@article {pmid40812662,
year = {2025},
author = {Yang, X and Shi, H and Zimba, BL and Xu, P and Wang, H and Huang, Y and Fu, Q and Zhang, X and Chen, W and Zhou, L and Xu, C and Chen, J and Wang, S},
title = {Smart bacteriophage release from bovine β-lactoglobulin fibrils based hydrogel with anti-MRSA, biofilm degradation and hair follicle regeneration functions for infected burn wound.},
journal = {International journal of biological macromolecules},
volume = {322},
number = {Pt 2},
pages = {146855},
doi = {10.1016/j.ijbiomac.2025.146855},
pmid = {40812662},
issn = {1879-0003},
abstract = {The emergence and spread of antibiotic-resistant bacteria negatively impact the effectiveness of antibiotics in treating burn wound infections, which significantly hinders the healing process. To address this, a β-lactoglobulin fibrils/oxidized dextran/phage (BLGFs/ODEX/Phage) hydrogel dressing has been developed to treat burn wounds that are infected with methicillin-resistant Staphylococcus aureus (MRSA). First, a highly efficacious MRSA phage is isolated and purified, and it exhibits excellent bactericidal efficiency, storage stability, biofilm degradation ability, and biocompatibility. Then, a pH-responsive BLGFs/ODEX hydrogel is developed to load MRSA phage by utilizing a Schiff base reaction between BLGFs and ODEX. The hydrogel possesses excellent mechanical properties, self-healing ability, cell adhesion promotion, and biocompatibility. The results of burn wound model experiments indicate that the MRSA phages encapsulated within the hydrogel can be released in a controlled manner dependent on bacterial growth, effectively mitigating the accumulation of excessive bacterial toxins. Therefore, the BLGFs/ODEX/Phage hydrogel dressing would display superior abilities in regulating inflammation, promoting collagen deposition, facilitating angiogenesis, and enhancing hair follicle regeneration, thereby accelerating wound healing. Overall, this work demonstrates that the combination of bacteriophage therapy and hydrogel dressing would be a promising and innovative strategy for the treatment of infected burn wounds, presenting expansive prospects for clinical application.},
}

@article {pmid40812437,
year = {2025},
author = {Patel, RR and Mishra, A and Singh, SK and Nath, G and Singh, M},
title = {Mechanistic evaluation of Clerodendrum serratum anti-biofilm potency against Mycobacterium species.},
journal = {Microbial pathogenesis},
volume = {208},
number = {},
pages = {107985},
doi = {10.1016/j.micpath.2025.107985},
pmid = {40812437},
issn = {1096-1208},
abstract = {Biofilm formation in Mycobacterium species significantly contributes to their pathogenicity and resistance to conventional antimicrobial therapies, posing a major challenge in clinical management. Plant-derived phytoconstituents have emerged as promising alternatives due to their diverse biological activities, including anti-biofilm properties. Clerodendrum serratum, a medicinal plant known for antimicrobial potential, offers a rich source of such bioactive compounds. This study aimed to first optimize robust biofilm formation in three Mycobacterium species (M. smegmatis, M. fortuitum, and M. marinum) using specific nutritional supplements, followed by the evaluation of the anti-biofilm efficacy of C. serratum leaf extract against pre-formed mature biofilms. Finally, potential molecular targets and mechanisms of action of key phytoconstituents were investigated through in silico analysis. Enhanced biofilm formation was achieved by supplementing Middlebrook 7H9 broth with KH2PO4, (NH4)2SO4, Acicase, and DTT, with KH2PO4 showing the most pronounced effect. Disruption of pre-formed biofilms by C. serratum leaf extract was quantified using the crystal violet microtiter plate assay and confirmed via Atomic Force Microscopy (AFM), which revealed significant alterations in biofilm architecture. In silico molecular docking of five major phytoconstituents (Hispidulin, Luteolin, Salvigenin, Pectolinarigenin, and Uncinatone) with critical biofilm-associated targets (FadD32, InhA, and MmpL3) showed strong binding affinities of docking score up to -11.0 kcal/mol, indicating potential mechanisms of biofilm disruption. This study presents the optimization of enhanced biofilm formation in Mycobacterium species and demonstrates the anti-biofilm potential of C. serratum leaf extract. The integrated experimental and computational approach offers new insights into targeting resilient mycobacterial biofilms using plant-based therapeutics.},
}

@article {pmid40811911,
year = {2025},
author = {Larsson, Y and Nikolausz, M and Kisielius, V and Møller, HB and Gosewinkel, U and Bester, K},
title = {Metabolic pathways for biotransformation of benzalkonium compounds in fungal- and bacteria-based biofilm reactors.},
journal = {Journal of hazardous materials},
volume = {496},
number = {},
pages = {139494},
doi = {10.1016/j.jhazmat.2025.139494},
pmid = {40811911},
issn = {1873-3336},
abstract = {Benzalkonium compounds (BACs) are quaternary ammonium biocides widely used in healthcare and industrial animal production, resulting in high concentrations in manure and wastewater. While BAC biodegradation has been demonstrated in wastewater and soil, little is known about the relevant biodegradation pathways in multi-species biofilm systems. This study investigated BAC degradation pathways in three aerobic moving bed biofilm reactor (MBBR) systems: (1) a fungi (Trametes versicolor) inoculated reactor, and heterotrophic biofilm in (2) water, and (3) manure. We characterised transformation products and elucidated their fragmentation mechanisms to explain the spectra. ω-oxidation and β/α-oxidation dominated in the water-MBBR, whereas formation of benzyldimethylamine (BDMA), was observed only in the manure-MBBR, which was the only system hosting Pseudomonas and Aeromonas species. The water-MBBR microbial community showed a significant increase in abundance of Rhodococcus, known for ω- and β-oxidation, suggesting a role in BAC degradation. None of the five ARGs (sul1, sul2, qnrD, tetM, tetA) tested in this study were elevated in the systems by exposure to BAC. These results demonstrate that BACs are degradable in different aerobic biofilm systems via diverse pathways, influenced by microbial composition and matrix complexity. However, the different metabolites indicate towards complex risk assessments and bioremediation strategies.},
}

@article {pmid40807491,
year = {2025},
author = {Liu, B and Bai, M and Tu, W and Shen, Y and Liu, J and Yang, Z and Bao, H and Dong, Q and Liu, Y and Wang, R and Zhang, H and Hu, L},
title = {The Virulence Factor LLO of Listeria monocytogenes Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect.},
journal = {Foods (Basel, Switzerland)},
volume = {14},
number = {15},
pages = {},
pmid = {40807491},
issn = {2304-8158},
support = {32373098//National Natural Science Foundation of China/ ; 2024YFE0198800//National Key Research and Development Program of China/ ; BZ2023004//Jiangsu Provincial Science and Technology Plan Special Fund/ ; },
abstract = {Listeria monocytogenes is a life-threatening bacterial foodborne pathogen that can persist in food-processing facilities for years. Although phages can control L. monocytogenes during food production, phage-resistant bacterial subpopulations can regrow in phage-treated environments. In this study, an L. monocytogenes hly defective strain, NJ05-Δhly, was produced, which considerably regulated the interactions between L. monocytogenes and phages. Specifically, we observed a 76.92-fold decrease in the efficiency of plating of the defective strain following infection with the Listeria phage vB-LmoM-NJ05. The lytic effect was notably diminished at multiplicities of infection of 1 and 10. Furthermore, the inactivation of LLO impaired biofilm formation, which was completely suppressed and eliminated following treatment with 10[8] PFU/mL of phage. Additionally, phages protected cells from mitochondrial membrane damage and the accumulation of mitochondrial reactive oxygen species induced by L. monocytogenes invasion. Transcriptomic analysis confirmed these findings, revealing the significant downregulation of genes associated with phage sensitivity, pathogenicity, biofilm formation, and motility in L. monocytogenes. These results underscore the vital role of LLO in regulating the pathogenicity, phage susceptibility, and biofilm formation of L. monocytogenes. These observations highlight the important role of virulence factors in phage applications and provide insights into the potential use of phages for developing biosanitizers.},
}

@article {pmid40807479,
year = {2025},
author = {Urbaniak, M and Lechowicz, Ł and Gawdzik, B and Hodorowicz, M and Wielgus, E},
title = {Structure-Activity Relationships in Alkoxylated Resorcinarenes: Synthesis, Structural Features, and Bacterial Biofilm-Modulating Properties.},
journal = {Molecules (Basel, Switzerland)},
volume = {30},
number = {15},
pages = {},
pmid = {40807479},
issn = {1420-3049},
abstract = {In this study, a series of novel alkoxylated resorcinarenes were synthesized using secondary and tertiary alcohols under mild catalytic conditions involving iminodiacetic acid. Structural characterization, including single-crystal X-ray diffraction, confirmed the successful incorporation of branched alkyl chains and highlighted the influence of substitution patterns on molecular packing. Notably, detailed mass spectrometric analysis revealed that, under specific conditions, the reaction pathway may shift toward the formation of defined oligomeric species with supramolecular characteristics-an observation that adds a new dimension to the synthetic potential of this system. To complement the chemical analysis, selected derivatives were evaluated for biological activity, focusing on bacterial growth and biofilm formation. Using four clinically relevant strains (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Bacillus subtilis), we assessed both planktonic proliferation (OD600) and biofilm biomass (crystal violet assay). Compound 2c (2-pentanol derivative) consistently promoted biofilm formation, particularly in S. aureus and B. subtilis, while having limited cytotoxic effects. In contrast, compound 2e and the DMSO control exhibited minimal impact on biofilm development. The results suggest that specific structural features of the alkoxy chains may modulate microbial responses, potentially via membrane stress or quorum sensing interference. This work highlights the dual relevance of alkoxylated resorcinarenes as both supramolecular building blocks and modulators of microbial behavior.},
}

@article {pmid40803262,
year = {2025},
author = {Frantar, A and Seme, K and Gašperšič, R and Oblak, Č and Šuster, K},
title = {Isolation and characterization of two novel oral bacteriophages with anti-biofilm activity against Cutibacterium acnes.},
journal = {International journal of medical microbiology : IJMM},
volume = {320},
number = {},
pages = {151668},
doi = {10.1016/j.ijmm.2025.151668},
pmid = {40803262},
issn = {1618-0607},
abstract = {Bacteriophage therapy offers a promising solution to combat antibiotic-resistant infections, yet its potential against biofilm-associated pathogens in oral diseases remains underexplored. This study investigates the opportunistic bacterium Cutibacterium acnes, an overlooked contributor to dental implant and prosthetic joint infections. Biofilms formed by C. acnes are highly resilient and resistant to antibiotics, complicating treatment. Two novel lytic bacteriophages, Ristretto and Corretto, targeting C. acnes, were isolated from human saliva, with morphological analysis confirming their classification as siphoviruses. Their genome sequencing revealed no harmful antimicrobial resistance or virulence genes, making them suitable for therapeutic use. Remarkably, phage Corretto demonstrated a broad host range and achieved near-complete eradication of mature biofilms across multiple C. acnes strains, outperforming Ristretto in efficacy and strain coverage. The activity of these phages was dosage-dependent and varied across bacterial strains, revealing potential strain-specific resistance mechanisms within biofilms. These findings highlight bacteriophage therapy's potential to disrupt persistent biofilms where antibiotics fail, offering a new approach for treating biofilm-driven infections in dental and medical implantology. This study underscores the need for further research into phage-based strategies to address the growing global challenge of antimicrobial resistance and improve outcomes in biofilm-related diseases.},
}

@article {pmid40803092,
year = {2025},
author = {Shen, Y and Zhang, X and Feng, X and Ma, X and Wang, S and Li, J},
title = {Unraveling tetracycline-induced biofilm stability and resistance mechanisms in aerobic granular sludge via proteomics.},
journal = {Water research},
volume = {287},
number = {Pt A},
pages = {124385},
doi = {10.1016/j.watres.2025.124385},
pmid = {40803092},
issn = {1879-2448},
abstract = {Tetracycline (TC) stress disrupts microbial communities, yet its impact on aerobic granular sludge (AGS) formation and stability remain poorly understood. This study employs quantitative proteomics to unravel the AGS adaptive mechanisms under continuous TC exposure (1 mg/L). The results demonstrated that TC accelerated AGS granulation, achieving stable structure within 20 days, accompanied by improved pollutants removal (COD: 87.4%; TN: 84.1%; TP: 66.5%; TC: 83.0%) and settling performance (SVI: 50 mL/g). A critical granule size threshold of 3-4 mm was identified, beyond which AGS exhibited destabilization tendencies. Extracellular polymeric substances (EPS), particularly proteins, play pivotal role in maintaining AGS stability, with the α-helix / (β-sheet + random coil) ratio correlated with granule integrity. Proteomic analysis revealed the upregulation of outer membrane protein A (OmpA), facilitating biofilm formation, while TC-targeted ribosomes and bacterial chemotaxis were identified as central mechanisms for TC resistance and stress adaptation, respectively. These findings uncover molecular adaptations driving AGS stability under antibiotic pressure and offer insights for optimizing high-antibiotic wastewater treatment.},
}

@article {pmid40803023,
year = {2025},
author = {Yang, W and Zheng, X and Jia, D and Weng, A and Jia, Y and Chen, X and Zhang, Y and Yu, Q},
title = {Engineered nanoplatform with DNase-mimetic catalysis and photothermal ablation for synergistic biofilm eradication.},
journal = {Colloids and surfaces. B, Biointerfaces},
volume = {256},
number = {Pt 2},
pages = {115019},
doi = {10.1016/j.colsurfb.2025.115019},
pmid = {40803023},
issn = {1873-4367},
abstract = {Bacterial infections represent an increasing global health threat, exacerbated by the decline in antibiotic effectiveness due to widespread resistance. Biofilms, structured microbial communities embedded in extracellular polymeric substances (EPS), significantly hinder treatment by limiting antibiotic penetration and promoting bacterial persistence. With over 80 % of bacterial infections involving biofilms, there is an urgent need for antibiotic-free approaches that can disrupt these protective matrices. This study presents a dual-functional nanoplatform (Au-Ce NR) combining gold nanorods (Au NRs) as photothermal agents with synthetic deoxyribonuclease (DNase) mimics (Ce[4][+]/nitrilotriacetic acid (NTA) complexes) for synergistic biofilm eradication. The system was developed through covalent conjugation of Ce[4+]/NTA complexes onto polyethylene glycol-functionalized Au NRs. Within biofilm microenvironments, Ce[4+]/NTA complexes selectively degrade extracellular DNA (eDNA), a vital EPS component, thereby destabilizing the biofilm and facilitating nanoplatform penetration. Near-infrared irradiation subsequently induces localized hyperthermia via Au NRs, effectively eliminating dispersed bacteria while minimizing the risk of resistance development. In vitro experiments demonstrated efficient eDNA degradation in methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa biofilms, significantly enhancing bactericidal activity compared to unmodified Au NRs. In vivo assessments using an MRSA-infected wound model confirmed the therapeutic efficacy through reduced inflammation and accelerated wound healing. By combining enzymatic matrix disruption with photothermal ablation, this strategy addresses key limitations of conventional treatments for biofilm-associated infections in the post-antibiotic era.},
}

@article {pmid40802570,
year = {2025},
author = {Huo, S and Chang, L and Liu, Y and Xu, Z and Xue, M and Shi, C and Xu, G and Wang, K},
title = {Polyamine-Activated Carbonyl Stress Nanoplatform Synergistically Reverses Biofilm-Driven Immunosuppressive Microenvironment.},
journal = {ACS nano},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsnano.5c08038},
pmid = {40802570},
issn = {1936-086X},
abstract = {Polyamine metabolic dysregulation induced by implant-associated infections (IAIs) is a pivotal contributor to the formation of an immunosuppressive microenvironment. Excessive polyamines facilitate pathogen persistence by suppressing bacterial membrane lipid peroxidation (LPO) and enhancing DNA repair mechanisms. Simultaneously, polyamines promote biofilm formation via quorum sensing (QS) modulation and inhibit host immunity to facilitate immune escape. Herein, we developed a polyamine-responsive carbonyl stress nanoplatform MIL-100@PAO@PVP (MPP), which synergistically integrates metabolic intervention and chemodynamic therapy (CDT), addressing the limitations inherent to conventional oxidative damage-based therapies. Specifically, plasma amine oxidase (PAO) within MPP catalyzes polyamine degradation at infection sites, generating highly toxic acrolein and hydrogen peroxide (H2O2). The produced H2O2 markedly enhances MIL-100-mediated CDT, triggering a burst of hydroxyl radicals ([•]OH) that induces severe bacterial membrane LPO and DNA damage. Importantly, the generated acrolein further amplifies bacterial DNA damage via the induction of carbonyl stress. Additionally, bacterial debris resulting from MPP-induced cell death acts as endogenous antigens, effectively activating the antigen-presenting functions of macrophages and dendritic cells (DCs), thus reshaping the local immune response and reversing immunosuppression. Experimental results demonstrated robust antibiofilm efficacy and immunostimulatory effects of MPP in both in vitro and in vivo models, highlighting a promising therapeutic strategy for treating IAIs.},
}

@article {pmid40802474,
year = {2025},
author = {Conceição, K and de Andrade, VM and de Oliveira, VDM and Ramu, VG and Heras, M and Bardaji, ER and Castanho, MARB and Capella, AG},
title = {Unleashing IbKTP-NH2 a kyotorphin derivative, against bacterial and fungal multispecies biofilm adhesion and viability on materials.},
journal = {Journal of applied microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1093/jambio/lxaf205},
pmid = {40802474},
issn = {1365-2672},
abstract = {AIMS: Healthcare-Associated Infection (HAI) are a major global health concern, contributing to increased mortality and substantial economic burden on healthcare systems. This study aims to evaluate the efficacy of the peptide kyotorphin conjugated with ibuprofen (IbKTP-NH2) in inhibiting multispecies biofilms formed by Candida albicans, Pseudomonas aeruginosa, and Streptococcus pneumoniae, particularly in the context of biofilm-associated infections linked to implanted medical devices.

METHODS AND RESULTS: Multispecies biofilms were cultured on polymeric and metallic materials. Antimicrobial susceptibility testing was performed to determine the minimum biofilm inhibitory concentrations (MBIC) of IbKTP-NH2 against the tested strains. Additionally, scanning electron microscopy (SEM) was utilized to analyze biofilm architecture, focusing on extracellular matrix production, cell density, and morphology. The study found that the MBIC for bacterial strains ranged from 46.5 to 1 mM. SEM analysis revealed significant biofilm disruption, characterized by reduced extracellular matrix production, decreased cell numbers, and altered cell morphology, indicating effective antimicrobial activity of IbKTP-NH2.

CONCLUSION: The peptide IbKTP-NH2 demonstrates substantial inhibition of multispecies biofilms on both polymeric and metallic surfaces. These findings underscore its potential as an antimicrobial agent with possible antivirulence properties, and highlights IbKTP-NH2 as a promising candidate for the development of new therapeutic strategies aimed at preventing and controlling HAI and addressing chronic wound pathogens.},
}

@article {pmid40801958,
year = {2025},
author = {Soon, KL and Rashid, SA and Nazakat, R and Al Quwatli, L and Shee, CY and Azman, AS and Aziz, MA},
title = {Hidden risks of biofilm-forming and multidrug-resistant Gram-negative bacteria in water vending machines.},
journal = {Antonie van Leeuwenhoek},
volume = {118},
number = {9},
pages = {128},
pmid = {40801958},
issn = {1572-9699},
support = {LL2023FYP010//Universiti Malaya Sustainable Development Centre (UMSDC)/ ; },
abstract = {Water vending machines are widely utilised in various locations, including workplaces, universities and urban areas due to their convenience and accessibility. However, the microbial quality of the water dispensed from these machines, particularly in residential colleges, has not been thoroughly studied. This study aims to analyse the physicochemical and microbiological quality of drinking water collected from water vending machines located in 14 residential colleges around Universiti Malaya, Kuala Lumpur. Our study revealed that all drinking water samples met WHO acceptable limits for pH, TDS and electrical conductivity. However, dissolved oxygen and free chlorine levels were slightly below the acceptable range. No faecal contamination was detected in the drinking water samples, as indicated by the absence of enterococci and coliform bacteria, including faecal coliforms. Nevertheless, Gram-negative non-coliform bacteria were identified in water samples from 11 residential colleges. The identified bacteria included Pseudomonas spp., Acinetobacter spp., Bosea sp., Stenotrophomonas sp., Achromobacter sp., Roseatels sp. and Cupriavidus sp. Using CLSI guidelines as the standard, antibiotic sensitivity tests showed that all bacterial isolates were resistant to penicillin G (100%), while most were susceptible to ofloxacin (> 80%). Notably, Pseudomonas spp., Acinetobacter spp., Stenotrophomonas sp., Achromobacter sp. and Cupriavidus sp. were identified as multidrug-resistant bacteria, showing resistance to three or more categories of antimicrobial agents tested in this study. The biofilm assay confirmed that Bosea sp., Roseateles sp. and Acinetobacter sp. possess biofilm formation capacity. Overall, this study highlights that while the water from vending machines generally meets physicochemical standards and is free of faecal contamination, multidrug-resistant Gram-negative bacteria are still prevalent. Although these bacteria may not pose immediate health risks upon consumption, their presence poses long-term risks due to biofilm accumulation, resistance gene transfer and poor maintenance. This underscores the need for proper maintenance measures of water vending machines, including regular cleaning and effective disinfection, to ensure the microbial safety of drinking water.},
}

@article {pmid40801043,
year = {2025},
author = {Leiva-Sabadini, C and Berríos, P and Saavedra, P and Carrasco-Rojas, J and González-Aramundiz, JV and Vera, M and Tarifeño-Saldivia, E and Schuh, CMAP and Aguayo, S},
title = {Biofilm formation on collagen substrates modulates Streptococcus mutans bacterial extracellular nanovesicle production and cargo.},
journal = {Nanoscale advances},
volume = {},
number = {},
pages = {},
pmid = {40801043},
issn = {2516-0230},
abstract = {Streptococcus mutans is the major microbial etiological agent of dental caries and can adhere to surfaces such as type-I collagen, which is present in dentin and periodontal tissues. Recent studies have characterized planktonic S. mutans bacterial extracellular vesicles (bEVs) at the nanoscale range and demonstrated environmental-induced changes due to sugar presence or pH alterations. However, to date, no studies have explored whether surface-derived changes can modulate bEV production in the context of oral biofilm formation in the elderly. Therefore, this work aimed to determine the role of biofilm formation and collagen glycation on the nanoscale morphology and proteomic composition of S. mutans bEVs. For this, bEVs from S. mutans biofilms on native and glycated collagen surfaces were isolated, characterized, and compared to bEVs from planktonic cells. Nanoparticle tracking analysis (NTA), atomic force microscopy (AFM), and electron microscopy confirmed bEV production and showed that bEVs from biofilms are smaller in size and less abundant than those from planktonic cells. Furthermore, proteome analysis revealed that S. mutans biofilm formation on native and glycated collagen led to the enrichment of several key virulence proteins. Also, a shift towards proteins involved in metabolic processes was found in bEVs following biofilm formation on collagen surfaces, whereas glucan metabolism proteins were overexpressed in vesicles from the planktonic state. These results demonstrate that biofilm formation, as well as the glycation of collagen associated with aging and hyperglycaemia, can modulate bEV characteristics and cargo and could play a central role in S. mutans virulence and the development of diseases such as dental caries and periodontal disease.},
}

@article {pmid40801021,
year = {2025},
author = {Doll-Nikutta, K and Mikolai, C and Heine, N and Kurselis, K and Fadeeva, E and Debener, N and Legutko, B and Kreuzkamp, C and Prasanthan, V and Bahnemann, J and Chichkov, BN and Stiesch, M},
title = {Biocompatible liquid-infused titanium minimizes oral biofilm adhesion in flow chamber and 3D implant-tissue-biofilm in vitro models.},
journal = {Bioactive materials},
volume = {53},
number = {},
pages = {706-717},
pmid = {40801021},
issn = {2452-199X},
abstract = {Biomedical implants are susceptible to bacterial colonization, which can lead to challenging implant-associated infections. In particular, dental implant abutments - which are continuously exposed to bacteria within the oral cavity - stand to greatly benefit from strategies which inhibit the development of bacterial biofilms. Liquid-infused titanium surfaces have demonstrated excellent biofilm repellency, but to date have not been analyzed with substances suitable for medical device approval in terms of biocompatibility under conditions mimicking the environment of dental implant abutments. In this study, different medical-grade lubricants coated onto laser-structured titanium were screened for stability and water-repellency - with the results suggesting that unmodified structured titanium coated with silicone oil was the most promising combination of materials. When analyzing biofilm formation, the coated surfaces showed a statistically significant reduction in oral commensal Streptococcus oralis biofilms grown under static conditions as well as oral multispecies biofilms grown under salivation-resembling flow conditions. This biofilm-reducing effect was also observed when the coated surface interfaced with a 3D implant-tissue-oral-bacterial-biofilm (INTERbACT) in vitro model, which allows for the direct interaction of human tissue and oral multispecies biofilm at the implant interface. Importantly, this biofilm reduction was not due to toxicity of the coated surfaces, but is most likely attributable to inhibition of bacterial attachment. Additionally, the surfaces were not cytotoxic, without altering adjacent soft tissue or causing elevated pro-inflammatory cytokine secretion. These findings highlight the promise of biocompatible liquid-infused titanium surfaces as biofilm-repellent implant abutment modifications and provide the basis for further investigations in targeted pre-clinical studies.},
}

@article {pmid40796787,
year = {2025},
author = {Miranda, A and Brandquist, ND and Johnson, K and Muldiiarova, E and Fadeev, A and Ghanbari, M and Endres, JL and Bayles, KW and MacTaggart, J and Svechkarev, D and Sadykov, MR and Salkovskiy, Y},
title = {Quaternized chitosan derivatives inhibit growth and affect biofilm formation of Staphylococcus aureus.},
journal = {Scientific reports},
volume = {15},
number = {1},
pages = {29606},
pmid = {40796787},
issn = {2045-2322},
support = {51233//Nebraska Research Initiative/ ; 51233//Nebraska Research Initiative/ ; 51233//Nebraska Research Initiative/ ; 51233//Nebraska Research Initiative/ ; 51233//Nebraska Research Initiative/ ; P20GM152301/NH/NIH HHS/United States ; P20GM152301/NH/NIH HHS/United States ; P01-AI83211/NH/NIH HHS/United States ; P01-AI83211/NH/NIH HHS/United States ; P20GM152301/NH/NIH HHS/United States ; P01-AI83211/NH/NIH HHS/United States ; P01-AI83211/NH/NIH HHS/United States ; P20GM152301/NH/NIH HHS/United States ; },
abstract = {Antimicrobial resistance (AMR) poses a global health threat, severely impeding the effective treatment of bacterial infections and jeopardizing the safety of routine medical procedures. Methicillin-resistant Staphylococcus aureus (MRSA) is particularly problematic because of its resistance to beta-lactams and the ability to form resilient biofilms. Conventional antibiotics, including last-resort options, have serious side effects and may contribute to further resistance. Chitosan, a natural biopolymer, offers a promising alternative due to its biocompatibility and antimicrobial properties, though its effectiveness against biofilms is limited. Recent studies suggest that increasing the positive charge density and adding hydrophobic moieties to chitosan, can enhance its antimicrobial properties. In this work, the antibacterial activity of quaternized chitosan derivatives against AMR S. aureus strains was assessed. Quaternization of chitosan's amino group and introduction of hydrophobic side chains was found to significantly inhibit bacterial growth in both methicillin-sensitive (MSSA) and MRSA strains. Notably, nanofibrous materials composed of polyethylene oxide and hexyl-modified chitosan demonstrate alterations in S. aureus biofilm development, leading to significant accumulation of dead cells. Combined, these results highlight the potential of modified chitosan derivatives as effective antimicrobial agents for surface treatments and medical device coatings, particularly in applications where antibiotics are traditionally used, such as biofilm-prone environments.},
}

