within-plate and across-experiment coefficients of variation (CV%).

RESULTS: Tissue-culture treated 96-well plates consistently reduced variability relative to untreated plates in within-plate and across-experiment comparisons. Increasing well size in tissue-culture treated plates markedly improved reproducibility, with 24-well plates yielding the lowest CV% values and 96-well plates showing the greatest variability. Linear mixed-effects modeling confirmed these trends in effect direction and showed substantial experiment-to-experiment noise in small-volume plate formats. Mixed-effects variance partitioning across the fifteen isolate panel demonstrated that biological heterogeneity between isolates accounted for ∼20% of total within-plate CV% variance, whereas experiment-level variance was negligible, indicating high day-to-day assay stability under optimized conditions.

CONCLUSION: Crystal violet assay reproducibility for C. difficile biofilm quantitation is governed by technical parameters and isolate-specific biofilm physiology. Tissue-culture treated, 24-well plates can limit technical variability between experiments.},
}

@article {pmid42270263,
year = {2026},
author = {Langlois, JP and Jubinville, E and Goulet-Beaulieu, V and Jean, J},
title = {Occurrence and control challenges of biofilm-forming bacteria in industrial fresh produce processing.},
journal = {Food research international (Ottawa, Ont.)},
volume = {239},
number = {},
pages = {119551},
doi = {10.1016/j.foodres.2026.119551},
pmid = {42270263},
issn = {1873-7145},
abstract = {Microbial contamination remains a major challenge in industrial fresh produce processing, despite rigorous washing and sanitation procedures, and contribute to product spoilage and losses. The objective of this study was to characterize cultivable bacteria associated with industrial-scale processing of frozen fresh vegetables and to assess their biofilm-forming capability as well as their tolerance to commonly used disinfectants and to heat treatment under industry relevant conditions. Equipment surfaces, wash water and products were collected from operating commercial processing facility. A total of 153 bacterial isolates were identified by Sanger sequencing. Among these isolates, 27% were classified as strong producers of biofilm, 32% as moderate producers and 41% as weak producers. Susceptibility testing of strong biofilm-producing isolates showed that approximately 13% were susceptible to both sodium hypochlorite (70 ppm) and peracetic acid (0.05% w/v), 7% were susceptible to only one of the two disinfectants, and 80% did not achieve a more than 3-log reduction in viable counts under the conditions tested. In contrast, heat treatment representative of commercial blanching was effective at eliminating all tested strong biofilm producers in the planktonic state. Overall, these finding highlight the prevalence of biofilm-forming bacteria in industrial fresh produce processing environments and illustrate limitations of commonly used disinfectant against established biofilms. The results underscore the importance of optimizing sanitation strategies and biofilms-control approaches under industrial processing conditions. IMPORTANCE: Biofilms formed by bacteria in produce-processing environments pose a persistent challenge to sanitation and can contribute to product spoilage and losses. This study demonstrates that a substantial proportion of bacteria isolated from an industrial frozen fresh vegetable producing facility are capable of forming biofilms and exhibit limited susceptibility to commonly used disinfectants under industry-relevant conditions. The observation that heat treatment was effective against all tested isolates between biofilm-associated and free-living bacterial tolerance. By focusing on native bacterial isolates recovered directly from an operating processing facility, this work provides practical insight into biofilm persistence and sanitation limitations in industrial fresh produce processing. These findings are relevant for optimizing hygiene strategies and improving microbial control in commercial produce-processing environments.},
}

@article {pmid42270277,
year = {2026},
author = {Wang, S and Tian, L and Li, F and Wang, Q and Tan, Y and Ning, Y and Cao, X and Liu, Y and Shangguan, J and Li, H and Jiang, A and Zhao, M and Hyde, KD and Ren, A},
title = {Hypoxia-induced downregulation of cAMP drives Ganoderic acid biosynthesis and restricts biofilm development in Ganoderma lucidum.},
journal = {Food research international (Ottawa, Ont.)},
volume = {239},
number = {},
pages = {119570},
doi = {10.1016/j.foodres.2026.119570},
pmid = {42270277},
issn = {1873-7145},
abstract = {Hypoxia, as a widely prevalent environmental factor, profoundly influences the adaptability of filamentous fungi by regulating energy metabolism and developmental processes. Previous studies have shown that oxygen-limited conditions can induce biosynthesis of GAs. However, the regulatory network governing GA synthesis under hypoxia remains unclear. This study shows hypoxic stress decreases intracellular ATP and increases transcription of oxygen sensor ofd1, while inhibiting AC activity and reducing cAMP signaling. Reducing cAMP levels through pharmacological and genetic approaches enhances hypoxia-induced GAs accumulation, confirming cAMP's negative regulatory role in GA synthesis. Additionally, the study found that AC activation significantly accelerates biofilm formation and increases its thickness, indicating that cAMP signaling participates in hypoxic adaptation by positively regulating biofilm development. Thus, hypoxia coordinates GA biosynthesis and biofilm maturation through cAMP-dependent signaling in G. lucidum, revealing novel insights into the synergistic adaptation of metabolism and development under environmental stress.},
}

@article {pmid42270662,
year = {2026},
author = {Sachla, AJ and Piñeros, M and Helmann, JD},
title = {MeeY and YybP, two proteins regulated by manganese-sensing riboswitches, are required for Bacillus subtilis biofilm formation.},
journal = {NPJ biofilms and microbiomes},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41522-026-01045-0},
pmid = {42270662},
issn = {2055-5008},
support = {R35GM122461/NH/NIH HHS/United States ; },
abstract = {In the rhizosphere, Bacillus subtilis forms biofilms on plant roots and contributes to improved plant growth and disease resistance. Manganese stimulates biofilm formation in B. subtilis 3610, but the underlying mechanism is unknown. Here, we show that YybP and MeeY (formerly YkoY), two proteins regulated by the eponymous manganese-sensing yybP-ykoY riboswitches, are required for biofilm formation. MeeY is a TerC family membrane protein implicated in manganese export and YybP is of unknown function. Inactivation of either meeY or yybP leads to reduced synthesis of matrix-associated exopolysaccharide and a loss of biofilm-associated divalent cations.Deletion of sinR restores exopolysaccharide but does not restore normal metalation of the biofilm matrix. We conclude that the YybP and MeeY membrane proteins are previously unappreciated regulators of biofilm formation important for the production and metalation of exopolysaccharide.},
}

@article {pmid42273085,
year = {2026},
author = {Stevenson, D and MacPhee, CE and Stanley-Wall, N},
title = {Microplate-based quantification of poly-γ-glutamic acid levels in biofilm samples.},
journal = {Access microbiology},
volume = {8},
number = {6},
pages = {},
pmid = {42273085},
issn = {2516-8290},
abstract = {Poly-γ-glutamic acid (PGA) is a commercially useful biopolymer produced by many Bacillus species. PGA has a diverse range of applications across medicine and industry, generating significant interest in optimizing PGA production and enhancing yields. One approach to improve PGA recovery involves identifying high-yield PGA-producing strains and determining optimal production conditions, both of which require an appropriate screening method. Here, we present a sensitive and reproducible assay for quantifying PGA from Bacillus subtilis biofilms, whereby the spectral profile of methylene blue changes when bound to PGA. PGA was purified and lyophilized from NCIB 3610 ΔtasA liquid cultures grown at 50 °C, allowing production of protein-free PGA for use as a standard at known concentrations. Standard curves were generated from methylene blue absorbance readings at 564 and 664 nm, enabling subsequent quantification of PGA from biofilm extracts. We validated the quantification protocol and determined the treatment steps required to minimize interference. The assay has a 96-well plate format, enabling quantification of many samples at low sample volume, while minimizing waste of laboratory consumables. Overall, our method offers a sensitive, reproducible approach for PGA quantification in biofilm research and should facilitate comparative analyses across strains, treatments or environmental conditions.},
}

@article {pmid42274278,
year = {2026},
author = {Fan, S and Sun, S and An, H and Dai, W and Zhang, K and Yang, Z and Meng, X},
title = {Ag-Based Schottky-Engineered MOF Sonosensitizers Delivered via Dissolvable Microneedles for Sonodynamic Biofilm Eradication and Wound Healing.},
journal = {ACS applied materials & interfaces},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsami.6c08500},
pmid = {42274278},
issn = {1944-8252},
abstract = {Biofilms with dense, multibacterial communities formed at the infected wound represent a major pathological barrier to effective healing, which are also closely associated with persistent inflammation and therapeutic failure. Conventional strategies, including mechanical debridement and antibiotic therapy, often result in incomplete biofilm removal and accelerated development of antimicrobial resistance. In this work, we propose a synergistic sonodynamic-microneedle (SDT-MN) platform for efficient biofilm eradication and promoted wound healing. A zirconium-based metal-organic framework sonosensitizer (Ag@Zr-BT), in situ decorated with silver nanoparticles, is engineered to enhance charge separation via Schottky junction formation, thereby amplifying the generation of highly toxic reactive oxygen species (ROS) under ultrasound (US) activation. Ag@Zr-BT with optimal properties is integrated into dissolvable hyaluronic acid-based microneedle patches to facilitate direct drug delivery into biofilms. Under US irradiation, this SDT-MN system exhibits potent antibiofilm activity, efficiently destroying preformed Staphylococcus aureus (S. aureus) biofilms and inhibiting their regrowth. In an infected wound mouse model, SDT-MN-mediated delivery significantly enhances the sonosensitizer's penetration into biofilms and infected tissues, yielding improved therapeutic efficacy. This work establishes an antibiotic-independent therapeutic paradigm that integrates engineered MOF sonosensitizers with a microneedle-assisted delivery system, offering a promising strategy for treating biofilm-associated wound infections.},
}

@article {pmid42274649,
year = {2026},
author = {Musil, O and Klíma, K},
title = {Use of Micro/Nanorobots In Vivo for the Eradication of Bacterial Biofilm: A Review of Challenges and Strategies.},
journal = {Nanomaterials (Basel, Switzerland)},
volume = {16},
number = {11},
pages = {},
pmid = {42274649},
issn = {2079-4991},
support = {NW24-08-00473//Agency for Health Research of the Czech Republic/ ; },
abstract = {The term bacterial biofilm refers to a complex community of microorganisms embedded within a self-produced matrix of extracellular polymeric substances. This structural organization creates an environment that, when present in an infectious context within a living organism, limits the effectiveness of conventional antibiotic therapy. Consequently, such conditions substantially promote the development of antibiotic resistance. The decline in the discovery of novel antibiotic agents, coupled with a concurrent increase in the prevalence of multidrug-resistant microorganisms, has intensified the search for alternative strategies to combat such infections. At the same time, advances in nanoscience have stimulated substantial research into the use of micro/nanorobots for the eradication of bacterial biofilms. These devices, engineered at the micro- to nanoscale, are capable of targeted intervention in otherwise inaccessible sites. However, the development of such "microscopic therapeutic agents" is still at an early stage. To date, the vast majority of available data has been derived from in vitro studies, while evidence regarding their feasibility, safety, and therapeutic effects in living organisms remains limited. This review discusses their antimicrobial mechanisms and critically evaluates the current evidence concerning their in vivo applications.},
}

@article {pmid42274673,
year = {2026},
author = {Delle Fave, F and Froio, M and Cisternino, D and Jayaraman, S and Ashley, C and Medaglia, PG and Giorgi, F},
title = {Characterising the Antimicrobial Performance of Engineered Layered Double Hydroxide Surfaces for Biofilm Control.},
journal = {Nanomaterials (Basel, Switzerland)},
volume = {16},
number = {11},
pages = {},
pmid = {42274673},
issn = {2079-4991},
abstract = {Antimicrobial resistance (AMR) is a growing global health concern driven by bacterial biofilm formation, which increases tolerance to treatments. Developing surface-based strategies to limit biofilm formation is therefore critical. Layered Double Hydroxides (LDHs) are 2D brucite-like nanomaterials with tuneable physicochemical properties that may reduce bacterial colonisation. Their ease of synthesis, with scalability potential for industrial production, alongside their characteristic and tunable physicochemical properties, makes them a promising nanostructured coating for antimicrobial applications. This study evaluates LDH thin-film coatings as intrinsic antimicrobial surfaces, focusing on the combined effects of chemical composition, nanotopography, and wettability on biofilm formation in Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Four aluminium-based LDHs (ZnAl-NO3, ZnAl-Cl2, MgAl-NO3, MgAl-Cl2) were synthesised via coprecipitation or in situ growth on aluminium substrates. Materials were characterised by XRD, SEM, EDS, and contact angle measurements. Antimicrobial performance was assessed by quantifying colony-forming units (CFU mL[-1]) after bacterial exposure. ZnAl-LDH surfaces showed significant antimicrobial activity against E. coli and S. aureus, while MgAl-LDHs showed no effect and occasionally increased bacterial growth. None of the LDH surfaces tested exhibited significant antimicrobial activity against P. aeruginosa strain. The antimicrobial performance of ZnAl-LDH can be attributed to the concurrent effect of the surface chemistry, wettability, and sharp platelet-like nanotopography. The results obtained demonstrate that ZnAl-LDH-based coatings are promising antimicrobial materials with potential relevance for translational research in clinical antimicrobial surface development.},
}

