@article {pmid42322458,
year = {2026},
author = {Behera, S and Das, S},
title = {Biofilm formation in Streptomyces nigra strain KDS4 and characterization of extracellular polymeric substances (EPS) for biomaterial development.},
journal = {World journal of microbiology & biotechnology},
volume = {42},
number = {7},
pages = {},
pmid = {42322458},
issn = {1573-0972},
mesh = {*Streptomyces/metabolism/physiology/genetics/growth & development ; *Extracellular Polymeric Substance Matrix/chemistry/metabolism ; *Biofilms/growth & development ; *Biocompatible Materials/chemistry ; Hydrogels/chemistry ; Polypropylenes ; Spores, Bacterial/growth & development ; Bacterial Proteins/genetics/metabolism ; Biosurfactants ; },
abstract = {Biofilm formation in marine Streptomyces is a dynamic yet poorly understood process, that limits their functional exploitation. This study investigated the biofilm development and extracellular polymeric substances (EPS) synthesis by Streptomyces nigra strain KDS4, a promising marine Actinobacterium, aiming to optimize EPS yield and characterize its bio-functional properties. Biofilm and EPS formation began with spore germination and hyphal growth, maturing by 60 h with dense hyphal intertwining, sporulation, and EPS secretion, followed by dispersal at 84 h. The bacterium showed the highest biofilm height (~ 2.35 μm) over the polypropylene substrate. Upregulated expression of the cslA gene, associated with biofilm matrix production, was confirmed during biofilm development. Structural analysis of EPS revealed α- and β-glycosidic linkages, hydroxyl, and alkyne groups, along with an amorphous morphology and diverse elemental composition. EPS exhibited thermal transition up to 300 °C and antioxidant and emulsifying properties. Notably, EPS demonstrated hydrogel-forming capability, with 5% (wt/wt) EPS-based hydrogel exhibiting rapid gelation (73 s), high porosity and pore size (31.66 μm), excellent swelling (53.54%), and strong viscoelasticity (G' > G''). At 20% (wt/wt) EPS, the hydrogel achieved a compressive strength of 36.83 kPa, demonstrating its mechanical robustness. These findings highlight S. nigra strain KDS4 as a promising source of multifunctional EPS for sustainable environmental and biomedical applications. While the study provides detailed in vitro insights, evaluation of EPS functionality and biocompatibility remain to be explored. Future work should focus on scale-up production, structural-functional correlations, and validation of EPS-based hydrogels in environmental remediation and biomedical models.},
}

*Streptomyces/metabolism/physiology/genetics/growth & development
*Extracellular Polymeric Substance Matrix/chemistry/metabolism
*Biofilms/growth & development
*Biocompatible Materials/chemistry
Hydrogels/chemistry
Polypropylenes
Spores, Bacterial/growth & development
Bacterial Proteins/genetics/metabolism
Biosurfactants