@article {pmid40795671,
year = {2025},
author = {Zhang, Y and Xu, R and Zhu, X and Yang, Z and Bian, Y and He, X and Tang, Y},
title = {UV-Induced VBNC state formation and resuscitation in E. coli: ATP-based metabolic activation and biofilm-mediated recovery.},
journal = {Water research},
volume = {287},
number = {Pt A},
pages = {124380},
doi = {10.1016/j.watres.2025.124380},
pmid = {40795671},
issn = {1879-2448},
abstract = {In this study, we systematically investigated the induction mechanism, metabolic characterization, and resuscitation behavior of viable but non-culturable bacteria (VBNC) by ultraviolet (UV) disinfection under different conditions, using E. coli as our example. Flow cytometry (FCM), ATP metabolism analyses and genetic assays revealed that low-dose UV (4.5 mJ/cm[2]) significantly inhibited bacterial culturability while maintaining cell membrane integrity and activating metabolic activity (total ATP levels were elevated to 182 % of initial values). Higher doses of UV (14.1 mJ/cm[2]), on the other hand, resulted in metabolic inhibition (ATP decreased to 58 %), but did not completely inactivate the bacteria. Gene expression analysis showed that the DNA repair gene recA and the antioxidant gene katG were significantly up-regulated with increasing UV money dose, while the stress response gene rpoS decreased, suggesting that the SOS repair system dominated the maintenance of the VBNC state. Resuscitation tests showed that biofilm greatly sped up the recovery of VBNC bacteria by releasing tyrosine-like extracellular polymers and quorum sensing signaling molecules, with resuscitation time shortened to 4 h in LB medium and 17 h in sterile water. This study reveals that UV disinfection-induced VBNC bacteria are characterized by enhanced metabolic activity and elucidates the mechanism by which biofilm promotes their resuscitation by secreting tyrosine-like EPS and C14 signaling molecules.},
}

@article {pmid40795591,
year = {2025},
author = {Xu, H and Chen, X and Jia, Y and Gao, X and Guo, S and Jia, D and Zhang, Y and Yu, Q},
title = {A trifunctional coating for biofilm prevention: integrating antifouling, photothermal killing, and quorum sensing interference.},
journal = {Journal of colloid and interface science},
volume = {701},
number = {},
pages = {138628},
doi = {10.1016/j.jcis.2025.138628},
pmid = {40795591},
issn = {1095-7103},
abstract = {Bacterial biofilm-associated infections pose a significant challenge in biomedical fields, especially in the context of medical devices and implants, where conventional treatments often fail to combat mature biofilms. To overcome this limitation, an antibiofilm coating was developed, incorporating three functional components: carbon nanoparticles derived from candle soot (CS), hydrophilic and pH-responsive copolymer brushes of 2-hydroxyethyl methacrylate (HEMA) and 3-(acrylamido)-phenylboronic acid (APBA), and natural quorum sensing inhibitors (QSIs). The carbon nanostructure of the CS substrate provides inherent photothermal conversion properties, while its high surface area facilitates efficient chemical functionalization. The copolymer design strategically incorporates hydrophilic poly(HEMA) domains that form bacteria-repellent hydration layers, alongside APBA moieties that covalently bind plant-derived QSIs (quercetin or baicalein) through pH-sensitive boronate ester linkages. This combination of components enables the coating to suppress biofilm formation via three synergistic mechanisms: bacterial adhesion is initially inhibited by hydration barriers, surface-bound pathogens are subsequently eradicated through photothermal conversion upon near-infrared activation, and the release of QSIs triggered by the acidic microenvironment disrupts biofilm maturation. The coating demonstrated sustained antibiofilm efficacy against both Pseudomonas aeruginosa and Staphylococcus aureus, while preserving mammalian cell viability and substrate compatibility across various biomedical materials. By integrating physical barriers, thermal ablation, and quorum sensing interference through precise material engineering, this coating offers a promising solution for preventing biofilm-associated infections in clinical settings.},
}

@article {pmid40794222,
year = {2025},
author = {De Marque, MB and Silva, BG and Foresti, E},
title = {Ammonia diffusivity in biofilm reactors: impact of polyurethane foam thickness and pore clogging on simultaneous nitrification and denitrification.},
journal = {Bioprocess and biosystems engineering},
volume = {},
number = {},
pages = {},
pmid = {40794222},
issn = {1615-7605},
abstract = {Simultaneous nitrification and denitrification (SND) processes represent a promising approach for nitrogen removal from effluents characterized by a low COD/N ratio, especially when combined with fixed-bed reactors to ensure that slow-growing biomass (e.g., nitrifiers) is not washed out. In this reactor configuration, biofilms are formed, which promote mass transport of the substrates involved in SND. Therefore, understanding the effective diffusivity of ammonia through the biofilm is essential to improve nitrogen removal, as it is influenced by the thickness of the support media and biomass growth, particularly under counter-diffusion conditions. For this type of study, flow cells (units for study particularities of a bioreactor) are used, as they provide greater operational control of the process. To evaluate this issue, were operated three flow cells for 234 days, each one with different thicknesses of polyurethane foam (i.e., 2 mm, 5 mm and 10 mm) as a support media for SND adhered biomass. Within each flow cell, the foam serves to segregate the aerated and non-aerated zones, thereby inducing counter-diffusion. Throughout operation, tests were conducted to estimate the effective diffusivity factor (EDF) of ammonia in the biofilm using the AQUASIM software. Routine analyses demonstrated that the average removal of organic matter and ammoniacal nitrogen were 73%, 68%, 57%, and 66%, 54%, 34% in the 2, 5, and 10 flow cells, respectively. Furthermore, EDF estimation tests demonstrated a 95% reduction in ammonia diffusivity over operating time, attributable to pore clogging induced by heterotrophic biomass growth within the support media. The decline in EDF of ammonia exerted a substantial impact on the total nitrogen removal and, consequently, on the performance of the simultaneous nitrification and denitrification process. Thus, the importance of considering mass transport phenomena in reactor designs with support media and long operating times, i.e., with biofilm growth and establishment, becomes evident.},
}

@article {pmid40793764,
year = {2025},
author = {Wen, X and Tang, T and Bao, T and Xu, T and An, B and Zhang, Y and Han, J},
title = {gltB encoding glutamate synthase is involved in persister and biofilm formation and virulence in Staphylococcus aureus.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0051125},
doi = {10.1128/spectrum.00511-25},
pmid = {40793764},
issn = {2165-0497},
abstract = {Glutamate metabolism plays a pivotal role in linking the tricarboxylic acid cycle, arginine biosynthesis, and purine metabolism, and these pathways have been shown to be involved in persister formation. However, the relationship among glutamate metabolism, bacterial antibiotic tolerance, and virulence remains unclear. In this study, gltB, which encodes the large subunit of glutamate synthase in Staphylococcus aureus, was knocked out. The ∆gltB mutant in the stationary phase showed less tolerance to antibiotics and was killed completely after exposure to lethal doses of ampicillin and norfloxacin after 11 and 6 days, respectively, while the parent strain still had abundant viable bacteria. The gltB complemented strain restored antibiotic tolerance. Interestingly, exogenous glutamate supplementation of ∆gltB restored the tolerance to antibiotics. Moreover, ∆gltB is more susceptible to heat, carbon starvation, and oxidative stress. Furthermore, the ability of ∆gltB to coagulate plasma, produce staphyloxanthin, and form biofilms was significantly weakened. In addition, ∆gltB attenuated virulence in BALB/c mice, and its 50% lethal dose (LD50) (1.14 × 10[10] CFU/mL, 95% CI: 7.29 × 10[9]-2.75 × 10[10]) was higher than that of the parent strain (2.39 × 10[9] CFU/mL, 95% CI: 9.99 × 10[8]-4.42 × 10[9]). The expression levels of major virulence genes, including eta, hla, hlgA, hlgC, lukD, lukE, lukS, lukF, and sea, as well as staphyloxanthin synthesis-related genes, including crtM and crtQ, were significantly downregulated in ∆gltB. This study revealed that gltB is involved in both antibiotic tolerance and virulence in S. aureus and provides new insights into the mechanism of persister formation and virulence, with implications for the development of novel drugs.IMPORTANCEStaphylococcus aureus is a leading bacterial cause of death, and persister formation renders it tolerant to antibiotics and is associated with its persistent infections. Glutamate metabolism plays a critical role in linking the tricarboxylic acid cycle, arginine biosynthesis, and purine metabolism, and these pathways have been shown to be involved in persister formation. This work first discovered that gltB, the large subunit of glutamate synthase gene in S. aureus, is involved in tolerance to antibiotics and heat, carbon starvation, and oxidative stress. Furthermore, the gltB mutant attenuated virulence in mice, owing to the inhibition of glutamate synthesis, which significantly weakened the ability of S. aureus to coagulate plasma, produce staphyloxanthin, form biofilms, and express virulence factors. These findings confirm the important role of glutamate metabolism in the formation of persister and virulence in S. aureus and provide new targets for developing novel anti-persister and anti-virulence drugs.},
}

@article {pmid40792556,
year = {2025},
author = {Qi, Y and Borglin, S and Li, L and Dong, W and Bill, M and Hao, Z and Pallud, C and Gilbert, B},
title = {Methanogenesis and Acetogenesis in Hydrogenotrophy with Carbonate Minerals: Dependence on Mineral Surface Area, Biofilm Growth, and Microbial Community.},
journal = {Environmental science & technology},
volume = {},
number = {},
pages = {},
doi = {10.1021/acs.est.4c14291},
pmid = {40792556},
issn = {1520-5851},
abstract = {The production, storage, and use of hydrogen are anticipated to grow substantially to achieve energy and climate goals. Consequently, microbial communities in many terrestrial and subsurface Earth environments could be exposed to elevated hydrogen concentrations. Hydrogen stimulates metabolic processes that reduce aqueous chemical species, such as bicarbonate or sulfate, that can exchange with solid mineral phases, but the controls on microbial hydrogenotrophy with mineral sources of electron acceptors are not fully understood. Herein, we applied laboratory experiments and biogeochemical modeling to study the response of a natural microbial community to an elevated partial pressure of hydrogen in the presence of carbonate minerals of varying composition, solubility, and size. Experimental incubations and simulation results showed that hydrogen consumption by microbial communities was initially dominated by sulfate reduction and, subsequently, transitioned to acetogenesis and methanogenesis. The rates of acetogenesis and methanogenesis were not correlated with the solubility of carbonate minerals. Instead, we observed strong linear correlations between the rates and surface area of carbonate minerals. Methane and acetate production slowed down in all incubations after about 2 weeks of incubation, although biogeochemical modeling predicted that the metabolic processes were not thermodynamically limited. Electron microscopy and infrared spectroscopy showed that biofilms with diverse microorganisms grew on the carbonates during this period. The methane δ[13]C value significantly increased, consistent with slower growth at elevated pH. This work highlights that microbial communities form biofilm on carbonate mineral surfaces as a response to hydrogen and that biofilm formation could pose a strong kinetic limitation to hydrogenotrophic metabolism utilizing carbonate minerals.},
}

@article {pmid40792184,
year = {2025},
author = {Gao, Y and Wang, Z and Wang, X and Xu, Y and Luo, J and Li, S and Yang, Y and Yan, L and Zou, X},
title = {Impact of Clove Oil on Biofilm Formation in Candida albicans and Its Effects on Mice with Candida Vaginitis.},
journal = {Mycobiology},
volume = {53},
number = {5},
pages = {661-675},
pmid = {40792184},
issn = {1229-8093},
abstract = {In this study, the effects of Clove oil (CEO) on biofilm formation of Candida albicans (C. albicans) were investigated using XTT and crystal violet staining methods. The therapeutic efficacy of CEO was studied using ICR mouse model with candida vaginitis. The potential targets and pathways of the CEO's effect against C. albicans were predicted by network pharmacology. The relative gene levels of CDC42, Ste11, Hst7, CEK1, and CPH1 were evaluated by qPCR. Subsequently, the protein levels of CDC42, CEK1, and p-CEK1 were determined by Western blot. The results indicated CEO can inhibit C. albicans adhesion, mycelium formation and biofilm maturation process. By regulating the related genes and proteins, the CEO can inhibit C. albicans biofilm maturation process, and weaken the ability of C. albicans to cause disease. For candida vaginitis ICR mouse model, CEO can lower inflammatory cytokines levels, alleviated the symptoms of vaginitis, and play a good therapeutic effect.},
}

@article {pmid40792117,
year = {2025},
author = {Patil, S and Powar, GS and Harale, S and Galatage, ST and Liu, S and Lopes, BS and Sharma, DK and Patil, AR and Wen, F},
title = {Formulation and Evaluation of a Licorice-Resveratrol Lollipop for Targeting Streptococcus mutans Biofilm and Antimicrobial Resistance.},
journal = {Infection and drug resistance},
volume = {18},
number = {},
pages = {3933-3946},
pmid = {40792117},
issn = {1178-6973},
abstract = {BACKGROUND: Streptococcus mutans is a key pathogen in dental caries, and the development of novel antimicrobial formulations is crucial to combat its resistance. This study aimed to evaluate a licorice-resveratrol medicated lollipop formulation (LRML) for its antimicrobial and anti-biofilm activity against S. mutans.

METHODS: The LRML was developed using a heating and congealing method, incorporating licorice extract (5% w/w) and resveratrol (2% w/w) in a sucrose-based matrix. The physicochemical properties of the formulation, including hardness, drug content uniformity, moisture content, and dissolution profile, were evaluated. The antimicrobial activity was assessed through Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), and time-kill assays. Anti-biofilm activity was evaluated using a crystal violet assay. The stability of the formulation was determined under accelerated conditions.

RESULTS: The LRML formulation showed efficient drug release, with formulation number LRML-7 demonstrating 96.87% release within 45 minutes. The antimicrobial tests revealed significant bactericidal effects against S. mutans at concentrations above 0.2 µg/mL, with a notable reduction in bacterial growth in time-kill assays. The formulation also demonstrated substantial inhibition of biofilm formation at both MIC and Minimum Bactericidal Concentration (MBC) levels. Stability studies confirmed that the formulation retained its physicochemical properties over three months.

CONCLUSION: The LRML exhibited promising antimicrobial and anti-biofilm activities against S. mutans, suggesting its potential as a novel therapeutic option for managing dental infections. Further clinical studies are required to optimize the formulation's efficacy and clinical applicability.},
}

@article {pmid40790539,
year = {2025},
author = {Venkataiah, VS and Karobari, MI},
title = {Effectiveness of biofilm-targeted therapy in managing and preventing dental caries: protocol for a systematic review and meta-analysis.},
journal = {Systematic reviews},
volume = {14},
number = {1},
pages = {165},
pmid = {40790539},
issn = {2046-4053},
mesh = {Systematic Reviews as Topic ; *Dental Caries/prevention & control/therapy/microbiology ; Humans ; *Biofilms/drug effects ; Meta-Analysis as Topic ; Photochemotherapy ; Probiotics/therapeutic use ; },
abstract = {BACKGROUND: Dental caries is a biofilm-dependent disease that continues to challenge conventional preventive strategies, such as fluoride application and mechanical plaque removal. Biofilm-targeted therapies (BTT), including probiotics, photodynamic therapy, enzymatic treatments, and natural compounds, represent promising alternatives to disrupt pathogenic biofilms effectively. However, the available evidence remains fragmented, with inconsistent methodologies and limited clinical data hindering comprehensive conclusions.

METHODS: This systematic review and meta-analysis will synthesize evidence on the effectiveness of BTT in preventing and managing dental caries. A comprehensive search of PubMed (MEDLINE), Embase, and the Cochrane Library will identify randomized controlled trials (RCTs) and observational studies evaluating BTT interventions. To ensure methodological robustness, quantitative synthesis of intervention effectiveness will be restricted to RCTs, while observational studies will contribute to a structured narrative synthesis. Two independent reviewers will conduct study selection, data extraction, and risk of bias assessment using the RoB 2 tool for RCTs and ROBINS-I for observational studies. Where feasible, pairwise meta-analyses will be performed using random-effects models. If sufficient data are available, a network meta-analysis (NMA) will be considered. Subgroup and sensitivity analyses will explore the impact of age, caries risk level, and intervention type on treatment outcomes.

DISCUSSION: This review anticipates providing evidence on the efficacy of BTT in reducing bacterial load, altering biofilm composition, and preventing caries progression. Variability in study design and outcome measures is expected, underscoring the need for standardized methodologies and more robust clinical trials. The findings aim to deliver evidence-based insights into the clinical applicability of BTT, addressing current knowledge gaps and guiding future research toward more effective caries prevention and management strategies.

TRIAL REGISTRATION: Systematic review registration: PROSPERO CRD42024615568.},
}

Systematic Reviews as Topic
*Dental Caries/prevention & control/therapy/microbiology
Humans
*Biofilms/drug effects
Meta-Analysis as Topic
Photochemotherapy
Probiotics/therapeutic use

@article {pmid40790144,
year = {2025},
author = {Dubey, AA and Toprak, P and Pring, A and Rodriguez-Navarro, C and Mukherjee, A and Dhami, NK},
title = {Effect of substrate mineralogy, biofilm and extracellular polymeric substances on bacterially induced carbonate mineralisation investigated with in situ nanoscale ToF-SIMS.},
journal = {Scientific reports},
volume = {15},
number = {1},
pages = {29368},
pmid = {40790144},
issn = {2045-2322},
support = {Discovery project DP220101990 "Microbially induced calcium carbonate precipitation in different substrates"//Australian Research Council/ ; Discovery project DP220101990 "Microbially induced calcium carbonate precipitation in different substrates"//Australian Research Council/ ; Discovery project DP220101990 "Microbially induced calcium carbonate precipitation in different substrates"//Australian Research Council/ ; Discovery project DP220101990 "Microbially induced calcium carbonate precipitation in different substrates"//Australian Research Council/ ; Discovery project DP220101990 "Microbially induced calcium carbonate precipitation in different substrates"//Australian Research Council/ ; MCIN/ AEI /10.13039/501100011033//Spanish Government grant PID2021. 125305NB.I00/ ; A way of making Europe; Junta de Andalucía research group RNM-179//European Regional Development Fund/ ; Unidad Científica de Excelencia UCE-PP2016-05.//Universidad de Granada/ ; },
mesh = {*Calcium Carbonate/metabolism/chemistry ; *Biofilms/growth & development ; *Extracellular Polymeric Substance Matrix/metabolism/chemistry ; Spectrometry, Mass, Secondary Ion/methods ; *Bacillus subtilis/metabolism/physiology ; *Sporosarcina/metabolism ; },
abstract = {Bacterial mineralisation of calcium carbonates (CaCO3) has become a focal point of interest in the scientific community owing to their versatile applications as biomaterials. However, despite extensive research, the knowledge on factors influencing biogenic CaCO3 polymorph (calcite, vaterite or aragonite) selection in nature remains obscure. Bacterial mineralisation happens in nature on diverse substrates by different pathways, often in the presence of organic matter such as biofilm and extracellular polymeric substances (EPS) secreted with the regular metabolic activities of microbes. This study examines the bacterial CaCO3 mineralisation process by two distinct pathways on different natural substrates with advanced analytical techniques, including Time of Flight- Secondary Ions Mass Spectrometry (ToF-SIMS). A high EPS-producing microbe (Bacillus subtilis, BS) was compared with the standard ureolytic strain (Sporosarcina pasteurii, SP). Natural geological minerals, including apatite, calcite and quartz, were selected as substrates. This study demonstrates that SP favours the precipitation of rhombohedral calcite crystals (2 to 40 μm in size), regardless of the mineral substrate. In contrast, the EPS-producing BS culture induced the formation of significantly larger vaterite structures (20 to 100 μm in size) in spheroid and hexagonal shapes. The mineralogy of precipitates was confirmed with Raman spectroscopy. ToF-SIMS enabled the spatial tracking of organic macromolecules and the adsorption of calcium ions on them. The functional groups of the EPS involved in these interactions were characterised by Fourier Transform Infrared Spectroscopy (FTIR). This study reveals that microbial activity dominates over substrate mineralogy in selecting the phase and shaping the morphology of biogenic CaCO3, with EPS playing a crucial role in promoting the aggregation of small nanocrystals into large vaterite structures and their stabilisation.},
}

*Calcium Carbonate/metabolism/chemistry
*Biofilms/growth & development
*Extracellular Polymeric Substance Matrix/metabolism/chemistry
Spectrometry, Mass, Secondary Ion/methods
*Bacillus subtilis/metabolism/physiology
*Sporosarcina/metabolism

@article {pmid40789448,
year = {2025},
author = {Mohanty, D and Padhee, S and Sarangi, A and Das, BS and Priyadarshini, A and Dash, KT and Kerry, RG and Jena, S and Sahoo, A and Chandra Panda, P and Nayak, S and Ray, A},
title = {Cinnamomum zeylanicum essential oil inactivates Shigella flexneri by inhibiting biofilm formation, disrupting cell membrane integrity, and suppressing virulence gene expression.},
journal = {Microbial pathogenesis},
volume = {208},
number = {},
pages = {107977},
doi = {10.1016/j.micpath.2025.107977},
pmid = {40789448},
issn = {1096-1208},
abstract = {Shigella flexneri, a foodborne pathogen that causes shigellosis, poses a significant risk to human health and food safety. Cinnamomum zeylanicum essential oil (CZEO) is known to inhibit microbial growth, but its possible antibacterial and antibiofilm mechanisms against S. flexneri remain unclear. The chemical composition of CZEO was identified via GC-MS. The minimum inhibitory concentration of CZEO against S. flexneri was 200 μg/ml. CZEO-treated S. flexneri exhibited an extended lag phase, indicating delayed bacterial growth, and demonstrated potent bactericidal activity in fresh coconut water. CZEO has an antibacterial effect by disrupting cell membranes, leading to leakage of nucleic acids and proteins. Congo red agar, safranin staining and XTT assays confirmed the ability of CZEO to prevent biofilm formation and decrease the metabolic activity of S. flexneri cells. CZEO reduced hydrophobicity, aggregation, and S. flexneri motility, contributing to the antibiofilm effect. An annexin V-FITC/PI assay revealed that CZEO induced cell death in S. flexneri via apoptosis, whereas gene expression analysis revealed significant downregulation of biofilm-associated genes such as yebL, csgD, rcsB, rcsC, rfbA, rfbE, mdoC, SF2159, and waaG. Light, fluorescence, and scanning electron microscopy revealed that CZEO inhibited biofilms in a dose-dependent manner and was able to induce irreversible morphological damage to S. flexneri cells. Homology modelling and molecular docking demonstrated interactions between virulence proteins and major constituents of CZEO. The MTT assay revealed that CZEO was nontoxic to human embryonic kidney (HEK293) cells even at relatively high concentrations. The above results confirmed that CZEO could be used as a natural food additive to combat Shigella flexneri and its biofilm.},
}

@article {pmid40788981,
year = {2025},
author = {Iemini, RCA and Trinca, ALM and Benedetti, MD and Cordeiro, CF and Ferreira, AV and Dias, ALT and Caldas, IS and Hawkes, JA and Carvalho, DT and Franco, LL},
title = {Synthesis of New Glycometronidazole Compounds With Antifungal and Antifungal Biofilm Activity.},
journal = {Chemical biology & drug design},
volume = {106},
number = {2},
pages = {e70154},
pmid = {40788981},
issn = {1747-0285},
support = {403194/2023-7//Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)/ ; APQ-00686-18//Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)/ ; APQ-00352-18//Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)/ ; BPD-00154-22//Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)/ ; RED-00110-23//Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)/ ; APQ-00544-23//Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)/ ; APQ-01455-24//Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)/ ; },
mesh = {*Biofilms/drug effects ; *Antifungal Agents/pharmacology/chemical synthesis/chemistry ; *Metronidazole/pharmacology/chemistry/chemical synthesis/analogs & derivatives ; Click Chemistry ; Microbial Sensitivity Tests ; Triazoles/chemistry/pharmacology/chemical synthesis ; Glycosylation ; },
abstract = {Carbohydrates are well known to be one of the most abundant and structurally diverse natural organic compounds, and they are of great importance as an energy source and as structural components of cell walls in different organisms. They are involved in various biological and pathological processes, including homeostasis, cell-cell interaction, cell migration, cell development, bacterial and viral infection, inflammation, immunology, and cancer metastasis. The variety of these properties is a result of the structural diversity found in carbohydrates. The chemistry of carbohydrates involved in the diagnosis and treatment of diseases has attracted increasing attention from researchers, which is why they should be one of the main focuses in new drug discovery. This study focuses on the synthesis of new glycotriazole-metronidazole compounds as antifungal agents and antifungal biofilm agents, from the glycosylation of metronidazole with various carbohydrates (d-glucose, d-galactose, d-N-acetylglucosamine, and d-lactose). Our hypothesis is that the glycosides could be taken into fungal biofilms through recognition by glycoreceptors and transporters, carrying the active residue with them. In a low-oxygen environment, the nitro group would then undergo bioreduction leading to the formation of toxic radicals potentially resulting in the destruction or paralysis of biofilm formation-essentially functioning as a bioactive "Trojan horse." The compounds were obtained via a click chemistry reaction using a triazole connector, and the subsequent antifungal tests showed good results for a number of compounds. In silico studies demonstrated positive data for all synthesized compounds, and, in general, they present low toxicological risks.},
}

*Biofilms/drug effects
*Antifungal Agents/pharmacology/chemical synthesis/chemistry
*Metronidazole/pharmacology/chemistry/chemical synthesis/analogs & derivatives
Click Chemistry
Microbial Sensitivity Tests
Triazoles/chemistry/pharmacology/chemical synthesis
Glycosylation

@article {pmid40788469,
year = {2025},
author = {Singh, I and Agrawal, H and Maurya, S and Tanwar, H and Gupta, S and Singh, NP},
title = {Efficacy and safety of N-acetylcysteine vs. probiotics in in-vivo biofilm prevention on ureteral stents: a prospective randomized controlled pilot in vivo study.},
journal = {International urology and nephrology},
volume = {},
number = {},
pages = {},
pmid = {40788469},
issn = {1573-2584},
abstract = {OBJECTIVE: To evaluate the efficacy and safety of N-acetylcysteine (NAC) vs. probiotics in preventing biofilm formation on ureteral stents in selected patients. Ureteral stents are essential for managing urinary obstructions and renal stones and are prone to complications such as biofilm formation, which can lead to infections and stent malfunction. NAC is known for its mucolytic and antimicrobial properties, has shown potential in preventing biofilm formation on ureteral stents; however, clinical evidence is limited.

METHODS: This prospective, open-label, randomized controlled experimental study was conducted in a tertiary care teaching hospital between October 2022 and March 2024. Sixty patients undergoing ureteral stent placement were randomized to receive NAC (600 mg twice daily) in Group I vs. probiotics (containing Streptococcus faecalis, Clostridium butyricum, Bacillus mesentericus, and Lactic Acid Bacillus) in Group II for 5 weeks. Primary outcomes included routine urine microscopy, culture, stent culture, and biofilm detection on the stent surfaces.