@article {pmid42262105,
year = {2026},
author = {Sharma, S and Tiwari, V},
title = {Autophagy modulation by OmpA-targeted lead compounds restores autophagic flux, reduces biofilm, and enhances immune response in Acinetobacter baumannii-infected pulmonary cells.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0003626},
doi = {10.1128/spectrum.00036-26},
pmid = {42262105},
issn = {2165-0497},
abstract = {Acinetobacter baumannii is an opportunistic pathogen known for causing severe nosocomial infections. Autophagy, a key host defense mechanism against invading pathogens, is modulated by the virulence factor OmpA. OmpA is also responsible for bacterial adherence to host cells. Targeting OmpA provides a potential strategy to counteract these autophagic disruptions. In silico analysis (molecular mechanics and molecular dynamics simulations) and experimental validation of identified small molecule lead against OmpA were performed. These leads were tested in A549 cells infected with A. baumannii, revealing significant autophagic modulations. mRNA expression analysis showed accumulation of the autophagy marker LC3B and suppression of elongation-associated genes upon infection. Confocal microscopy of mCherry-GFP-LC3-transfected cells demonstrated a reduced mCherry/GFP ratio, indicating downregulated autophagic flux during infection. Treatment with the leads reversed these effects, restoring autophagic activity to levels seen in uninfected cells. Additionally, SEM analysis confirmed that the leads inhibited OmpA-mediated biofilm formation. They also reduced the internalization of A. baumannii in A549 cells. The leads also enhanced IFN-γ and IL-13 expression while reducing IL-10, promoting immune clearance of A. baumannii via autophagy modulation and suppression of Treg functions. These findings suggest that the identified leads hold promise as autophagy modulators, potentially regressing A. baumannii's ability to escape host defense.IMPORTANCEAutophagic escape of Acinetobacter baumannii is mediated by OmpA; therefore, targeting OmpA represents a promising strategy to counteract autophagy disruption. The identified lead effectively reverses autophagy modulation, inhibits OmpA-mediated biofilm formation and bacterial internalization, and promotes a pro-clearance immune response.},
}

@article {pmid42262127,
year = {2026},
author = {Qu, Y and Kaye, DM and McGiffin, D and Peleg, AY},
title = {Using biofilm-targeting local antibiotics to treat recalcitrant medical device-associated infections: driveline infection as a model.},
journal = {The Journal of antimicrobial chemotherapy},
volume = {81},
number = {7},
pages = {},
doi = {10.1093/jac/dkag203},
pmid = {42262127},
issn = {1460-2091},
support = {//Artificial Heart Frontier Program Grant/ ; },
abstract = {Driveline infection, exemplifying medical device-associated infection, is a difficult-to-treat complication of ventricular assist device implantation in which biofilm formation underlies both infection onset and recurrence. Although clinical experience remains limited, treatment success has been reported in cases where local antibiotics at high concentrations were used in combination with surgical debridement and/or device reimplantation. We recently identified the low metabolic state of biofilm cells as the central mechanism underlying the high antimicrobial resistance of staphylococcal biofilms and as a potential therapeutic target for driveline infections. We hypothesize that rationally combining weakly and strongly metabolism-dependent antibiotics at high concentrations for an extended period may eradicate staphylococcal biofilms, including the embedded tolerant and persister cells. If clinically validated, this strategy may hold the key to achieving long-term treatment success in many medical device-associated infections, including the driveline infection.},
}

@article {pmid42263057,
year = {2026},
author = {Beckman I V, RL and Victoria, B and Santiago, FZ and Echeverria, GN and Pinheiro, BV and D T Torres, M and Suits, L and Garcia, S and Wantuch, PL and de la Fuente-Nunez, C and Eswara, P and Rosen, DA and Fleeman, RM},
title = {Correction: Molecular response to the non-lytic peptide bac7 (1-35) triggers disruption of Klebsiella pneumoniae biofilm.},
journal = {PLoS pathogens},
volume = {22},
number = {6},
pages = {e1014298},
doi = {10.1371/journal.ppat.1014298},
pmid = {42263057},
issn = {1553-7374},
abstract = {[This corrects the article DOI: 10.1371/journal.ppat.1013437.].},
}

@article {pmid42263215,
year = {2026},
author = {Cibik, S and Duran, A},
title = {Effects of Lysozyme and Silver Anode Treatment on Pseudomonas aeruginosa Growth and Biofilm Formation in Raw Milk.},
journal = {Journal of food science},
volume = {91},
number = {6},
pages = {e71179},
pmid = {42263215},
issn = {1750-3841},
abstract = {Pseudomonas aeruginosa is an important spoilage‑associated and opportunistic pathogen in raw milk, producing heat‑stable enzymes and robust biofilms that resist conventional sanitation. The limitations of existing decontamination methods necessitate novel, nonthermal, and ecofriendly strategies. This study evaluated the antibacterial and anti‑biofilm effects of lysozyme combined with the silver anode technique (SAT) against P. aeruginosa ATCC 27853 in raw milk. The minimum inhibitory concentration (MIC) of lysozyme was estimated by disk diffusion (8 g/100 mL). Raw milk samples were assigned to eight experimental groups: untreated controls, lysozyme alone, SAT alone, and their combination. Bacterial counts (log CFU/mL) and biofilm formation (crystal violet assay) were monitored over 72 h of refrigerated storage, and silver ion migration was quantified by ICP‑MS. Lysozyme alone was predominantly bacteriostatic, while SAT alone produced a bactericidal reduction (up to 1.5-2 log). The combination reduced planktonic counts by up to 2.5 log and biofilm by up to 78%. These reductions were greater than either treatment alone, although the effect was additive, not strictly synergistic. Silver migration (3.88 µg/L) gave a daily intake of 0.055 µg/kg for an adult consuming 1 L, well below EPA (5 µg/kg/day) and EFSA (50 µg/kg food) limits. The combination offers an effective, non‑thermal, eco‑friendly dairy pretreatment strategy. It can extend shelf life, reduce biofilm contamination, and minimize chemical disinfectants, aligning with green technology.},
}

@article {pmid42263990,
year = {2026},
author = {Meng, Q and Zeng, W and Zhang, J and Liu, H and Li, S and Peng, Y},
title = {Efficient nutrient removal from low C/N municipal wastewater using a phototrophic biofilm system integrating simultaneous nitrification-denitrification and phosphorus removal (SND).},
journal = {Environmental research},
volume = {},
number = {},
pages = {124859},
doi = {10.1016/j.envres.2026.124859},
pmid = {42263990},
issn = {1096-0953},
abstract = {Microalgae-bacteria systems based on phosphorus-accumulating organisms (PAOs) offer low-energy and low-carbon-emission solutions for wastewater treatment, but their performance declines with low carbon-to-nitrogen (C/N) ratios municipal wastewater. In this study, a phototrophic biofilm system capable of coupling simultaneous nitrification-denitrification with phosphorus removal (P-SNDPRB) was developed to enhance low C/N ratios (3.32-4.11) municipal wastewater treatment. Before biofilm integration, total nitrogen (TN) removal was below 75%. After integration, TN removal increased to over 82%, while organic matter and phosphorus removal efficiencies remained at 85% and 90% in the P-SNDPRB system, respectively. Microalgae photosynthesis supplied oxygen to the biofilm, enabling denitrification. Chemometric and metagenomic analyses revealed denitrification and phosphorus accumulating metabolism (PAM) as key pathways for nitrogen and phosphorus removal. Flow cytometry sorting showed that biofilm spatial distribution promoted synergistic interactions among Accumulibacter, Competibacter, Nitrosomonas, Chlorella, and Cyanobacteria, further enhancing nitrogen and phosphorus removal. This study provides a low-energy and sustainable approach for the treatment of municipal wastewater with a low C/N ratio.},
}

@article {pmid42256692,
year = {2026},
author = {Requena-Mendizábal, M and Bocanegra-Arista, R and Maurtua-Torres, DJ and Rosales-Cifuentes, T and Collantes-Díaz, IE and Flores-León, DC and Valdez-Jurado, FR and Calla-Poma, R},
title = {Antibacterial activity of organic crude extract of Camellia sinensis on bi-species cariogenic biofilm.},
journal = {Revista cientifica odontologica (Universidad Cientifica del Sur)},
volume = {14},
number = {2},
pages = {e285},
pmid = {42256692},
issn = {2523-2754},
abstract = {OBJECTIVE: The study aimed to determine the antibacterial inhibitory activity and anti-biofilm effect of the ethanolic crude extract of Camellia sinensis leaves on bi-species oral bacterial biofilms.

MATERIALS AND METHODS: An ethanolic crude extract of Camellia sinensis leaves was used. Streptococcus mutans ATCC 25175 and Streptococcus gordonii ATCC 51656 were cultivated and used under anaerobic conditions for biofilm formation. Three groups were formed: Camellia sinensis extract, 0.12% chlorhexidine, and dimethyl sulfoxide. The inhibitory effect was determined by using the Kirby-Bauer Disk Diffusion Agar method, measuring the zone of inhibition formed around each of them. Additionally, the minimum inhibitory concentration was identified, considering the concentration where no growth of bacteria was observed. Immunofluorescence analysis was performed to evaluate cell viability in biofilms. Student's t-test for independent samples was used to compare groups.

RESULTS: A zone of inhibition of 17.4±0.4 mm for S. mutans, and 13.1 ± 0.3 mm for S. gordonii was produced by Camellia sinensis, while chlorhexidine produced 23.6 ± 0.3 mm for S. mutans, and 13.2 ± 0.5 mm for S. gordonii. The minimum inhibitory concentration was 3.12 mg/dL for S. mutans and 1.56 mg/dL for S. gordonii. Analysis of cell viability after 48 hours of incubation showed that the Camellia sinensis extract reduced the viability of biofilm cells.

CONCLUSION: Despite the study's limitations, Camellia sinensis shows antibacterial activity against multispecies cariogenic biofilms, with sensitivity in the tested bacteria.},
}

@article {pmid42257234,
year = {2026},
author = {Karahutová, L and Čobanová, K and Bujňáková, D},
title = {Virulence profiling and biofilm-forming capacity of different E. coli pathotypes and phylogroups isolated from calf diarrhoea.},
journal = {Veterinary and animal science},
volume = {33},
number = {},
pages = {100710},
pmid = {42257234},
issn = {2451-943X},
abstract = {Different Escherichia coli (E. coli) pathotypes are known to cause recurring and widespread calf diarrhoea, leading to substantial economic losses in the livestock industry. This study aimed to evaluate the prevalence of E. coli in calves with diarrhoea and to describe the molecular characteristics of 100 obtained isolates. The isolates were tested for virulence genes, such as papC, eaeA, stI, stII, ltI, stx1, stx2, cnf1 and cnf2, and genes for phylogenetic classification (chuA, yjaA, arpA, TspE4.C2 DNA fragment, arpAgpE, and trpAgpC). Additionally, their biofilm-forming ability was evaluated. The study also examined and compared the effectiveness of two forms of zinc oxide (ZnO) - classical inorganic and nanoparticle - at five different concentrations against strong biofilm producers. The detected virulence gene frequencies were: papC (44%), stx2 (20%), cnf1 (16%), eaeA (13%), cnf2 (13%), stx1 (4%) and st1 (1%). The isolates were divided into seven phylogroups, with the most common being D (32%), followed by B1 (25%), B2 (12%), C (10%), E (10%), A (9%) and F (2%). About 87% of the isolates could form biofilms, with 32% classified as strong biofilm producers. Both forms of ZnO inhibited biofilm formation at all tested concentrations, with nanoparticles showing slightly better efficacy. These findings demonstrate that calves with diarrhoea harbour various E. coli pathotypes, including STEC, NTEC, EPEC, ETEC and EHEC, many of which can form biofilms.},
}

@article {pmid42259706,
year = {2026},
author = {Takeda, TTS and Borges, MHR and Malheiros, SS and Costa, RC and Bertolini, MME and Nagay, BE and Souza, JGS and Barão, VAR},
title = {Effect of probiotics on oral biofilm formation on titanium and SLA implant surfaces: An in vitro and in situ study.},
journal = {The Journal of prosthetic dentistry},
volume = {},
number = {},
pages = {},
doi = {10.1016/j.prosdent.2026.05.032},
pmid = {42259706},
issn = {1097-6841},
abstract = {STATEMENT OF PROBLEM: Peri-implant biofilm formation is a key etiologic factor in peri-implant disease. Although probiotic supplementation has been proposed as an adjunct strategy to modulate oral biofilms, its effect on early biofilm development on implant-related titanium implant surfaces remains unclear.

PURPOSE: The purpose of this study was to evaluate whether exposure to Lactobacillus reuteri influenced early biofilm formation, architecture, and antimicrobial susceptibility on machined and airborne-particle abraded, large-grit, acid-etched (SLA) titanium implant surfaces.