@article {pmid42322466,
year = {2026},
author = {da Silva, JSSC and da Mata, LC and Monteiro, VN and Ribeiro, EL and Dias, LD and Naves, PLF},
title = {Curcumin-mediated antimicrobial photodynamic therapy prevents Candida biofilm formation in a species-dependent manner.},
journal = {Lasers in medical science},
volume = {41},
number = {1},
pages = {},
pmid = {42322466},
issn = {1435-604X},
mesh = {*Curcumin/pharmacology ; *Biofilms/drug effects/radiation effects/growth & development ; *Photochemotherapy/methods ; *Candida/drug effects/physiology ; *Photosensitizing Agents/pharmacology ; Humans ; },
abstract = {This study aimed to systematically evaluate the antibiofilm efficacy of curcumin-mediated antimicrobial photodynamic therapy (aPDT) activated by 450 nm LED light against biofilm formation and mature biofilms of clinically relevant Candida species, with particular emphasis on preventive potential and species-dependent susceptibility; Biofilm formation and maturation were investigated in five standard Candida strains. Biofilm formation (preventive approach) and established biofilms were treated with curcumin followed by irradiation with 450 nm LED light. Biofilm biomass was quantitatively assessed using the crystal violet assay. Comparative analyses were performed to determine species-dependent responses to aPDT. Data normality was assessed using the Shapiro-Wilk test, and intergroup comparisons were performed using the Kruskal-Wallis test followed by Dunn's post hoc test when appropriate. All Candida strains demonstrated a pronounced ability to form biofilms, being classified as strong biofilm producers, except Candida dubliniensis, which exhibited moderate biofilm formation. Curcumin-mediated aPDT significantly inhibited biofilm development across all tested species (p < 0.05), achieving reductions of 86.82% for C. metapsilosis, 85.05% for C. orthopsilosis, 83.33% for C. parapsilosis, 75.34% for C. dubliniensis, and 68.19% for C. albicans. This preventive antibiofilm effect is likely associated with reactive oxygen species generation, resulting in oxidative damage to essential cellular structures and impairment of early extracellular matrix establishment. In contrast, aPDT activity against mature biofilms was significantly attenuated and highly species-dependent, with reductions ranging from 49.63% (C. parapsilosis) to 0.64% (C. dubliniensis); however, these reductions were not statistically significant (p > 0.05). Curcumin-mediated aPDT exerted a significant preventive antibiofilm effect against Candida biofilm formation but showed limited and non-significant activity against mature biofilms. These findings position curcumin-mediated aPDT as a promising strategy for preventing Candida biofilm establishment, while indicating that improved photosensitizer delivery and optimized irradiation protocols are required to enhance activity against established biofilms.},
}

*Curcumin/pharmacology
*Biofilms/drug effects/radiation effects/growth & development
*Photochemotherapy/methods
*Candida/drug effects/physiology
*Photosensitizing Agents/pharmacology
Humans

@article {pmid42323344,
year = {2026},
author = {Ledger, EVK and Horgan, NE and Lynch, D and Bateman, LM and Massey, RC},
title = {Human serum triglycerides promote Staphylococcus aureus biofilm formation and antibiotic tolerance.},
journal = {NPJ biofilms and microbiomes},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41522-026-01064-x},
pmid = {42323344},
issn = {2055-5008},
support = {GA004851//Microbiology Society/ ; 21/FFP-A/9704//Research Ireland/ ; 212258/Z/18/Z/WT_/Wellcome Trust/United Kingdom ; },
abstract = {Biofilm formation and antibiotic tolerance are major contributors to the persistence of Staphylococcus aureus infections, yet how the host environment affects these phenotypes remains poorly understood. Here, we show that incubation in human serum primes S. aureus to form robust biofilms and tolerate vancomycin and daptomycin, last resort antibiotics for the treatment of antibiotic-resistant staphylococcal infections. Mechanistically, we demonstrate that the staphylococcal Geh lipase is essential for serum-induced biofilm formation by liberating glycerol from host lipids, which is then used to promote increased synthesis of D-alanylated wall teichoic acids, driving biofilm development. Inhibition of the Geh lipase or wall teichoic acid synthesis markedly reduces biofilm formation and restores antibiotic susceptibility, highlighting clinically achievable strategies to inhibit host-induced biofilm formation and prevent the associated antibiotic tolerance. Together, our findings reveal a host-driven mechanism of biofilm-associated antibiotic tolerance in S. aureus and provide rational targets for therapeutic intervention.},
}