RESULTS: Group I showed significantly reduced (pyuria, positive urine cultures, and biofilm formation on stents) to group II. Biofilm was absent in group I but was detected in 6.7% of group II. No significant treatment-emergent adverse effects (TEAEs) were observed in either group.

CONCLUSION: NAC demonstrated a strong potential in preventing biofilm formation on ureteral stents, which correlated with improved outcomes in terms of reduced stent-associated bacterial growth, bacteriuria, and pyuria. However, further studies with larger sample sizes and longer follow-up periods are needed to confirm these findings and assess the precise utility and long-term safety of NAC in clinical settings for ureteral stenting and UTIs.

DRUG TRIAL REGISTRY: TCTR20250511001 dt 11th May '2025, https://www.thaiclinicaltrials.org/show/TCTR20250511001.},
}

@article {pmid40788302,
year = {2025},
author = {Walton, B and Abbodante, S and Marshall, ME and Dobruchowska, JM and Alvi, A and Gallagher, LA and Vallikat, N and Zhang, Z and Wozniak, DJ and Yu, EW and Boons, GJ and Pearlman, E and Rietsch, A},
title = {A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptor.},
journal = {eLife},
volume = {13},
number = {},
pages = {},
pmid = {40788302},
issn = {2050-084X},
support = {R01AI169865/NH/NIH HHS/United States ; R01EY14362/NH/NIH HHS/United States ; R01AI145069/NH/NIH HHS/United States ; },
mesh = {*Biofilms/growth & development ; *Pseudomonas aeruginosa/virology/physiology ; Animals ; *Pseudomonas Phages/physiology/metabolism/genetics ; Mice ; *Polysaccharides, Bacterial/metabolism ; Pseudomonas Infections/microbiology/therapy ; *Receptors, Virus/metabolism ; },
abstract = {Bacteria in nature can exist in multicellular communities called biofilms. Biofilms also form in the course of many infections. Pseudomonas aeruginosa infections frequently involve biofilms, which contribute materially to the difficulty to treat these infections with antibiotic therapy. Many biofilm-related characteristics are controlled by the second messenger, cyclic-di-GMP, which is upregulated on surface contact. Among these factors is the exopolysaccharide Psl, which is a critically important component of the biofilm matrix. Here, we describe the discovery of a P. aeruginosa bacteriophage, which we have called Clew-1, that directly binds to and uses Psl as a receptor. While this phage does not efficiently infect planktonically growing bacteria, it can disrupt P. aeruginosa biofilms and replicate in biofilm bacteria. We further demonstrate that the Clew-1 can reduce the bacterial burden in a mouse model of P. aeruginosa keratitis, which is characterized by the formation of a biofilm on the cornea. Due to its reliance on Psl for infection, Clew-1 does not actually form plaques on wild-type bacteria under standard in vitro conditions. This argues that our standard isolation procedures likely exclude bacteriophage that are adapted to using biofilm markers for infection. Importantly, the manner in which we isolated Clew-1 can be easily extended to other strains of P. aeruginosa and indeed other bacterial species, which will fuel the discovery of other biofilm-tropic bacteriophage and expand their therapeutic use.},
}

*Biofilms/growth & development
*Pseudomonas aeruginosa/virology/physiology
Animals
*Pseudomonas Phages/physiology/metabolism/genetics
Mice
*Polysaccharides, Bacterial/metabolism
Pseudomonas Infections/microbiology/therapy
*Receptors, Virus/metabolism

@article {pmid40788144,
year = {2025},
author = {Shafeeq, S and Pannanusorn, S and Dainat, J and Dadvar, A and Tellgren-Roth, C and Sennblad, B and Nystedt, B and Römling, U},
title = {Genome sequences of six clinical isolates of Candida parapsilosis exhibiting different degrees and temporal regulation of biofilm formation.},
journal = {Microbiology resource announcements},
volume = {},
number = {},
pages = {e0130024},
doi = {10.1128/mra.01300-24},
pmid = {40788144},
issn = {2576-098X},
abstract = {Candida parapsilosis is a major pathogen causing central venous catheter-associated bloodstream infections with biofilm formation as virulence factor. We sequenced the genomes of six C. parapsilosis isolates from bloodstream infections displaying no, low, and high biofilm under conditions mimicking the clinical setting.},
}

@article {pmid40785722,
year = {2025},
author = {Shadvar, N and Yousefi, F and Barazesh, A and Tajbakhsh, S},
title = {Investigation of virulence factors and genes associated with biofilm and protease in Stenotrophomonas maltophilia isolates in Bushehr, Iran.},
journal = {Iranian journal of microbiology},
volume = {17},
number = {4},
pages = {559-568},
pmid = {40785722},
issn = {2008-3289},
abstract = {BACKGROUND AND OBJECTIVES: This study aimed to investigate the virulence factors and genes associated with biofilm and protease in Stenotrophomonas maltophilia in Bushehr, Iran.

MATERIALS AND METHODS: Eighty-seven S. maltophilia isolates (67 clinical and 20 environmental isolates) were studied. The isolates were assessed for the production of virulence factors including several enzymes and biofilm. To detect rmlA, rpfF, spgM, smf-1, StmPr1 868 bp, StmPr1 1621 bp, and StmPr2 genes, PCR and sequencing were performed.

RESULTS: All isolates (100%) produced DNase, hemolysin, protease, lipase, and hyaluronidase. Seventy-eight (89.7%) isolates were gelatinase producers, and 85 (97.7%) isolates were lecithinase producers. All isolates were biofilm producers: 79 (90.8%) isolates produced strong biofilm, 5 (5.7%) isolates produced moderate biofilm, and 3 (3.5%) isolates produced weak biofilm. The frequency of smf-1, rmlA, rpfF, and spgM was 93.1%, 86.2%, 26.4%, and 59.8%, respectively. The frequency of protease genes including StmPr1 868 bp, StmPr1 1621 bp, and StmPr2 was 12.6%, 41.4%, and 18.4%, respectively.

CONCLUSION: Our findings revealed a high frequency of isolates that produce DNase, hemolysin, protease, gelatinase, lipase, lecithinase, hyaluronidase, and biofilm. All isolates that harbored spgM or rpfF or both genes were strong biofilm producers. Notably, the presence of isolates that lacked spgM and rpfF genes but produced strong biofilm indicates that in addition to these two genes, other genes or factors may play a role in the production of strong biofilm. Based on this research, S. maltophilia in our area possesses the capability to produce several factors that could play roles in pathogenicity.},
}

@article {pmid40785719,
year = {2025},
author = {Khashei, S and Fazeli, H and Rahimi, F and Karbasizade, V},
title = {Exploring the genetic diversity and the association of drug resistance and biofilm production in Acinetobacter baumannii strains isolated from burn wound infections.},
journal = {Iranian journal of microbiology},
volume = {17},
number = {4},
pages = {549-558},
pmid = {40785719},
issn = {2008-3289},
abstract = {BACKGROUND AND OBJECTIVES: Acinetobacter baumannii is considered a troublesome cause of infection in burn units, where its capability to form biofilm and resist antibiotics significantly hampers therapeutic success. This study explored the correlations between antimicrobial resistance profiles, biofilm-producing capacity, and genetic diversity of A. baumannii strains from patients with burn wound infection in Isfahan, Iran.

MATERIALS AND METHODS: Ninety-six isolates were analyzed for antibiotic resistance using the disk diffusion technique and for biofilm formation through the microtiter dish assay. The prevalence of ten biofilm-related genes was investigated using specific primers. Clonal relatedness among bacterial strains was defined by Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR).

RESULTS: A vast majority of isolates (99%) exhibited resistance to meropenem, ciprofloxacin, ceftriaxone, cefotaxime, piperacillin-tazobactam, and imipenem, qualifying them as extensively drug-resistant (XDR). Twenty-five percent of the strains were strong biofilm formers, while 68% demonstrated moderate or weak biofilm formation. The most commonly identified biofilm-related genes included bfmR (100%), ompA (100%), and bap (99%). A significant association was found between the production of biofilm, resistance to aminoglycosides, and the presence of csuE and bap genes. ERIC-PCR typing showed the presence of 3 clonal types and 7 single types, with biofilm producers predominantly clustering to clonal type 2.

CONCLUSION: This work highlights a notable prevalence of biofilm-producing XDR A. baumannii in burn patients, underscoring the need for continuous surveillance and enhanced infection control strategies.},
}

@article {pmid40785718,
year = {2025},
author = {Ahmadbeigi, Y and Soleimani, N and Azizmohseni, F and Amini-Bayat, Z},
title = {Biofilm formation and eradication of Staphylococcus aureus: a study of culture conditions and endolysin ZAM-CS effect.},
journal = {Iranian journal of microbiology},
volume = {17},
number = {4},
pages = {586-592},
pmid = {40785718},
issn = {2008-3289},
abstract = {BACKGROUND AND OBJECTIVES: Staphylococcus aureus significantly contributes to healthcare-associated infections, with biofilm formation causing chronic, antibiotic-resistant cases. Because biofilms show high resistance to conventional antibiotics, endolysins have emerged as a promising alternative for treating antibiotic-resistant, biofilm-associated infections. This study evaluated the effects of four culture media and different incubation times on biofilm formation in methicillin-sensitive (MSSA) and methicillin-resistant (MRSA) S. aureus strains and assessed the anti-biofilm efficacy of a novel chimeric endolysin called ZAM-CS (catalytic domain of SAL-1 endolysin and binding domain of lysostaphin).

MATERIALS AND METHODS: Biofilms were grown for 24, 48, and 72 hours in Mueller-Hinton broth (MHB), Luria broth (LB), terrific broth (TB), and tryptic soy broth (TSB). The crystal violet assay was used to assess the biomass of the biofilm. The optimal biofilm conditions were then used to test ZAM-CS's activity at different concentrations.

RESULTS: MSSA formed the strongest biofilms in TB. MRSA formed stable, high-biomass biofilms in TSB, TB, and LB, while MHB was the least supportive medium for both strains. ZAM-CS significantly reduced biofilm biomass in both MSSA and MRSA (up to 77%).

CONCLUSION: ZAM-CS's rapid and potent anti-biofilm activity at low concentrations highlights its potential as a promising treatment against antibiotic-resistant S. aureus biofilm infections.},
}

@article {pmid40785712,
year = {2025},
author = {Shabanzadeh, M and Rezaei-Matehkolaei, A and Zarei Mahmoudabadi, A},
title = {The efficacy of luliconazole and caspofungin on planktonic and biofilm of Candida albicans from different sources.},
journal = {Iranian journal of microbiology},
volume = {17},
number = {4},
pages = {653-659},
pmid = {40785712},
issn = {2008-3289},
abstract = {BACKGROUND AND OBJECTIVES: The ability of Candida albicans to produce biofilm is considered an important pathogenic factor. In addition, the low sensitivity of biofilms to antifungal drugs is a challenge for patients, clinicians, and laboratory workers. We aimed to investigate the effectiveness of luliconazole and caspofungin on the planktonic and biofilm types of C. albicans strains.

MATERIALS AND METHODS: Fifty C. albicans from vaginitis, candiduria, gastrointestinal candidiasis, and saliva were examined for antifungal susceptibility against caspofungin and luliconazole using the CLSI M27 guideline. Moreover, the susceptibility of biofilms was detected using 96 well microplates and the MTT method.

RESULTS: The capacity of the isolates to produce biofilm within 2, 6, and 24 h was different, however, all tested isolates produced biofilm after 24 h. Vaginal and esophagitis isolates had a high and low ability for biofilm production during 24-hour incubation. In our study, 90% of isolates were sensitive to caspofungin, while 7.5 and 2.5% of them were intermediate and resistant. The MIC range of all isolates against luliconazole was 0.01562-1 μg/mL.

CONCLUSION: The MICs of biofilms were 15.6, and 171.3 higher than that of planktonic cells for caspofungin and luliconazole, respectively. Moreover, paradoxical and trailing effects occurred at 4 and 32 μg/mL of caspofungin and luliconazole, respectively.},
}

@article {pmid40784500,
year = {2025},
author = {Rahbardar, MG and Kesharwani, P and Sahebkar, A},
title = {Resveratrol in anti-biofilm therapy: Mechanisms, molecular derivatives, and emerging drug delivery strategies.},
journal = {Fitoterapia},
volume = {},
number = {},
pages = {106800},
doi = {10.1016/j.fitote.2025.106800},
pmid = {40784500},
issn = {1873-6971},
abstract = {Microbial biofilms serve an important function in various aspects of human health, including infections, chronic diseases, and industrial processes, but their development poses significant challenges due to inherent resistance to traditional antimicrobial agents. Natural compounds are increasingly being investigated as potential anti-biofilm therapy. Resveratrol, a polyphenolic molecule found in many plants, has sparked widespread interest since it has numerous physiological properties, namely antibacterial characteristics. The purpose of this paper is to conduct a thorough analysis of the effect of resveratrol on microbial biofilm. We will elaborate on how resveratrol reduces biofilm formation, disrupts pre-formed biofilms, and modulates gene expression related to biofilm development. Furthermore, we discuss the effects of resveratrol on the structural integrity, composition, and virulence factors associated with biofilms. Additionally, we explore the potential synergistic interactions between resveratrol and other antimicrobial agents, while highlighting the challenges and future prospects of utilizing resveratrol as an effective anti-biofilm agent. Overall, the present investigation offers details on promising benefits of resveratrol in eliminating microbial biofilms, emphasizing its efficacy as an alternative treatment for biofilm-associated illnesses and their consequences.},
}

@article {pmid40783906,
year = {2025},
author = {Kumari, A and Verma, B and Chattopadhyay, S},
title = {Isolation and Characterisation of Biofilm-Forming Thermophilic Bacteria for Congo Red Decolorization.},
journal = {Current microbiology},
volume = {82},
number = {9},
pages = {445},
pmid = {40783906},
issn = {1432-0991},
mesh = {*Congo Red/metabolism ; *Biofilms/growth & development ; Biodegradation, Environmental ; RNA, Ribosomal, 16S/genetics ; *Coloring Agents/metabolism ; Soil Microbiology ; India ; *Bacillus/isolation & purification/genetics/metabolism/classification/physiology ; Temperature ; *Bacteria/isolation & purification/metabolism/genetics/classification ; Textile Industry ; Phylogeny ; },
abstract = {In the present research, we isolate and characterize the thermophilic biofilm-forming bacteria of the textile industry that are responsible for the degradation of azo dye. The thermophilic bacteria that form biofilm grow at a temperature range of 45-80 ℃ and play an important role in bioremediation. Thermo-stable biofilm-producing bacteria are a unique class of microorganisms that flourish well in extreme environments. The study aims to screen and characterize thermophilic biofilm-producing bacteria isolated from the soil of the textile industry in Hindpiri, Ranchi, Jharkhand, India, to uncover their potential for Congo red decolorization. Two isolates, A1 and A2, which are able to form biofilm when grown in TSA, were used for further studies. After 3 days of incubation, maximum biofilm developed at 45 °C with pH 7-9. Biofilms were grown on glass coverslips and incubated with Congo red dye to evaluate their dye decolorization ability. Isolate A1 exhibited a high decolorization potential, 80% Congo red dye in 18 h, whereas isolate A2 had a lower efficiency, decolorizing only 60% of the dye under 45 ℃ at static conditions. 16S rRNA-based molecular characterization identified isolate A1 as Bacillus licheniformis A1 (PV362359). Analytical techniques like FESEM and FTIR analysis showed significant changes in bacterial cells and in functional groups, with a marked reduction or disappearance of peaks, respectively. Peaks at 1745 cm[-1], 2859 cm[-1], 2931 cm[-1], and 1154 cm[-1] give the alterations in protein and polysaccharides associated functional groups. These demonstrate the breakdown of the azo linkages and sulfonate groups. The present study indicates the potential of thermophilic biofilm-producing bacteria for azo-dye bioremediation.},
}

*Congo Red/metabolism
*Biofilms/growth & development
Biodegradation, Environmental
RNA, Ribosomal, 16S/genetics
*Coloring Agents/metabolism
Soil Microbiology
India
*Bacillus/isolation & purification/genetics/metabolism/classification/physiology
Temperature
*Bacteria/isolation & purification/metabolism/genetics/classification
Textile Industry
Phylogeny

@article {pmid40783805,
year = {2025},
author = {Ong Watts, J and Epperson, RT and Kawaguchi, B and Olsen, A and Tam, J and Pasquina, P and Isaacson, B and Gililland, J and Williams, D},
title = {Assessing the Potential of Antimicrobial Blue Light (aBL) to Manage Biofilm Burden at the Skin-Implant Interface of Percutaneous Osseointegrated Implants.},
journal = {Journal of orthopaedic research : official publication of the Orthopaedic Research Society},
volume = {},
number = {},
pages = {},
doi = {10.1002/jor.70039},
pmid = {40783805},
issn = {1554-527X},
support = {//This study was supported by the Department of Defense under Award #HU00011920009./ ; },
abstract = {Percutaneous osseointegrated implants offer significant benefits to amputees, including improved mechanical feedback and reduced skin irritation. However, their percutaneous nature increases susceptibility to biofilm-associated infections. This study evaluated the potential efficacy of antimicrobial blue light (aBL) as a preventive strategy to manage biofilm burden in a 12-week ovine model wherein two treatment cycles were implemented. In this study, aBL was applied proactively to reduce surface colonization and prevent infection progression, rather than to treat confirmed or deep infections. Sheep were inoculated with Staphylococcus aureus biofilm and treated with aBL or systemic antibiotics, with or without a washing protocol. aBL with washing achieved the greatest bacterial reduction, lowering CFU levels to 3.0 ± 0.3 log10 during the second treatment period, significantly outperforming washing alone, which resulted in 5.6 ± 0.4 log10 (p < 0.05). The study also showed a lower observed infection rate (14% vs. 43%), when comparing aBL with washing to washing only based on histologic, microCT, and gross assessments, though no statistical comparison was performed. Despite these benefits, the aBL treatments induced notable tissue changes, including increased epithelial thickening, rete ridge elongation, and accelerated skin/tissue downgrowth. Systemic antibiotics were effective during the initial treatment period but failed to reduce biofilm burden during the second treatment cycle. Altogether, while aBL combined with washing shows promise as a localized preventive therapy for reducing biofilm burden and implant-related infection, the observed tissue remodeling and accelerated downgrowth highlight the need for further investigation to ensure long-term safety.},
}

@article {pmid40782980,
year = {2025},
author = {Pradhan, P and Singh, B},
title = {Antibiotic Resistance, Biofilm Formation, and Virulence Gene Profiling of Multidrug-Resistant Pseudomonas aeruginosa Isolates from a Multi-Speciality Hospital in Sikkim, India.},
journal = {Microbial pathogenesis},
volume = {},
number = {},
pages = {107976},
doi = {10.1016/j.micpath.2025.107976},
pmid = {40782980},
issn = {1096-1208},
abstract = {UNLABELLED: Effective antibiotic stewardship and monitoring of antimicrobial resistance are crucial for mitigating the emergence and spread of opportunistic nosocomial pathogens, such as Pseudomonas aeruginosa. This study aimed to identify antibiotic resistance patterns, the ability to form biofilms, and the presence of virulence-associated genes in multidrug-resistant (MDR) P. aeruginosa isolates from a multi-speciality government hospital in Sikkim, India.

METHODS: Forty clinical isolates of P. aeruginosa were characterised through antimicrobial susceptibility testing (AST). MDR isolates were evaluated for biofilm formation and the presence of carbapenemase genes (blaKPC, blaGES, blaIMP, blaVIM, blaNDM, blaOXA-48), quorum sensing (lasR), efflux pump (mexAB-oprM), and various virulence-associated genes (toxA, lasA, lasB, popB, aprA, pvdA, phzM, phzS, algD) using PCR.

RESULTS: The ecfX gene specific to P. aeruginosa was detected in all isolates, confirming their identification. Among the isolates, 37.5% were MDR, and 7.5% were extensively drug-resistant (XDR), with a multiple antibiotic resistance (MAR) index ranging from 0.210 to 0.894. Some MDR isolates demonstrated resistance to meropenem (MIC 15.62 to 125 μg/mL), indicating carbapenem resistance. All isolates showed a MIC of ≤ 2 μg/mL to colistin. These MDR isolates formed biofilms and harboured lasR, mexAB-oprM, along with the majority of virulence-related genes (toxA, lasA, popB, phzM, and algD). However, only two carbapenemase genes were detected: blaGES (66.66%) and blaIMP (73.33%).

CONCLUSION: This study highlights the occurrence of biofilm-forming, MDR, and carbapenem-resistant clinical isolates of P. aeruginosa. These isolates harbour several virulence-associated genes and are fully susceptible to colistin, suggesting its potential role in treating such infections. The findings emphasise the need for continued surveillance and targeted antimicrobial stewardship in hospital settings.},
}

@article {pmid40782784,
year = {2025},
author = {Ye, F and Chu, K and Hua, Z and Liu, Y and Yang, X and Qing, S and Gu, L},
title = {Distribution and fate of antibiotics in water-sediment-biofilm phases and their potential risks in urban lakes within a large-scale city in eastern China.},
journal = {Journal of hazardous materials},
volume = {496},
number = {},
pages = {139421},
doi = {10.1016/j.jhazmat.2025.139421},
pmid = {40782784},
issn = {1873-3336},
abstract = {Urban lakes often serve as the key nodes of material and energy flows in the river network ecosystems in large-scale cities. A comprehensive understanding of the multimedium distribution and fate of antibiotics in the lacustrine water-sediment-biofilm system is the critical prerequisite for their accurate ecological risk assessment and regional control and remediation. This study presents a comprehensive investigation of 21 antibiotics across six families in the water, sedimentary and biofilm phases of six typical urban lakes in Nanjing, a large-scale city in eastern China. The fates of these pollutants in multiple environmental compartments and their risks were assessed under real urban lake scenarios. The results indicated that the target antibiotics were ubiquitously present in the lakes. Quinolones (QNs) dominated all the lacustrine matrices, i.e., 10.63-38.78 ng/L in water, 0.4-12.77 ng/g in sediments, and not detected (ND) ∼73.30 ng/g in biofilms; wherein biofilms exhibit a much greater adsorption capacity to the chemicals than sediments, could serve as pollution markers owing to the significantly positive correlation between the concentration of QNs in the biofilm phase and the intensity of human activity (r = 0.92, p < 0.01). Notable differences in the cooccurrence patterns of antibiotics, metals and environmental factors among the biofilm, water and sedimentary phases were detected, of which the network in the biofilms was the most robust. An innovative model was developed to calculate the fugacity fractions of antibiotics in water, sediments and biofilms simultaneously, and the results suggested that biofilm accumulation would be the main fate of the target antibiotics in urban lakes. Risk assessment indicated that NOR posed a high ecological risk to algae in Xuanwu Lake, whereas most antibiotics posed low to no risk to aquatic organisms in urban lakes. Moreover, low antibiotic resistance development risks were obtained, except for ENR in Xuanwu Lake, which exhibited a moderate resistance risk. This study bridges a gap in knowledge of the distribution and environmental fate of antibiotics in the water, sediments and biofilms of urban lakes in large cities.},
}

@article {pmid40782482,
year = {2025},
author = {Ando, D and Lwin, HY and Aoki-Nonaka, Y and Matsugishi-Nasu, A and Minato, Y and Warita, Y and Takahashi, N and Tabeta, K},
title = {Ferulic acid suppresses Porphyromonas gingivalis biofilm formation via the inhibition of autoinducer-2 production and receptor activity.},
journal = {Archives of oral biology},
volume = {178},
number = {},
pages = {106365},
doi = {10.1016/j.archoralbio.2025.106365},
pmid = {40782482},
issn = {1879-1506},
abstract = {OBJECTIVE: This study aimed to clarify the antibacterial and antibiofilm effects of ferulic acid against periodontal pathogenic bacteria.

DESIGN: The cytotoxicity of ferulic acid was examined using the MTT assay on the human oral epithelial cell line Ca9-22. To determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration, Porphyromonas gingivalis ATCC 33277, Fusobacterium nucleatum ATCC 25586, Prevotella intermedia ATCC 25611, Aggregatibacter actinomycetemcomitans JP2, and Streptococcus mitis ATCC 903 were treated with ferulic acid. The inhibition of biofilm formation was evaluated by crystal violet staining. The inhibition of P. gingivalis autoinducer-2 (AI-2) production and receptor activity was evaluated by luminescence measurements using the sensor strain Vibrio harveyi BB170.

RESULTS: Ferulic acid did not exhibit any cytotoxicity on human oral epithelial cells. The MICs of ferulic acid against P. gingivalis and A. actinomycetemcomitans were 1000 and 500 µg/mL, respectively. It did not show antibacterial activity against F. nucleatum, P. intermedia, and S. mitis, indicating the weak antibacterial activity of ferulic acid. However, ferulic acid significantly inhibited P. gingivalis biofilm formation at low concentrations below 1/8 MIC. It specifically inhibited AI-2 production from P. gingivalis below 1/8 MIC and suppressed the receptor activity of AI-2.