MATERIAL AND METHODS: A 2-part study was conducted. In vitro polymicrobial biofilms were formed on machined and SLA titanium implant surface disks for 2 or 24 hours under control (untreated), placebo (oil vehicle), or probiotic conditions. Viable counts, metabolic activity, and biofilm biomass were assessed. In situ, 4-day biofilms were developed intraorally using a randomized, double-blind, crossover palatal appliance model under identical treatments. Microbial counts, metabolic activity, biofilm architecture, and susceptibility to 0.12% chlorhexidine were evaluated. Data were analyzed using 2-way ANOVA and Tukey HSD tests (α=.05).

RESULTS: In vitro, probiotic exposure increased early viable counts and metabolic activity (P<.05), particularly on machined surfaces. After 24 hours, probiotic-treated groups showed higher viable counts than the control and placebo groups, while biofilm biomass was surface-dependent (P<.05). In the intraoral model, L. reuteri exposure did not increase lactobacilli, anaerobes, or Candida albicans counts. Untreated SLA surfaces exhibited higher streptococcal and total viable counts, whereas placebo- and probiotic-treated SLA surfaces demonstrated reduced levels comparable to machined titanium. Probiotic-treated biofilms displayed a less compact architecture. Chlorhexidine susceptibility was unchanged.

CONCLUSIONS: Lactobacillus reuteri modulated early peri-implant biofilm organization, increasing microbial activity and viable counts in vitro, without increasing pathogenic burden or altering chlorhexidine susceptibility in situ, with effects more evident on SLA surfaces.},
}

@article {pmid42259815,
year = {2026},
author = {Liang, W and Guan, S and Bing, J and Du, H and Zheng, Q and Tao, L and Li, B and Huang, G and Chu, H},
title = {The Wor2 phenotypic switching regulator controls biofilm formation in Candida auris.},
journal = {NPJ biofilms and microbiomes},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41522-026-01043-2},
pmid = {42259815},
issn = {2055-5008},
support = {24PJD124//Shanghai Pujiang Program/ ; 32170193//National Natural Science Foundation of China/ ; 32370202//National Natural Science Foundation of China/ ; 32530005//National Natural Science Foundation of China/ ; 2025YFE0205500//National Key Research and Development Program of China/ ; },
abstract = {The emerging fungal pathogen Candida auris is a serious global public health threat due to its ability to persist in healthcare environments and on human skin. Here, we report a prevalent cluster of clinical C. auris strains with enhanced biofilm formation, a contributor to environmental and skin persistence. Genomic analyses show that the clinical isolates with enhanced biofilm formation ability carry a loss-of-function mutation in the WOR2 locus, which encodes a white-opaque switching regulator in Candida species. Analysis of 13,314 published genomes revealed that 3104 strains (23.3%) harbor nonsense or frameshift mutations in WOR2, indicating frequent clinical occurrence. Deletion of WOR2 in a clinical strain markedly increased biofilm formation, whereas reintroduction of an intact WOR2 significantly attenuated biofilm development. Further analyses show that Wor2 inactivation upregulates GFC1, ALS4, and multiple biofilm-associated genes. Together, these findings reveal a key regulatory mechanism underlying biofilm development, environmental persistence, and transmission of C. auris.},
}

@article {pmid42260342,
year = {2026},
author = {Yang, SB and Ku, D and Kwack, KH and Jang, EY and Jun, SK and Moon, JH and Lee, JH},
title = {Complete genome sequence of Streptococcus lingualis isolated from subgingival biofilm.},
journal = {BMC genomic data},
volume = {},
number = {},
pages = {},
doi = {10.1186/s12863-026-01450-6},
pmid = {42260342},
issn = {2730-6844},
support = {RS-2024-00404660//Korea Basic Science Institute/ ; RS-2021-NR061563//National Research Foundation of Korea/ ; RS-2025-16066503//National Research Foundation of Korea/ ; RS-2025-20512973//National Research Foundation of Korea/ ; },
abstract = {OBJECTIVE: Streptococcus lingualis is a recently described oral streptococcal species; however, genomic information for this species remains limited, with only a single complete genome currently available for the type strain S5ᵀ. The objective of this study was to generate a high-quality complete genome sequence of a subgingival biofilm-derived S. lingualis isolate and expand the genomic resources available for this species.

DATA DESCRIPTION: The S. lingualis strain KHUD_012 was isolated from subgingival biofilm of a healthy adult and sequenced using the PacBio Sequel IIe platform. A single SMRT cell produced 144,821 HiFi reads, which were assembled de novo using the IPA HiFi genome assembler v2.0. The complete genome consists of a single circular chromosome of 2,115,610 bp with a G + C content of 42.0%, encoding 1,956 protein-coding genes, four rRNA operons, and 62 tRNAs. BUSCO analysis indicated 99.5% completeness. Pairwise whole-genome comparisons with closely related type strains showed that KHUD_012 was most closely related to S. lingualis S5ᵀ, with an OrthoANI value of 96.44%, and Parsnp-based core-genome SNP analysis supported this phylogenetic placement. Comparative genomic analysis with S5ᵀ identified 1,717 shared core gene clusters and 198 unique gene clusters in KHUD_012, many of which were functionally uncharacterized.},
}

@article {pmid42248931,
year = {2026},
author = {Kim, HS and Kim, GY and Park, JW and Lee, JH and Yerke, AM and Yun, B and Hwang, S and Moon, KH},
title = {A Korean native halophyte extract attenuates the virulence of methicillin-resistant Staphylococcus aureus by inhibiting biofilm formation.},
journal = {Scientific reports},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41598-026-53460-0},
pmid = {42248931},
issn = {2045-2322},
support = {No. 2021R1C1C1004734, and 2022R1F1A1065328//National Research Foundation of Korea/ ; },
abstract = {Since the COVID-19 outbreak, global attention toward infectious diseases has intensified, and many experts anticipate that the next pandemic may stem from multidrug-resistant (MDR) bacteria, often termed "Superbugs". Among them, methicillin-resistant Staphylococcus aureus (MRSA) represents one of the most widespread MDR bacterium responsible for severe nosocomial infections. As the emergence of new resistant strains accelerates due to overuse and misuse of antibiotics, development of anti-pathogenic therapeutics has gained significant interest. In this study, we explored the anti-biofilm activity of Artemisia fukudo (Af), a halophyte abundant in bioactive metabolites. The n-butanol (n-BuOH) fraction of Af markedly inhibited MRSA biofilm formation, independent of bacterial growth inhibition or biofilm degradation. Transcriptional profiling by qRT-PCR revealed that expression of adhesion-related genes was notably downregulated. In A549 cell line infection assay, Af n-BuOH fraction treatment significantly reduced MRSA attachment and internalization. Furthermore, in a Caenorhabditis elegans infection model, Af n-BuOH fraction exposure extended host life-span, suggesting attenuation of bacterial virulence. Taken together, our findings demonstrate that Af-derived compounds interfere with initial adhesion process crucial for MRSA biofilm development and host colonization. This study highlights the therapeutic potential of halophyte-derived natural products as promising anti-virulence alternatives to conventional antibiotics for controlling infections caused by multidrug-resistant pathogens.},
}

@article {pmid42248932,
year = {2026},
author = {Jin, HW and Jung, SY and Eom, YB},
title = {Linalool exhibits potent antibacterial and anti-biofilm activity against carbapenem-resistant Acinetobacter baumannii.},
journal = {Scientific reports},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41598-026-53720-z},
pmid = {42248932},
issn = {2045-2322},
support = {SCH-20130328//Soonchunhyang University/ ; RS-2023-NR076438 (NRF-2023R1A2C1003486)//Ministry of Science and ICT, South Korea/ ; },
abstract = {Acinetobacter baumannii poses an urgent clinical challenge due to its extensive antimicrobial resistance and enhanced capacity for biofilm development. These biofilm structures act as protective barriers that impede antibiotic penetration and shield bacterial cells from immune-mediated clearance, often leading to persistent and hard-to-treat infections. This investigation explored the therapeutic potential of linalool, a naturally occurring monoterpene alcohol, against clinical carbapenem-resistant Acinetobacter baumannii (CRAB) isolates. Linalool demonstrated robust inhibitory effects on both early-stage biofilm formation and the disruption of preformed biofilms, as evidenced through crystal violet staining and live/dead viability imaging via confocal microscopy. Even at sub-inhibitory concentrations where planktonic growth was not fully suppressed, the growth kinetics changed in a dose-dependent manner. In addition, metabolic activity within the biofilms was markedly suppressed following linalool exposure, as determined by XTT reduction assays. At the molecular level, qPCR analysis revealed that linalool downregulated critical virulence- and resistance-associated determinants, including bap, bfmR, csuA/B, ompA, and blaOXA-23. These cumulative findings indicate that linalool exerts a multi-targeted mechanism of action and may serve as a viable candidate for adjunctive therapies targeting CRAB in the context of rising antibiotic resistance.},
}

@article {pmid42249399,
year = {2026},
author = {Wu, W and Zhao, H and Hao, S and Li, G and Xue, X and Li, J and Cui, W and Jiang, Y and Ma, Y and Qiao, X},
title = {Correction: Distribution of Staphylococcus aureus drug resistance genes, biofilm formation and cell wall characteristics in dairy cattle from dairy farms in Northeast China.},
journal = {BMC veterinary research},
volume = {22},
number = {1},
pages = {},
pmid = {42249399},
issn = {1746-6148},
}

@article {pmid42251870,
year = {2026},
author = {Kao, CY and Hidrosollo, JH and Longjam, N and Africa, AE and Lu, JJ},
title = {Biofilm-forming ability of Staphylococcus lugdunensis clinical isolates associates with sequence type and virulence in a Galleria mellonella model.},
journal = {Microbial pathogenesis},
volume = {217},
number = {},
pages = {108619},
doi = {10.1016/j.micpath.2026.108619},
pmid = {42251870},
issn = {1096-1208},
abstract = {Staphylococcus lugdunensis, one of the most virulent members of coagulase-negative staphylococci, exhibits a strong biofilm-forming capacity, raising the question of whether this trait directly contributes to its virulence in vivo. Here, we used crystal violet staining to assess the biofilm-forming capacity of 202 S. lugdunensis and examined its association with sample source, multilocus sequence types (STs), agr types, oxacillin susceptibility, and virulence in a larval infection model. Among the 202 isolates, 135 (66.8%) formed strong biofilms, 46 (22.8%) moderate, and 21 (10.4%) weak biofilms. Most S. lugdunensis isolates were obtained from blood, pus, and deep tissue samples, accounting for 47.5%, 15.3%, and 12.4% of cases, respectively, with only a weak association between sample source and biofilm-forming ability. Bacteremia isolates were classified as primary infections (n = 36) or contaminants (n = 40), with primary infection isolates forming significantly stronger biofilms. Biofilm formation was not strongly associated with oxacillin susceptibility, SCCmec, or agr types; however, ST3 isolates exhibited higher biofilm formation than ST6 and ST27. In addition, gene annotation of whole-genome sequences from 19 S. lugdunensis isolates revealed that biofilm-associated genes are highly conserved across all strains regardless of their biofilm phenotype. Larvae infected with extremely weak biofilm formers showed 100% survival up to 7 days, whereas strong biofilm formers caused markedly reduced survival, with 75% showing <50% survival compared to 30% among weak biofilm formers. Therefore, despite the phenotypic variation in biofilm formation, our work demonstrated an association between biofilm phenotype, bacteremia classification, sequence types, and in vivo virulence.},
}

@article {pmid42254362,
year = {2026},
author = {Yang, J and Feng, W and Xie, M and Yu, D and Yang, W and Ma, Y and Xi, Z},
title = {The roles of SAP2, STP1, and MRR2 on biofilm formation and itraconazole resistance through autophagy in Candida albicans.},
journal = {Frontiers in medicine},
volume = {13},
number = {},
pages = {1817039},
pmid = {42254362},
issn = {2296-858X},
abstract = {BACKGROUND: Candida albicans biofilm formation and azole antifungal resistance are major obstacles to clinical treatment of candidiasis. STP1 and MRR2 are implicated in C. albicans virulence and stress responses, while SAP2 plays critical roles in biofilm formation and drug resistance. However, the regulatory mechanisms of STP1 and MRR2 on SAP2 expression, biofilm formation, and itraconazole (ITR) resistance, particularly via autophagy, remain unclear.

METHODS: We compared biofilm-forming ability (via crystal violet assay and microscopy), ITR susceptibility, and autophagic responses among standard ATCC11006, STP1∆/∆, MRR2∆/∆, ITR-sensitive, and ITR-resistant strains. Gene expression levels of SAP2, and key autophagy-related genes were quantified using RT-qPCR under planktonic and biofilm conditions. The role of autophagy was further investigated using the inducer rapamycin and nitrogen starvation assays.