@article {pmid42324599,
year = {2026},
author = {Kunyeit, L and Rao, R},
title = {Biofilm overproduction enhances gastrointestinal stress tolerance and intestinal fitness in Bacillus subtilis.},
journal = {Gut microbes},
volume = {18},
number = {1},
pages = {2684066},
doi = {10.1080/19490976.2026.2684066},
pmid = {42324599},
issn = {1949-0984},
mesh = {*Bacillus subtilis/physiology/genetics/growth & development ; Animals ; Caenorhabditis elegans/microbiology ; *Biofilms/growth & development ; *Stress, Physiological ; Probiotics ; *Gastrointestinal Tract/microbiology ; *Intestines/microbiology ; Microbial Viability ; Saccharomyces boulardii/physiology/growth & development ; Lacticaseibacillus rhamnosus/physiology ; },
abstract = {Microorganisms with health-promoting potential often experience substantial losses in viability and function due to stresses encountered during manufacturing and gastrointestinal transit. In this study, we investigate whether biofilm can be leveraged to enhance microbial resilience and functional performance. Using Bacillus subtilis as a model biofilm-forming bacterium, we examined strains with defined biofilm phenotypes: a biofilm-deficient mutant (tasA eps), a biofilm-overproducing mutant (sinR), and an isogenic wild-type control. These strains were evaluated across multiple functional benchmarks, including survival in simulated gastric and bile juices, thermotolerance, and intestinal bacterial colonization in the Caenorhabditis elegans model. Commercially available strains Lactobacillus rhamnosus GG and Saccharomyces boulardii were included as reference comparators. The biofilm-overproducing B. subtilis sinR strain demonstrated markedly enhanced survival under simulated gastrointestinal conditions and showed increased colonization within the C. elegans intestine. In contrast, the biofilm-deficient tasA eps mutant exhibited severe sensitivity to gastric stress and reduced the intestinal bacterial load. Furthermore, we demonstrate that cell-free B. subtilis biofilm can function as an effective bioencapsulation matrix. When used to encapsulate multiple probiotic strains, the biofilm matrix significantly improved their survival under acidic gastric conditions by neutralizing the environmental pH, indicating its broad potential for probiotic formulations and targeted gastrointestinal delivery. Overall, biofilms are traditionally studied for their roles in infection and antimicrobial resistance; however, their protective and adaptive traits may be repurposed for beneficial use. As an example of this concept, our findings show that B. subtilis biofilms enhance multiple functional and technological traits and highlight biofilm-based strategies as a promising platform for improving beneficial microbial robustness and the delivery of live biotherapeutics.},
}

*Bacillus subtilis/physiology/genetics/growth & development
Animals
Caenorhabditis elegans/microbiology
*Biofilms/growth & development
*Stress, Physiological
Probiotics
*Gastrointestinal Tract/microbiology
*Intestines/microbiology
Microbial Viability
Saccharomyces boulardii/physiology/growth & development
Lacticaseibacillus rhamnosus/physiology

@article {pmid42325454,
year = {2026},
author = {Alsuwat, MA and Shah, AA and Ullah, S and Khan, RU and Alissa, M and Khan, MS},
title = {Microbial Biofilm Formation to Mitigate Foodborne Pathogens Strategies and Control Measures.},
journal = {Indian journal of microbiology},
volume = {66},
number = {3},
pages = {496-511},
pmid = {42325454},
issn = {0046-8991},
abstract = {Foodborne pathogens pose a serious public health threat, causing widespread illness and severe consequences. A major challenge in their control is the formation of biofilms on surfaces in food production environments, enhancing bacterial survival, antimicrobial resistance, and persistence. This review investigates biofilm formation strategies employed by key pathogens like Salmonella, Escherichia coli, and Listeria monocytogenes, emphasizing the pivotal role of biofilm management in addressing food safety concerns. The study explores the genetic, molecular, and environmental factors influencing biofilm development, which are crucial for devising effective control measures. Strategies employed by bacteria, such as quorum sensing, adhesion mechanisms, and extracellular polymeric substance production, are detailed. This review also discusses current control measures, including chemical and physical interventions, novel approaches like bacteriophages and biofilm-disrupting enzymes, and considerations in surface material design to minimize biofilm formation. In conclusion, a comprehensive understanding of biofilm formation strategies and effective control measures is essential for ensuring food safety. This review provides insights into managing biofilm-associated risks in the food industry, contributing to innovative and sustainable approaches for mitigating the impact of foodborne pathogens.},
}