CONCLUSIONS: Although ferulic acid showed weak antibacterial activity against periodontopathogenic bacteria, it had low cytotoxicity and inhibited P. gingivalis biofilm formation. Ferulic acid inhibited AI-2 production and receptor activity, suggesting that ferulic acid is an efficient quorum-sensing inhibitor for controlling P. gingivalis biofilm formation.},
}

@article {pmid40782449,
year = {2025},
author = {Li, H and Feng, Y and Lin, B and Zhang, S and Ren, Y and Yue, J},
title = {Polyurea-based multimodal interaction nanogels for synergistic bacterial biofilm eradication and prevention of re-colonization.},
journal = {Biomaterials},
volume = {325},
number = {},
pages = {123607},
doi = {10.1016/j.biomaterials.2025.123607},
pmid = {40782449},
issn = {1878-5905},
abstract = {Bacterial biofilm eradication and prevention of re-colonization are critical for effective treatment of biofilm-associated infections. Although significant progress has been made in nanovehicle-assisted antimicrobial platforms for biofilm eradication, strategies to address re-colonization remain underdeveloped. In this study, we constructed a versatile antimicrobial delivery platform based on multimodal interaction polyurea nanogels (MIPN). MIPN demonstrated excellent biocompatibility and could effectively load various antimicrobials with high capacity due to the multiple intermolecular interactions between the antimicrobials and nanocarriers, including hydrogen bonding, electrostatic, and hydrophobic interactions. By incorporating self-synthesized quorum sensing inhibitors (QSI) within MIPN, bacteria re-colonization was successfully prevented by blocking the quorum sensing pathway and disrupting surface-associated bacterial motilities. Furthermore, MIPN coloaded with QSI- and antibiotics showed a synergistic effect on biofilm eradication and re-colonization prevention, significantly enhancing the healing of biofilm-associated infections in chronic wounds.},
}

@article {pmid40778760,
year = {2025},
author = {Kalinina, I and Wilson, D},
title = {Candida albicans Goliath cells pioneer biofilm formation.},
journal = {mBio},
volume = {},
number = {},
pages = {e0342524},
doi = {10.1128/mbio.03425-24},
pmid = {40778760},
issn = {2150-7511},
abstract = {Candida albicans is a fungal pathogen that can cause life-threatening bloodstream infections. Indwelling medical devices, such as catheters, play a critical role in their progression, as C. albicans can colonize these devices and form biofilms, which can then seed the bloodstream, resulting in recurrent candidemia. Under conditions of zinc restriction, C. albicans forms enlarged Goliath cells. Here, we show that Goliath cells exhibit enhanced adhesion to host tissue under static conditions, particularly to vaginal and oral epithelial cell monolayers. Moreover, Goliath cells are fully capable of forming hyphae and damaging host tissue. Notably, the adhesion of Goliath cells to abiotic surfaces surpasses their adhesion to epithelia or endothelia, suggesting that Goliath cells may possess a unique advantage in colonizing indwelling medical devices, such as catheters within the circulatory system. We also show that the cell wall of Goliath cells has increased hydrophobicity compared with yeasts. In microfluidic channels mimicking the shear stress present in the circulatory system, yeast cells exhibit negligible adhesion to plastic surfaces, even at minimal flow rates. In contrast, Goliath cells demonstrate robust adherence to abiotic surfaces, maintaining attachment even under supraphysiological shear stress conditions. We show that biofilms formed by Goliath cells are more metabolically active and structurally denser in terms of the volume of hyphae. A high number of adhesive "pioneer" Goliath cells in the base of the biofilm significantly affects their architecture and anchors them to abiotic surfaces. These insights into Goliath cells' pathogenicity shed light on their potential role in colonization of medical devices.IMPORTANCEGoliath cells, known for their large size and stickiness to plastic, are triggered by limited access to the essential mineral zinc. They are a specialized cell type of Candida albicans, a fungus that causes skin, oral, and vaginal yeast infections as well as severe bloodstream infections. Medical devices like catheters can make these bloodstream infections worse because Candida forms biofilms on them, which results in continuous seeding of fungal cells into the bloodstream. We found that Goliath cells adhere to vaginal and oral tissue better than normal yeast cells. Surprisingly, they stick even better to abiotic surfaces than to mammalian cells. This suggests they might have an advantage in attaching to catheters during Candida bloodstream infection. When we simulated blood flow, regular yeast cells did not stick to plastic surfaces, but Goliath cells attach even at very high flow rates. This allowed them to form much thicker and active biofilms on these surfaces. Understanding how Goliath cells work can help us figure out better ways to prevent and treat infections caused by Candida albicans, especially those related to medical devices like catheters.},
}

@article {pmid40778403,
year = {2025},
author = {Zuo, R and Fu, L and Pang, W and Kong, L and Weng, L and Sun, Z and Li, R and Qu, S and Li, L},
title = {Self-assembled and intestine-targeting florfenicol nano-micelles effectively inhibit drug-resistant Salmonella typhimurium, eradicate biofilm, and maintain intestinal homeostasis.},
journal = {Journal of pharmaceutical analysis},
volume = {15},
number = {7},
pages = {101226},
pmid = {40778403},
issn = {2214-0883},
abstract = {Antimicrobial resistance (AMR) is a growing public health crisis that requires innovative solutions. Emerging multidrug resistant (MDR) Salmonella typhimurium has raised concern for its effect on pathogenic infection and mortality in humans caused by enteric diseases. To combat these MDR Salmonella typhimurium pathogens, highly effective and broad-spectrum antibiotics such as flufenicol (FFC) need to be evaluated for their potent antibacterial activity against Salmonella typhimurium. However, the low solubility and low oral bioavailability of flufenicol need to be addressed to better combat AMR. In this work, we develop a novel nano-formulation, flufenicol nano-micelles (FTPPM), which are based on d-α-tocopherol polyethylene glycol 1,000 succinate (TPGS)/poloxamer 188 (P188), for the targeted treatment of biofilms formed by drug-resistant Salmonella typhimurium in the intestine. Herein, FTPPM were prepared via a thin film hydration method. The preparation process for the mixed micelles is simple and convenient compared with other existing nanodrug delivery systems, which can further decrease production costs. The optimized FTPPM demonstrated outstanding stability and sustained release. An evaluation of the in vivo anti-drug-resistant Salmonella typhimurium efficacy demonstrated that FTPPM showed a stronger efficacy (68.17 %) than did florfenicol-loaded TPGS polymer micelles (FTPM), flufenicol active pharmaceutical ingredients (FFC-API), and flufenicol commercially available medicine (FFC-CAM), and also exhibited outstanding biocompatibility. Notably, FTPPM also inhibited drug-resistant Salmonella typhimurium from forming biofilms. More importantly, FTPPM effectively restored intestinal flora disorders induced by drug-resistant Salmonella typhimurium in mice. In summary, FTPPM significantly improved the solubility and oral bioavailability of florfenicol, enhancing its efficacy against drug-resistant Salmonella typhimurium both in vitro and in vivo. FTPPM represent a promising drug-resistant Salmonella typhimurium treatment for curbing bacterial resistance via oral administration.},
}

@article {pmid40772334,
year = {2025},
author = {Wang, X and Wang, M and Yang, L and Li, G and Xu, L},
title = {Green-Synthesized Multifunctional Carbon Dots from a Natural Nucleoside for Synergistic Antibacterial and Anti-Biofilm Therapies.},
journal = {Small (Weinheim an der Bergstrasse, Germany)},
volume = {},
number = {},
pages = {e04348},
doi = {10.1002/smll.202504348},
pmid = {40772334},
issn = {1613-6829},
support = {2025D04J0052//Science and Technology Planning Project of Guangzhou City/ ; 202002030089//Science and Technology Planning Project of Guangzhou City/ ; 241104218034882//Science and Technology Planning Project of Shaoguan City/ ; 31971314//National Science Foundation of China/ ; },
abstract = {The pursuit of sustainable antimicrobial strategies to combat biofilm infections has become imperative in addressing the global crisis of antibiotic resistance. Here, a green-chemistry paradigm is reported for fabricating multicolor-emissive carbon dots (A-CDs) through a solvent-free, single-step solvothermal process using adenosine. Under dual-light irradiation, A-CDs displayed enhanced antimicrobial activity against both bacteria and fungi through a triple-modal mechanism. RNA sequencing reveals that A-CDs combined with dual light irradiation disrupt bacterial metabolism, antioxidant systems, and cell wall integrity, effectively inhibiting bacterial proliferation and pathogenicity. A-CDs significantly inhibit biofilm formation and eliminate mature biofilms by destroying key components of extracellular polymeric substances and downregulating the key biofilm markers under laser light exposure. Its antibiofilm efficacy is further validated in a S. aureus biofilm-infected mouse model. Additionally, A-CDs exhibit high biocompatibility and minimal potential for resistance development over 25 days, making them a promising sustainable alternative to conventional antibiotics. These findings suggest that A-CDs can serve as an environmentally friendly, antibiotic-free solution for treating biofilm-related infections, offering broad clinical applications.},
}

@article {pmid40771684,
year = {2025},
author = {Huang, J and Fu, Q and Shao, X and Li, Y},
title = {Ultrasonic strategies for mitigating microbial adhesion and biofilm formation on medical surfaces: a mini review.},
journal = {Frontiers in microbiology},
volume = {16},
number = {},
pages = {1558035},
pmid = {40771684},
issn = {1664-302X},
abstract = {Biofilm formation on medical surfaces poses significant challenges, leading to compromised device functionality and an increased risk of infections. Addressing this issue requires effective strategies that balance efficacy with safety. This mini-review examines the application of ultrasound as a promising approach for biofilm control in medical contexts. Drawing from recent studies, it explores the mechanisms by which ultrasound disrupts biofilms, highlighting its ability to break down extracellular polymeric matrices and enhance the efficacy of antimicrobials. The review also discusses practical considerations, including ultrasound parameter optimization, biocompatibility, and integration with other anti-biofilm strategies. While ultrasound has demonstrated potential in disrupting biofilms, further research is essential to refine these approaches, improve treatment outcomes, and ensure compatibility with medical applications. By advancing our understanding and application of ultrasonic techniques, this field holds promise for improving patient safety and enhancing medical device longevity.},
}

@article {pmid40769254,
year = {2025},
author = {Zhuang, J and Ai, W and Wang, Q and Wang, H and Wu, B and Xu, X and Huang, T},
title = {Sludge reflux promotes biofilm formation in upflow membrane aerated biofilm reactor for partial nitritation: insight from computational fluid dynamics stimulation and experiments.},
journal = {Bioresource technology},
volume = {437},
number = {},
pages = {133077},
doi = {10.1016/j.biortech.2025.133077},
pmid = {40769254},
issn = {1873-2976},
abstract = {Achieving partial nitritation (PN) process using membrane aerated biofilm reactor (MABR) is a novel approach to provide nitrite for mainstream anammox, gained increasing interests. However, insufficient microbial-membrane attachment raises a challenge for biofilm formation in MABR-PN. This study employed computational fluid dynamics modeling to evaluate sludge volume fraction in biofilm formation zones, using liquid reflux and sludge reflux as comparative strategies for enhancing biomass attachment. The optimal performance was obtained in sludge reflux (5 min/4 h), with 40 % sludge volume fraction and the uniformity of sludge distribution in the biofilm formation zone was 80 %. Two bench-scale MABR were developed with liquid reflux (R1:400 % reflux) and sludge reflux (R2:5 min/4 h) under influent NH4[+]-N concentration of 50 ± 2 mg/L. The biofilm formation time in R2 was significantly shortened 30 % compared to R1, and the nitrite accumulation rate of R2 was significantly higher than that of R1 by 18.5 %. 16S rRNA gene sequencing coupled with observation revealed that the extracellular polymeric substance (EPS)-producing microorganism was enriched in biofilm of R2, meanwhile the abundance of the Nitrosomonas in R2_biofilm was significantly higher than flocs. Moreover, in R2, the abundance of key synthesis genes for EPS in biofilm was 2.4-fold higher than in flocs. The EPS concentration determination showed that R2_biofilm > R1_flocs > R1_biofilm > R2_flocs, confirmed that the high EPS-producing capacity in R2 biofilms could promote biofilm formation. Sludge reflux could promote the formation of MABR biofilm and enable rapid start-up of the PN process. Additionally, the energy consumption of R2 was only 2.5 % of R1.},
}

@article {pmid40769248,
year = {2025},
author = {Sarosh, S and Ramaswami, S},
title = {The salinity tipping point: determining the impact of stepwise increase in salinity on nitrification and biofilm community structure in nitrifying packed bed biofilm reactors.},
journal = {Bioresource technology},
volume = {},
number = {},
pages = {133089},
doi = {10.1016/j.biortech.2025.133089},
pmid = {40769248},
issn = {1873-2976},
abstract = {Nitrification is a crucial process in wastewater treatment and is generally inhibited at high salinities. Although packed-bed biofilm reactors (PBBRs) are well-known to facilitate high nitrification rates (NRs ≥ 1 kg N·m[-3]·d[-1]), the influence of salinity on nitrifying PBBRs remains understudied. This study aims to investigate the effect of stepwise increment of salinity on nitrification process and biofilm community structure in high-rate nitrifying PBBRs. Five PBBRs operated at five different water up-flow velocities (1-20 m/h) and ammonia loading rate of about 1600 g N·m[-3]·d[-1] were systematically subjected to incremental salinity shifts from 0 to 35 ‰ over 340 days. All reactors exhibited remarkable resilience and efficiency, achieving nearly complete conversion of ammonia to nitrate, with ammonia-removals consistently >98 % and NRs ≈ 1570 g N·m[-3]·d[-1] for 0-20 ‰ salinities. However, a transient accumulation of NH4[+] and NO2[-] was observed at intermediate salinity levels of 25 ‰ and 30 ‰, with NRs dropping by about 40 %. Nevertheless, at 35 ‰, all reactors demonstrated exceptional nitrification performance, with efficiencies exceeding 95 % and NRs ≈ 1527 g N·m[-3]·d[-1]. Nitrosomonas emerged as the dominant ammonia-oxidizing genus, increasing in abundance from 2-7 % at 10 ‰ salinity to 9-21.3 % at 35 ‰. The nitrite-oxidizing community underwent a shift in dominance from Nitrospira (3-8 % at 10 ‰ to 0.65-2.3 % at salinity 35 ‰) to Phycisphaerales genus (which was undetected at 10-20 ‰ salinity, to 4.2-6.2 % and 15 % at higher salinity levels of 30 and 35 ‰). This study shows the possibility for realising efficient high-rate nitrification up to seawater-salinities using PBBRs, which is desirable across several sectors.},
}

@article {pmid40769225,
year = {2025},
author = {Liu, Q and Cao, X and Chen, M and Yue, X and Liu, L and Yang, R and Xu, B and Deng, H},
title = {Isolation, whole-genome analysis, and anti-biofilm activity of carbapenem-resistant Enterobacteriaceae phage KP-BW9 and its lysin Lys-BW9.},
journal = {Microbial pathogenesis},
volume = {208},
number = {},
pages = {107960},
doi = {10.1016/j.micpath.2025.107960},
pmid = {40769225},
issn = {1096-1208},
abstract = {Carbapenem-resistant Enterobacteriaceae (CRE) have been classified as "urgent priority pathogens" due to their high drug resistance and wide spread. Among them, carbapenem-resistant Escherichia coli (CREco) and carbapenem-resistant Klebsiella pneumoniae (CRKP) are the main types. Phages are naturally occurring viruses that can specifically infect and lyse host bacteria, making them an effective alternative to antibiotics. This study isolated and identified a CREco phage, KP-BW9. The sequencing of phage KP-BW9 showed a genome size of 59665 bp, a GC content of 56.67 %, and encoded 79 predicted proteins. It has a latent period and a lysis period, with strong adaptability to the environment, and a lysis rate of 41.67 % for CREco. Both phage KP-BW9 and lysin Lys-BW9 could effectively lyse Escherichia coli and Klebsiella pneumoniae, and significantly inhibit and eradicate the biofilm of their individual and mixed bacteria. As a novel anti-CRE phage, KP-BW9 offers an innovative solution for controlling multidrug-resistant bacterial infections.},
}

@article {pmid40768400,
year = {2025},
author = {Hsu, CR and Huang, YL and Hsu, PH and Huang, CH and Huang, YC and Kao, MT and Ye, YW},
title = {A novel klebsiella pneumoniae diguanylate cyclase contributes to intestinal cell adhesion, biofilm formation, iron utilization, and in vivo virulence by gastrointestinal infection.},
journal = {Virulence},
volume = {},
number = {},
pages = {2544882},
doi = {10.1080/21505594.2025.2544882},
pmid = {40768400},
issn = {2150-5608},
abstract = {Klebsiella pneumoniae is responsible for various infections such as bacteraemia, urinary tract infections, pneumonia, and liver abscesses. Multidrug-resistant K. pneumoniae infections pose a critical public health threat, often associated with high mortality rates. The emergence of hypervirulent K. pneumoniae has also raised global health concerns due to its invasive disease potential. Clinical studies suggest intestinal colonization by K. pneumoniae as a risk factor for subsequent infections but underlying mechanisms remain unclear. Cyclic di-GMP (c-di-GMP), a bacterial signalling molecule synthesized by diguanylate cyclases (DGCs), controls various cellular processes and is absent in higher organisms, making it an attractive target for antimicrobial development. In this study, we identified a novel DGC, designated as DgcG, in K. pneumoniae, which plays a pivotal role in gastrointestinal colonization and pathogenesis. Using genetic deletion and complementation analyses in a bacteraemia and liver abscesses-inducing strain Ca0437, we observed that DgcG promoted intestinal adherence, biofilm formation, iron utilization, and in vivo virulence. RT-qPCR showed that DgcG regulated genes involved in type 3 fimbrial expression and iron transport. In a gastrointestinal infection model of female BALB/cByl mice, dgcG deletion significantly reduced host mortality and bacterial load in the liver, spleen, and intestines, underscoring its role in enhancing bacterial survival and dissemination. Additionally, dgcG gene was found highly conserved and prevalent among diverse K. pneumoniae isolates. These findings provide new insights into c-di-GMP-mediated virulence regulation in K. pneumoniae and highlight DgcG as a potential therapeutic target for controlling K. pneumoniae infections, especially amidst the growing global antimicrobial resistance crisis.},
}

@article {pmid40768344,
year = {2025},
author = {Zou, Y and Liang, C and Tang, Z and Huang, Y and Liang, Q and Chen, L and Chen, J and Zhu, X},
title = {Biofilm-Responsive Nano-Antibiotics for Degradation of Extracellular Polymeric Substance Matrix and Reduction of Pathogenicity against Drug-Resistant Bacterial Infections.},
journal = {ACS nano},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsnano.5c09764},
pmid = {40768344},
issn = {1936-086X},
abstract = {Methicillin-resistant Staphylococcus aureus (MRSA) poses a major threat to global public health, particularly due to its biofilm-associated refractory infections. The dense three-dimensional structure of the extracellular polymeric substances matrix (EPS) is a key factor contributing to the challenge of eradicating biofilm-associated infections. Therefore, in this study, ultrasmall cobalt selenide nanoparticles (CoSe NPs) were synthesized and modified with dextran (Dex), successfully obtaining dextran-functionalized cobalt selenide nanoparticles (Dex@CoSe NPs). In vitro experimental results showed that Dex@CoSe NPs not only disperse biofilm by degrading components of the extracellular polymeric substance matrix such as amyloid fibers and polysaccharides but also generate reactive oxygen species (ROS) to promote bacterial cell membrane rupture for enhanced bactericidal effects. Additionally, Dex@CoSe NPs reduce MRSA pathogenicity by down-regulating genes related to the quorum sensing system and ABC transporters. More importantly, Dex@CoSe NPs were shown to effectively alleviate both chronic wound infections and systemic infections caused by MRSA. This study thus provides strategic insights for developing ultrasmall nanoparticles specifically designed to combat biofilm-associated infections.},
}

@article {pmid40767295,
year = {2025},
author = {Karhunen, PJ and Pessi, T and Hörkkö, S and Karhunen, V and Goebeler, S and Louhelainen, AM and Martiskainen, M and Haapaniemi, T and Lappeteläinen, J and Ijäs, T and Lyytikäinen, LP and Raitoharju, E and Sioris, T and Tuomisto, S and Huhtala, H and Wang, C and Monaco, C and Oksala, N and Lehtimäki, T and Laaksonen, R},
title = {Viridans Streptococcal Biofilm Evades Immune Detection and Contributes to Inflammation and Rupture of Atherosclerotic Plaques.},
journal = {Journal of the American Heart Association},
volume = {},
number = {},
pages = {e041521},
doi = {10.1161/JAHA.125.041521},
pmid = {40767295},
issn = {2047-9980},
abstract = {BACKGROUND: Bacterial DNA from the oral cavity, respiratory tract, gut, and skin has been detected in atherosclerotic plaques, suggesting a role in chronic inflammation linked to atherosclerosis. Chronic bacterial infections often form biofilms resistant to antibiotics and immune detection, giving rise to a new generation of virulent bacteria in suitable conditions. This study explores the role of the immune system in bacterial-induced inflammation of atherosclerotic plaques.

METHODS: Coronary plaques from 121 sudden death victims and endarterectomy samples from 96 surgical patients were analyzed using bacterial real-time quantitative polymerase chain reaction, immunohistochemistry, and genome-wide expression analysis. TLR (toll-like receptor) signaling was examined in bacterial-activated TLR cell lines.

RESULTS: Of the bacteria detected, oral viridans group streptococcal DNA was the most common, being found in 42.1% of coronary plaques and 42.9% of endarterectomies. Immunopositivity for viridans streptococci correlated with severe atherosclerosis (P<0.0001) in both series and death from coronary heart disease (P=0.021) or myocardial infarction (P=0.042). Viridans streptococci colonized the core of the atheroma as a biofilm unrecognized by macrophages of the innate immune system. In contrast, immunopositive streptococci that appeared to have originated from the biofilm infiltrated the ruptured fibrous cap of the atheroma in endarterectomy samples and coronary plaques and were detected by pattern-recognizing receptors and coexpressed with the adaptive immune response. Among the viridans streptococcal strains, TLR2 was the most activated bacterial-signaling pathway. Genome-wide expression analysis of endarterectomy samples showed upregulation of bacterial recognition pathways.

CONCLUSIONS: Latent chronic bacterial inflammation evades immune detection and may contribute to the pathogenesis of complicated atherosclerotic plaques and fatal myocardial infarction.},
}

@article {pmid40766083,
year = {2025},
author = {Palma, F and Díaz-Navarro, M and Visedo, A and Sanz-Ruíz, P and Brandi, G and Schiavano, GF and Guembe, M},
title = {Assessment of the anti-biofilm effect of UV-C irradiation (254 nm) against healthcare associated infections related microorganisms.},
journal = {Frontiers in microbiology},
volume = {16},
number = {},
pages = {1570334},
pmid = {40766083},
issn = {1664-302X},
abstract = {INTRODUCTION: Biofilm-related Multidrug Resistance (MDR) is a major problem in healthcare-associated infections (HAI). Hospital surface decontamination is essential to ensure the safety of patients and to eliminate the dissemination of MDR pathogens. New eco-friendly decontamination technologies, such as UV-C irradiation, are only gaining popularity now, but their use against the biofilm of common microorganisms causing HAI has not been properly assessed. We aimed to assess the efficacy of UV-C irradiation (254 nm) in a 2-phase study by assessing its anti-biofilm effect against sessile cells from microorganisms of hospital interest.

METHODS: The following strains were tested: methicillin-susceptible Staphylococcus aureus (MSSA) (ATCC 29213), methicillin-resistant Staphylococcus aureus (MRSA) (ATCC 43300), Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 15442, and Candida albicans (ATCC 14053), and a clinical strain of methicillin-resistant Staphylococcus epidermidis. First, the tested strains' UV-susceptibility was evaluated through irradiation tests on plates using different UV doses, considering both planktonic and 24 h-biofilm states. Second, the anti-biofilm effect of UV-C was evaluated on stainless steel discs contaminated with a 24 h-biofilm of each strain.

RESULTS: With a UV dose of 946.7 mJ/cm[2], the UV-C irradiation on MSSA ATCC 29213, MRSA ATCC 43300, and MRSE biofilm showed a log10 reduction of 4.34 ± 0.70, 4.70 ± 0.60, and 4.85 ± 0.98, respectively, while C. albicans ATCC 14053 showed higher UV-resistance in 24 h-biofilm state, being the log10 reduction of 3.17 ± 0.08. Against Gram negative bacteria biofilm, a UV dose of 467.8 mJ/cm[2] was enough to achieve a microbial titer <1 CFU/mL. Regarding the 24 h-biofilm on discs, a log10 reduction >3 logs was achieved with all microorganisms applying a UV dose of 946.7 mJ/cm[2].

CONCLUSION: The application of UV-C irradiation could be a valid additional approach in the management of biofilm HAI.},
}

@article {pmid40765761,
year = {2025},
author = {Daneshyar, F and Alikhani, MY and Tayebi, S and Abedi Firouzjaei, D},
title = {Evaluation of the Antibacterial and Anti-Biofilm Activity of Chitosan-Arginine Nanoparticles and Sodium Fluoride against Streptococcus mutans.},
journal = {International journal of molecular and cellular medicine},
volume = {14},
number = {2},
pages = {637-645},
pmid = {40765761},
issn = {2251-9637},
abstract = {Dental caries is among the most prevalent chronic diseases. It arises from bacterial biofilm formation on tooth surfaces due to metabolic activity. Streptococcus mutans (S. mutans) is a key pathogen implicated in the development of dental caries. As bacterial resistance to conventional treatments increases, there is a growing interest in using novel compounds that possess antibacterial and antibiofilm properties. This study evaluated the effect of chitosan-arginine nanoparticles (CS-Arg NPs) and sodium fluoride (NaF) on inhibiting S. mutans' growth. After synthesizing CS-Arg NPs, their size, morphology, and chemical structure were evaluated. The broth microdilution method determined the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CS-Arg NPs and NaF. The combined antibacterial and antibiofilm effect of CS-Arg NPs and NaF was assessed using the checkerboard method. The CS-Arg NPs had an average size of 269.9 nm with a zeta potential of +38.3 mV. The MIC of S. mutans for CS-Arg NPs and NaF was 312 µg/mL and 625 µg/mL, respectively, and the MBC for these NPs and NaF was 625 µg/mL and 2500 µg/mL, respectively. The fractional inhibitory concentration index (FICI) of the combination of CS-Arg NPs and NaF showed an additive effect (FICI = 1). The inhibitory effect of different concentrations of CS-Arg NPs and NaF, alone or in combination, on biofilm formation in the studied strain ranged from approximately 12% to 81%. This study demonstrated that CS-Arg NPs have antibacterial and antibiofilm properties against S. mutans, and their combination with NaF can enhance these antibacterial effects. These findings suggest that CS-Arg NPs and NaF, as a novel combination, could effectively develop oral hygiene products.},
}

@article {pmid40765753,
year = {2025},
author = {Yalçın, S and Özel, S and Çamyar, D and Bonabian, EF},
title = {Candida albicans Impact on the Progression, Morphology, and Cellular Integrity of Biofilm Formation on the Surfaces of Implants; Current Knowledge and Future Perspectives.},
journal = {International journal of molecular and cellular medicine},
volume = {14},
number = {2},
pages = {620-636},
pmid = {40765753},
issn = {2251-9637},
abstract = {The capacity of Candida albicans to adhere to diverse oral substrates constitutes a pivotal preliminary phase in the formation of a pathogenic fungal biofilm. Yeast cells demonstrate a considerable ability to bind to host tissues, encompassing dental structures and mucosal surfaces, in addition to synthetic, non-biological materials such as dental appliances. Biomaterials utilized for the restoration of oral functionality are prone to biofilm formation, which can detrimentally affect oral health. Oral microorganisms can adhere to both hydrophobic and hydrophilic surfaces; however, in vivo investigations indicate that hydrophobic surfaces tend to accumulate minimal biofilm due to differential shear forces. Rough surfaces are observed to retain more biofilm compared to their smooth counterparts. The presence of biofilms on composite materials and glass-ionomer cement types results in surface degradation, consequently fostering additional biofilm development. While the leaching of residual monomers from composites has been shown to influence biofilm proliferation in vitro, the effect in vivo appears to be less consequential, likely attributable to the dilution and continual renewal of saliva. Furthermore, research has produced inconsistent findings regarding the influence of fluoride release from glass-ionomer cement types. A comparative analysis is conducted between biomaterial-associated infections in implants and devices situated in other anatomical regions and the formation of oral biofilms. The discourse critically evaluates alterations to biomaterials aimed at diminishing biofilm formation on implants and devices, taking into account their prospective applications within dentistry. The conclusion reached is that for dental applications, antimicrobial coatings that exterminate fungi upon contact are deemed more efficacious than those that gradually release antimicrobial agents.},
}

@article {pmid40765099,
year = {2025},
author = {Bhattacharya, D and Gupta, J and Gupta, A and Nag, M and Lahiri, D and Mitra, D},
title = {In silico Analysis to Search for an Alternative Inhibitor of Biofilm Production by Helicobacter pylori.},
journal = {Current drug discovery technologies},
volume = {},
number = {},
pages = {},
doi = {10.2174/0115701638369494250723071458},
pmid = {40765099},
issn = {1875-6220},
abstract = {INTRODUCTION: Antibiotic Resistance is a growing concern in the current world. Regarding this, Helicobacter pylori is known to be present in the digestive tracts of human beings. In some people, the infection leads to stomach cancer. With the increasing problem of Antibiotic resistance, reports show that drugs are no longer working with the same efficacy rate as they used to. The present work is a study of the two-component Acid-Response System (ArsRS) protein, one of the key pro-teins involved in biofilm formation by H. pylori.

METHODS: In this study, an overall bioinformatics analysis was conducted of the bioactive compound, along with some FDA-approved drugs, to identify potential novel treatments for diseases associated with H. pylori. The molecular docking studies have been done with common drugs that are used for the treatment of the infection caused by H. pylori as Amoxicillin, Clarithromycin, Tetracycline, Levofloxacin, Metronidazole, Omeprazole, and Rabeprazole. Another aspect of the current study was to investigate the potential for national compounds such as Quercetin, α-mangostin, Phytol, Liquiri-tin, and D-mannitol as an alternative therapeutic agent. ADMET Analysis and toxicity assessment were done to check the pharmacokinetics of the bioactive compounds. An in silico investigation of the H. pylori protein revealed its stability and compactness.