RESULTS: ITR-resistant strains exhibited significantly stronger biofilm-forming ability than sensitive strains. STP1 deletion enhanced biofilm formation, while MRR2 deletion impaired it. SAP2, STP1, and MRR2 expression was significantly higher in resistant strains than in sensitive strains under both planktonic and biofilm states, with a significant positive correlation between SAP2 and MRR2 under planktonic conditions (r = 0.659, p = 0.002). Notably, expression of all tested autophagy-related genes was significantly upregulated in biofilms and in ITR-resistant strains, but was markedly downregulated in both STP1Δ/Δ and MRR2Δ/Δ strains. Rapamycin induced autophagosome formation and upregulated SAP2 and ATG gene expression, but this response was blunted in the knockout strains. Furthermore, both STP1Δ/Δ and MRR2Δ/Δ strains displayed increased sensitivity to nitrogen starvation.

CONCLUSION: Our findings demonstrate that STP1 and MRR2 are critical regulators of biofilm formation and ITR resistance in Candida albicans. They exert these effects, at least in part, by modulating the expression of SAP2 and, importantly, by governing the autophagic pathway.},
}

@article {pmid42254524,
year = {2026},
author = {Zhang, T and Xiang, J and Yu, N and Tang, Y and Qu, J and Liu, Y and Li, X},
title = {Study on the mechanism of hinokitiol in inhibiting the biofilm activity of Staphylococcus epidermidis.},
journal = {Frontiers in microbiology},
volume = {17},
number = {},
pages = {1811623},
pmid = {42254524},
issn = {1664-302X},
abstract = {Mastitis in dairy cows is one of the most prevalent infectious diseases in the dairy industry, leading to reduced milk production, increased production costs, and substantial economic losses. Antibiotics are commonly used to prevent and treat mastitis; however, persistent biofilm formation by Staphylococcus epidermidis (S. epidermidis) has contributed to rising antibiotic resistance and poses significant challenges for complete eradication. Therefore, the development of effective therapeutic strategies is urgently needed. This study investigated the antibacterial and anti-biofilm activities of hinokitiol, both alone and in combination with Ceftiofur sodium, against biofilm-producing S. epidermidis. The checkerboard assay revealed a significant synergistic effect between hinokitiol and Ceftiofur sodium (FICI ≤ 0.5). Importantly, hinokitiol exhibited negligible cytotoxicity toward mammalian cells and low hemolytic activity against sheep erythrocytes at therapeutic concentrations. Mechanism studies demonstrated that hinokitiol disrupted bacterial membrane permeability, as evidenced by increased NPN uptake, PI staining, and significant intracellular protein leakage. Furthermore, hinokitiol induced the accumulation of intracellular reactive oxygen species (ROS), leading to oxidative stress-mediated cell death. Transcriptomic analysis and RT-qPCR validation showed that hinokitiol significantly down-regulated the expression of key genes involved in biofilm formation (icaA, fnbA, aap), quorum sensing (agrA, luxS), and global regulation (sarA, sigB), while up-regulating the repressor icaR. These findings suggest that hinokitiol exerts its antibacterial and anti-biofilm effects by damaging the cell membrane, inducing oxidative stress, and modulating multi-target regulatory pathways, making it a promising candidate for treating S. epidermidis infections. However, this study is limited by the absence of in vivo mastitis models and pharmacokinetic data. Future research should focus on validating these findings in animal models, optimizing administration routes (e.g., intramammary formulations), and assessing clinical efficacy in field conditions to facilitate the translation of hinokitiol into a viable therapeutic option for refractory S. epidermidis mastitis.},
}

@article {pmid42256255,
year = {2026},
author = {Wu, X and Xiao, H and Chen, Y and Wu, C and Xie, Q and Gu, Y and Qin, X},
title = {Blueberry leaf polyphenols suppress biofilm formation and restore oral microbial homeostasis for caries control.},
journal = {Journal of oral microbiology},
volume = {18},
number = {1},
pages = {2681284},
pmid = {42256255},
issn = {2000-2297},
abstract = {BACKGROUND: Dental caries is a dysbiotic biofilm-induced disease driven by Streptococcus mutans (S. mutans). Blueberry leaf polyphenols (BLP) are plant-derived polyphenols exhibiting antimicrobial properties. However, their effects on S. mutans remain incompletely understood.

METHODS: The composition of BLP was determined by ultrahigh-pressure liquid chromatography-triple quadrupole tandem mass spectrometer (UPLC-MS/MS). The minimum inhibitory concentration (MIC) and acid suppression activity were determined. In addition, the effect on biofilm was investigated by adhesion and aggregation assay, crystal violet assay, glycosyltransferases (GTF) activity, extracellular polysaccharides (EPS) quantification, scanning electron microscope (SEM) images and qRT-PCR. Furthermore, we carried out a rat caries model, and microbiome analyzes were performed by 16S rRNA sequencing.

RESULTS: UPLC-MS/MS analysis identified a range of compounds in BLP, including quercetin and chlorogenic acid. The MIC of BLP was 500 μg/mL, and it inhibited the acid production of S. mutans. Importantly, BLP decreased the sucrose-dependent adhesion and its auto-aggregation, inhibited the activity of GTF, resulting in reduced EPS and down-regulated genes like gtfB, comA and gbpB. In addition, it inhibited dental caries in rats and ameliorated the microbiota dysbiosis associated with caries.

CONCLUSIONS: BLP combats caries by inhibiting S. mutans growth, biofilm formation and restoring the oral microbiota.},
}

@article {pmid42256333,
year = {2026},
author = {Grøn, LV and Muñoz-Duarte, L and Marshall, IPG and Johnsen, NK and Eser, BE and Ma, Y and Eken, M and Kofoed, MVW and Koren, K and Philips, J},
title = {Adaptive laboratory evolution increased biofilm formation by Sporomusa ovata through a mutation in galU.},
journal = {Biofilm},
volume = {11},
number = {},
pages = {100368},
pmid = {42256333},
issn = {2590-2075},
abstract = {Acetogenic bacteria are attractive biocatalysts for the conversion of CO2 with H2 into acetate, as in gas fermentation. Gas fermentation reactors may benefit from biofilm formation, but attachment by acetogens is often limited. This study indeed found that the acetogen Sporomusa ovata 2663 was mainly planktonic and aimed to increase its biofilm formation through adaptive laboratory evolution. The adaptation strategy consisted of growing S. ovata on plastic (HDPE) carriers in bottles with a H2:CO2 headspace and transferring two carriers to a bottle with fresh carriers over eight serial transfers. This procedure resulted in the evolved S. ovata 2663-BF, which had a consistent increased propensity to attach. In heterotrophic growth conditions, four times more cells attached to the bottom of well plates in comparison to the wild type, while the adapted S. ovata produced 1.8 times more extracellular polysaccharides. Moreover, twice as many cells adhered to carriers when grown on H2:CO2. This improved attachment, however, did not lead to higher acetate production rates in simple trickle bed reactors, as the experimental setup rather stimulated planktonic growth. Only medium replacement sometimes favored the rate of the adapted strain. Interestingly, the evolved S. ovata had a point mutation in the gene galU, which likely increased the activity of the encoding UDP-glucose pyrophosphorylase, i.e. an enzyme involved in the synthesis of extracellular polysaccharides. Overall, this study demonstrates that cell attachment by S. ovata was increased through adaptive laboratory evolution, offering the prospect of investigating the importance of biofilm formation in biofilm-based gas fermentation reactors.},
}

@article {pmid42248538,
year = {2026},
author = {Elawady, R and Gaballah, A},
title = {eDNA: A dual nongenetic gamechanger in biofilm development and depletion.},
journal = {Microbial pathogenesis},
volume = {217},
number = {},
pages = {108612},
doi = {10.1016/j.micpath.2026.108612},
pmid = {42248538},
issn = {1096-1208},
abstract = {Biofilms, first described by Anthony van Leeuwenhoek (1632-1723), are surface-attached, organized microbial communities. They form when single cells embed within an extracellular matrix (ECM) composed mainly of extracellular polymeric substances (EPS), enabling a multicellular lifestyle. Extracellular DNA (eDNA), a major EPS component, makes up a substantial portion of biofilm biomass. It interacts with polysaccharides, proteins, and nucleic acids to create a stable network that reinforces biofilm structure. eDNA, present in all biofilm ECM, plays a paradoxical role in bacterial survival. It is vital for structural integrity, guiding the biofilm from initial attachment through maturation and dispersion, yet this dependence also creates a critical vulnerability. Recent research highlights that while eDNA supports biofilm growth, disrupting its release, interactions, or structure destabilizes the matrix and drives depletion. Recognizing eDNA as both a structural scaffold and a therapeutic target enables more effective strategies to inhibit and disperse resilient biofilm communities. This review highlights recent advances in eDNA-releasing mechanisms, its multifunctional roles in biofilm support, and how its structures and interactions regulate development. We further show that modulating eDNA provides a targeted strategy for antibiofilm therapy and clinical dispersal. Notably, eDNA itself may also act as a cue for biofilm inhibition.},
}

@article {pmid42240610,
year = {2026},
author = {Fritz, BG and Christensen, MH and Heiland, L and Tolker-Nielsen, T and Jakobsen, TH and Jensen, PØ and Bjarnsholt, T},
title = {Shaken batch cultures of Pseudomonas aeruginosa contain biofilm-like heterogeneity.},
journal = {Microbiology (Reading, England)},
volume = {172},
number = {6},
pages = {},
pmid = {42240610},
issn = {1465-2080},
abstract = {Two potential sources of differentially tolerant subpopulations of Pseudomonas aeruginosa in batch cultures are aggregation and growth rates. Here, we dissected these effects by treating batch cultures with tobramycin, colistin and a combination after various time intervals. Tobramycin was significantly more effective than colistin at early time points, while both showed similar efficacy at 24 and 48 h. Combination therapy was most effective at later stages, indicating the emergence of tolerant subpopulations. Reactivating growth of stationary-phase cultures by treating in fresh media altered antibiotic tolerance, further supporting the role of growth rate in tolerance. To examine the effect of aggregation, we separated batch cultures using filtration as well as examined the WT against a hyper-aggregating ΔwspF mutant. Aggregates demonstrated lower growth rates than single cells. However, microcalorimetry and colony enumeration after treatment revealed that aggregation only partially explained the variance associated with antibiotic treatment. Given that tobramycin and colistin target metabolically distinct subpopulations, this suggests that overall metabolic rate plays a larger role than aggregation regarding antibiotic tolerance in batch cultures.},
}

@article {pmid42241129,
year = {2026},
author = {James, V and Prasad, RV and Charanya, C and Balagopal, S and Mahalaxmi, S},
title = {Biocompatibility of Nanocurcumin-Nanopiperine Gels and Their Efficacy Against Streptococcus Mutans - Lactobacillus Rhamnoses Biofilm Model.},
journal = {Indian journal of dental research : official publication of Indian Society for Dental Research},
volume = {},
number = {},
pages = {},
doi = {10.4103/ijdr.ijdr_576_25},
pmid = {42241129},
issn = {1998-3603},
abstract = {CONTEXT: The rise of antibiotic-resistant bacterial strains and adverse effects associated with synthetic drugs have spurred interest in herbal alternatives. Co-administration of curcumin with piperine and its nano-formulation incorporation can significantly improve its bioavailability and drug release.

AIMS: To evaluate the biocompatibility and assess the anti-biofilm activity of gel integrating nanocurcumin with nanopiperine.

METHODS AND MATERIALS: Nanocurcumin, nanopiperine, and a nanocurcumin-nanopiperine (1:0.2) combination gel were formulated using carboxymethyl cellulose. Human gingival fibroblasts were exposed to concentrations ranging from 100 to 1000 μg/mL, and cell viability was assessed using the MTT assay at 570 nm. The anti-biofilm efficacy of the combination gel (0.9-500 μg/mL) against a dual-species biofilm of Streptococcus mutans and Lactobacillus rhamnosus was evaluated using a crystal violet assay and compared with calcium hydroxide, with absorbance measured at 600 nm.

STATISTICAL ANALYSIS USED: The data were analysed using descriptive statistics. For inter-group comparison, one-way analysis of variance followed by Tukey's post-hoc test for multiple pairwise comparisons was used with P < 0.05.

RESULTS: MTT assay showed dose-dependent cytotoxicity: nanocurcumin maintained >70% viability (71.1-100%), nanopiperine showed 69.7-92%, and the combination showed 37% at 1000 μg/mL but >71% at ≤700 μg/mL. Crystal violet assay showed maximum biofilm inhibition at 500 μg/mL.