@article {pmid42325458,
year = {2026},
author = {Dixit, S and Rai, S and Gupta, D and Sharma, S},
title = {Biofilm Formation on Different Fabrics in the Presence of Sweat.},
journal = {Indian journal of microbiology},
volume = {66},
number = {3},
pages = {587-595},
pmid = {42325458},
issn = {0046-8991},
abstract = {UNLABELLED: The colonization of textiles by axillary skin bacteria produces an unpleasant odour due to the rapid growth of a selective community of bacteria. Such colonized textiles subsequently act as vectors for transmitting nosocomial infections among healthcare workers and patients. An in-depth understanding of bacterial behaviour on soft surfaces like fabrics is necessary to mitigate the transmission of infections. This study examined the effect of artificial human sweat on biofilm formation by Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, and Pseudomonas aeruginosa, on three fabrics, viz. polyester, cotton, and polyester-cotton (70:30) blend. Artificial sweat was constituted to replicate the natural human sweat on textiles. Using atomic force microscopy, the three-dimensional topography of the biofilm was determined, and scanning electron microscopy was employed to visualise the biofilm that had developed on the fabrics. All bacterial strains showed maximum growth on polyester fabric in the presence of sweat. P. aeruginosa and S. aureus were found to be strong biofilm producers, whereas E. coli and E. faecalis were moderate producers. The ability of the four bacterial strains to form biofilm was related to their production of extracellular polymeric substances (EPS). P. aeruginosa produced viscous EPS in contrast to the EPS produced by other bacterial strains. In conclusion, this study corroborates that sweat plays a major role in the colonization of textiles by bacteria. Regular practice of fabric hygiene, and the development of modified fabrics with anti-pathogen properties, could potentially reduce the prevalence of nosocomial infections in healthcare settings.

SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-024-01409-0.},
}

@article {pmid42325464,
year = {2026},
author = {Subramanian, A},
title = {Molecular Characterization of Cariogenic, Biofilm-forming, Multi-drug-resistant Pseudomonas aeruginosa and Detection of NDM-1 and blaVIM-1 Genes.},
journal = {Indian journal of microbiology},
volume = {66},
number = {3},
pages = {782-792},
pmid = {42325464},
issn = {0046-8991},
abstract = {The prevalence of cariogenic bacteria in dental caries is attributed to acidification of the oral microenvironment. Cariogenic Pseudomonas aeruginosa strains were isolated from dental caries. A total of 101 P. aeruginosa strains were isolated from 55 samples (n = 55). The isolates were classified based on their acid tolerance and acidogenic properties. The isolated bacterial strains were cultured in soy peptone (2%) and maintained in a simulated oral environment. The survival rate of the isolates varied from 40.2 ± 2.1 to 74.7 ± 1.8%, and only 11 strains had a significant survival rate (> 60%). Among these DS47 strain exhibited strong biofilm production, with an optical density value significantly higher than that of the other isolates (p < 0.01). This strain was further tested to determine its demineralization effect on teeth. The biofilm-forming bacteria DS47 released Ca[2+] ions in a time-dependent manner (p < 0.01), indicating its potential role in enamel demineralization. Additionally, the DS47 isolate was found to be multidrug-resistant, showing resistance to tobramycin, cefotaxime, ceftazidime, amoxicillin, trimethoprim, ciprofloxacin, levofloxacin, amikacin, ceftazidime, cefoxitin, and gentamicin. The New Delhi metallo β-lactamase-1 (NDM-1) gene and blaVIM-1 genes were detected in the drug-resistant P. aeruginosa DS47. The biofilm-producing P. aeruginosa showed cariogenicity and contributed to dental caries. These findings suggest that biofilm-producing P. aeruginosa plays a significant role in cariogenicity and contributes to dental caries.},
}

@article {pmid42328219,
year = {2026},
author = {Baidya, D and Pathivada, L and Garg, N and Karthik, KM and Jain, AK and Bhyri, P},
title = {Effect of Three Different Topical Fluoride Varnishes on Streptococcus mutans Count in Biofilm Samples of Children Aged 6-10 Years: A Randomized Controlled Trial.},
journal = {International journal of clinical pediatric dentistry},
volume = {19},
number = {3},
pages = {343-349},
pmid = {42328219},
issn = {0974-7052},
abstract = {OBJECTIVE: This study aims to check how well three fluoride varnishes work. The varnishes are GC MI Varnish, Ultradent Enamelast, and Ivoclar Fluor Protector. They are tested to see if they can lower Streptococcus mutans levels in biofilm samples from children aged 6-10 years.