RESULTS: A Higher number of intra-protein interactions increases the stability of H. pylori protein. Liquiritin emerged as an active molecule that can be used for inhibiting H. pylori biofilms. The FDA-approved drug Clarithromycin showed the highest binding energy among the synthetic group.

DISCUSSIONS: The target protein's structural and sequential analysis demonstrated how the correct number of amino acids boosts the protein's stability. The ability of bioactive chemicals to function as medications is indicated by their drug-likeness characteristics. According to toxicity evaluation, they do not have any serious effects when they come into contact with people through the environment. Liquiritin and Clarithromycin were the two top compounds that appeared as the best inhibitors.

CONCLUSIONS: Bioactive compounds that can be used as drugs. This work will also be beneficial for the development of synthetic drugs.},
}

@article {pmid40764584,
year = {2025},
author = {Kimijima, M and Narisawa, N and Hama, Y and Nakagawa-Nakamura, T and Ito, T},
title = {Isofloridoside: a novel inhibitor of Streptococcus mutans biofilm formation and glucosyltransferase activity.},
journal = {BMC research notes},
volume = {18},
number = {1},
pages = {343},
pmid = {40764584},
issn = {1756-0500},
support = {22K02143//the Grants-in-Aid for the Development of Scientific Research from the Ministry of Education, Science, and Culture of Japan./ ; },
mesh = {*Biofilms/drug effects/growth & development ; *Streptococcus mutans/drug effects/enzymology/physiology/growth & development ; *Glucosyltransferases/antagonists & inhibitors/metabolism ; Sucrose/pharmacology ; Melibiose/pharmacology ; Xylitol/pharmacology ; Galactose/pharmacology ; },
abstract = {OBJECTIVE: Isofloridoside, a galactose-containing heteroside derived from marine red algae, has potential applications as a sweetener because it can activate the sweetness receptors T1R2/T1R3. The purpose of this study was to investigate the effects of isofloridoside on the growth and sucrose-dependent biofilm formation of the cariogenic bacterium Streptococcus mutans, to evaluate its potential as a caries-preventing agent.

RESULTS: The results showed that S. mutans did not grow when isofloridoside was the sole carbon source. Isofloridoside inhibited sucrose-dependent biofilm formation by S. mutans in a concentration-dependent manner, similar to galactose and glucose, but unlike melibiose and galactose-containing disaccharides. Biofilm inhibition induced by isofloridoside was associated with inhibition of glucosyltransferase activity. Isofloridoside exhibited biofilm inhibition comparable to xylitol, an alternative sugar known to inhibit biofilm formation. The differential effects of isofloridoside and melibiose on biofilm formation may result from the structural differences that affect their interactions with S. mutans enzymes. These findings highlight the potential of galactose and its polysaccharides as regulators of S. mutans biofilm formation, and suggest that isofloridoside is a promising alternative sweetener for caries prevention.},
}

*Biofilms/drug effects/growth & development
*Streptococcus mutans/drug effects/enzymology/physiology/growth & development
*Glucosyltransferases/antagonists & inhibitors/metabolism
Sucrose/pharmacology
Melibiose/pharmacology
Xylitol/pharmacology
Galactose/pharmacology

@article {pmid40764534,
year = {2025},
author = {Tamrat, E and Asmare, Z and Geteneh, A and Sisay, A and Getachew, E and Kassanew, B and Dessale, M and Gashaw, Y and Jemal, A and Gashaw, M and Bazezew, A and Gedfie, S and Kassahun, W and Abebe, W and Dejazmach, Z and Misganaw, T and Ashagre, A and Nigatie, M and Damtie, AA and Alemu, BB and Tefera, Z and Mezgebu, B and Kumie, G and Kiros, M and Reta, MA},
title = {The global prevalence of biofilm-forming Enterococcus faecalis in clinical isolates: a systematic review and meta-analysis.},
journal = {BMC infectious diseases},
volume = {25},
number = {1},
pages = {981},
pmid = {40764534},
issn = {1471-2334},
mesh = {*Enterococcus faecalis/physiology/isolation & purification ; *Biofilms/growth & development ; Humans ; Prevalence ; *Gram-Positive Bacterial Infections/epidemiology/microbiology ; Global Health ; Cross Infection/microbiology/epidemiology ; },
abstract = {BACKGROUND: Enterococcus faecalis (E. faecalis) is a major cause of healthcare-associated infections (HAIs). It exhibits a strong biofilm-forming ability, which contributes to treatment resistance and persistence. Despite its clinical relevance, the global prevalence of biofilm-forming E. faecalis remains poorly defined. This study aimed to estimate the pooled prevalence of biofilm-forming E. faecalis in clinical isolates worldwide.

METHODS: Following PRISMA 2020 guidelines, we systematically searched PubMed, Scopus, ScienceDirect, Google Scholar, and institutional repositories for studies published between 2015 and 2024. A total of 56 studies involving 3,739 clinical isolates met the inclusion criteria. We used a random-effects model to estimate pooled prevalence and conducted subgroup analyses based on WHO region, continent, publication year, specimen type, and biofilm detection method. Meta-regression and sensitivity analyses assessed heterogeneity and robustness. Publication bias was evaluated using Egger's test and corrected with trim-and-fill analysis.

RESULTS: The global pooled prevalence of biofilm-forming E. faecalis was 68.68% (95% CI: 61.33-76.02%), with significant heterogeneity (I² = 99.30%). By WHO region, prevalence ranged from 57.93% (95% CI: 41.01-71.85%) in South-East Asia to 73.66% (95% CI: 63.40-83.92%) in the Eastern Mediterranean. By continent, South America (all from Brazil) showed the highest prevalence at 89.79% (95% CI: 73.02-106.56%). Studies from 2021 to 2024 reported higher prevalence (76.18%, 95% CI: 66.25-86.11%) than those from 2015 to 2020. Among specimens, urine showed the highest prevalence (80.47%, 95% CI: 61.17-99.77%). Among biofilm-positive isolates, 47.92% (95% CI: 39.34-56.51%) were strong producers. Meta-regression identified WHO region (p = 0.005) and specimen type (p = 0.043) as significant sources of heterogeneity. Egger's test indicated publication bias (p = 0.0066), but trim-and-fill analysis yielded a consistent adjusted prevalence of 68.08%.

CONCLUSION: Biofilm formation is highly prevalent in E. faecalis clinical isolates globally, with substantial regional and specimen-based variation. These findings highlight the urgent need for standardized biofilm detection protocols, improved infection prevention and control, tailored antibiotic stewardship, and the development of anti-biofilm therapies to mitigate biofilm-associated resistance and enhance patient outcomes.},
}

*Enterococcus faecalis/physiology/isolation & purification
*Biofilms/growth & development
Humans
Prevalence
*Gram-Positive Bacterial Infections/epidemiology/microbiology
Global Health
Cross Infection/microbiology/epidemiology

@article {pmid39858311,
year = {2025},
author = {Honegger, AL and Schweizer, TA and Achermann, Y and Bosshard, PP},
title = {Antimicrobial Efficacy of Five Wound Irrigation Solutions in the Biofilm Microenvironment In Vitro and Ex Vivo.},
journal = {Antibiotics (Basel, Switzerland)},
volume = {14},
number = {1},
pages = {},
pmid = {39858311},
issn = {2079-6382},
support = {INOV00149//USZ Innovationspool/ ; INOV00153//USZ Innovationspool/ ; },
abstract = {Background/Objectives: Periprosthetic joint infections (PJI) are difficult to treat due to biofilm formation on implant surfaces and the surrounding tissue, often requiring removal or exchange of prostheses along with long-lasting antibiotic treatment. Antiseptic irrigation during revision surgery might decrease bacterial biofilm load and thereby improve treatment success. This in vitro study investigated and compared the effect of five advanced wound irrigation solutions to reduce bacterial burden in biofilm microenvironment. Methods: We treated in vitro biofilms grown on titanium alloy implant discs with clinical bacterial strains isolated from patients with PJIs, as well as abscess communities in a plasma-supplemented collagen matrix. The biofilms were exposed for 1 min to the following wound irrigation solutions: Preventia[®], Prontosan[®], Granudacyn[®], ActiMaris[®] forte ('Actimaris'), and Octenilin[®]. We measured the bacterial reduction of these irrigation solutions compared to Ringer-Lactate and to the strong bactericidal but not approved Betaseptic solution. Additionally, ex vivo free-floating bacteria isolated directly from clinical sonication fluids were treated in the same way, and regrowth or lack of regrowth was recorded as the outcome. Results: Irrigation solutions demonstrated variable efficacy. The mean CFU log10 reduction was as follows: Octenilin, 3.07, Preventia, 1.17, Actimaris, 1.11, Prontosan, 1.03, and Granudacyn, 0.61. For SACs, the reduction was: Actimaris, 8.27, Octenilin, 0.58, Prontosan, 0.56, Preventia, 0.35, and Granudacyn, 0.24. Conclusions: AAll solutions achieved complete bacterial eradication in all tested ex vivo sonication fluids, except Granudacyn, which was ineffective in 33% of the samples (2 out of 6). Advanced wound irrigation solutions have the potential to reduce bacterial burden in the biofilm microenvironment. However, their efficacy varies depending on bacterial species, growth state, and the composition of the irrigation solution. While Octenilin should be avoided for deep tissue irrigation due to its potential to cause tissue necrosis, the clinical benefit of wound irrigation solutions in infection prevention warrants further investigation in prospective clinical trials.},
}

@article {pmid40762829,
year = {2025},
author = {de Sá Barreto Maia Leite, DP and de Oliveira, PRF and Oliveira, FCM and Melo-Santos, G and Maricato, G and Beraldo, KRF and Rodrigues, ALF and Damasceno, AAP and Oliveira, RHT and Fraga-Silva, TFC and Juliano, MA and Monteiro, FOB and Silva-Júnior, A and Mota, RA},
title = {Antimicrobial resistance and biofilm production by Staphylococcus spp. and Mammaliicoccus Sciuri in Inia araguaiaensis: surveillance under a one health perspective in the Amazon.},
journal = {Veterinary research communications},
volume = {49},
number = {5},
pages = {271},
pmid = {40762829},
issn = {1573-7446},
mesh = {Animals ; *Biofilms/growth & development/drug effects ; Brazil/epidemiology ; *Staphylococcus/drug effects/physiology ; *Anti-Bacterial Agents/pharmacology ; *Drug Resistance, Bacterial ; Microbial Sensitivity Tests ; *Staphylococcal Infections/veterinary/microbiology/epidemiology ; },
abstract = {Antimicrobial resistance in natural environments poses an increasing threat to public health and biodiversity conservation. This study investigated antimicrobial resistance and biofilm formation in bacteria isolated from Inia araguaiaensis, an Amazonian endemic dolphin species. Skin lesion samples were collected from eight individuals in the Mocajuba region, Pará, Brazil. Bacterial isolation was performed on Brain Heart Infusion agar and Mannitol Salt agar. Species identification was carried out using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and antimicrobial susceptibility testing was conducted by disk diffusion following. Resistance genes were detected by polymerase chain reaction, and biofilm formation was assessed using the crystal violet staining method. A total of 19 bacterial isolates were recovered, including Staphylococcus spp. (n = 14) and Mammaliicoccus sciuri (n = 5). The most frequently identified species were Staphylococcus aureus (4/19; 21.1%), S. warneri (4/19; 21.1%), and S. epidermidis (4/19; 21.1%). A high prevalence of penicillin resistance was observed (18/19; 94.7%), followed by resistance to erythromycin, tetracycline, and sulfamethoxazole-trimethoprim (each 4/19; 21.1%), and gentamicin (3/19; 15.8%). Four isolates (4/19; 21.1%) exhibited a multidrug-resistant (MDR) profile. Genotypic screening revealed the presence of the blaZ gene in 8/19 isolates (44.4%), followed by mecA, msrA, and norC (each 2/19; 10.5%). The tetracycline resistance genes tetL and tetM were detected in 3/19 (15.8%) and 1/19 (5.3%) isolates, respectively. Biofilm formation was detected on 15/19 (78.9%) of the isolates, of which 4/15 (26.7%) were classified as strong producers, 1/15 (6.7%) as moderate, and 10/15 (66.7%) as weak producers. These findings support the recognition of river dolphins as potential sentinels for antimicrobial resistance in aquatic ecosystems and reinforce the importance of integrating microbiological monitoring into broader conservation efforts under the One Health perspective.},
}

Animals
*Biofilms/growth & development/drug effects
Brazil/epidemiology
*Staphylococcus/drug effects/physiology
*Anti-Bacterial Agents/pharmacology
*Drug Resistance, Bacterial
Microbial Sensitivity Tests
*Staphylococcal Infections/veterinary/microbiology/epidemiology

@article {pmid40762578,
year = {2025},
author = {Shahrour, H and Alves Ferreira, D and Fitzgerald-Hughes, D and O'Gara, JP and Coffey, A and O'Neill, E},
title = {Use of the bacteriophage-derived endolysin CHAPK-SH3blys as a potent novel treatment for biofilm-associated Staphylococcus aureus wound infections.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0071625},
doi = {10.1128/spectrum.00716-25},
pmid = {40762578},
issn = {2165-0497},
abstract = {The pathogenesis of Staphylococcus aureus acute and chronic wound infections is associated with biofilm formation. This study investigated the antimicrobial and antibiofilm potential of CHAPK-SH3blys, a bacteriophage-derived endolysin, against S. aureus strains implicated in chronic wound infections. CHAPK-SH3blys exhibited potent antibacterial activity with a minimum inhibitory concentration (MIC) of 3.9 µg/mL. Biofilm assays conducted under static and dynamic wound-like conditions showed a concentration-dependent reduction in biofilm metabolic activity, with up to a 4-log reduction in viable cells at concentrations 50 times the MIC. Further investigations revealed that CHAPK-SH3blys showed biocompatibility with human cell lines and did not induce an inflammatory response. Notably, prolonged exposure to sub-MIC levels did not lead to resistance development in S. aureus. These findings highlight CHAPK-SH3blys as a promising therapeutic agent for the treatment of biofilm-associated S. aureus wound infections.IMPORTANCESuccessful treatment of wound infections, which is most commonly caused by the bacteria Staphylococcus aureus, is compromised by bacterial colonization resulting in slime layer or biofilm formation in the wound bed. Associated antimicrobial resistance (AMR) and limited treatment options can also impair wound healing and further highlight the need for the development and use of novel antimicrobial therapies. This study explores the antibiofilm and antimicrobial effectiveness of a bacteriophage-derived endolysin named CHAPK-SH3blys against S. aureus strains implicated in chronic wound infections. Bacteriophage-derived endolysin represents novel antimicrobial agents, and in this study, we demonstrate the effectiveness of this agent in treating wound and biofilm- related infections under wound-like conditions in the laboratory without affecting the immune response or inducing antimicrobial resistance. Due to the urgent public health need to develop alternative antimicrobial agents to combat biofilm-associated infections, our study highlights CHAPK-SH3blys as a promising and exciting candidate for further therapeutic development.},
}

@article {pmid40762463,
year = {2025},
author = {Le, VH and King, T and Wuerzberger, B and Bauer, OR and Carver, MN and Chan, TS and Henson, AL and Hubbard, GK and Kopadze, T and Patterson, CF and McGraw, SM and O'Hara, A and Yarkosky, EJ and LaMontagne, MG and Hotze, EM and Ferreira, RBR},
title = {Metabolites derived from bacterial isolates of the human skin microbiome inhibit Staphylococcus aureus biofilm formation.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0130625},
doi = {10.1128/spectrum.01306-25},
pmid = {40762463},
issn = {2165-0497},
abstract = {UNLABELLED: The human skin microbiome is a diverse ecosystem that can help prevent infections by producing biomolecules and peptides that inhibit growth and virulence of bacterial pathogens. Staphylococcus aureus is a major human pathogen responsible for diseases that range from acute skin and soft tissue infections to life-threatening septicemia. Its ability to form biofilms is a key virulence factor contributing to its success as a pathogen as well as to its increased antimicrobial resistance. Here, we investigated the ability of bacterial skin commensals to produce molecules that inhibit S. aureus biofilm formation. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) identified 77 human skin microbiome bacterial isolates from Staphylococcus and Bacillus genera. Metabolites from cell-free concentrated media (CFCM) from 26 representative isolates were evaluated for their ability to inhibit biofilm formation by both methicillin-resistant (MRSA) and methicillin-sensitive (MSSA) S. aureus strains. CFCM, derived from most of the isolates, inhibited biofilm formation to varying extents but did not inhibit planktonic growth of S. aureus. Size fractionation of the CFCM of three S. epidermidis isolates indicated that they produce different bioactive molecules. Cluster analysis, based on either MALDI-TOF mass spectra or whole-genome sequencing draft genomes, did not show clear clusters associated with levels of biofilm inhibition among S. epidermidis strains. Finally, similar biosynthetic gene clusters were detected in all S. epidermidis strains analyzed. These findings indicate that several bacterial constituents of the human skin microbiome display antibiofilm in vitro activity, warranting further investigation on their potential as novel therapeutic agents.

IMPORTANCE: The skin is constantly exposed to the environment and consequently to numerous pathogens. The bacterial community that colonizes healthy skin is thought to play an important role in protecting us against infections. S. aureus is a leading cause of death worldwide and is frequently involved in several types of infections, including skin and soft tissue infections. Its ability to adhere to surfaces and produce biofilms is considered an important virulence factor. Here, we analyzed the activity of different species of bacteria isolated from healthy skin on S. aureus biofilm formation. We found that some species of Staphylococcus and Bacillus can reduce S. aureus biofilm formation, although a generally lower level of inhibitory activity was observed compared to S. epidermidis isolates. Among S. epidermidis isolates, strength of activity was dependent on the strain. Our data highlight the importance of mining the skin microbiome for isolates that could help combat skin pathogens.},
}

@article {pmid40762038,
year = {2025},
author = {Wang, S and Liu, H and Zhong, H and Li, S and Yu, X and Lu, Y and Tang, Y and Wang, C and Yuan, Z},
title = {Manufacture and application of antibacterial biofilm microneedles.},
journal = {Journal of materials chemistry. B},
volume = {},
number = {},
pages = {},
doi = {10.1039/d5tb01029b},
pmid = {40762038},
issn = {2050-7518},
abstract = {Bacterial biofilms in chronic wounds establish drug-impermeable barriers through extracellular polymeric substance (EPS) matrix formation. Versus surgical debridement and systemic approaches, microneedle systems have evolved as minimally invasive platforms exhibiting marked progress in biofilm management. Microneedles enhance contact area with bacterial biofilms by >13.74% versus traditional wound dressings, improving treatment outcomes. This review systematically examines recent therapeutic innovations in microneedle-mediated biofilm intervention between 2018 and 2025. Firstly, this paper systematically reviews the action mechanisms of solid, coated, hollow, soluble, and hydrogel microneedles against bacterial biofilms. It then discuss the advanced fabrication methods for antibiofilm systems. Antimicrobial mechanisms are innovatively classified into three modalities, physical disruption, chemical targeting, and bioactive regulation. The following sections present preclinical applications corresponding to each mechanistic category. These results provide a roadmap for clinical translation of microneedle antimicrobial technologies.},
}

@article {pmid40761286,
year = {2025},
author = {Zhang, X and Zhang, D and Zhou, D and Zheng, S and Li, S and Hou, Q and Li, G and Han, H},
title = {A comprehensive review of the pathogenic mechanisms of Pseudomonas aeruginosa: synergistic effects of virulence factors, quorum sensing, and biofilm formation.},
journal = {Frontiers in microbiology},
volume = {16},
number = {},
pages = {1619626},
pmid = {40761286},
issn = {1664-302X},
abstract = {Pseudomonas aeruginosa (P. aeruginosa) is a ubiquitous opportunistic pathogen and a major cause of nosocomial infections worldwide. It can provoke a spectrum of clinical manifestations-ranging from postoperative wound infections, pressure ulcers, abscesses, and otitis media to life-threatening bacteremia and sepsis, especially in burn patients. Over the past decade, extensive research has elucidated its complex virulence repertoire, including exotoxins, proteases, and siderophores; the hierarchical Quorum Sensing (QS) networks; and its robust capacity for biofilm formation. In this review, we consolidate significant studies published since 2015 to develop a comprehensive framework elucidating the virulence mechanisms of P. aeruginosa. Beyond cataloging individual factors, we highlight how QS regulators coordinate toxin production and biofilm maturation, and how these processes converge to facilitate immune evasion. We further examine cross-talk between QS circuits (Las, Rhl, and Pqs), their response to environmental cues, and the modulatory role of host signals. Despite these advances, significant gaps remain: the spatiotemporal interplay among different virulence modules; the precise molecular triggers of biofilm dispersal; and the dynamics of bacterial-host immune interactions in vivo. Notably, targeting QS with small-molecule inhibitors has shown promise in attenuating pathogenicity, yet translating these findings into clinical therapies requires more nuanced understanding of resistance emergence and host microbiome effects. We propose that future investigations prioritize (1) the structural biology of QS receptors to guide rational inhibitor design; (2) single-cell and organ-on-a-chip models to dissect biofilm heterogeneity; (3) dual-omics approaches to map host-pathogen signaling crosstalk; and (4) environmental modulators-such as iron availability and shear stress-that fine-tune virulence expression. Such multidisciplinary efforts will underpin the development of next-generation anti-virulence therapies, ultimately improving prevention and treatment of P. aeruginosa infections and safeguarding public health.},
}

@article {pmid40761048,
year = {2025},
author = {Sanda, ZS and Vural, DG and Birinci, A},
title = {Investigation of biofilm formation in methicillin resistant Staphylococcus aureus isolates by genotypic and phenotypic methods and effect of vancomycin and teicoplanin on biofilm inhibition.},
journal = {Pakistan journal of pharmaceutical sciences},
volume = {38},
number = {4},
pages = {1133-1140},
doi = {10.36721/PJPS.2025.38.4.REG.13543.1},
pmid = {40761048},
issn = {1011-601X},
mesh = {*Biofilms/drug effects/growth & development ; *Methicillin-Resistant Staphylococcus aureus/drug effects/genetics/physiology/isolation & purification ; *Teicoplanin/pharmacology ; *Anti-Bacterial Agents/pharmacology ; Microbial Sensitivity Tests ; *Vancomycin/pharmacology ; Genotype ; Phenotype ; Humans ; Bacterial Proteins/genetics ; Penicillin-Binding Proteins ; },
abstract = {The pathogenicity factors of Staphylococcus aureus include biofilm production. In this study, the biofilm production abilities of methicillin-resistant S. aureus (MRSA) strains were investigated using genotypic and phenotypic methods. Additionally, the effect of glycopeptides on biofilm was examined. This study included 130 MRSA isolates. Biofilm was detected by the microtiter plate method. The minimum inhibitory concentration (MIC) of glycopeptides was evaluated through the broth microdilution method. The biofilm inhibitor concentration (BIC) values were investigated in isolates with strong biofilm production. The mecA (methicillin resistance gene), icaA, and icaD (biofilm-associated genes) were amplified by polymerase chain reaction techniques. Eighty-one isolates (62.31%) formed biofilms, while thirty isolates (23.08%) exhibited strong biofilm formation. Thirty isolates had higher BIC90 values than MIC90 values. The mecA gene was confirmed in 125 (96.15%) isolates, the icaA gene in 96 (73.85%) isolates, and the icaD gene in 100 (76.92%) isolates. There was statistical significance between ica genes and the biofilm produced (p<0.05). In conclusion, increased biofilm formation due to the effect of ica genes increases the concentration values at which antibiotics act.},
}

*Biofilms/drug effects/growth & development
*Methicillin-Resistant Staphylococcus aureus/drug effects/genetics/physiology/isolation & purification
*Teicoplanin/pharmacology
*Anti-Bacterial Agents/pharmacology
Microbial Sensitivity Tests
*Vancomycin/pharmacology
Genotype
Phenotype
Humans
Bacterial Proteins/genetics
Penicillin-Binding Proteins

@article {pmid40758696,
year = {2025},
author = {Thavorasak, T and Santajit, S and Tunyong, W and Kong-Ngoen, T and Reamtong, O and Ampawong, S and Saelim, N and Srisai, T and Aiumurai, P and Pumirat, P and Chaicumpa, W and Indrawattana, N},
title = {CRISPR/Cas9-targeted smpB mutation revealing roles in biofilm formation, motility, and antibiotic susceptibility in Acinetobacter baumannii.},
journal = {PloS one},
volume = {20},
number = {8},
pages = {e0329638},
pmid = {40758696},
issn = {1932-6203},
mesh = {*Biofilms/drug effects/growth & development ; *Acinetobacter baumannii/genetics/drug effects/pathogenicity/physiology ; *CRISPR-Cas Systems/genetics ; *Bacterial Proteins/genetics/metabolism ; *Anti-Bacterial Agents/pharmacology ; Mutation ; Animals ; Microbial Sensitivity Tests ; Virulence/genetics ; Gene Editing ; Acinetobacter Infections/microbiology/drug therapy ; },
abstract = {BACKGROUND: Acinetobacter baumannii is a multidrug-resistant pathogen and a major cause of hospital-acquired infections worldwide. Its ability to survive in harsh environments and evade antibiotic treatments underscores the urgent need for new therapeutic targets. Emerging evidence suggests that the small protein B (SmpB) may also play broader roles in bacterial virulence, including regulation of biofilm formation, motility, and stress adaptation. However, the specific contributions of SmpB to these pathogenic traits in A. baumannii remain poorly defined. Addressing this knowledge gap is essential for evaluating SmpB as a potential antimicrobial target and developing new strategies to combat multidrug-resistant infections.

METHODS: CRISPR/Cas9-mediated gene editing was used to generate a targeted smpB mutant in A. baumannii. The smpB mutant was assessed for growth, biofilm formation, motility, antibiotic susceptibility, and virulence. Biofilm was quantified via crystal violet staining and microscopy, while motility was examined using swimming, swarming, and twitching assays. Antibiotic susceptibility was evaluated using disk diffusion. Virulence was tested in the Galleria mellonella infection model. Proteomic analysis was performed to identify changes in protein expression associated with smpB disruption.

RESULTS: CRISPR/Cas9-mediated editing successfully introduced a C212T nucleotide substitution in the smpB gene, resulting in an A89G amino acid change. Growth curve analysis showed no significant difference between the wild-type and smpB mutant strains under nutrient-rich conditions. However, the mutant exhibited a significant reduction in biofilm formation (p = 0.0079) and impaired twitching motility, while swimming and swarming motility remained unaffected. Antibiotic susceptibility testing revealed increased sensitivity to ceftizoxime, piperacillin/tazobactam, and gentamicin, alongside decreased susceptibility to cefepime, tetracycline, and spectinomycin. In the G. mellonella infection model, the smpB mutant showed reduced virulence, with 84% larval survival compared to 72% in the wild type (p = 0.4183). Proteomic analysis revealed downregulation of key stress response and virulence-associated proteins, including GroEL, DnaK, RecA, and PirA, while proteins involved in ribosome maturation and transcription, such as RimP and RpoA, were upregulated. STRING network analysis supported the broad regulatory role of SmpB in biofilm formation, motility, stress adaptation, and pathogenesis.