CONCLUSIONS: The novel curcumin-piperine nanogel formulation exhibits excellent biocompatibility on human gingival fibroblast cells at concentrations below 700 μg/ml. Remarkable biofilm inhibition against the combined biofilm model at 500 μg/ml and lesser concentration was also achieved.},
}

@article {pmid42243405,
year = {2026},
author = {Su, H and Xu, T and Hu, W and Wang, H and Pei, Z and Lu, W},
title = {A multi-strain biofilm consortium enhances gut microbiota resilience and restores post-antibiotic homeostasis.},
journal = {World journal of microbiology & biotechnology},
volume = {42},
number = {6},
pages = {},
pmid = {42243405},
issn = {1573-0972},
support = {32172216//National Natural Science Foundation of China/ ; JUSRP202504013//Fundamental Research Funds for the Central Universities/ ; },
abstract = {Antibiotics can significantly disrupt gut microbiota homeostasis, reducing microbial diversity and causing dysbiosis associated with health issues. Gut biofilms play a critical role in resilience and stress tolerance of the intestinal ecosystem. Mucosal microbial communities also help restore the gut microbiota after interventions like probiotics, antibiotics, or fecal transplants. Previously, we developed a core bacterial consortium with strong in vitro biofilm-forming and stress-resilient properties, but its colonization ability and in vivo function remained unclear. In this study, we first validated the in vivo biofilm formation of the microbial consortium using a germ-free (GF) mouse model, then introduced single-, dual-, and multi-strain combinations with varying biofilm-forming abilities into specific-pathogen-free mice to assess their potential for recovering antibiotic-disrupted gut microbiota. Our findings indicate that the robust, in vitro-selected consortium continued to form substantial biofilms in GF mouse intestines. 16 S rRNA sequencing showed that, compared to single- or dual-strain treatments, administering the core consortium significantly increased microbial richness and diversity. The gut microbiota of consortium-treated mice more closely resembled healthy controls, suggesting the core consortium has superior potential to restore healthy gut microbiota. Overall, our research demonstrates the core consortium markedly improves gut microbiota resilience to antibiotic-induced disruptions in mice, accelerates restoration of health-associated taxa, and reestablishes gut homeostasis. This approach could transform probiotic interventions from passive supplementation to active ecological engineering, providing a theoretical and experimental basis for next generation of engineered probiotics and microbiome restoration therapies.},
}

@article {pmid42243673,
year = {2026},
author = {Fenclova, D and Hrazdilova, K and Coufalova, M and Ter Beek, J and Berntsson, RP and Zurek, L and Cihalova, K},
title = {Prolonged zinc exposure modulates biofilm metabolic activity and conjugation in Enterococcus faecalis.},
journal = {BMC microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1186/s12866-026-05250-x},
pmid = {42243673},
issn = {1471-2180},
support = {IGA24-AF-IP-032//Ministerstvo Školství, Mládeže a Tělovýchovy/ ; 2023-02423//Svenska Forskningsrådet Formas/ ; },
abstract = {BACKGROUND: Zinc oxide (ZnO), including its nanoparticulate form (ZnONPs), is widely used in agriculture and accumulates in the environment, where it may impose sustained selective pressure on microbial communities. However, the impact of prolonged zinc exposure on horizontal gene transfer and conjugation dynamics in Enterococcus faecalis remains poorly understood.

RESULTS: We exposed Enterococcus faecalis OG1RF:pCF10 (donor) and OG1SSp (recipient) to prolonged zinc exposure (20 serial passages) and analyzed phenotypic and transcriptional changes associated with conjugation and virulence-related traits. Chronic exposure to ZnO and ZnONPs was associated with pronounced aggregation in the plasmid-carrying donor strain, reduced optical density values, and significantly lower recoverable CFU/mL at 24 h, although extensive clumping likely affected CFU recovery. Zinc exposure was also associated with increased metabolic activity within established biofilms, while gelatinase production and antibiotic susceptibility remained unchanged. ZnONP-adapted recipient cells showed a significant increase in conjugation frequency, whereas ZnO-adapted recipients and zinc-adapted donors showed non-significant upward trends. Notably, transcription of genes within the plasmid-encoded prgQ conjugation operon was increased even in the absence of exogenous pheromone stimulation. In contrast, short-term zinc exposure did not enhance plasmid transfer, indicating that increased conjugation required long-term adaptation rather than acute stress.

CONCLUSIONS: These findings indicate that prolonged zinc exposure is associated with altered aggregation, biofilm-associated metabolic activity, and conjugation dynamics in E. faecalis. However, the underlying mechanisms remain unresolved and may involve a combination of physiological, regulatory, and genetic adaptations arising from long-term exposure.},
}

@article {pmid42247360,
year = {2026},
author = {Conner, C and AbdulAzees, PA and Chen, XD and Yeh, CK and Garcia-Godoy, F},
title = {Laboratory evaluation of mixed-species biofilm formation by Streptococcus mutans and Lactobacillus casei on three dental resin-based restorative materials.},
journal = {American journal of dentistry},
volume = {39},
number = {3},
pages = {117-120},
pmid = {42247360},
issn = {0894-8275},
abstract = {PURPOSE: To evaluate the biofilm-forming potential of mixed-species bacteria on three commercially available dental restorative composites and compare their resistance to biofilm accumulation. Given the increasing emphasis on bioactive and fluoride-releasing materials in caries prevention, the study specifically assessed the difference between restorative with and without fluoride content.

METHODS: Three restorative materials, Activa (bioactive fluoride releasing), Filtek Bulk Fill (non-fluoride releasing) and TPH (non-fluoride containing) were tested in the laboratory. Standardized resin-discs were fabricated, saliva-coated to simulate oral conditions, and inoculated with a 1:1 mixed culture of Streptococcus mutans and Lactobacillus casei. Biofilm development was assessed at 24- and 48-hours using colony forming unit (CFU) quantification. Statistical analysis was performed using one-way ANOVA and Tukey's post hoc test (α= 0.05).

RESULTS: Significant differences in biofilm accumulation were observed among materials at both points (P< 0.05). TPH exhibited the highest CFU counts at 24 hours (0.24 ± 0.03 x 10⁷ CFU/mL) and 48 hours (1.07 ± 0.12 x 10⁷ CFU/mL). In contrast, Activa and Filtek showed significantly lower biofilm formation at both points (24 hours: 0.093 ± 0.05 x 10⁷ CFU/mL; 48 hours: 0.47 ± 0.31 and 0.53 ± 0.12 x 10⁷ CFU/mL, respectively), with no significant difference between them.

CLINICAL SIGNIFICANCE: The restorative materials tested demonstrated varying degrees of resistance to biofilm formation. Activa and Filtek showed reduced biofilm accumulation compared to TPH at both early and mature stages of growth. The selection of restorative materials should consider their potential to resist bacterial colonization, especially in high-caries-risk patients and restorations placed near gingival margins.},
}

@article {pmid42234481,
year = {2026},
author = {Akbar, JH and Karched, M},
title = {Impact of Printing Orientation on the Surface Properties and Microbial Biofilm Formation of 3D-Printed Denture Resins.},
journal = {The International journal of prosthodontics},
volume = {0},
number = {0},
pages = {1-26},
doi = {10.11607/ijp.9754},
pmid = {42234481},
issn = {1942-4426},
abstract = {PURPOSE: This study aimed to characterize the adhesion and biofilm formation of key denture-associated microbes-Candida albicans, Porphyromonas gingivalis, and Streptococcus mutans-on a 3D-printed photopolymer resin (FotoDent®).

MATERIALS AND METHODS: Discs were fabricated at three distinct printing orientations (0°, 45°, and 90°) to assess the impact of layer deposition on surface properties and microbial colonization. Conventionally polymerized ProBase® cold resin discs served as controls. All specimens underwent comprehensive surface characterization, including measurement of areal surface roughness, static contact angle with polar and apolar liquids, and surface free energy components. Adherent microbial cells were quantified using species-specific quantitative real-time PCR (qPCR) following standardized biofilm assays.

RESULTS: Microbial adhesion to the 3D-printed resin was highly dependent on print orientation, with cell counts for all three species decreasing significantly (p<0.05) from the 0° to the 90° orientation. This trend was paralleled by an increase in surface roughness (Ra) with higher print angles. A strong, statistically significant positive correlation was observed between the surface roughness of FotoDent® discs and biofilm cell numbers for C. albicans (r=0.815), P. gingivalis (r=0.883), and S. mutans (r=0.903). In contrast, adhesion to the control ProBase® material was consistently higher and showed no significant correlation with any measured surface characteristic-roughness, wettability, or surface energy.

CONCLUSION: These findings indicate that for 3D-printed dental resins, printing parameters, especially orientation, critically determine the final surface topography, which in turn is a affects microbial adhesion and biofilm formation.},
}

@article {pmid42235080,
year = {2026},
author = {Mian, AA and Hussain, A and Saba, S and Amin, A and Cheema, MT and Sohail, M and Saleem, HGM and Khurshid, M},
title = {Biofilm Disruption and Gene Expression Alteration by Phages against Multi-Drug-Resistant Pseudomonas aeruginosa.},
journal = {Canadian journal of microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1139/cjm-2026-0082},
pmid = {42235080},
issn = {1480-3275},
abstract = {Pseudomonas aeruginosa is a major opportunistic pathogen responsible for severe human infections and is increasingly associated with multidrug resistance and limited treatment options. In this study, we isolated and characterized bacteriophages targeting drug-resistant P. aeruginosa strains and evaluated their capacity to disrupt biofilms and modulate biofilm-associated gene expression. Phages recovered from sewage showed broad lytic activity against P. aeruginosa and were classified morphologically into the families Podoviridae and Myoviridae. Two phages, A2 and A4, exhibited the highest lytic activity against the resistant strains examined. Both phages significantly degraded preformed biofilms, with phage A4 showing comparatively stronger antibiofilm activity. In addition, phage exposure altered the expression of several biofilm-associated genes, including pelA, htpB, bifA, psl, fimW, and wspA. A notable finding was the reversal of bifA expression from downregulation at 12 h to upregulation at 24 h following treatment with phage A4. Collectively, these results demonstrate that phages A2 and A4 possess strong lytic and antibiofilm activities and are capable of modulating biofilm-associated transcriptional responses in drug-resistant P. aeruginosa. The findings support the potential application of bacteriophages as alternative therapeutic agents against drug-resistant biofilm-associated infections.},
}

@article {pmid42235129,
year = {2026},
author = {Wang, L and Guan, M and Sun, D and Yu, H and Jiang, X},
title = {Biofilm-induced modification of passive film and corrosion resistance of TC4 alloy by Bacillus safensis, Pseudoalteromonas nigrifaciens and Chlorella marina.},
journal = {Bioelectrochemistry (Amsterdam, Netherlands)},
volume = {172},
number = {},
pages = {109353},
doi = {10.1016/j.bioelechem.2026.109353},
pmid = {42235129},
issn = {1878-562X},
abstract = {TC4 (Ti-6Al-4 V) titanium alloy resists seawater corrosion by forming a TiO2 passive film, yet its integrity is strongly affected by biofilm-induced interfacial heterogeneity. This work examined passive film modification under five conditions: sterile artificial seawater, Bacillus safensis, Pseudoalteromonas nigrifaciens, Chlorella marina, and a bacterial-algal symbiotic system. Electrochemical impedance spectroscopy, polarization curves, Mott-Schottky analysis, X-ray photoelectron spectroscopy, and cell quantification were combined to correlate biofilm features with semiconductor defects and corrosion performance. Bacterial and mixed biofilms developed extracellular polymeric substance (EPS) barriers that stabilized n-type TiO2 with oxygen-vacancy defects, raising electron escape and improving protection. P. nigrifaciens produced a compact, viscous EPS layer that yielded the best corrosion resistance, while B. safensis showed dynamic evolution from early protection to mid-stage defect increase and late partial self-repair. In contrast, the porous algal film of C. marina generated p-type defects, facilitating Cl[-] ingress and poorer resistance. The symbiotic system balanced these effects through concurrent O2 generation and EPS shielding. The overall corrosion-resistance order was P. nigrifaciens > ASW ≈ B. safensis > Symbiotic > C. marina. These findings reveal how biofilm structural properties mediates passive film semiconductor properties and suggest an EPS-based interfacial design to improve the durability of marine titanium components.},
}

@article {pmid42237056,
year = {2026},
author = {Xing, Y and He, W and Chen, W and Gao, C and Zhang, M and Wu, Y and Qu, C and Dai, K and Huang, Q and Cai, P},
title = {Cadmium Stress Favours Biofilm Cooperation and Polysaccharide-Enriched Matrix Remodelling in Bacterial Consortia.},
journal = {Environmental microbiology},
volume = {28},
number = {6},
pages = {e70347},
doi = {10.1111/1462-2920.70347},
pmid = {42237056},
issn = {1462-2920},
support = {42225706//National Natural Science Foundation of China/ ; 42407174//National Natural Science Foundation of China/ ; 460324005//Natural Science Foundation of Hubei Province of China/ ; BX20230139//China National Postdoctoral Program for Innovative Talents/ ; 202405AF140079//Science and Technology Talents and Platform Program (Workstation for Academicians and Experts)/ ; 2023AFA009//Innovative Research Group Project of Hubei Provincial Natural Science Foundation/ ; },
abstract = {Environmental stressors trigger complex adaptations in microbial communities, yet the associations between social network dynamics and metabolic strategies remain poorly understood, limiting our ability to design robust synthetic microbiomes. Here, we show that Cd stress was associated with structural simplification of soil bacterial co-occurrence networks, characterized by a 48.5% reduction in connectivity. In vitro assays further revealed a 93% increase in cooperative interactions within biofilms under Cd exposure. This shift in biofilm-associated cooperation coincided with altered physiological patterns, including a 45.9% increase in EPS synthesis and a 13.7% decrease in community CO2 emissions. Crucially, our analysis suggests that the intensity of cooperative interactions was associated with specific extracellular matrix allocation patterns. High-intensity cooperation was linked to a higher proportion of polysaccharide-rich matrices, which improved the stability and efficiency of Cd sequestration in vitro. Leveraging these interaction-informed patterns, we assembled a synthetic core consortium that modulated the rhizosphere microbiome in a proof-of-concept hydroponic system and reduced Cd accumulation in rice leaves by 52.9%. These findings support a conceptual framework in which biofilms may act as sites of social integration and altered extracellular matrix production, offering an interaction-informed basis for assembling stress-resilient microbiomes.},
}

@article {pmid42237335,
year = {2026},
author = {Khalefa, HS and Ali, AM and Farag, HS and Kamel, MS and Mosleh, AAE and Bakry, NM},
title = {Phenotypic and genotypic characterization of biofilm-forming Escherichia coli from subclinical bovine mastitis and association with antimicrobial resistance.},
journal = {BMC veterinary research},
volume = {},
number = {},
pages = {},
doi = {10.1186/s12917-026-05598-2},
pmid = {42237335},
issn = {1746-6148},
abstract = {BACKGROUND: Bovine mastitis is one of the most significant and costly diseases in the dairy industry, and Escherichia coli (E. coli) is a common etiological agent of subclinical mastitis. This study aimed to phenotypically and genotypically characterize biofilm-forming E. coli isolated from subclinical bovine mastitis and to investigate their association with antimicrobial resistance. A total of 254 composite milk samples were collected from cows with subclinical mastitis in five dairy farms in Egypt.