MATERIALS AND METHODS: A randomized controlled trial was conducted with 144 children divided into three groups. Samples of plaque were gathered initially and then again at intervals of 1, 3, and 6 months following the application of fluoride varnish. The levels of S. mutans were measured using culture methods and reported as colony-forming units per milliliter (CFU/mL).

RESULTS: All three fluoride varnishes significantly reduced S. mutans counts over the 6-month period. GC MI Varnish showed the greatest reduction, particularly at 1 and 3 months (p < 0.0001 and p = 0.01, respectively). The mean CFU/mL for GC MI Varnish decreased from 7.64 ± 0.18 at baseline to 5.76 ± 2.05 at 6 months. Ultradent Enamelast and Fluor Protector S also demonstrated significant reductions but to a lesser extent than GC MI Varnish.

CONCLUSION: GC MI Varnish containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) demonstrated superior antimicrobial efficacy against S. mutans compared to Ultradent Enamelast and Fluor Protector S. All three fluoride varnishes were effective in reducing S. mutans counts, with GC MI Varnish showing the most significant and consistent reductions over time.

HOW TO CITE THIS ARTICLE: Baidya D, Pathivada L, Garg N, et al. Effect of Three Different Topical Fluoride Varnishes on Streptococcus mutans Count in Biofilm Samples of Children Aged 6-10 Years: A Randomized Controlled Trial. Int J Clin Pediatr Dent 2026;19(3):343-349.},
}

@article {pmid42328873,
year = {2026},
author = {Solano Morales, AK and Cazano, E and Pirani, C and Jones, G and Goode, A and Walker, AR and Sperduto, A and Dwivedi, B and Bantha, P and Peter, SD and McLellan, LK and Alam, MA and Shields, RC},
title = {Genomic instability and biofilm determinants in Streptococcus mutans: insights from a sequence-defined arrayed transposon library.},
journal = {Journal of bacteriology},
volume = {},
number = {},
pages = {e0017426},
doi = {10.1128/jb.00174-26},
pmid = {42328873},
issn = {1098-5530},
abstract = {UNLABELLED: Streptococcus mutans is a primary architect of dental caries, utilizing complex genetic networks to build resilient, acid-producing biofilms. While pooled screens (Tn-seq) have identified important fitness factors, they often fail to capture extracellular or moderate-effect determinants due to community-level masking. Therefore, to study biofilm phenotypes, we constructed a comprehensive arrayed library of 9,216 mutants and used Cartesian Pooling-Coordinate Sequencing (CP-CSeq) to establish a sequence-defined resource covering 51% of non-essential genes. By screening the entire collection in isolation, we identified several novel biofilm determinants, including the putative metal transporter SMU_635 and the glycosylation-associated protein SMU_2160. However, systematic whole-genome sequencing (WGS) of our hits revealed an interesting level of genomic instability: 25% of biofilm-defective mutants had undergone spontaneous recombination at the gtfBC locus, while 7% had lost the TnSmu1 element, an excision rate 1,000-fold higher than previously reported. While targeted mutagenesis confirmed that TnSmu1 loss does not impact biofilm integrity, the gtfBC deletions directly accounted for the most severe phenotypes, highlighting a systemic risk of misattributing gene functions to primary transposon insertions. Our findings provide a powerful new genetic resource for the S. mutans community while establishing a critical new standard: an arrayed library is only as defined as its underlying genome, making systematic genomic verification an essential prerequisite for accurate functional genomics.