CONCLUSION: This study demonstrates that SmpB is a key regulator of biofilm formation, twitching motility, antibiotic response, and virulence in A. baumannii. While not essential for growth under optimal conditions, smpB disruption impairs multiple pathogenic traits and alters stress-related proteomic pathways. These findings highlight the potential of SmpB as a novel antimicrobial target, offering a promising strategy to weaken bacterial virulence without promoting resistance. Targeting the trans-translation system may pave the way for innovative therapies against multidrug-resistant A. baumannii.},
}

*Biofilms/drug effects/growth & development
*Acinetobacter baumannii/genetics/drug effects/pathogenicity/physiology
*CRISPR-Cas Systems/genetics
*Bacterial Proteins/genetics/metabolism
*Anti-Bacterial Agents/pharmacology
Mutation
Animals
Microbial Sensitivity Tests
Virulence/genetics
Gene Editing
Acinetobacter Infections/microbiology/drug therapy

@article {pmid40758621,
year = {2025},
author = {Castagnini, D and Palma, K and Jara-Wilde, J and Navarro, N and González, MJ and Toledo, J and Canales-Huerta, N and Scavone, P and Härtel, S},
title = {Super-resolution Imaging of Proteus mirabilis Biofilm by Expansion Microscopy.},
journal = {Journal of visualized experiments : JoVE},
volume = {},
number = {221},
pages = {},
doi = {10.3791/67932},
pmid = {40758621},
issn = {1940-087X},
mesh = {*Proteus mirabilis/physiology ; *Biofilms/growth & development ; *Microscopy/methods ; },
abstract = {Accessing detailed visual information and quantitative data from microbiological samples using conventional optical microscopy is limited by the diffraction barrier. One solution to enhance resolution is Expansion Microscopy (ExM), an innovative and cost-effective super-resolution technique that physically enlarges samples by approximately four times their original size. For successful expansion, it is essential to homogenize the mechanical properties of the biological material. Biofilms are bacterial communities, adhering to a surface and embedded in an extracellular matrix they produce; they require ExM protocols to be adapted to accommodate their unique structural components. This article presents Proteus mirabilis biofilm ExM (PmbExM), a specialized variant of ExM that enables super-resolution visualization of P. mirabilis biofilms grown for 48 h. The protocol focuses on the targeted degradation of key structural components of samples through serial enzymatic digestions, optimized near their theoretical conditions. PmbExM utilizes a combination of enzymes, including α-amylase, cellulase, and lyticase glycoside-hydrolases for polysaccharide hydrolysis; mutanolysin for peptidoglycan hydrolysis; and proteinase K for protein hydrolysis. These digestion procedures are independent of the gelation process, allowing modifications to meet specific homogenization requirements in different biofilm models. This adaptability offers great potential for application across various bacterial species and growth conditions. ExM has been applied to different biofilm species with overall suboptimal expansion factors. In contrast, PmbExM achieves the theoretical maximum expansion factor of the standard acrylamide-acrylate ExM hydrogel, without significant distortion of morphology or topology. The aim of this work is to provide an accessible super-resolution protocol for visualizing the architecture, assembly, and cellular and intracellular features of P. mirabilis biofilms.},
}

*Proteus mirabilis/physiology
*Biofilms/growth & development
*Microscopy/methods

@article {pmid40757308,
year = {2025},
author = {Peri̇z, ÇD and Ulusoy, S and Kaya Kinaytürk, N},
title = {Molecular Docking and In Vitro Evaluation of Violacein-Alginate Beads for Targeted Inhibition of Biofilm Formation.},
journal = {ACS omega},
volume = {10},
number = {29},
pages = {31827-31839},
pmid = {40757308},
issn = {2470-1343},
abstract = {Violacein, produced by , exhibits significant antibacterial, antiviral, antifungal, and antioxidant properties. However, its poor aqueous solubility substantially limits its bioavailability. To overcome this constraint, violacein was encapsulated in sodium alginate beads, and its antibiofilm efficacy against was evaluated. Violacein-loaded alginate beads (VIABs) were synthesized and characterized using Fourier-transform infrared spectroscopy, scanning electron microscopy, and ultraviolet-visible spectrophotometry. Molecular docking analysis was further conducted to examine the interactions between violacein and the icaADBC proteins (UniProt accession: Q9RQP9, Q9RQP8, Q9RQP7, andQ9RQP6), which play a critical role in polysaccharide intercellular adhesin production during biofilm formation. The crude violacein extract displayed antibacterial activity, generating an inhibition zone of 12.3 ± 0.5 mm against . The average particle sizes of dry alginate beads and violacein-loaded alginate beads were 0.97 ± 0.16 and 0.66 ± 0.11 mm, respectively. VIABs (20-5 mg/mL) significantly suppressed biofilm formation by 77.4, 67.4, and 46.8%. Molecular docking analysis demonstrated strong binding affinities between violacein and the target proteins. Furthermore, violacein adhered to Lipinski's Rule of Five, suggesting favorable pharmacokinetic properties. These findings highlight the potential of VIABs as a promising therapeutic and food preservative agent due to their potent antibiofilm activity.},
}

@article {pmid40756206,
year = {2025},
author = {Niazy, AA and Alrashed, MM and Niazy, AA},
title = {Effect of 5-fluorouracil on Pseudomonas aeruginosa: impact on virulence, biofilm formation, and bacterial growth.},
journal = {Frontiers in microbiology},
volume = {16},
number = {},
pages = {1584479},
pmid = {40756206},
issn = {1664-302X},
abstract = {Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen that poses a significant threat to public health due to its inherent and acquired resistance to multiple antibiotics. This mini-review explores the antimicrobial potential of 5-fluorouracil (5-FU), a chemotherapeutic agent normally used in oncology, and currently under investigation for its bacteriostatic and antibiofilm effects against P. aeruginosa. 5-FU functions by inhibiting thymidylate synthase, disrupting nucleotide metabolism, and interfering with essential bacterial processes, such as quorum sensing. Studies in vitro have demonstrated its ability to inhibit biofilm creation and decrease virulence, but findings about its impact on preformed biofilms have been contradictory. Synergistic interactions between 5-FU and antibiotics, especially gentamicin, have shown promise in enhancing antibacterial effectiveness. The aim of this mini-review was to consolidate current findings, pinpoint research gaps, and suggest future directions for potentially repurposing 5-FU as an adjunctive treatment for P. aeruginosa. By placing the current evidence in context, we hope to guide further studies toward determining the clinical viability of 5-FU as a treatment option against this formidable pathogen.},
}

@article {pmid40755812,
year = {2025},
author = {Jones, L and Hanrahan, N and Salta, M and Skovhus, TL and Thomas, K and Illson, T and Wharton, J and Webb, J},
title = {Evaluating biocide efficacy in mixed-species biofilms: insights from a dual anaerobic biofilm reactor.},
journal = {Npj Materials degradation},
volume = {9},
number = {1},
pages = {97},
pmid = {40755812},
issn = {2397-2106},
abstract = {Understanding biocide performance in mixed-species biofilms is critical to mitigating microbiologically influenced corrosion (MIC). In this study, a novel dual anaerobic biofilm reactor was used to evaluate glutaraldehyde efficacy under environmentally relevant conditions, using a complex microbial consortium from marine sediment. Despite biocide dosing, biofilms persisted and induced localized corrosion, indicating incomplete mitigation. Each biocide application led to an electronegative shift in E corr and a reduction in H 2 S concentration, suggesting partial suppression of microbial activity. Raman spectroscopy and profilometry revealed differences in corrosion product composition and pit morphology between biotic and abiotic systems. 16S rRNA sequencing showed enrichment of stress-tolerant genera, including Exiguobacterium and Serpentinicella, consistent with increased chemical tolerance. These findings highlight the limitations of conventional biocide strategies and demonstrate the need for adaptive, community-informed treatment approaches. The dual-reactor model provides a robust platform for future MIC standardization efforts and mechanistic investigation of biofilm resilience under anoxic conditions.},
}

@article {pmid40755022,
year = {2025},
author = {Kulshrestha, A and Gupta, P},
title = {Control of Candida albicans and Staphylococcus aureus dual species interkingdom biofilm development via SAP5 inhibition.},
journal = {Future microbiology},
volume = {},
number = {},
pages = {1-13},
doi = {10.1080/17460913.2025.2539011},
pmid = {40755022},
issn = {1746-0921},
abstract = {AIM: Microbial biofilms, particularly polymicrobial ones formed by Candida albicans and Staphylococcus aureus, exhibit increased resistance to conventional treatment. This study investigates the inhibitory effect of 4-hydroxyphenylacetic acid on Secreted Aspartyl Protease 5, a key virulence factor involved in hyphal development and biofilm formation in Candida albicans, to investigate its therapeutic potential in inhibiting polymicrobial biofilms developed with Staphylococcus aureus.

MATERIALS & METHODS: Biofilm formation was studied using a Biofilm Infection Simulator System and polymethylmethacrylate coupons. Biofilms were analyzed under untreated and 4-HPA-treated conditions. SAP5 proteinase activity on BSA was quantified in treated & un-treated cultures.

RESULTS: 4-HPA treatment significantly restricted C. albicans hyphal growth, reducing S. aureus attachment. The biofilm decreased by over 78% in mono-species and 81% in polymicrobial cultures. The inhibition of SAP5 protease activity was 79% in mono-species and 65% in mixed cultures.

CONCLUSIONS: 4-HPA effectively reduces biofilm formation and SAP5 activity, highlighting its potential as a biofilm-specific therapeutic for polymicrobial infections.},
}

@article {pmid40752855,
year = {2025},
author = {Wei, J and Xia, T and Xia, J and Gao, T and Zhang, G and Huang, P and Li, X},
title = {Biofilm-triggered interfacial assembly of dual-porphyrin heterojunctions for chemo-/sonodynamic treatment of pyomyositis.},
journal = {Acta biomaterialia},
volume = {},
number = {},
pages = {},
doi = {10.1016/j.actbio.2025.07.069},
pmid = {40752855},
issn = {1878-7568},
abstract = {Sonodynamic therapy (SDT) indicates advantages in combating antibiotics-resistant bacteria and deep tissue infections, but challenges remain in the development of highly efficient, infection-responsive, and biocompatible sonosensitizers. Herein, tetrakis (4-hydroxyphenyl) porphyrin (TH) and tetrakis (4-carboxyphenyl) zinc porphyrin (ZnTC) are proposed to construct dual-porphyrin heterojunctions (TH/ZnTC), and their matched interface and strong interfacial electric field (IEF) enhance charge density and transfer for selective and efficient SDT. Specifically, metal organic frameworks were constructed through coordination of Fe[3+] and ZnTC and simultaneously loading TH to prepare TH@FeM. In infection sites with elevated glutathione, Fe[3+] is reduced to Fe[2+], triggering TH@FeM decomposition and TH/ZnTC self-assembly through π-π stacking and electrostatic interactions. IEF from ZnTC to TH drives the formation of S-scheme TH/ZnTC heterojunctions and greatly promotes efficient separation and transfer of the generated charges at the matched interface for efficient generation of reactive oxygen species. Meanwhile, GSH-reductive releases of Fe[2+] enable high Fenton reaction activity for chemodynamic therapy. After intravenous injection into a mouse pyomyositis model, the enhanced penetration and retention in the infected muscles implements up to 3.1-folds higher fluorescence intensities than those of the major tissues. Ultrasonication of TH@FeM fully destructs bacteria, downregulates inflammatory factor levels, promotes angiogenesis, and accelerates healing of infected muscles without significant pathological and functional changes in the main organs, leading to continuous decreases in clinical scores and full survival of pyomyositis mice. Thus, the concise design represents the first attempt to explore biofilm-responsive heterojunction formation for synergistic chemo-/sonodynamic therapies of bacterial infections. STATEMENT OF SIGNIFICANCE: Nearly 80% of chronic infections are linked to biofilm formation on living tissues. Extracellular polysaccharides produced by biofilms confer protection, making bacteria 10-1000 times more resistant to antibiotics compared to their planktonic counterparts, thus complicating treatment. Sonodynamic therapy (SDT) offers promising advantages in addressing antibiotics-resistant bacteria and deep tissue infections, but challenges remain in the development of highly efficient, infection-responsive, and biocompatible sonosensitizers. Herein, we propose biofilm-responsive generation of dual-porphyrin heterojunctions with matched interface and strong interfacial electric field, which enhance charge density and transfer for selective and efficient SDT. The glutathione-responsive formation and charge transfer mechanisms were both theoretically calculated and experimentally validated. Furthermore, target accumulation and treatment efficacy were demonstrated in a pyomyositis model.},
}

@article {pmid40751169,
year = {2025},
author = {Yang, X and Yao, M and Li, P and van der Hoek, JP and Zhang, L and Liu, G},
title = {Granular activated carbon (GAC)-driven microbial electron shuttle boosts denitrification and mitigates N2O in cold and carbon-limited biofilm system.},
journal = {Microbiome},
volume = {13},
number = {1},
pages = {178},
pmid = {40751169},
issn = {2049-2618},
support = {52200077//National Natural Science Foundation of China/ ; 52261145702//National Natural Science Foundation of China/ ; GZB20230691//Postdoctoral Fellowship Program (Grade B) of China Postdoctoral Science Foundation/ ; 2024HWYQ-040//Shandong Excellent Young Scientists Fund Program (Overseas)/ ; tsqn202408074//Taishan Scholars Young Expert Program/ ; },
mesh = {*Denitrification ; *Biofilms/growth & development ; *Nitrous Oxide/metabolism ; *Carbon/metabolism ; Wastewater/microbiology ; Electron Transport ; Cold Temperature ; *Charcoal/chemistry ; *Bacteria/metabolism/genetics/classification ; Water Purification/methods ; },
abstract = {BACKGROUND: Denitrification in wastewater treatment is severely limited under low-temperature and low-carbon ("dual-low") conditions, hindering sustainable nitrogen removal. Biofilm systems, though energy-efficient, suffer from reduced efficiency in such environments due to impaired interspecies electron transfer (IET). Granular activated carbon (GAC), a conductive mediator, offers potential to enhance IET between electroactive microorganisms (EAMs) and denitrifiers, yet its role in dual-low systems remains underexplored. This study investigates GAC's capacity to optimize biofilm functionality and mitigate greenhouse gas (GHG) emissions under these constraints.

RESULTS: Under dual-low conditions (4-6°C, C/N = 4), GAC increased denitrification efficiency by 19.4-21.9% and reduced N2O emissions by 10.6-22.9%. Metatranscriptomes revealed upregulation of denitrifying genes (e.g., nosZ) and electron transport pathways (e.g., omcB in Geobacter). FISH/SEM confirmed GAC-driven coacervates of EAMs and denitrifiers, linked by nanowires, enhancing direct electron transfer. Microbial diversity decreased, but functional redundancy improved, with Pseudomonas fluorescens and Geobacter sulfurreducens dominating. TOC removal rose under low temperatures, indicating enhanced carbon utilization.

CONCLUSIONS: GAC fosters synergistic EAM-denitrifier partnerships, enabling efficient denitrification and GHG mitigation in cold and carbon-limited ("dual-low") biofilm systems, advancing sustainable wastewater management. Video Abstract.},
}

*Denitrification
*Biofilms/growth & development
*Nitrous Oxide/metabolism
*Carbon/metabolism
Wastewater/microbiology
Electron Transport
Cold Temperature
*Charcoal/chemistry
*Bacteria/metabolism/genetics/classification
Water Purification/methods

@article {pmid40748998,
year = {2025},
author = {Sahl, C and Andersson, A and Larsson, N and Paulsson, M and Shannon, O and Påhlman, LI},
title = {The cystic fibrosis pathogen Achromobacter xylosoxidans inhibits biofilm formation of Pseudomonas aeruginosa.},
journal = {Journal of medical microbiology},
volume = {74},
number = {8},
pages = {},
pmid = {40748998},
issn = {1473-5644},
mesh = {*Biofilms/growth & development ; *Pseudomonas aeruginosa/physiology/growth & development/isolation & purification ; *Achromobacter denitrificans/physiology/isolation & purification/growth & development ; Humans ; *Cystic Fibrosis/microbiology/complications ; *Pseudomonas Infections/microbiology ; *Gram-Negative Bacterial Infections/microbiology ; *Antibiosis ; },
abstract = {Background. Achromobacter xylosoxidans and Pseudomonas aeruginosa are two pathogens that cause persistent airway infections in individuals with cystic fibrosis (CF). The persistence of P. aeruginosa is partly due to a high capacity to form biofilms and the ability to exert antagonism against other bacteria. Loss of microbial diversity in conjunction with chronic P. aeruginosa colonization is strongly correlated with low lung function in CF. A. xylosoxidans and P. aeruginosa are frequently co-isolated in CF airway cultures. This study aims to investigate the reciprocal effects on growth inhibition and biofilm formation between P. aeruginosa and A. xylosoxidans in vitro.Method. Six isolates of A. xylosoxidans, isolated from three CF patients in early and late stages of a chronic infection, were cultured together with a CF isolate of P. aeruginosa. Biofilm formation was assessed using a microtiter assay and crystal violet staining. Quantitative PCR was used to quantify species proportions in biofilms. Growth curves were performed to compare planktonic growth rates.Results. Three A. xylosoxidans isolates, all of which were from early-stage infections, inhibited biofilm formation of P. aeruginosa. The inhibition was concentration-dependent and required the interaction of live bacteria during the early stages of biofilm development. The inhibitory effect was not caused by nutrient depletion of the planktonic cells. The selected A. xylosoxidans isolate had a stronger capacity to adhere to plastic surfaces compared to the P. aeruginosa isolate.Conclusions . A. xylosoxidans can inhibit P. aeruginosa biofilm formation in vitro. The observed effect requires active interactions between live cells during the attachment stage of biofilm formation, possibly due to differences in adhesion capacity.},
}

*Biofilms/growth & development
*Pseudomonas aeruginosa/physiology/growth & development/isolation & purification
*Achromobacter denitrificans/physiology/isolation & purification/growth & development
Humans
*Cystic Fibrosis/microbiology/complications
*Pseudomonas Infections/microbiology
*Gram-Negative Bacterial Infections/microbiology
*Antibiosis

@article {pmid40748066,
year = {2025},
author = {Fung, BL and Mullins, C and Rusch, DB and Musto, EG and van Kessel, JC and Visick, KL},
title = {Genetic and biochemical analyses reveal direct interactions between LitR and genes important for Vibrio fischeri physiology, including biofilm production.},
journal = {Journal of bacteriology},
volume = {},
number = {},
pages = {e0004225},
doi = {10.1128/jb.00042-25},
pmid = {40748066},
issn = {1098-5530},
abstract = {Bacteria can link gene expression to population density to promote group behaviors using quorum sensing. Quorum sensing controls a multitude of bacterial processes, such as virulence, motility, and biofilm formation. In Vibrio fischeri, the quorum-sensing-dependent transcription factor LitR inhibits biofilm formation. A previous study showed that LitR inhibits transcription (~1.4-fold) of the bcs locus, which comprises the genes responsible for producing the cellulose polysaccharide. However, beyond that, the mechanism of LitR-mediated inhibition of biofilm formation was unknown. Here, we find that LitR transcriptionally activates pdeV, which encodes a c-di-GMP phosphodiesterase that indirectly promotes cleavage of the large adhesive protein LapV from the surface of V. fischeri, leading to biofilm dispersal. LitR also induces transcription of the gene for sensor kinase VF_A1016, which we determined to be important for biofilm inhibition. Like the loss of LitR, the loss of VF_A1016 increased bcs transcription (~1.6-fold). Through chromatin immunoprecipitation sequencing (ChIP-seq), we found that LitR directly binds to the VF_A1016 and pdeV regulatory region. In total, we identified 147 LitR-binding sites in the genome and confirmed transcriptional control over a subset of these putative regulatory targets. Specifically, we determined that LitR induces transcription of the genes encoding the diguanylate cyclase VF_1200 and the glyoxylate shunt protein AceB and inhibits expression of the putative transcription factor TfoY. These data expand our understanding of LitR-mediated regulation of genes involved in biofilm formation and the physiology of V. fischeri.IMPORTANCEBacteria can coordinate their behaviors on a population level using quorum sensing, a process that results in altered gene regulation. In Vibrio fischeri, the quorum-sensing-regulated transcription factor LitR inhibits the formation of biofilms, communities of attached and protected bacteria, by diminishing the production of cellulose. Here, we determined that LitR controls additional known or putative biofilm factors. We also identified other possible targets of LitR regulation by high-throughput chromatin immunoprecipitation sequencing. This work furthers our understanding of the established connection between quorum sensing and biofilm formation in V. fischeri strain ES114. These findings also have the potential to translate to known pathways in other Vibrios where quorum sensing and biofilm production are linked.},
}

@article {pmid40747489,
year = {2025},
author = {Li, Y and Yu, Y and Cui, Y and Jiang, L and Luo, Y and Xie, H and Hong, H},
title = {Oleum ocimi gratissimi as a promising natural preservative against fish spoilage bacteria through i006Ehibition of planktonic growth and biofilm formation.},
journal = {Food chemistry: X},
volume = {29},
number = {},
pages = {102816},
pmid = {40747489},
issn = {2590-1575},
abstract = {Strategies to control spoilage bacteria and biofilm formation are essential for minimizing food losses. Although Oleum ocimi gratissimi (OG) effectively preserves chill-stored fish, its specific inhibitory mechanisms against fish-derived spoilage bacteria remain unclear. This study investigated the antibacterial activity of OG against Aeromonas sobria, Pseudomonas versuta, and Shewanella putrefaciens. OG exhibited minimum inhibitory concentrations (MIC) of 2, 0.75, and 0.375 mg/mL and minimum bactericidal concentrations (MBC) of 3, 4, and 3 mg/mL against these bacteria, respectively. At MIC levels, OG inhibited planktonic growth by disrupting cell membranes, inducing adenosine triphosphate (ATP) leakage, reducing ATPase activity, and altering cellular morphology. Biofilm formation was completely suppressed, with OG further diminishing metabolic activity and extracellular polysaccharide/protein. These findings elucidate the dual role of OG in inhibiting bacterial growth and biofilm formation, highlighting its potential to enhance fish safety and shelf life by targeting biochemical and physiological pathways in spoilage bacteria.},
}

@article {pmid40747426,
year = {2025},
author = {Huang, X and Prasad, R and Saluja, S and Yang, Y and Yan, Q and Shuster, SO and Olson, R and Lin, C and Davis, CM and Jiang, X and Zhou, HX and Yan, J},
title = {Conformations and sequence determinants in the lipid binding of an adhesive peptide derived from Vibrio cholerae biofilm.},
journal = {bioRxiv : the preprint server for biology},
volume = {},
number = {},
pages = {},
doi = {10.1101/2025.07.14.664771},
pmid = {40747426},
issn = {2692-8205},
abstract = {UNLABELLED: Surface adhesion is critical to the survival of pathogenic bacteria both in natural niches and during infections, often via forming matrix-embedded communities called biofilms. We previously identified a 57-amino acid peptide (Bap1-57aa) as a key contributor to biofilm adhesion of the pandemic pathogen Vibrio cholerae to various surfaces including lipid membranes. Here, we combine biophysical, computational, and genetic approaches to elucidate the molecular mechanism. A central aromatic-rich motif anchors the peptide to lipid bilayers while peripheral pseudo repeats enhance binding through avidity. Surprisingly, the core motif undergoes a lipid-induced conformational transition into a β-hairpin, enabling robust membrane insertion. Moreover, the biofilm-derived peptide, conserved in several other Vibrio species, can adhere to model host surfaces and is sensitive to membrane curvature. Our results provide molecular insight into biofilm adhesion and may lead to new strategies for targeted biofilm removal and the design of bioinspired underwater adhesives.

TEASER: A short peptide from Vibrio cholerae binds lipids using a unique β-hairpin motif and contributes to host colonization.},
}

@article {pmid40746958,
year = {2025},
author = {Wang, Y and Zhang, R and Wang, P and Zhang, W and Li, Z and Pang, X and Huang, F and Wang, S and Liu, X and Zhang, H},
title = {Biofilm-mediated resistance to berberine in Escherichia coli.},
journal = {Frontiers in cellular and infection microbiology},
volume = {15},
number = {},
pages = {1565714},
pmid = {40746958},
issn = {2235-2988},
mesh = {*Biofilms/drug effects/growth & development ; *Berberine/pharmacology ; *Escherichia coli/drug effects/physiology/genetics ; Microbial Sensitivity Tests ; *Drug Resistance, Bacterial ; *Anti-Bacterial Agents/pharmacology ; Gene Expression Regulation, Bacterial/drug effects ; Escherichia coli Proteins/genetics ; Gene Expression Profiling ; RNA, Messenger/analysis ; Real-Time Polymerase Chain Reaction ; },
abstract = {OBJECTIVE: To investigate the mechanism of biofilm-mediated resistance to berberine in Escherichia coli.

METHODS: The resistance of berberine against E. coli was induced by 1/2 MIC (minimum inhibitory concentration). Biofilm formation was detected by crystal violet staining. The mRNA level was detected by RT-qPCR, and the gene csgD was determined. the csgD-overexpressed strain was constructed. We measured the MIC of berberine against E. coli, as well as biofilm formation and the expression of mRNA.

RESULTS: The MIC after berberine induction was more than 32 times than the MIC before induction. the biofilm was significantly increased at 24, 48 and 72 hours (p<0.01) after berberine induction. In addition, the amount of biofilm production at 24, 48 and 72 hours was 1.3, 1.51 and 1.98 times after berberine induction than that before induction, respectively. The expression of csgD gene was significantly increased (p=0.016) after induction compared with that before induction. the MIC of csgD-overexpressed strain was about 5.8 times that before induction. The expression of csgD gene was significantly increased (p=0.016), which was 5.8 times higher than that before induction. The MIC of csgD-overexpressed strain was 100 μg/mL. Biofilm formation in csgD-overexpressed strain was 2.9 times higher than that of the control. The expression of biofilm-related genes, bcsA, luxS and csgD, was 45, 22.5 and 1628 times higher than that of the control, respectively.

CONCLUSION: Berberine might increase biofilm formation by inducing the expression of csgD gene, which might result in drug resistance in E. coli.},
}

*Biofilms/drug effects/growth & development
*Berberine/pharmacology
*Escherichia coli/drug effects/physiology/genetics
Microbial Sensitivity Tests
*Drug Resistance, Bacterial
*Anti-Bacterial Agents/pharmacology
Gene Expression Regulation, Bacterial/drug effects
Escherichia coli Proteins/genetics
Gene Expression Profiling
RNA, Messenger/analysis
Real-Time Polymerase Chain Reaction

@article {pmid40746480,
year = {2025},
author = {Turbatmath, K and Sharma, S},
title = {Comparative evaluation of antimicrobial and biofilm inhibition effects of royal jelly, chlorhexidine, and calcium hydroxide - An in vitro study.},
journal = {Journal of conservative dentistry and endodontics},
volume = {28},
number = {7},
pages = {607-612},
pmid = {40746480},
issn = {2950-4708},
abstract = {INTRODUCTION: Pulp capping is a viable treatment option for management of deep carious lesions in pulp preservation with reduced remaining thickness of dentin. Traditionally, calcium hydroxide (Ca(OH)2) and currently contemporary bioceramic pulp capping agents have been well researched in the dental domain and evidenced varying but promising success rates. However, there is a growing interest in exploring indigenous, innovative, alternative natural products derived from apiarian sources, which possess characteristics and properties which can potentiate predictable treatment outcome including antimicrobial properties. Hence, this study aims to compare the efficacy of indigenously developed apiarian products like royal jelly (RJ) with contemporary pulp capping agents in eradicating Streptococcus mutans biofilm, a predominant cariogenic microorganism.