RESULTS: The overall prevalence of E. coli was 26.7% (68/254), with the highest isolation rates in Giza (37.1%) and Cairo-Alexandria (30.2%) regions. Molecular screening of diarrheagenic E. coli (DEC) virulence genes revealed the presence of st (10.3%), stx2 (5.9%), and eae (4.4%), whereas all isolates were negative for stx1. All isolates harbored the biofilm-associated luxS gene, and 80.9% of isolates also carried fimH. Based on virulence gene profiles, 7.4%, 4.4%, 2.9%, and 2.9% of isolates were classified as ETEC, EPEC, STEC, and hybrid ETEC-STEC (ETST), respectively, while 82.4% were DEC-negative. Phylogrouping showed a predominance of group B1 (51.9%), followed by group A (35.3%). Antimicrobial susceptibility testing revealed high resistance rates to amoxicillin/clavulanic acid (60.3%) and cefuroxime (58.8%), whereas all isolates were susceptible to cefepime, and most were susceptible to enrofloxacin (95.6%) and nalidixic acid (94.1%). Multidrug resistance was detected in 52.9% of isolates, mainly involving resistance to amoxicillin/clavulanic acid, ampicillin, tetracycline, and sulpha/trimethoprim. Biofilm formation assessed using the microtiter plate assay showed that 44.1% of isolates were moderate and 38.2% were strong biofilm producers, with significant variation between farms (p < 0.0001). Hierarchical clustering and chi-square analyses indicated that biofilm strength was associated with resistance to selected beta-lactams (amoxicillin/clavulanic acid, ampicillin, and cefuroxime), phylogenetic background (especially groups A and B1), and farm origin, whereas MDR status and most virulence genes showed weak or no association.

CONCLUSION: These findings highlight the important contribution of environmental and commensal E. coli lineages to subclinical mastitis in Egypt and underscore the need for improved farm hygiene and prudent antimicrobial use to limit the spread of biofilm-forming and antimicrobial-resistant E. coli in dairy herds.},
}

@article {pmid42237920,
year = {2026},
author = {Vreugde, S and Ambachew, S and Psaltis, AJ and Wormald, PJ},
title = {Reply to Stratified Anti-Biofilm Therapy: Translating Staphylococcal Exoprotein Toxicity Hierarchies Into Clinical Practice.},
journal = {International forum of allergy & rhinology},
volume = {},
number = {},
pages = {},
doi = {10.1002/alr.70199},
pmid = {42237920},
issn = {2042-6984},
}

@article {pmid42237937,
year = {2026},
author = {Li, P and Kang, T},
title = {Stratified Anti-Biofilm Therapy: Translating Staphylococcal Exoprotein Toxicity Hierarchies Into Clinical Practice.},
journal = {International forum of allergy & rhinology},
volume = {},
number = {},
pages = {},
doi = {10.1002/alr.70198},
pmid = {42237937},
issn = {2042-6984},
}

@article {pmid42238662,
year = {2022},
author = {Cieplik, F and Aparicio, C and Kreth, J and Schmalz, G},
title = {Development of standard protocols for biofilm-biomaterial interface testing.},
journal = {JADA foundational science},
volume = {1},
number = {},
pages = {100008},
pmid = {42238662},
issn = {2772-414X},
abstract = {The oral biofilm is associated with the most common oral diseases such as caries, periodontitis, and peri-implantitis. It is also linked to failures of dental treatment approaches (eg, direct or indirect restorations because of adjacent caries). Therefore, the development of materials with antibacterial properties is desirable. However, the design of meaningful tests to confirm such properties faces severe problems because of the complexity of the interaction of materials with the oral biofilm. Furthermore, owing to practical reasons, such tests need to be performed in vitro. In contrast, there is a need for predictive data that are comparable between different laboratories. Therefore, standardization of such tests has been advocated. The International Organization for Standardization (ISO) with its Technical Committee 106-Dentistry may be the relevant platform for this purpose. A standard (ISO 3990) is being developed for testing the antibacterial properties of dental restorative materials. This standard defines basic requirements for sample preparation, selection of bacterial strains, test methods and assessment, and reporting of results. It is considered to be the first step, and regular revisions are planned as new scientific evidence emerges. The support of the scientific communities providing multidisciplinary input is needed.},
}

@article {pmid42238883,
year = {2026},
author = {Pizarro, J and López, L and Díaz, M and Gansbiller, M and Álvarez, SA and Schmid, J and Vera, M},
title = {Sugar supplementation enhances biofilm formation and extracellular polysaccharides production in Sulfobacillus acidophilus.},
journal = {Frontiers in microbiology},
volume = {17},
number = {},
pages = {1838761},
pmid = {42238883},
issn = {1664-302X},
abstract = {Sulfobacillus is an acidophilic bacterial genus that plays a key role in bioleaching of metal sulfides at moderately thermophilic temperatures (45-55 °C), owing to its mixotrophic metabolism and capacity to form biofilms. Biofilm development depends on an extracellular polymeric substances (EPS) matrix, which is largely composed of polysaccharides. However, the composition and function of polysaccharides in sulfobacilli remain poorly characterized. A genome-wide survey identified a diverse repertoire of putative sugar uptake transporters, belonging to three distinct families. We demonstrate that supplementation with glucose, fructose, xylose or sucrose increases biofilm formation and extracellular polysaccharides production in Sulfobacillus acidophilus DSM 10332ᵀ. Enhanced biofilm formation was quantified using large-scale, in-depth image analysis. Fluorescent lectin binding analysis revealed a concomitant increase in extracellular polysaccharides abundance, while HPLC-UV-ESI-MS confirmed a quantitative rise in secreted polysaccharides under sugar-supplemented conditions. Detailed compositional analysis showed that the extracellular polysaccharides produced by S. acidophilus [T] are predominantly composed of glucose and mannose residues. Overall, our findings indicate that sugar supplementation stimulates biofilm development in S. acidophilus, likely by enhancing extracellular polysaccharides biosynthesis. This study provides the first detailed compositional characterization of S. acidophilus [T] extracellular polysaccharides and establishes Sulfobacillus as a promising model for investigating sugar uptake and carbon cycling in bioleaching systems, as members of this genus can also grow without the supplementation of sugars.},
}

@article {pmid42238891,
year = {2026},
author = {De Gregorio, E and Dé, E and Pompilio, A and Khan, AU and Zarrilli, R},
title = {Editorial: Breaking the biofilm barrier: analysis of molecular mechanisms underlying biofilm formation and identification of novel antimicrobial approaches.},
journal = {Frontiers in microbiology},
volume = {17},
number = {},
pages = {1856687},
doi = {10.3389/fmicb.2026.1856687},
pmid = {42238891},
issn = {1664-302X},
}

@article {pmid42238977,
year = {2023},
author = {Head, D and Marsh, PD and Devine, D and Tenuta, LMA},
title = {In silico study of hyposalivation and sugar exposure on biofilm dysbiosis.},
journal = {JADA foundational science},
volume = {2},
number = {},
pages = {100019},
pmid = {42238977},
issn = {2772-414X},
abstract = {BACKGROUND: Dental caries develops under actively sugar-fermenting dental biofilms, but the most successful control methods available only target mineral loss. Reduced salivary flow rates (hyposalivation) significantly exacerbate caries progression by lessening sugar and acid clearance near tooth surfaces. Maintaining dental biofilm symbiosis (health) under hyposalivation requires knowledge of the impact of acid inhibition under given dietary regimens.

METHODS: An individual-based mathematical model was used to predict biofilm dysbiosis under normal or hyposalivatory conditions by regulating the frequency of sugar intake and inhibiting microbial glycolysis, reducing the acid challenge to the tooth mineral. The impact of pH-dependent (stronger inhibition at lower pH [eg, fluoride]) and pH-independent (general percentage reduction in acid production) strategies on pH near the tooth surface during sugar intake, and the corresponding compositional changes in the biofilm, were quantified.

RESULTS: Under normal saliva flow, reducing the frequency of sugar intake and increasing the inhibition of acid production by pH-dependent or pH-independent strategies could prevent bacterial dysbiosis and prevent the biofilm from having a caries-associated (dysbiotic) to a health-associated (symbiotic) composition. However, under hyposalivatory conditions, dysbiosis occurred beyond 2 sugar intakes per day, and the degree of inhibition of glycolysis required to prevent dysbiosis was not feasible with available therapeutics.

CONCLUSIONS: Model data predict that to counteract the drastic effect of hyposalivation on biofilm dysbiosis, it will be essential to significantly reduce the frequency of fermentable sugar intake and any direct inhibition of bacterial metabolism.},
}

@article {pmid42239032,
year = {2026},
author = {Martins, GF and Guardiola-Flores, KA and Zaman, L and Horowitz, JM and Hallinen, KM and Wood, K},
title = {Cooperative antibiotic response in coupled biofilm and planktonic E. faecalis communities.},
journal = {bioRxiv : the preprint server for biology},
volume = {},
number = {},
pages = {},
doi = {10.64898/2026.05.18.725849},
pmid = {42239032},
issn = {2692-8205},
abstract = {Bacterial communities grow as dynamic populations that respond to their environments. A clinically relevant example is the inactivation of beta-lactam antibiotics by intracellular beta-lactamase in E. faecalis resistant strains. In these populations, resistant bacteria act as antibiotic sinks, detoxifying the environment and allowing sensitive bacteria to survive treatment through a cooperative interaction. In this work, we study strongly coupled planktonic and biofilm populations of mixed sensitive-resistant E. faecalis bacteria under antibiotic stress using fluorescent microscopy. The presence of resistant bacteria in the system benefits both resistant and sensitive cells, leading to mixed planktonic and biofilm populations at super-inhibitory drug concentrations. We show that a beta-lactam antibiotic with or without the addition of a beta-lactam inhibitor can lead to a population inversion effect, characterized by a non-monotonic relation between initial and final fractions of resistant bacteria. The effect is observed in both the planktonic and biofilm populations and is modulated by the total initial cell density. A well-mixed model with competition mediated by resource sharing and cooperation from global degradation of toxins predicts the experimentally observed behavior. These observations suggest underlying population-level mechanisms that are largely independent of biofilm spatial structure.},
}

@article {pmid42239163,
year = {2026},
author = {Mohamed, N and Lam, DM and Abdikarin, M and Mohammed-Abraham, R and Davies, DG and Cook, LC and McKenney, PT},
title = {Biofilm dispersion in Enterococcus faecalis is mediated by nutrient step-change and intra-species signaling.},
journal = {bioRxiv : the preprint server for biology},
volume = {},
number = {},
pages = {},
doi = {10.64898/2026.05.20.724677},
pmid = {42239163},
issn = {2692-8205},
abstract = {UNLABELLED: Enterococcus faecalis is a Gram-positive intestinal commensal and opportunistic pathogen capable of causing serious infections, including urinary tract infections, endocarditis, and wound infections. A major contributor to its persistence during infection is the ability to form biofilms on host tissues and medical devices. Biofilm cells have higher phenotypic tolerance to antimicrobial treatment than planktonic bacteria. While mechanisms governing biofilm assembly in E. faecalis have been widely studied, the processes that regulate biofilm dispersion, the final stage of the biofilm life cycle, remain poorly understood. In this study, we found that dispersion is triggered by a tenfold step-change increase in nutrient availability and by cell free supernatant (CFS) of E. faecalis OG1RF cultures. Cells released from biofilms regain sensitivity to antibiotics similar to planktonic cells but maintain a high potential for adherence. We characterized the glycosyltransferase epaOX , which contributes to the structure of the enterococcal polysaccharide antigen as necessary for nutrient step-change induced dispersion, CFS induced dispersion, and adhesion of dispersed cells. Supplementation of epaOX mutant CFS with galactose and N-acetylgalactosamine was sufficient to restore CFS induced dispersion. Together these data suggest that dispersion in OG1RF occurs with fast kinetics, affects antibiotic sensitivity and is regulated in part by known virulence factors.