IMPORTANCE: Streptococcus mutans causes dental caries through resilient, acidogenic biofilm formation. While pooled screens often overlook extracellular or moderate-effect determinants due to community masking, this study presents a sequence-defined arrayed mutant library to dissect individual gene functions in isolation. Beyond known machinery, we identified novel biofilm determinants, including metal transporter SMU_635 and glycosylation-associated protein SMU_2160. Crucially, we uncovered pervasive genomic instability at the gtfBC and TnSmu1 loci. This reveals a systemic functional genomics risk: misattributing phenotypes to primary mutations when backgrounds undergo large-scale rearrangements. By establishing whole-genome verification as a necessary standard, this research ensures that future therapeutic target identification is built upon a verified genetic foundation.},
}

@article {pmid42328874,
year = {2026},
author = {Meyer, MD and Bergkessel, M and DePas, WH},
title = {Activation of the antibiotic resistance factor WhiB7 can stimulate aggregate biofilm formation in stationary phase Mycobacterium smegmatis by reinitiating translation.},
journal = {Journal of bacteriology},
volume = {},
number = {},
pages = {e0007626},
doi = {10.1128/jb.00076-26},
pmid = {42328874},
issn = {1098-5530},
abstract = {There is a growing understanding that slow growth and dormancy due to nutrient deprivation are very common physiological states exhibited by bacterial communities in a myriad of environments. However, very little is known about the role of slow growth and dormancy in biofilm regulation. Here, we utilize tractable dormancy and aggregation assays in nontuberculous mycobacteria (NTM) to ask the fundamental question of how growth arrest impacts the processes of aggregation and dispersal. First, we show that the well-conserved dormancy regulator DosSR affects biofilm formation in Mycobacterium smegmatis, as a dosR deletion mutant undergoes spurious re-aggregation and dispersal during aerobic late stationary phase. Identification of a suppressor mutation blocking re-aggregation in the ΔdosR mutant allowed us to determine a role for the antibiotic resistance factor WhiB7 in driving re-aggregation in M. smegmatis. We utilized BioOrthogonal NonCanonical Amino acid Tagging (BONCAT), qPCR, and quantitative aggregation assays to build a model wherein reductive stress in ΔdosR potentiates stationary phase translation in a WhiB7-dependent manner, permitting aggregation in dormant stationary phase cells. In addition, during stationary phase, WhiB7-activating reducing agents and antibiotics could trigger re-aggregation in both wild-type M. smegmatis and clinical isolates of the opportunistic NTM pathogen Mycobacterium abscessus. Finally, we determined that, in contrast to aerobic stationary phase, M. smegmatis does not aggregate or disperse in response to chemical cues or antibiotics under the Wayne model of hypoxic dormancy. Our work reveals a regulatory interaction between dormancy and aggregation that could have broad implications for treating and preventing NTM biofilms.IMPORTANCEMycobacteria aggregate to form multicellular biofilms that provide protection from external stressors and increase antibiotic tolerance. Understanding the pathways regulating biofilm formation can aid the identification of useful targets for developing new drugs. With a growing appreciation that pathogens are often in a slow growth/dormant state during infection, we investigate how dormancy affects biofilm formation and dispersal in two nontuberculous mycobacteria (NTM) species: Mycobacterium smegmatis and the opportunistic pathogen Mycobacterium abscessus. We find that activation of the WhiB7-mediated antibiotic response permits biofilm formation in aerobic stationary phase by reinitiating protein synthesis; however, cells under hypoxic dormancy are unresponsive. Our work provides important context to combatting biofilm formation in infection sites, informing future studies and aiding design of biofilm dispersal agents.},
}