MATERIALS AND METHODS: RJ was prepared at 4% concentration by dissolving 4 g of pure RJ in 100 mL dimethyl sulfoxide (DMSO). Ca(OH)2 was prepared by mixing 2 g of Dycal (Dentsply) in 100 mL of sterile water. Chlorhexidine (CHX) was used as a 2% gel formulation (Gluco-Chex, Cerkamed). DMSO was used as a solvent to ensure proper dispersion. Agar diffusion results: RJ (97.89%), CHX (72.95%), and Ca(OH)2 (78.19%). Biofilm assay: RJ (97.89%), CHX (72.95%), and Ca(OH)2 (78.19%). Evaluation of the antimicrobial activity was conducted with various tests such as zone of inhibition and biofilm viability assay. Data collection was done and statistical analysis was done using G*Power (P < 0.05).

RESULTS: Agar diffusion: RJ showed the highest antimicrobial effect (97.89%), followed by CHX (72.95%) and Ca(OH)2 (78.19%). Biofilm assay: 4% RJ exhibited the highest inhibition (97.89%), outperforming CHX (72.95%) and Ca(OH)2 (78.19%).

CONCLUSION: The findings of this comparative evaluation support the potential of the innovation of indigenously developed apiarian pulp capping agent as an effective antimicrobial agent against S. mutans cariogenic biofilm. Further research is warranted to explore the optimal inhibitory concentration against dental biofilm and further evaluate their long-term clinical efficacy of this indigenous innovation and safety in dental practice as a pulp capping agent.},
}

@article {pmid40746247,
year = {2025},
author = {Ferreira, TL and Sá, LGDAV and Cabral, VPF and Rodrigues, DS and Moreira, LEA and Souza, BO and Cavalcanti, BC and Magalhães, HIF and Lima, ISP and Leitão, AC and Moraes, MO and Andrade Neto, JB and Nobre Júnior, HV and da Silva, CR},
title = {Antibacterial activity of diazepam against planktonic and biofilm strains of methicillin-resistant Staphylococcus aureus.},
journal = {Biofouling},
volume = {},
number = {},
pages = {1-11},
doi = {10.1080/08927014.2025.2540534},
pmid = {40746247},
issn = {1029-2454},
abstract = {The aim was to evaluate the antibacterial activity of diazepam against methicillin-susceptible (MSSA) and methicillin-resistant (MRSA) strains of Staphylococcus aureus and its possible mechanism of action. The broth microdilution assay was used to determine the minimum inhibitory concentration (MIC) of diazepam. A checkerboard assay was used to evaluate the interaction of diazepam with different antibiotics. Colorimetric assays with MTT were used to evaluate the effect of diazepam against the biofilms by MSSA and MRSA. Flow cytometry and fluorescence microscopy were used to evaluate the possible mechanism of action of diazepam against MRSA. Diazepam had a MIC of 256 µg/mL. It only had indifferent interactions with the analyzed antibiotics. Diazepam significantly reduced the viability of MSSA and MRSA biofilms. Diazepam caused fragmentation of bacterial DNA and carbonylation of proteins, resulting in reduced cell viability. Therefore, diazepam has in vitro antimicrobial activity against planktonic and biofilm strains of MRSA and MSSA.},
}

@article {pmid40745039,
year = {2025},
author = {Gheitasi, R and Weiss, D and Müller, MM and Sommer, K and Röll, D and Mosig, A and Pletz, MW and Makarewicz, O},
title = {Staphylococcus aureus biofilm-associated component PNAG stimulates the secretion of the immunomodulatory chemokine CXCL10 via Dectin-1 signaling.},
journal = {Communications biology},
volume = {8},
number = {1},
pages = {1136},
pmid = {40745039},
issn = {2399-3642},
support = {861323//EC | Horizon 2020 Framework Programme (EU Framework Programme for Research and Innovation H2020)/ ; },
mesh = {*Biofilms ; *Chemokine CXCL10/physiology ; *Lectins, C-Type/metabolism ; Leukocytes, Mononuclear/physiology ; *Polysaccharides, Bacterial/physiology ; *Staphylococcus aureus/pathogenicity/physiology ; Healthy Volunteers ; Humans ; Syk Kinase ; CARD Signaling Adaptor Proteins ; },
abstract = {Staphylococcus aureus is a common human pathogen associated with many infections. The key factor contributing to the virulence of S. aureus is its ability to form difficult-to-treat and recalcitrant biofilms. One of the major staphylococcal biofilms matrix compounds is poly-N-acetylglucosamine (PNAG). In previous study, we observed an increased secretion of various cytokines and chemokines when immune cells were stressed by S. aureus biofilms. In this study, we aimed to analyze the effect of PNAG on the secretion of the CXCL10 chemokine subfamily by peripheral blood mononuclear cells and monocytes and studied the connection to the Dectin-1-Syk-CARD9 signaling pathway, as Dectin-1 is the major pattern recognized by polysaccharide structures. We showed that, in contrast to the major virulence factor surface protein A, PNAG primarily elevates the secretion of CXCL10. This secretion was interrupted by blocking the Dectin-1 receptor or tyrosine kinase Syk. PNAG exposure resulted in increased Dectin-1 and CARD9 expression as well as increased NF-κB and CXCL10 expression, which may be related to the long-term memory processes of T cells. We also showed that PNAG induces the formation of CD14 + CXCL10+ monocytes that can migrate to the site of infection, triggering an innate immune response against S. aureus. This study provides insights into the complex interaction of the staphylococcal biofilms matrix with immune chemotaxis and shows that immunologic processes leading to bacterial infections should be viewed in a more differentiated manner, as biofilms are the preferred formation of microorganisms.},
}

*Biofilms
*Chemokine CXCL10/physiology
*Lectins, C-Type/metabolism
Leukocytes, Mononuclear/physiology
*Polysaccharides, Bacterial/physiology
*Staphylococcus aureus/pathogenicity/physiology
Healthy Volunteers
Humans
Syk Kinase
CARD Signaling Adaptor Proteins

@article {pmid40744290,
year = {2025},
author = {Janež, N and Ladányi, M and Zaveršek, N and Čotar, P and Sebastijanović, A and Štrancar, J and Sabotič, J and Pajk, S},
title = {Synthesis and evaluation of lipophilic fluorescent probes for the labelling of Listeria and their impact on biofilm formation.},
journal = {Journal of microbiological methods},
volume = {236},
number = {},
pages = {107206},
doi = {10.1016/j.mimet.2025.107206},
pmid = {40744290},
issn = {1872-8359},
abstract = {Imaging bacterial biofilms using confocal fluorescence microscopy is used to study their structures, but its wider application is constrained by the limited availability of effective labelling tools. Small chemical fluorescent probes offer a versatile alternative to heterologous expression of fusion or reporter proteins, but data on their effects on biofilm formation are lacking. In this study, we synthesized a series of new lipophilic fluorescent probes based on Nile blue, Nile red and coumarin scaffold. We investigated them for the labelling of Listeria biofilms and determined their effects on the growth and biofilm biomass formation. The Nile red probe SP-AM 7 and the coumarin probe PAG 31 inhibited biofilm development and showed a strong bactericidal effect. The Nile blue probe PAG 19 had the least effect on the tested parameters, but labelled slowly, while the fast-labelling Nile red probe SP-AM 8 promoted biofilm formation. Both are suitable for use during biofilm growth, resulting in less variation in biomass-related measurements than probes added prior to imaging. In the 3D imaging-based measurements for selected probes, we found no difference in the total biomass formed compared to the control dye, but a redistribution of biomass in the 3D layers was observed. Other probes were found to be slow to label, leave traces of unused probes or interfere with attachment to the surface. Our results show that fluorescent probe labelling should be evaluated from chemical, physical and biological points of view to understand their reliability and credibility.},
}

@article {pmid40744271,
year = {2025},
author = {Xu, Q and Tian, F and Wang, X and Lian, J and Zhang, X and Lin, X and Liu, Y},
title = {Proteomics reveals the role of the EamB transporter from Aeromonas hydrophila LP-2 in biofilm formation.},
journal = {Journal of proteomics},
volume = {321},
number = {},
pages = {105510},
doi = {10.1016/j.jprot.2025.105510},
pmid = {40744271},
issn = {1876-7737},
abstract = {Biofilms play a pivotal role in the survival and persistence of microorganisms, endowing them with heightened resistance to environmental stressors and antimicrobial agents. The EamB protein, which encodes an inner membrane transporter, acted as a negative regulator of biofilm formation, and the gene eamB deletion in the pathogen Aeromonas hydrophila LP-2 resulted in a significant increase in biofilm formation. Proteomic analysis revealed a total of 616 differentially abundant proteins between the ΔeamB and wild-type (WT) strains, with 308 downregulated and 308 upregulated. RT-qPCR was employed to verify the stability and accuracy of the proteomics data. Bioinformatic analysis indicated that EamB is involved in critical bacterial biological processes, including flagellar assembly, amino acid metabolism, and fatty acid degradation. Biofilm formation assays further revealed that supplementation with exogenous lysine significantly inhibited biofilm formation in the ΔeamB strain, conversely, exogenous cysteine and O-acetylserine obviously increased biofilm formation in the ΔeamB strain. These findings demonstrated that EamB may modulate bacterial biofilm formation in A. hydrophila through the regulation of amino acid metabolism. This finding provides novel insights into the regulatory mechanism underlying biofilm formation and highlights potential targets for the development of future antibacterial strategies. SIGNIFICANCE STATEMENT: This study elucidates the critical role of the eamB gene in Aeromonas hydrophila, a significant aquatic pathogen, by demonstrating its impact on biofilm formation and physiological traits. Through comparative proteomic analysis, we identified 616 differentially abundant proteins in the ΔeamB mutant, revealing its involvement in key metabolic pathways such as amino acid metabolism, flagellar assembly, and fatty acid degradation. Notably, eamB deletion enhanced biofilm formation, while exogenous amino acids like cysteine and O-acetylserine obviously increased biofilm formation in the ΔeamB strain. These findings highlight EamB as a regulator of biofilm formation, offering novel molecular insights into bacterial pathogenicity. This research advances our understanding of biofilm-associated antibiotic resistance and provides potential targets for developing strategies to mitigate infections caused by A. hydrophila in aquaculture and public health.},
}

@article {pmid40744180,
year = {2025},
author = {Rashid, Z and Ali, SMA and Shahid, R and Rahman, A and Imran, M},
title = {Nano-active carboxymethyl cellulose hydrogels infused with sodium deoxycholate functionalized permeosomes to combat biofilm producing resistant pathogens associated with chronic wounds.},
journal = {International journal of biological macromolecules},
volume = {321},
number = {Pt 3},
pages = {146383},
doi = {10.1016/j.ijbiomac.2025.146383},
pmid = {40744180},
issn = {1879-0003},
abstract = {During the past decade, the incidence of chronic wounds has continued to rise due to the growing threat of multidrug-resistant (MDR) biofilm-producing bacteria residing inside the convoluted architecture of skin. Herein, we have developed sodium deoxycholate functionalized liposomes (nano-permeosomes) loaded with tea tree oil (TTO) infused inside carboxymethyl cellulose hydrogels (CMC-TTO-LP). Nano-permeosomes exhibited high encapsulation efficiency of TTO (91 % ± 0.36 %). SEM revealed the smooth and ultra-deformable morphology of nano-permeosomes. Nano-permeosomes exhibited an average size of 178.5 ± 6.42 nm with a zeta potential of -53.2 ± 7.03 mV. FTIR spectroscopy confirmed the presence of weak electrostatic interactions, including hydrophobic forces, among nano-permeosomes, TTO, and hydrogels. CMC-TTO-LP exhibited significant antibacterial and antibiofilm activity against MDR Pseudomonas aeruginosa and Staphylococcus aureus. CMC-TTO-LP exhibited high antioxidant activity (68 % ± 2.8 %), hemocompatibility, and cytocompatibility. Nano-permeosomes prevented the burst release and allowed sustained release kinetics of TTO from CMC-TTO-LP. Moreover, under ex vivo conditions, nano-active hydrogels exhibited high apparent permeability, average flux (42.9 ± 2.42 μL/cm[2]/h), and skin retention (24.94 ± 0.45 μL/cm[2]) of TTO across the dermal barrier of rabbit skin. Hence, these advanced nano-active hydrogels (CMC-TTO-LP) can potentially deliver TTO via the transdermal route and act as an appealing candidate in controlling chronic wound infections persisting due to MDR pathogens.},
}

@article {pmid40743952,
year = {2025},
author = {Bilal, H and Zhang, CX and Choudhary, MI and Shah, MR and Dej-Adisai, S and Liu, Y and Chen, ZF},
title = {Synthesis of thiosemicarbazide-based zinc complexes and evaluation of their inhibition of bacterial biofilm formation via targeting extracellular proteins.},
journal = {Journal of inorganic biochemistry},
volume = {272},
number = {},
pages = {113014},
doi = {10.1016/j.jinorgbio.2025.113014},
pmid = {40743952},
issn = {1873-3344},
abstract = {Three new zinc(II) complexes with N-benzyl-2-((2-hydroxynaphthalen-1-yl)methylene) hydrazine-1-carbothioamide-methanol solvate (L), 1,10-phenanthroline (phen), and pyridine (py) as ligands, were synthesized and fully characterized. They are named as [Zn2(L)2Cl2)].CH3OH (1), [Zn(L)(phen)].CH3OH (2), and [Zn2(L)2(py)2] (3). Complex 1 and 3 are dinuclear structures. Complex 1 contains two L ligands; complex 3 also contains two L ligands in addition of two py as co-ligands, and complex 2 is mononuclear with one L and one phen as co-ligand. The in vitro antibacterial activities of complexes 1-3 were tested on five bacterial strains with minimum inhibitory concentrations (MICs) of 1.56 to 64 μg/mL. Complexes 1 and 2 exhibited significantly stronger antibacterial activity against Enterococcus faecalis (E. faecalis) and methicillin-resistant Staphylococcus aureus (MRSA) (S. aureus) than L, complex 3, and Vancomycin. The complexes 1 and 2 inhibited bacterial biofilm formation at concentrations from 0.5 to 10 μg/mL, higher than L, complex 3 and Vancomycin. Complexes 1 and 2 strongly interacted with extracellular proteins (ECPs) of bacterial biofilms. Furthermore, molecular docking (MD) studies have validated the interactions of L and complexes 1-3 with biofilm-associated proteins (Baps) and SARS-CoV-2 receptors, positioning these zinc(II) complexes as promising candidates for developing antibacterial and anti-biofilm agents.},
}

@article {pmid40743484,
year = {2025},
author = {Li, X and Li, Q and He, A and Dang, M and Zhang, Y and Wang, M and Sun, Q and Dai, Z and Ding, M and Zheng, J and Mou, Y and Xiu, W and Dong, H},
title = {Disrupting Biofilm Tolerance by Ionic Microbubble-Mediated Copper Ion Surge for Infection Clearance.},
journal = {ACS nano},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsnano.5c08035},
pmid = {40743484},
issn = {1936-086X},
abstract = {Bacterial infections caused by drug-resistant bacteria persist due to biofilm-mediated tolerance, which limits the efficacy of both antimicrobial agents and host immune defenses. Here, we develop ionic microbubbles (MB-CuTA) self-assembled by Fe3O4@CuTA nanoparticles to enhance copper ion-mediated antibiofilm therapy. Upon ultrasound activation, MB-CuTA undergoes inertial cavitation, disrupting biofilm integrity and generating a localized surge of copper ions. This process achieves a dual therapeutic effect: (1) disruption of bacterial metabolic homeostasis, thereby overcoming the intrinsic resistance of biofilms to conventional antimicrobial agents, and (2) activation of cellular immunity to effectively counteract bacterial immune evasion mechanisms. By breaking biofilm tolerance through both metabolic and immunological pathways, our strategy enables deep copper ion penetration in biofilms and effective infection clearance in both mouse implant infection and peritonitis infection models. Our approach introduces a biofilm tolerance disruption method through inducing bacterial cuproptosis-like death and cellular immunity activation, offering a promising strategy against biofilm infections.},
}

@article {pmid40742327,
year = {2025},
author = {Sun, X and Xu, Z and Hu, G and Xie, J and Li, Y and Tao, L and Zhang, N and Xun, W and Miao, Y and Zhang, R and Shen, Q and Kost, C and Kovács, ÁT},
title = {Presence of a biofilm beneficiary alters the evolutionary trajectory of a biofilm former.},
journal = {The ISME journal},
volume = {},
number = {},
pages = {},
doi = {10.1093/ismejo/wraf160},
pmid = {40742327},
issn = {1751-7370},
abstract = {Biofilm evolution is typically studied in monocultures or in communities displaying mutualistic or exploitative interactions. However, in communities with fluctuating interactions, the influence of biofilm-beneficiary bacteria on the evolution of biofilm-founder bacteria remains less understood. Biofilm-beneficiary bacteria cannot form robust biofilms independently but can incorporate into the biofilm of biofilm-formers, thereby gaining the ability to colonize given niche. In this study, we demonstrate that the biofilm-former Bacillus velezensis SQR9 reproducibly diversified into biofilm-enhanced slimy and biofilm-weakened rough types, both in the presence and absence of a biofilm-beneficiary Stutzerimonas degradans XL272 (formerly Pseudomonas stutzeri), but with variable frequencies under the two conditions. The exopolysaccharide producer slimy types dominated B. velezensis populations in monoevolution, whereas the exploiter rough types, which exploit the exopolysaccharides produced by the slimy types, dominate in coculture evolution. Phenotypic changes in B. velezensis were linked to mutations in specific genes that regulate biofilm formation and sporulation, including ywcC, comA, comP, degS, degQ, and spo0F. A frameshift mutation in the cpsA gene of Saccharophagus degradans increased its exopolysaccharide production in the dual-species biofilm, which served as shared resources and allow the B. velezensis exploiter (i.e., rough type) to outcompete the producer (i.e., slimy type) during coculture evolution. Additionally, longitudinal population sequencing and "replay" evolution experiments with the S. degradans mutant revealed that the cpsA mutation accelerated the fixation of the rough type within B. velezensis populations. In conclusion, this research demonstrates that interspecific interactions can adaptively favor exploiters within biofilm populations.},
}

@article {pmid40741807,
year = {2025},
author = {Regalado Guerrero, MF and Iñiguez Gutiérrez, L and Gómez Sandoval, JR and Ramírez López, IG and Gutiérrez Silerio, GY and Chávez Maciel, JM and Guerrero Velázquez, C and Rodríguez Montaño, R},
title = {Dental biofilm in children with normal weight, overweight and obesity: a pilot study.},
journal = {Acta odontologica latinoamericana : AOL},
volume = {38},
number = {1},
pages = {49-58},
pmid = {40741807},
issn = {1852-4834},
mesh = {Humans ; Pilot Projects ; *Biofilms ; *Dental Plaque/microbiology ; Child ; Male ; Female ; *Overweight ; Body Mass Index ; *Pediatric Obesity ; Adolescent ; *Obesity ; },
abstract = {UNLABELLED: Microorganisms attached to surfaces form intricate colonies known as biofilms. Dental plaque is the biofilm formed on the tooth surface, including the gingival sulcus. Plaque staining makes it easier to see which areas need more brushing time, and which have higher risk of periodontal disease or caries. Dental plaque is largely influenced by diet, becoming more and more dependent on diet related carbohydrates as it ages. Inadequate dental care and bad eating habits are frequently associated with the presence of visible bacterial plaque.

AIM: The aim of this study is to evaluate the percentage of dental plaque according to body mass index (BMI) and to identify whether the plaque is newly deposited, mature or acidified.

MATERIALS AND METHOD: Twenty-five patients from the Pediatric Dentistry Clinics at the University of Guadalajara were evaluated. The percentage and type of plaque were identified using a disclosing gel. The percentage of plaque was compared among children with normal weight, overweight and obesity.

RESULTS: 14 children were normal weight, six were overweight, four were obese, and one was underweight. Average percentage of total plaque was 70.92%. The most predominant plaque was newly deposited (pink staining), followed by mature plaque (purple staining), and a lower percentage of acidified plaque (light blue staining).

CONCLUSIONS: The high percentage of plaque indicates lack or inefficacy of tooth brushing, highlighting the importance of using plaque disclosure for diagnostic and educational purposes for children and parents.},
}

Humans
Pilot Projects
*Biofilms
*Dental Plaque/microbiology
Child
Male
Female
*Overweight
Body Mass Index
*Pediatric Obesity
Adolescent
*Obesity

@article {pmid40741671,
year = {2025},
author = {Biswas, K and Mercy, DJ and Girigoswami, A and Girigoswami, K},
title = {Exploring the inhibitory potentials of metal-doped graphene in combating biofilm formation: a review.},
journal = {Zeitschrift fur Naturforschung. C, Journal of biosciences},
volume = {},
number = {},
pages = {},
pmid = {40741671},
issn = {1865-7125},
abstract = {The emergence of anti-microbial resistance (AMR) has become a pressing need for the design of novel and potential anti-microbial and anti-biofilm agents. Nano-scaled materials have been designed in the research domain to curb the populous spread of microbial biofilms. In the context of nano-scaled materials, metal nanoparticles are of unique importance and have been studied in detail for the treatment of biofilm propagation and menace. The crucial aspects of metal nanoparticles and their complexation with graphene have been extensively understood at the molecular level in this review article. The molecular interplay between the various external stimuli, like pH, temperature, sound, mechanical stimuli, and different external factors, on biofilm regulation has been studied and accentuated in the paper. The effect of such external factors succumbing to the biofilm-producing microbes, to the potential inhibitory activities of such novel nano-composites has been unraveled in the present review. Understanding the methodology behind nanoscaled treatments and their molecular mechanisms could pave the way for novel biofilm targeting strategies.},
}

@article {pmid40741325,
year = {2025},
author = {Benes, K and Liguori, M and Velikaneye, CJ and Kispert, S and Pishnyuk, A and Luzik, E and Sun, H and Xiao, D and Gu, H},
title = {Harnessing Biofilm-Mediated Plastic Biodegradation: Innovating Smart Material Design.},
journal = {ACS applied engineering materials},
volume = {3},
number = {7},
pages = {1915-1926},
pmid = {40741325},
issn = {2771-9545},
abstract = {The persistent environmental challenges posed by synthetic plastics, particularly petroleum-derived petropolymers, such as polyethylene (PE), polypropylene (PP), and polystyrene (PS), have intensified the need for innovative recycling methods. Traditional recycling techniques often rely on harsh conditions, raising environmental and economic concerns. Biofilm-mediated biodegradation has emerged as a promising alternative, operating under mild conditions such as room temperature, neutral pH, and atmospheric pressure. However, the interactions between biofilm-forming microorganisms and synthetic plastics and the roles of secreted enzymes in these processes remain incompletely understood. This review explores the current understanding of biofilm-mediated biodegradation(?)biodeterioration, biofragmentation, bioassimilation, and mineralization(?)and the biochemical and physical interactions that control these processes. We highlight the latest findings on the enhancement of petropolymer degradation by biofilms, focusing on the roles of oxidative and attachment enzymes and the environmental factors influencing degradation efficiency. Understanding these complex interactions can inform the design of next-generation enzyme-responsive polymers that are not only easier to degrade but can also serve as smart materials for diverse applications, such as antifouling coatings on metals. This perspective bridges critical knowledge gaps and provides insights into harnessing biofilm-mediated processes for sustainable material innovation.},
}

@article {pmid40739230,
year = {2025},
author = {Cyris, M and Hach, G and Dörfer, CE and El-Sayed, KF and Graetz, C},
title = {Professional biofilm management during supportive periodontal therapy-a longitudinal observational study.},
journal = {BMC oral health},
volume = {25},
number = {1},
pages = {1277},
pmid = {40739230},
issn = {1472-6831},
mesh = {Humans ; *Biofilms ; Male ; Longitudinal Studies ; Female ; Middle Aged ; *Periodontitis/therapy ; Aged ; *Dental Polishing/methods ; Adult ; },
abstract = {BACKGROUND: Professional mechanical biofilm reduction represents the cornerstone measure during supportive periodontal therapy (SPT). Conventionally, rotating polishing rubber cups (RCs) and brushes with polishing paste or air-polishing (AP) devices using low-abrasive powders can be used. This study aimed to evaluate the effectiveness of both methods in periodontitis patients in a university SPT setting.

METHODS: Patients diagnosed with periodontitis who attended regular SPT at the Department of Conservative Dentistry and Periodontology at the University Hospital Schleswig-Holstein, Kiel campus, at least once a year between 2018 and 2023 were included. Clinical parameters such as number of teeth and pocket probing depth (PPD) were recorded at T1 (first documented SPT session) and T2 (last documented SPT session).

RESULTS: A total of 430 patients (AP/RC: n = 152/n = 278) with an average age of 60.7(11.5) years, were included. Most patients had Stage III (AP/RC: 56.6%/56.8%) and Grade B (AP/RC: 52%/64.4%) periodontitis. The treatment time was 77.9(21.0) min and did not differ between groups (p = 0.378). No significant differences were found in the number of sites with PPD ≤ 4 mm at T1 or T2 (p > 0.05). Sites with PPD ≥ 5 mm differed significantly at T1 (AP: 8 [4-16], RC: 6 [3-12]; p = 0.002) but not at T2 (AP: 6 [3-13], RC: 5.5 [3-11]; p = 0.104). No significant intergroup differences were notable regarding stability, improvement, or deterioration of sites with PPD ≥ 5 mm over time. However, the AP group had significantly more multirooted teeth with Stage III furcation involvement regardless of bleeding on probing (BOP) at T1 (AP: 2 [1-5], RC: 1 [1-3]; p = 0.046) but not at T2. AP demonstrated a significant advantage in preventing deterioration of PPD ≥ 5 mm in molars (AP: 48 [45.3%], RC: 62 [33.3%]; p = 0.027).

CONCLUSIONS: Both methods of professional biofilm removal are similarly effective in terms of stabilizing or improving periodontal sites with PPD ≥ 5 mm when performed regularly. However, in molars with furcation involvement, RC intervention showed more favorable outcomes compared to AP, particularly in preventing the deterioration of sites with PPD ≥ 5 mm, in this study cohort treated in a specialized university-based SPT setting.