IMPORTANCE: E. faecalis causes difficult to treat infections at numerous body sites in human patients. E. faecalis biofilms are adherent populations that require high levels of antibiotics for treatment. Biofilms undergo a disassembly process named dispersion that allows individual cells to leave the biofilm and colonize new locations. Dispersed cells in other species are killed by lower amounts of antibiotics than biofilm cells. Here we showed that dispersion occurs in E. faecalis and lowers the level of antibiotics needed to kill dispersed cells. Dispersion triggers could be used in the future to design treatments that increase the effectiveness of antibiotics.},
}

@article {pmid42239287,
year = {2026},
author = {Elpers, L and Schmeisser, B and Felgner, P and Koettermann, M and Drauch, V and Hess, C and Koepp, N and Lueken, L and Hess, M and Gal-Mor, O and Hensel, M},
title = {Novel pESI-encoded autotransporter adhesin PeaP of epidemic Salmonella strains mediates adhesion, atypical biofilm formation, and poultry colonization.},
journal = {bioRxiv : the preprint server for biology},
volume = {},
number = {},
pages = {},
doi = {10.64898/2026.05.20.725250},
pmid = {42239287},
issn = {2692-8205},
abstract = {Salmonella enterica serovar Infantis (SIN) has rapidly become the dominant serovar in poultry worldwide, a success largely linked to the acquisition of the 285 kb megaplasmid pESI. While pESI-encoded antibiotic-resistance and iron-uptake systems are well characterized, pESI-mediated adhesion mechanisms remain poorly understood. Here we identify a novel pESI-encoded monomeric autotransporter adhesin, designated PeaP (pESI-encoded autotransporter protein), and demonstrate its pivotal role in atypical biofilm formation, interference with motility, and colonization of the chicken host. Biofilm assays revealed that pESI-harboring strain SIN 119944 forms robust biofilms at 37 °C and 42 °C, temperatures at which CsgD-dependent biofilm formation is negligible. Deletion of csgD did not impair this phenotype, whereas deletion of peaP abolished high-temperature biofilm development and restored motility to wild-type levels. Proteomic profiling of sessile versus planktonic cells highlighted PeaP as the most abundant pESI-derived protein in the biofilm fraction. AlphaFold-based modelling and negative-stain transmission electron microscopy showed that PeaP comprises a C-terminal β-barrel and a 1,500 aa passenger domain with three tandem repeats, projecting filamentous appendages ~37 nm from the outer membrane. Antibody blockade of PeaP reduced surface adhesion >6-fold, confirming its adhesive function. In an infection model of 2 day-old chicken, the peaP mutant displayed significantly lower colonization, indicating PeaP-mediated adhesion in vivo. Collectively, pESI-positive SIN deploys PeaP for CsgD-independent, temperature-tolerant biofilm formation and enhanced gastrointestinal colonization, providing a mechanistic basis for the epidemic spread of this multidrug-resistant pathogen in poultry.},
}

@article {pmid42239704,
year = {2024},
author = {Elsharkasi, MM and Eckert, GJ and Gregory, RL},
title = {Effect of silver diamine fluoride on nicotine-induced Streptococcus mutans biofilm.},
journal = {JADA foundational science},
volume = {3},
number = {},
pages = {100039},
pmid = {42239704},
issn = {2772-414X},
abstract = {BACKGROUND: Silver diamine fluoride (SDF) reagent is used to prevent and arrest caries lesions. However, the mechanism of its action is not fully reported. The antimicrobial effect of SDF was determined on an established nicotine-induced Streptococcus mutans biofilm by measuring colony-forming units (CFUs), different application times, and extracellular polysaccharide (EPS) synthesis.

METHODS: S mutans biofilm was established with and without 2 mg/mL of nicotine in tryptic soy broth supplemented with sucrose. Nicotine and nonnicotine-induced biofilm groups were treated with 38% SDF for 1 minute. Non-SDF-treated groups were used as a control. The biofilm was plated on blood agar plates, and CFUs were determined. In addition, different SDF application times (30 seconds and 1, 2, and 3 minutes) were tested. For EPS production, S mutans were incubated with and without nicotine in tryptic soy broth supplemented with sucrose. SDF groups were treated with SDF for 1 minute. A phenol-sulfuric acid assay was used to measure the total carbohydrate produced.

RESULTS: There was a significant effect of SDF on reducing CFUs for both nicotine and nonnicotine groups. The different application times of SDF reduced CFUs for all tested groups. EPS production was significantly reduced with the SDF application.

CONCLUSIONS: The use of SDF with different application times disrupted established S mutans biofilms. In addition, EPS production was reduced by the application of SDF. This confirms the ability of a shorter SDF treatment time to be effective and suggests a possible mechanism of action for the inhibition of caries development.},
}

@article {pmid42229743,
year = {2026},
author = {Saini, Y and Wani, MY and Hameed, S},
title = {Antifungal Peptides for Biofilm Disruption: Mechanisms, Design Strategies, and Translational Outlook.},
journal = {Microbial pathogenesis},
volume = {},
number = {},
pages = {108601},
doi = {10.1016/j.micpath.2026.108601},
pmid = {42229743},
issn = {1096-1208},
abstract = {Invasive fungal infections in intensive care units are a serious concern, especially when they are associated with biofilm formation. These infections often lead to high mortality because biofilms make the fungi more resistant to antifungal drugs and harder for the immune system to clear. Pathogens such as Candida auris, Candida albicans, and Aspergillus fumigatus are particularly problematic, as they are known to develop multidrug resistance and cause persistent infections in critically ill patients. These biofilms often show a much higher tolerance to standard antifungal drugs and can escape the host body's immune defenses. This makes the infections they cause more persistent and very difficult to treat in clinical practice. Antifungal peptides (AFPs), whether derived from natural host-defense molecules or designed through rational engineering, are emerging as promising options for tackling fungal biofilms. They act through several mechanisms, such as disrupting the fungal cell membrane, blocking early adhesion and morphogenesis, and weakening the extracellular matrix. Importantly, they may also work in synergy with existing antifungal drugs, making treatment more effective. Recent progress in peptide engineering and delivery methods, such as nanocarriers and hydrogel-based systems has enhanced the stability, selectivity, and ability of peptides to target fungal biofilms in experimental models. At the same time, there are important challenges that remain, including their tendency to break down due to proteolytic enzymes, possible cytotoxic effects, difficulties in large scale manufacturing, and regulatory hurdles linked to peptide-based therapies. Overall, AFPs represent a promising and fast developing area of research, but their use in clinical practice is limited. More studies are needed to confirm their safety, effectiveness and practical feasibility for managing biofilm-associated fungal infections.},
}

@article {pmid42229762,
year = {2026},
author = {de Maceda, FG and Lopes-Olhê, FC and Torres-Carrillo, AJS and da Silva Goulart, R and Petean, IBF and de Castro-Vasconcelos, GA and Silva-Sousa, AC and Estrela, C and Mazzi-Chaves, JF and Silva-Sousa, YTC and Pitondo-Silva, A and Sousa-Neto, MD},
title = {Effect of ultrasonic irrigant activation and negative apical pressure in removing mature biofilm from the isthmus of curved root canal: A 3D-printed model study.},
journal = {Journal of microbiological methods},
volume = {},
number = {},
pages = {107567},
doi = {10.1016/j.mimet.2026.107567},
pmid = {42229762},
issn = {1872-8359},
abstract = {INTRODUCTION: Although the iVac® system has shown promising cleaning performance, its effectiveness in biofilm removal from anatomically complex areas remains untested in standardized 3D models, highlighting the relevance of the present study to evidence-based irrigation strategies.

METHODS: A standardized 3D mandibular molar model with curvature and an isthmus was created, incorporating dentin blocks and a 21-day E. faecalis biofilm. Samples (n = 48) were randomized into four groups: syringe-needle irrigation, iVac System, ultrasonic irrigant activation (UIA), and control. All groups received NaOCl, EDTA, PBS, and sodium thiosulfate. Biofilm removal was assessed by CFU counting and SEM.

RESULTS: ANOVA of CFUs showed that UIA and iVac significantly reduced bacterial counts compared to the control (p < 0.001), with no difference between them (p = 0.342). Syringe and needle irrigation showed results similar to the control (p = 0.124) and iVac (p = 0.087), but had higher CFUs than UIA (p = 0.017). These reductions corresponded to substantial decreases in bacterial load. SEM revealed that iVac and UIA groups exhibited exposed dentinal tubules with few scattered bacteria, while syringe and needle irrigation showed a dense amorphous matrix with E. faecalis-like cells covering most of the dentin, with limited exposed areas.

CONCLUSIONS: Active irrigant agitation with UIA or iVac was more effective in reducing mature biofilm and promoting its disruption in the isthmus of curved root canals than syringe and needle irrigation.},
}

@article {pmid42230424,
year = {2026},
author = {Passamani, GB and de Oliveira Pereira, L and Ferreira, TA and da Silva, PV and Backes, L and de Freitas Santos, J and Sardi, JO and de Almeida, TDD and Campos, LT and Borba, AM and Romário-Silva, D},
title = {Association of oral and non-oral microorganisms in a multispecies biofilm model formed on orthodontic appliances: an in vitro study.},
journal = {Odontology},
volume = {},
number = {},
pages = {},
pmid = {42230424},
issn = {1618-1255},
abstract = {Orthodontic devices can promote dental biofilm formation and increase the risk of oral diseases when hygiene is inadequate. Invisible aligners, although removable and easier to clean, are frequently exposed to external environments, which may allow contamination by non-oral microorganisms. This study aimed to evaluate the association and viability of oral and non-oral microorganisms in mono- and multispecies biofilm models formed on orthodontic appliances. Methods: Stimulated human saliva was used to form the acquired pellicle on specimens of invisible aligners, conventional brackets, and self-ligating brackets. The samples were exposed to Streptococcus mutans, Candida albicans, Staphylococcus aureus, and Pseudomonas aeruginosa, and incubated for 48 h under mono- and multispecies conditions. After ultrasonic detachment, viable cells were quantified by CFU/mL enumeration, and extracellular polymeric substance (EPS) content was determined. Statistical analysis was performed using one-way ANOVA followed by Tukey's post hoc test for monospecies biofilms, and two-way ANOVA followed by Bonferroni's post hoc test for multispecies biofilms (p < 0.05). Higher CFU/mL values were observed for S. aureus and P. aeruginosa compared to C. albicans and S. mutans (p < 0.05). In multispecies biofilms, C. albicans showed higher values than S. mutans (p < 0.05). CFU/mL increased over time, with no differences among orthodontic devices (p > 0.05). EPS content was higher in conventional brackets, intermediate in self-ligating brackets, and lower in aligners (p < 0.05). Oral and non-oral microorganisms remained viable and were recovered from biofilm models formed on orthodontic appliances. These findings suggest that such devices may act as substrates for the persistence of non-oral microorganisms under biofilm-forming conditions, highlighting the importance of appropriate hygiene and storage practices.},
}

@article {pmid42230493,
year = {2026},
author = {Minko, AG and Danilova, TA and Lunin, VG and Gromova, MS and Danilina, GA and Adzhieva, AA and Zhukhovitsky, VG},
title = {The Effect of Streptococcal Pyrogenic Exotoxin Type A (SpeA) on the Formation of Streptococcus pyogenes Biofilm.},
journal = {Bulletin of experimental biology and medicine},
volume = {},
number = {},
pages = {},
pmid = {42230493},
issn = {1573-8221},
abstract = {The effect of recombinant streptococcal (Streptococcus pyogenes) exotoxin type A (rSpeA) on biofilm formation by S. pyogenes was studied using a laboratory strain of serotype M30 and five non-typeable clinical strains of S. pyogenes. The inhibition of bacterial growth was observed during both the initial formation stage and within established biofilms. It was shown that under the action of rSpeA on biofilm formation, the growth of strain M30 was inhibited by 1.6 times and the growth of strains 1, 2, and 3 was inhibited by 2.3, 1.3, and 1.4 times, respectively. For strains 4 and 5, the inhibition was practically absent. Application of SpeA to the formed biofilm of streptococcal strains produced a more pronounced inhibitory effect: 2.5-fold for strain M30, and 5.5-, 3.4-, and 4.2-fold for strains No. 2, 3, and 5, respectively. These findings suggest that SpeA being a superantigen can also destroy streptococcal biofilms promoting the spread of bacteria in the body and leading to generalization of the infectious process.},
}