@article {pmid42329318,
year = {2026},
author = {Qiao, Z and Luo, S and Mao, Y and Wang, H and Huang, J},
title = {Genomic, Probiotic, and Safety Characterization of Lactiplantibacillus Plantarum L-1 and the Anti-Biofilm Activity of its Bacteriocin Against Listeria Monocytogenes.},
journal = {Probiotics and antimicrobial proteins},
volume = {},
number = {},
pages = {},
pmid = {42329318},
issn = {1867-1314},
support = {262300422148//Natural Science Foundation of Henan Province/ ; QNZX202501//Zhumadian City Science and Technology Innovation Youth Special Project/ ; XKPY-2022005//Science and technology incubation fund of Huanghuai University/ ; 501621701002//Youth Backbone Teacher Project of Huanghuai University/ ; },
abstract = {Listeria monocytogenes biofilms persist in food processing environments and pose a serious threat to food safety. This study aimed to comprehensively characterize Lactiplantibacillus plantarum L-1, a strain isolated from traditional Chinese Jiangshui, and to evaluate the anti-biofilm activity of its crude bacteriocin against L. monocytogenes. The strain exhibited promising probiotic attributes, including high survival rates under simulated gastrointestinal conditions (68.9% at pH 2.0, 85.0% in the presence of 0.3% bile salts) and a satisfactory safety profile (antibiotic susceptibility, γ-hemolysis, and no toxicity in mice). Whole-genome sequencing identified genetic determinants for stress tolerance and a gene cluster encoding multiple bacteriocins, including pln EF, pln J, pln N, and a putative bacteriocin. LC-MS identified three expressed bacteriocins: Plantaricin E, F, and N. The crude bacteriocin showed high stability under a range of temperatures (60-121 °C) and pH (2.0-12.0), with a MIC of 2.2 mg/mL against L. monocytogenes. At sub-inhibitory concentrations that did not affect planktonic growth (1/32×, 1/16×, 1/8× MIC), it significantly inhibited biofilm formation in a concentration-dependent manner, achieving 89.5% inhibition at 1/4 × MIC. The bacteriocin suppressed metabolic activity, reduced exopolysaccharide (EPS) production, and inhibit the integrity of biofilm structure, and downregulated the expression of key biofilm-related genes without affecting bacterial growth. These findings highlight the potential of L. plantarum L-1 as a dual-functional probiotic and a natural biocontrol agent against L. monocytogenes biofilms in the food industry.},
}

@article {pmid42320152,
year = {2026},
author = {Bell, A and Ergas, SJ and Main, K and Rhody, N and Guttman, L},
title = {Performance of moving bed biofilm, periphyton, and halophyte biofilters in marine multi-trophic aquaculture systems.},
journal = {The Science of the total environment},
volume = {1045},
number = {},
pages = {181943},
doi = {10.1016/j.scitotenv.2026.181943},
pmid = {42320152},
issn = {1879-1026},
abstract = {Combining multi-trophic recirculating aquaculture system (MT-RAS) biofilter types leverages the strengths of different ecological biomes, benefits water treatment, resource recovery, economics, and environmental sustainability. The overall goal of this study was to determine the effects of different aquaculture biofilter combinations on MT-RAS. Three duplicate biofilter combinations were tested in a pilot scale MT-RAS with red drum (Sciaenops ocellatus): 1) periphyton with halophytes (P + H), 2) periphyton with moving bed biofilm reactors (P + M), and 3) periphyton only (P[2]). Experiments were performed in two trials (spring and summer) with four replicates. Water quality tests validated that NH3/NH4[+], NO2[-], NO3[-], and CO2 were below fish toxic limits for all biofilter combinations. Fish mortalities were low, with food conversion ratios between 1.1 and 2.0. In all trials, periphyton added dissolved oxygen (DO) to the water (at an average of +3.95 ± 6.52 mg/(L*d)), thus reducing energy costs. Periphyton was also found to include valuable lipid content (4.55 ± 2.24% of dry weight) with the detection of Ω-3 fatty acids. The P[2] trials maintained a stable alkalinity and pH balance. The M + P trials removed NH3/NH4[+] at a high rate; however, they also required more energy for DO. Edible sea purslane growth rates (1.0431 ± 0.3361 g/day/plant) were efficient in all P + H trials. The microbiome revealed abundance of Ignavibacterium bacteria, Navicula and Chlorella algae, Nitrospira, Nitrospirae, Nitrosospharota, and Nitrosoarchaeum nitrogen cyclers. Overall, periphyton biofilter combinations nitrify, denitrify, stabilize pH, photosynthesize, and produce oxygen and a value-added product.},
}