TRIAL REGISTRATION: The study was retrospectively registered in the DRKS-German Clinical Trials Register (https://www.drks.de) with the registration-ID DRKS00037021 (22/05/2025).},
}

Humans
*Biofilms
Male
Longitudinal Studies
Female
Middle Aged
*Periodontitis/therapy
Aged
*Dental Polishing/methods
Adult

@article {pmid40738923,
year = {2025},
author = {Mekky, AE and Saied, E and Al-Habibi, MM and Shouaib, ZA and Hasaballah, AI and Rashed, ME and Khaled, AF and Alshammari, AN and Youssef, FS and Al-Shahat, AM and Alfaifi, MY and Elbehairi, SEI and Aufy, M and Selim, TA},
title = {Eco‑friendly biosynthesis of gold nanoparticles from Amphimedon compressa with antibacterial, antioxidant, anti-inflammatory, anti-biofilm, and insecticidal properties against diseases vectors.},
journal = {Scientific reports},
volume = {15},
number = {1},
pages = {27845},
pmid = {40738923},
issn = {2045-2322},
mesh = {Animals ; *Metal Nanoparticles/chemistry/ultrastructure ; *Gold/chemistry/pharmacology ; *Antioxidants/pharmacology/chemistry ; *Insecticides/pharmacology/chemistry ; *Anti-Inflammatory Agents/pharmacology/chemistry ; *Anti-Bacterial Agents/pharmacology/chemistry ; *Porifera/chemistry/metabolism ; Biofilms/drug effects ; Green Chemistry Technology ; Disease Vectors ; },
abstract = {Gold nanoparticles (AuNPs) are increasingly recognized for their potential in biology due to their excellent drug delivery capabilities and ease of synthesis. To create AuNPs using marine sponge Amphimedon compressa, we used several techniques, including ultraviolet-visible (UV-visible) spectrophotometry, Fourier transform infrared (FTIR) spectrophotometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and x-ray diffraction (XRD). The UV-visible spectroscopy results demonstrated the formation of stable AuNPs at a pH of 7, with a peak absorption at 564 nm. FTIR spectroscopy indicated that secondary metabolites featuring -OH functional groups acted as reducing agents in the production of AuNPs. Morphological analysis showed that the AuNPs were spherical, consistently shaped particles averaging 10-40 nm in diameter, with proven stability over time. The inhibition zones for the bacteria tested with the synthesized AuNPs varied from 26 to 31 mm. Both the AuNPs and the A. compressa extract displayed significant antioxidant activity, achieving DPPH radical scavenging percentages of 70.73% and 85.62%, respectively. In terms of anti-inflammatory activity, the AuNPs showed dose-dependent anti-inflammatory activity, with hemolytic inhibition percentages of 4.8%, 10.2%, 12.8%, 14.9%, 19.5%, and 22.4% at increasing concentrations. Furthermore, both the crude extract and the synthesized AuNPs exhibited adulticidal activity against the house fly (Musca domestica) and the mosquito (Culex pipiens). The LC50 and LC90 values for the crude extract were 34.988 and 62.836 ppm for M. domestica, and 9.258 and 17.399 ppm for C. pipiens. For the AuNPs, the corresponding values were 8.545 and 15.157 ppm for M. domestica, and 7.573 and 14.074 ppm for C. pipiens. Adult mortality caused by the AuNPs extract reached 100.00% for both Musca domestica and Culex pipiens at a concentration of 6 ppm. Overall, M. domestica and C. pipiens were more sensitive to AuNPs than to the crude extract. Both the synthesized AuNPs and the crude extract caused a significant, dose-dependent reduction in fecundity and hatchability in M. domestica and C. pipiens. In conclusion, the marine sponge A. compressa serves as an effective biological source for the synthesis of AuNPs, which demonstrate significant antibacterial, antioxidant, anti-inflammatory, anti-biofilm, and insecticidal activities, highlighting their potential in biomedical and environmental fields.},
}

Animals
*Metal Nanoparticles/chemistry/ultrastructure
*Gold/chemistry/pharmacology
*Antioxidants/pharmacology/chemistry
*Insecticides/pharmacology/chemistry
*Anti-Inflammatory Agents/pharmacology/chemistry
*Anti-Bacterial Agents/pharmacology/chemistry
*Porifera/chemistry/metabolism
Biofilms/drug effects
Green Chemistry Technology
Disease Vectors

@article {pmid40737320,
year = {2025},
author = {Sonani, RR and Liu, Y and Xiang, J and Cvirkaite-Krupovic, V and Du, S and Chen, X and Krupovic, M and Egelman, EH},
title = {Tat-dependent bundling pilus of a halophilic archaeon assembles by a strand donation mechanism and facilitates biofilm formation.},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
volume = {122},
number = {31},
pages = {e2514980122},
doi = {10.1073/pnas.2514980122},
pmid = {40737320},
issn = {1091-6490},
support = {GM122510//HHS | NIH | National Institute of General Medical Sciences (NIGMS)/ ; ANR-21-CE11-0001//Agence Nationale de la Recherche (ANR)/ ; },
mesh = {*Biofilms/growth & development ; *Archaeal Proteins/metabolism/genetics/chemistry ; *Fimbriae Proteins/metabolism/genetics/chemistry ; *Twin-Arginine-Translocation System/metabolism ; *Halobacteriaceae/metabolism/physiology/genetics ; },
abstract = {Diverse extracellular filaments present on the surface of archaea mediate multiple key processes, such as motility, adhesion, and biofilm formation. Although several archaeal filament types have been characterized in considerable detail, many remain understudied, particularly those utilizing noncanonical secretion systems. Here, we describe the Tafi bundling pilus that facilitates biofilm formation in the haloarchaeon Natrinema sp. J7-2. Unlike previously characterized archaeal pili, Tafi is secreted via the twin-arginine translocation (Tat) pathway, which transports fully folded proteins across the cytoplasmic membrane. Structural analysis reveals that although Tafi pili assemble via a canonical strand-donation mechanism, the pilin subunit (TafE) adopts a distinct structural topology that sets it apart from the previously characterized Sec-dependent pilins that form bundling pili in archaea. Sequence analyses show that TafE homologs are also present in thermophilic archaea from different phyla, but Tat-signal sequences are exclusive to pilins of halophilic archaea. Nevertheless, we find that Tat signal peptides in haloarchaeal TafE-like pili were exchanged back to the Sec signal peptides on multiple independent occasions. These findings expand our understanding of the diversity and evolution of archaeal extracellular filaments and highlight the Tat pathway as a route for pilus assembly in halophilic archaea.},
}

*Biofilms/growth & development
*Archaeal Proteins/metabolism/genetics/chemistry
*Fimbriae Proteins/metabolism/genetics/chemistry
*Twin-Arginine-Translocation System/metabolism
*Halobacteriaceae/metabolism/physiology/genetics

@article {pmid40736269,
year = {2025},
author = {Li, Y and Koley, D},
title = {Analytical methods to study the complex dynamics of biofilm-biomaterial interfaces.},
journal = {Biointerphases},
volume = {20},
number = {4},
pages = {},
doi = {10.1116/6.0004613},
pmid = {40736269},
issn = {1559-4106},
mesh = {*Biofilms/growth & development ; *Biocompatible Materials/chemistry ; Humans ; Electrochemical Techniques/methods ; *Bacteria ; Surface Properties ; },
abstract = {Biofilm-biomaterial interfaces have an important role in biofilm development and pose a critical challenge in healthcare, contributing to device failures and chronic infections that affect patient outcomes and healthcare economics. This review explores the complex dynamics of these interfaces, from initial protein adsorption through mature biofilm development, highlighting how bacteria and materials are involved in bidirectional interactions that determine both infection progression and material degradation. It also examines different advanced analytical methods for characterizing these dynamic biofilm-biomaterial interactions, with particular emphasis on the recent developments in electrochemical techniques (ion-selective electrodes, electrochemical impedance spectroscopy, and scanning electrochemical microscopy) that enable real-time monitoring of critical parameters such as pH, oxygen gradients, and metabolic activities, providing unique insights into biofilm heterogeneity and localized chemical changes. In addition, the review explores future developments in sensor technology and standardized protocols needed to accelerate biomaterial innovation, potentially transforming our approach to implant-associated infections through responsive surfaces that adapt to microbial challenges.},
}

*Biofilms/growth & development
*Biocompatible Materials/chemistry
Humans
Electrochemical Techniques/methods
*Bacteria
Surface Properties

@article {pmid40736253,
year = {2025},
author = {Holley, CL and Dhulipala, V and Le Van, A and Balthazar, JT and Oliver, VJ and Jerse, AE and Shafer, WM},
title = {Gentamicin induction of the gonococcal hicAB toxin-antitoxin-encoding system and impact on gene expression influencing biofilm formation and in vivo fitness in a strain-specific manner.},
journal = {mBio},
volume = {},
number = {},
pages = {e0159525},
doi = {10.1128/mbio.01595-25},
pmid = {40736253},
issn = {2150-7511},
abstract = {UNLABELLED: The continued emergence of Neisseria gonorrhoeae (Ng) isolates resistant to first-line antibiotics has focused efforts on understanding how alternative therapies, such as the expanded use of gentamicin (Gen), might counteract this global public health problem. Focusing on Gen as a viable alternative antibiotic for the treatment of gonorrheal infections, we used RNA-seq to determine if sub-lethal levels of Gen might impact gonococci on a transcriptional level. We found that sub-lethal Gen levels altered the expression of 23 genes in Ng strain FA19. Many of the differentially regulated genes were associated with known stress responses elaborated by Ng under different harmful conditions. We found that the transcripts of the hicAB operon, which encodes a putative HicA-HicB toxin-antitoxin system that is encoded by tandem genes with the prophage Ngo φ3, were increased in response to Gen. Although the loss of hicAB did not impact gonococcal susceptibility to a variety of antimicrobial agents or harmful environmental conditions, it did reduce biofilm formation in Ng strains F62, FA1090, WHO X, and CDC200 but not that of strain FA19. Furthermore, in strain F62, but not FA19, loss of hicAB reduced the in vivo fitness of Ng during experimental lower genital tract infection of female mice. Furthermore, we found that expression of hicAB can influence levels of the norB transcript, which encodes the nitrate reductase shown previously to be upregulated in gonococcal biofilms. We propose that sub-lethal Gen has the capacity to influence gonococcal pathogenesis through the action of the HicAB toxin-antitoxin system.

IMPORTANCE: During antibiotic treatment, bacteria can be exposed to sub-lethal levels that could serve as a stress signal, resulting in changes in gene expression. The continued emergence of multi-drug-resistant strains of Ng has rekindled interest in expanded use of gentamicin (Gen) for the treatment of gonorrheal infections. We report that sub-lethal levels of Gen can influence levels of Ng transcripts, including that of the gonococcal hicAB-encoded toxin-antitoxin (TA) locus, which is embedded within an integrated prophage. Although the loss of this TA locus did not impact Ng susceptibility to Gen, it reduced the biofilm-forming ability of four of five Ng strains. Furthermore, in an examined strain in this group, we found that Ng fitness during experimental infection was negatively impacted. We propose that levels of the hicA-hicB transcripts can be increased by sub-lethal levels of an antibiotic used in the treatment of gonorrhea and that this could influence pathogenicity.},
}

@article {pmid40733297,
year = {2025},
author = {Di Girolamo, D and Badalamenti, N and Castagliuolo, G and Ilardi, V and Varcamonti, M and Bruno, M and Zanfardino, A},
title = {South Tyrol (Italy) Pastinaca sativa L. subsp. sativa Essential Oil: GC-MS Composition, Antimicrobial, Anti-Biofilm, and Antioxidant Properties.},
journal = {Molecules (Basel, Switzerland)},
volume = {30},
number = {14},
pages = {},
pmid = {40733297},
issn = {1420-3049},
support = {CUP UNIPA B73C22000790001//- NextGenerationEU; Project code CN_00000033/ ; },
mesh = {*Oils, Volatile/pharmacology/chemistry ; *Antioxidants/pharmacology/chemistry ; Gas Chromatography-Mass Spectrometry ; *Biofilms/drug effects ; *Anti-Infective Agents/pharmacology/chemistry ; Italy ; *Apiaceae/chemistry ; Microbial Sensitivity Tests ; Anti-Bacterial Agents/pharmacology/chemistry ; },
abstract = {Pastinaca L. is a small genus belonging to the Apiaceae family, traditionally used for both nutritional and medicinal purposes. Pastinaca sativa L. subsp. sativa is a biennial plant widely distributed in Europe and Asia, with recognized ethnopharmacological relevance. In this study, the essential oil (EO) obtained from the aerial parts of P. sativa subsp. sativa, collected in Alto Adige (Italy)-a previously unstudied accession-was analyzed by GC-MS, and the volatile profile has been compared with that of EOs previously studied in Bulgaria and Serbia. The EO was found to be rich in octyl acetate (38.7%) and octyl butanoate (26.7%), confirming that this species biosynthesizes these natural esters. The EO and its main constituents were tested to evaluate their antimicrobial properties. Furthermore, their biological potential was evaluated through antimicrobial, antibiofilm and antioxidant assays. This research work, in addition to evaluating possible chemotaxonomic differences at the geographical level of EOs of Pastinaca sativa subsp. sativa, has been extended to the determination of the biological properties of this accession never investigated before, with the aim of acquiring a broader vision of biofilm and antibacterial properties.},
}

*Oils, Volatile/pharmacology/chemistry
*Antioxidants/pharmacology/chemistry
Gas Chromatography-Mass Spectrometry
*Biofilms/drug effects
*Anti-Infective Agents/pharmacology/chemistry
Italy
*Apiaceae/chemistry
Microbial Sensitivity Tests
Anti-Bacterial Agents/pharmacology/chemistry

@article {pmid40732795,
year = {2025},
author = {Alghuthaymi, MA},
title = {Antifungal Nanocomposites from Honeybee Chitosan and Royal Jelly-Mediated Nanosilver for Suppressing Biofilm and Hyphal Formation of Candida albicans.},
journal = {Polymers},
volume = {17},
number = {14},
pages = {},
pmid = {40732795},
issn = {2073-4360},
abstract = {Candida albicans complications challenged researchers and health overseers to discover effectual agents for suppressing such yeast growth, biofilm formation and conversion to hyphal form. The nanomaterials and their composites provided extraordinary bioactivities and functionalities as antimicrobial preparations. The extraction of chitosan (BCt) from honeybee corpuses was achieved as an innovative biopolymer for nanocomposite formation. The green (bio)synthesis of nanosilver (AgNPs) was promisingly performed using royal jelly (RJ) as a mediator of synthesis. The RJ-synthesized AgNPs had an average diameter of 3.61 nm and were negatively charged (-27.2 mV). The formulated nanocomposites from BCt/RJ/AgNPs at 2:1 (F1), 1:1 (F2), and 1:2 (F3) ratios had average diameters of 63.19, 27.65, and 52.74 nm, where their surface charges were +33.8, +29.3, and -11.5 mV, respectively. The infrared (FTIR) analysis designated molecules' interactions, whereas the transmission microscopy emphasized the homogenous distribution and impedance of AgNPs within the biopolymers' nanocomposites. Challenging C. albicans strains with nanomaterials/composites pinpointed their bioactivity for suppressing yeast growth and biofilm formation; the F2 nanocomposite exhibited superior actions, with the lowest inhibitory concentrations (MICs) of 125-175 mg/L, whereas the MIC ranges were 150-200 and 175-225 mg/L for F3 and F1, respectively. The different BCht/RJ/AgNP nanocomposites could entirely suppress the biofilm formation of all C. albicans strains. The scanning microscopy reflected the nanocomposite efficiency for C. albicans cell destruction and the complete suppression of hyphal formation. The application of generated BCht/RJ/AgNP nanocomposites is strongly recommended as they are effectual, natural and advanced materials for combating C. albicans pathogens.},
}

@article {pmid40732790,
year = {2025},
author = {Krivoruchko, A and Nurieva, D and Luppov, V and Kuyukina, M and Ivshina, I},
title = {The Lipid- and Polysaccharide-Rich Extracellular Polymeric Substances of Rhodococcus Support Biofilm Formation and Protection from Toxic Hydrocarbons.},
journal = {Polymers},
volume = {17},
number = {14},
pages = {},
pmid = {40732790},
issn = {2073-4360},
support = {122031400671-1//Ministry of Science and Higher Education of the Russian Federation/ ; 124020500028-4//Ministry of Science and Higher Education of the Russian Federation/ ; FSNF-2025-0013//Ministry of Science and Higher Education of the Russian Federation/ ; },
abstract = {Extracellular polymeric substances (EPS) are multifunctional biopolymers that have significant biotechnological potential. In this study, forty-seven strains of Rhodococcus actinomycetes were screened for EPS production and the content of its main components: carbohydrates, lipids, proteins, and nucleic acids. The Rhodococcus strains produced lipid-rich EPS (15.6 mg·L[-1] to 71.7 mg·L[-1]) with carbohydrate concentrations varying from 0.6 mg·L[-1] to 58.2 mg·L[-1] and low amounts of proteins and nucleic acids. Biofilms of R. ruber IEGM 231 were grown on nitrocellulose filters in the presence of n-hexane, n-hexadecane, or diesel fuel. The distribution of β-polysaccharides, glycoconjugates, and proteins between cells and the extracellular matrix was examined using fluorescence microscopy. The observed release of β-polysaccharides into the biofilm matrix in the presence of n-hexane and diesel fuel was regarded as an adaptation to the assimilation of these toxic hydrocarbons by Rhodococcus cells. Atomic force microscopy of the dried EPS film revealed adhesion forces between 1.0 and 20.0 nN, while some sites were highly adhesive (Fa ≥ 20.0 nN). EPS biosynthetic genes were identified, with two glycosyltransferases correlating with an increase in carbohydrate production. The production of EPS by Rhodococcus cells exhibited strain-specific rather than species-specific patterns, reflecting a high genetic diversity of these bacteria.},
}

@article {pmid40732751,
year = {2025},
author = {Hegazy, EE and ElNaghy, WS and Shalaby, MM and Shoeib, SM and Abdeen, NSM and Fouda, MH and Elshora, OA and Elnaggar, MH and Elrefaey, W and Hagag, RY and Elhadidy, AA and Elsebaey, MA and Eltomey, MA and El Nakib, AM and Ageez, MN and Elnady, MS},
title = {Study of Class 1, 2, and 3 Integrons, Antibiotic Resistance Patterns, and Biofilm Formation in Clinical Staphylococcus aureus Isolates from Hospital-Acquired Infections.},
journal = {Pathogens (Basel, Switzerland)},
volume = {14},
number = {7},
pages = {},
pmid = {40732751},
issn = {2076-0817},
mesh = {*Biofilms/growth & development/drug effects ; Humans ; *Staphylococcal Infections/microbiology ; Anti-Bacterial Agents/pharmacology ; *Cross Infection/microbiology ; *Staphylococcus aureus/drug effects/genetics/isolation & purification/physiology ; *Integrons/genetics ; Microbial Sensitivity Tests ; *Drug Resistance, Bacterial/genetics ; Methicillin-Resistant Staphylococcus aureus/drug effects/genetics/isolation & purification ; Bacterial Proteins/genetics ; Penicillin-Binding Proteins ; },
abstract = {Antibiotic resistance and biofilm formation complicate Staphylococcus aureus infections, raising concerns for global health. Understanding antimicrobial resistance and biofilm formation in these pathogens is essential for effective infection management. The current research aimed to assess antibiotic resistance patterns, biofilm formation, and the occurrence of integron classes 1, 2, and 3 in clinical S. aureus isolates. The disc diffusion method tested antibiotic susceptibility. MRSA strains were identified by cefoxitin disc diffusion, and the mecA gene by PCR. The D-test also assessed macrolide-lincosamide-streptogramin B. A microtiter plate assay assessed biofilm formation. By PCR, integron classes were examined. Of the 63 S. aureus isolates, 25 were MSSA and 38 were MRSA. Pus (39.5%) was the most prevalent clinical source of MRSA isolates, while blood (24%) was the predominant source of MSSA isolates. MRSA isolates were more resistant to clindamycin, ciprofloxacin, ofloxacin, levofloxacin, tetracycline, and doxycycline than MSSA isolates. In total, 76.2% of the isolates produced biofilm. Biofilm-producing isolates were more resistant to cefoxitin and clindamycin. The isolates had 33.3% cMLSB resistance. The intI1 gene was found in 21 S. aureus isolates (33.3%), whereas the intI2 or intI3 genes were not detected. Our findings demonstrate the need for strict infection control to prevent the spread of resistant bacteria.},
}

*Biofilms/growth & development/drug effects
Humans
*Staphylococcal Infections/microbiology
Anti-Bacterial Agents/pharmacology
*Cross Infection/microbiology
*Staphylococcus aureus/drug effects/genetics/isolation & purification/physiology
*Integrons/genetics
Microbial Sensitivity Tests
*Drug Resistance, Bacterial/genetics
Methicillin-Resistant Staphylococcus aureus/drug effects/genetics/isolation & purification
Bacterial Proteins/genetics
Penicillin-Binding Proteins

@article {pmid40732174,
year = {2025},
author = {Austin, KM and Frizzell, JK and Neighmond, AA and Moppel, IJ and Ryno, LM},
title = {L-Arabinose Alters the E. coli Transcriptome to Favor Biofilm Growth and Enhances Survival During Fluoroquinolone Stress.},
journal = {Microorganisms},
volume = {13},
number = {7},
pages = {},
doi = {10.3390/microorganisms13071665},
pmid = {40732174},
issn = {2076-2607},
support = {MCB-2226953//U.S. National Science Foundation/ ; DBI-1828041//U.S. National Science Foundation/ ; N/A//Oberlin College/ ; },
abstract = {Environmental conditions, including nutrient composition and temperature, influence biofilm formation and antibiotic resistance in Escherichia coli. Understanding how specific metabolites modulate these processes is critical for improving antimicrobial strategies. Here, we investigated the growth and composition of Escherichia coli in both planktonic and biofilm states in the presence of L-arabinose, with and without exposure to the fluoroquinolone antibiotic levofloxacin, at two temperatures: 28 and 37 °C. At both temperatures, L-arabinose increased the growth rate of planktonic E. coli but resulted in reduced total growth; concurrently, it enhanced biofilm growth at 37 °C. L-arabinose reduced the efficacy of levofloxacin and promoted growth in sub-minimum inhibitory concentrations (25 ng/mL). Transcriptomic analyses provided insight into the molecular basis of arabinose-mediated reduced susceptibility of E. coli to levofloxacin. We found that L-arabinose had a temperature- and state-dependent impact on the transcriptome. Using gene ontology overrepresentation analyses, we found that L-arabinose modulated the expression of many critical antibiotic resistance genes, including efflux pumps (ydeA, mdtH, mdtM), transporters (proVWX), and biofilm-related genes for external structures like pili (fimA) and curli (csgA, csgB). This study demonstrates a previously uncharacterized role for L-arabinose in modulating antibiotic resistance and biofilm-associated gene expression in E. coli and provides a foundation for additional exploration of sugar-mediated antibiotic sensitivity in bacterial biofilms.},
}

@article {pmid40732122,
year = {2025},
author = {Carvalho, PLB and Pessan, JP and do Amaral, B and Troncha, AC and Sousa, SC and Monteiro, DR and Hosida, TY and Delbem, ACB and Sampaio, C},
title = {Impact of Different Regimens of Fluoridated Dentifrice Application on the pH and Inorganic Composition in an Oral Microcosm Biofilm Model.},
journal = {Microorganisms},
volume = {13},
number = {7},
pages = {},
doi = {10.3390/microorganisms13071612},
pmid = {40732122},
issn = {2076-2607},
support = {001//Coordination for the Improvement of Higher Education Personnel (CAPES)/ ; },
abstract = {This study evaluated the pH, fluoride (F), and calcium (Ca) concentrations in saliva-derived microcosm biofilms following treatments with dentifrices applied at different amounts and F concentration. Human saliva was inoculated into McBain culture medium, and treatments were applied at 72/78/96 h (1 min). Fluoridated dentifrices containing 550 or 1100 ppm F (550F and 1100F, respectively) were used at the following combinations (intensities): (i-1) 550F/0.08 g or 1100F/0.04 g; (i-2) 550F/0.16 g or 1100F/0.08 g; (i-3) 550F/0.32 g or 1100F/0.16 g. A negative control (fluoride-free dentifrice-PLA) was also included. Biofilm F and Ca were measured with an ion-selective electrode and colorimetrically, respectively, while pH in the culture medium was measured with a pH electrode. Data were subjected to ANOVA and Student-Newman-Keuls' test (p < 0.05). F-dentifrices did not significantly alter pH compared to PLA, except for 1100F at i-3. Biofilm F levels at i-1 and i-2 were comparable, for both 550F and 1100F, while 1100F at i-3 led to the highest biofilm F concentration. All F-groups showed significantly higher Ca levels than PLA, especially at i-2 and i-3. In conclusion, the interplay between dentifrice amount and F concentration was more influential on the biofilm's inorganic composition and pH than either variable alone.},
}

@article {pmid40735245,
year = {2024},
author = {Bhattacharya, R and Cornell, K and Browning, J},
title = {Simulation of a Radio-Frequency Wave Based Bacterial Biofilm Detection Method in Dairy Processing Facilities.},
journal = {Applied sciences (Basel, Switzerland)},
volume = {14},
number = {11},
pages = {},
pmid = {40735245},
issn = {2076-3417},
support = {P20 GM148321/GM/NIGMS NIH HHS/United States ; },
abstract = {This paper describes the principles behind the radio-frequency (RF) sensing of bacterial biofilms in pipes and heat exchangers in a dairy processing plant using an electromagnetic simulation. Biofilm formation in dairy processing plants is a common issue where the absence of timely detection and subsequent cleaning can cause serious illness. Biofilms are known for causing health issues and cleaning requires a large volume of water and harsh chemicals. In this work, milk transportation pipes are considered circular waveguides, and pasteurizers/heat exchangers are considered resonant cavities. Simulations were carried out using the CST studio suite high-frequency solver to determine the effectiveness of the real-time RF sensing. The respective dielectric constants and loss tangents were applied to milk and biofilm. In our simulation, it was observed that a 1 μm thick layer of biofilm in a milk-filled pipe shifted the reflection coefficient of a 10.16 cm diameter stainless steel circular waveguide from 0.229 GHz to 0.19 GHz. Further sensitivity analysis revealed a shift in frequency from 0.8 GHz to 1.2 GHz for a film thickness of 5 μm to 10 μm with the highest wave reflection (S11) peak of ≈-120 dB for a 6 μm thick biofilm. A dielectric patch antenna to launch the waves into the waveguide through a dielectric window was also designed and simulated. Simulation using the antenna demonstrated a similar S11 response, where a shift in reflection coefficient from 0.229 GHz to 0.19 GHz was observed for a 1 μm thick biofilm. For the case of the resonant cavity, the same antenna approach was used to excite the modes in a 0.751 m × 0.321 m × 170 m rectangular cavity with heat exchange fins and filled with milk and biofilm. The simulated resonance frequency shifted from 1.52 GHz to 1.54 GHz, for a film thickness varying from 1 μm to 10 μm. This result demonstrated the sensitivity of the microwave detection method. Overall, these results suggest that microwave sensing has promise in the rapid, non-invasive, and real-time detection of biofilm formation in dairy processing plants.},
}

@article {pmid40732107,
year = {2025},
author = {Zheng, J and He, L and Wei, Y and Lu, J and Liu, X and Li, W},
title = {Cold Shock Proteins Balance Biofilm-Associated Antibiotic Resistance and Oxidative Vulnerability in Mycobacteria.},
journal = {Microorganisms},
volume = {13},
number = {7},
pages = {},
doi = {10.3390/microorganisms13071597},
pmid = {40732107},
issn = {2076-2607},
support = {32470034//National Natural Science Foundation of China Grants/ ; },
abstract = {Cold Shock Proteins (Csps) are multifunctional regulators critical for bacterial stress adaptation. While Csps are known to regulate biofilm formation and low-temperature growth in some species, their roles in mycobacteria remain unclear. Here, we explored the functions of three Csps (CspA1, CspA2, and CspB) in Mycobacterium smegmatis. We found that CspA1 promotes biofilm formation and isoniazid (INH) resistance but negatively affects oxidative stress resistance. In contrast, CspB promotes biofilm formation, whereas CspA2 appears functionally redundant in this process. Notably, CspB and CspA2 do not contribute redundantly to oxidative stress resistance. Proteomic analysis revealed that CspA1 significantly modulates the expression of key metabolic and stress-response proteins, including WhiB3 and KatG. Our findings establish CspA1 as a key regulatory factor in mycobacteria, linking metabolic adaptation to biofilm-associated drug resistance and oxidative defense.},
}