@article {pmid42231706,
year = {2026},
author = {Wang, S and Rong, L and Chen, Y and He, W and Wen, X and Deng, Y and Lai, S},
title = {Deciphering Augmented Dual-ROS-Driven Biofilm Eradication by Facilitating Long-Range Spatial Charge Decoupling in Polymer Carbon Dots.},
journal = {Angewandte Chemie (International ed. in English)},
volume = {},
number = {},
pages = {e2382848},
doi = {10.1002/anie.2382848},
pmid = {42231706},
issn = {1521-3773},
support = {22305098//National Natural Science Foundation of China/ ; 32571536//National Natural Science Foundation of China/ ; 32271392//National Natural Science Foundation of China/ ; 2026NSFSC0995//Natural Science Foundation of Sichuan/ ; 2026YFHZ0080//Natural Science Foundation of Sichuan/ ; },
abstract = {Developing carbon dots (CDs) with reactive oxygen species (ROS) production capability provides an attractive approach to address the dilemma of biofilm eradication caused by the robust extracellular polymeric substance (EPS) matrix. The challenge for the exploration of highly potent CDs is to circumvent the severe thermodynamic and kinetic paradox to transform surrounding substrates into ultra-reactive ROS. To address this conundrum, we propose a long-pathway electron-accepting strategy promoting the absolute spatial charge decoupling by the elaborate marriage of carbonized core and polynaphthalenediimide (PNDI) network, which significantly boosts the superoxide anion (·O2 [-]) and hydroxyl radical (·OH) dual-ROS generation of the constructed polymer CDs. Systematic mechanism exploration reveals that ultrafast intramolecular charge transfer after photoirradiation enables energetic long-life electrons to migrate along the PNDI highway for abundant ·O2 [-] production. Intriguingly, this profound separation firmly anchors uncompensated highly oxidative holes at the extraordinarily deep highest occupied molecular orbital level of the carbon core, successfully unlocking the thermodynamic threshold for direct ·OH generation. This tailored dual-ROS storm induces catastrophic EPS matrix degradation and massacres the embedded pathogens, achieving near-complete (∼99.9%) eradication of Escherichia coli and Staphylococcus aureus biofilms. This work establishes a potent nanoplatform and provides profound mechanistic insights for tackling global biofilm-associated threats.},
}

@article {pmid42232210,
year = {2026},
author = {Udawatte, NS and Liu, C and Staples, R and Han, P and Kumar, PS and Arumugam, TV and Ivanovski, S and Seneviratne, CJ},
title = {Transient restructuring of the active oral resistome during probiotic Streptococcus salivarius K12 colonization in a 3D polymicrobial biofilm model.},
journal = {Journal of oral microbiology},
volume = {18},
number = {1},
pages = {2680793},
pmid = {42232210},
issn = {2000-2297},
abstract = {BACKGROUND: The oral cavity harbours a complex and transcriptionally active antibiotic resistance gene (ARG) reservoir shaped by polymicrobial biofilm ecology. Whether probiotic-mediated ecological modulation can remodel the active resistome without promoting horizontal gene transfer remains poorly understood.

OBJECTIVE: To investigate the impact of Streptococcus salivarius K12 (Ssk12) colonisation on active resistome dynamics within saliva derived polymicrobial biofilms and determine whether probiotic driven ecological restructuring transiently alters resistance-associated transcriptional signatures.

DESIGN: Saliva-derived polymicrobial biofilms were established on three-dimensional melt electrowritten poly(ε-caprolactone) (MEW-mPCL) scaffolds and exposed to Ssk12. Metatranscriptomic profiling was performed across four time points (Baseline, Day 4, Day 7, and Day 10), complemented by quantitative PCR validation and ARG-mobile genetic element (MGE) co-localisation analysis to characterise resistome restructuring during probiotic colonisation and decolonisation.

RESULTS: Baseline biofilms contained 27 ARGs spanning 16 antibiotic classes, predominantly ermB, tet(M), and tet(W). During peak Ssk12 colonisation (Days 4-7), total ARG abundance declined to approximately 17% of baseline levels, with marked reductions in efflux-associated and β-lactam/fluoroquinolone resistance-associated transcripts. Partial resistome recovery occurred by Day 10 (~32% of baseline), indicating reversible ecological modulation rather than permanent dysbiotic restructuring. ARG dynamics were primarily reshaped by ARG-bearing taxa rather than enrichment of high-confidence putatively mobile resistance determinants.

CONCLUSIONS: S. salivarius K12 transiently remodelled the transcriptionally active oral resistome within structured polymicrobial biofilms without evidence of enhanced putative horizontal resistance gene mobilisation. These findings support a proof-of-concept model in which probiotic driven ecological restructuring may create a transient resistome state potentially associated with altered responsiveness to selected antibiotic classes.},
}

@article {pmid42233349,
year = {2026},
author = {Yu, Y and Bauer, RM and Mahdi, R and Piepenbrink, KH},
title = {Insight into the relationship between type IV pilus function and biofilm formation.},
journal = {Biochemical Society transactions},
volume = {54},
number = {6},
pages = {633-650},
doi = {10.1042/BST20250151},
pmid = {42233349},
issn = {1470-8752},
support = {P20-GM113126//HHS | NIH | National Institute of General Medical Sciences (NIGMS)/ ; MCB-2310647//National Science Foundation (NSF)/ ; },
abstract = {Type IV pili (T4P) are protein nanofibers that can be extended and retracted from the surfaces of many bacterial taxa. They are involved in many aspects of bacterial physiology that differ between bacterial species, including surface motility, DNA uptake, and host-cell adherence, but genetically and structurally distinct type IV pilus systems from distantly related bacterial species have also been found to promote the formation of bacterial biofilms. The molecular mechanisms underpinning the promotion of biofilm remain an area of active investigation and may be both manifold and variable between type IV pilus systems. Two areas of recent interest are interactions between T4P and extracellular DNA and the relationship between surface-adhered biofilms and suspended aggregates. In the present review, we critically discuss the current state of knowledge of type IV pilus function and how these structures may interact with other biomolecules to influence the formation of multicellular bacterial communities. We examine the evidence for how alterations in DNA-binding, pilus retraction, and pilus composition have downstream effects on the formation of bacterial biofilms.},
}

@article {pmid42233673,
year = {2026},
author = {Fulman-Levy, H and Sinberger, LA and Geva, P and Lellouche, J and Navon-Venezia, S},
title = {Klebsiella pneumoniae biofilm formation predicts its survival in human serum.},
journal = {mBio},
volume = {},
number = {},
pages = {e0075226},
doi = {10.1128/mbio.00752-26},
pmid = {42233673},
issn = {2150-7511},
abstract = {Klebsiella pneumoniae is a prominent pathogen causing life-threatening bloodstream infections. Although biofilm formation and resistance to human serum are well-recognized virulence traits, their interrelatedness during K. pneumoniae bloodstream infections remains unclear. Here, we hypothesize that biofilm production is related to K. pneumoniae's ability to thrive in human serum and, therefore, may predict the strains' ability for serum survival. We analyzed 57 clinical, genetically diverse classical K. pneumoniae strains and characterized their survival and biofilm-producing ability in human serum. Serum survival patterns revealed three serum resistance categories-Low, Mid, and High. In addition, the biofilm biomass produced by the strains correlated with their serum resistance level (P < 0.001), and 3D biofilm visualization using confocal microscopy further confirmed that biofilm extracellular polysaccharide substances and biomass patterns were consistent with the serum resistance categories. Moreover, we revealed a direct correlation between the level of biofilm formation and the strain's serum survival level (R[2] = 0.696), a prerequisite for systemic K. pneumoniae dissemination. As biofilm formation in serum reflects both survival and biofilm-forming ability, we assessed biofilm formation in defined modified basal medium (BM2), to rule out serum-mediated killing, and discovered a strong and significant association between the serum resistance category and BM2 biofilm biomass (P < 0.0001). By applying regression models, we discovered that biofilm formation serves as a significant predictor for bacterial survival in serum. Overall, our findings establish biofilm production in K. pneumoniae as a biomarker of serum survival and may open a new avenue for predicting bloodstream infection risk in clinical settings.IMPORTANCEBloodstream infections caused by Klebsiella pneumoniae are devastating life-threatening infections worldwide. Understanding the survival strategies of K. pneumoniae in the bloodstream is critical for elucidating key aspects of bacterial pathogenicity and developing new diagnostic and therapeutic modalities. Although serum survival is a recognized virulence trait necessary to thrive in the bloodstream, the relationship between serum resistance and biofilm formation, a multicellular organization that may protect bacteria from bloodstream stressors, remains poorly understood. In this article, we demonstrate biofilm production in human serum by clinical classical K. pneumoniae strains for the first time and discovered a direct correlation between the level of biofilm biomass formation and the degree of serum survival in human serum and in defined modified basal medium. These findings offer insights into the importance of biofilm production in K. pneumoniae serum resistance and may be used to develop future therapeutic strategies targeting bloodstream infections.},
}

@article {pmid42234182,
year = {2026},
author = {Pabousi Sadatmahale, A and Nouhi Kararoudi, A and Zahmatkesh, H and Haghdoust Kooyshahi, H and Nezamivand Chegini, S and Garakoui, SR and Shahriarinour, M and Nikpassand, M and Ranji, N},
title = {Anti-biofilm and Efflux Pump Inhibitory Function of Fe3O4@SBA-3@Curcumin Nanoparticles on drug resistant isolates of Pseudomonas aeruginosa and Molecular Docking Analysis.},
journal = {Current microbiology},
volume = {83},
number = {7},
pages = {},
pmid = {42234182},
issn = {1432-0991},
abstract = {Increased biofilm formation and efflux pumps activity are two major contributors to the antimicrobial resistance in clinical isolates of Pseudomonas aeruginosa (P. aeruginosa). In the present study, curcumin-functionalized Fe3O4 nanoparticles were synthesized and evaluated for their anti-biofilm and efflux pump inhibitory effects against P. aeruginosa. The nanoparticles, designated Fe3O4@SBA-3@Curcumin, were prepared via co-precipitation method followed by surface functionalization. Comprehensive physicochemical characterization was performed using thermal analysis and multiple spectroscopic techniques. The antimicrobial efficacy of Fe3O4@SBA-3@Curcumin alone and in combination with ciprofloxacin was assessed using fractional inhibitory concentration (FIC) analysis, biofilm formation assays, and pyocyanin production measurements. FT-IR spectroscopy confirmed successful curcumin functionalization without structural degradation. FE-SEM and TEM images demonstrated nanoparticle size of 52.12 nm and 32.20 nm, respectively. Dynamic light scattering (DLS) analysis revealed a mean particle diameter of 101.8 nm and excellent colloidal stability, as indicated by a zeta potential of -86.8 mV. Combination therapy exhibited a synergistic effect and significantly reduced biofilm formation and pyocyanin production. Furthermore, combined treatment with Fe3O4@SBA-3@Curcumin and ciprofloxacin resulted in downregulation of efflux pump genes (mexA, mexB, and oprM) and biofilm-associated genes (algD and pelA). Molecular docking analyses predicted favorable binding interactions between curcumin and key biofilm-related proteins involved in exopolysaccharide synthesis (algD and pelD), as well as efflux pump components associated with antibiotic resistance (mexB, mexA, and oprM) in P. aeruginosa. Collectively, these findings support the potential role of curcumin functionalization in attenuating biofilm formation and efflux pump activity. Overall, Fe3O4@SBA-3 nanoparticle may serve as an effective nanocarrier for targeted drug delivery into bacterial cells.},
}

@article {pmid42234242,
year = {2026},
author = {Temel, A and Ateş, A and Aksoyalp, ZŞ},
title = {Effects of empagliflozin and metformin on biofilm formation and pathogenicity factors of urinary Escherichia coli isolates.},
journal = {Folia microbiologica},
volume = {},
number = {},
pages = {},
pmid = {42234242},
issn = {1874-9356},
abstract = {Escherichia coli, a major cause of urinary tract infections (UTIs), forms biofilms that contribute to antimicrobial resistance. Antidiabetic medications have gained attention for their potential antimicrobial effects, though data remain limited. This study investigated the inhibitory effects of empagliflozin and metformin against urinary E. coli isolates. Minimum inhibitory concentrations (MICs) were determined via broth microdilution, and synergistic interactions were assessed using the checkerboard method. Biofilm inhibition at sub-inhibitory drug concentrations was evaluated spectrophotometrically, and gene expression of fimH and luxS, were analyzed using RT-qPCR. Empagliflozin and metformin inhibited bacterial growth, with MICs ranging from 3.12-6.25 mg/mL and 25-50 mg/mL, respectively. A synergistic effect was observed in two isolates. Both drugs significantly reduced biofilm formation (51.8-72.9%) and downregulated fimH and luxS gene expression (p < 0.01). This study showed that empagliflozin and metformin could have inhibitory effects against urinary E. coli isolates, supporting their potential in drug repurposing strategies. Empagliflozin and metformin demonstrated significant dose-dependent in vitro antivirulence and antibiofilm activities, further supported by the downregulation of key virulence-associated genes (fimH and luxS). To the best of our knowledge, this is the first report investigating the in vitro effects of empagliflozin against urinary E. coli isolates, and further investigation is required to determine the impact of antidiabetic medications on E. coli.},
}