@article {pmid42320320,
year = {2026},
author = {Vargová, M and Bujňáková, D and Galambošiová, T and Zigo, F and Zahumenská, J and Hisira, V},
title = {Biofilm production, molecular attributes and antibiotic resistance of Staphylococcus aureus isolated from bovine milk.},
journal = {Research in veterinary science},
volume = {210},
number = {},
pages = {106303},
doi = {10.1016/j.rvsc.2026.106303},
pmid = {42320320},
issn = {1532-2661},
abstract = {The formation of biofilm and resistance to certain antibiotics by staphylococci are acknowledged as a significant virulence factors. We focused on monitoring phenotype antibiotic resistance and biofilm production, including some genes encoding for cell wall-anchored surface proteins of Staphylococcus aureus (S. aureus) from bovine milk with subclinical mastitis. A total of 230 lactating cows were selected for the detection of mastitis. Of the tested dairy cows, 82 (35.7%) were positive on the California Mastitis Test (CMT). From a total of 105 CMT-positive quarter milk samples, 87 cases were identified as bacterial pathogens of the mammary gland. The most common udder pathogens among all infected quarters were coagulase-negative staphylococci (50.6%) and S. aureus (20.7%). Of all isolates of S. aureus (18), 10 isolates showed resistance to ampicillin, followed by resistance to erythromycin (10/18), to piperacillin + tazobactam, clindamycin (9/18), to oxacillin, ampicillin + sulbactam and to cefoxitin (8/18). One strain was resistant to chloramphenicol. Strong biofilm production was detected in 5 S. aureus; half of the strains were weak producers of biofilm, and 4 isolates had moderate formation capacity. The genes isdA, isdB and agrI were detected in all examined isolates from cows with subclinical mastitis. In our study, fnbpA was present in 17 isolates. The majority of S. aureus exhibited antibiotic resistance, biofilm production and the presence of virulence genes, hence constituting a potential public health threat.},
}

@article {pmid42320403,
year = {2026},
author = {Kigo, Y and Furuno, S and Miura, H and Kobayashi, M and Takahashi, S and Terada, A},
title = {Enhanced oxygen transfer, stable nitrifying biofilm, and low N2O emissions in a pilot-scale hybrid MABR incorporating gear-structured gas-permeable membranes.},
journal = {Water research},
volume = {304},
number = {},
pages = {126260},
doi = {10.1016/j.watres.2026.126260},
pmid = {42320403},
issn = {1879-2448},
abstract = {This study demonstrates the feasibility and performance of a pilot-scale hybrid membrane-aerated biofilm reactor (MABR) equipped with an innovative gear-structured polyethylene membrane. Over 210 days of continuous operation, the integrated system-comprising an anaerobic tank, a hybrid MABR tank, containing 2-16 cassettes within a membrane module, and an aerobic tank-achieved removal efficiencies of 88 ± 12% for BOD, 40 ± 14% for total nitrogen, and 90 ± 14% for ammonia. The total sludge yield was 0.20 kg-TSS/kg-CODMn, significantly lower than that of conventional activated sludge systems. Spatial and temporal variations in oxygen transfer rate (OTR) and efficiency (OTE) were observed across membrane cassettes. The average OTR ranged from 3.51 to 9.63 g-O2/(m[2]·day) and was influenced by operating time and membrane surface area, with lower surface areas enhancing OTR. These results highlight the importance of hydrodynamics and cassette configuration in optimizing oxygen supply. Campaign-based assessments revealed low N2O emission factors of 0.058% on day 100 (29.2 °C) and 0.074% on day 190 (18.5 °C). At 18.5 °C, N2O emissions were primarily attributed to the aerobic tank, while contributions from the MABR tank decreased over time, likely due to biofilm maturation. Combined microbial analyses revealed dense populations of ammonia-oxidizing bacteria within the membrane biofilm, dominated by comammox Nitrospira, which likely contributed to the observed low N2O emissions. Overall, gear-structured membranes promoted resilient biofilm formation, reduced sludge production, and minimized N2O emissions. These findings provide important insights into the design and operation of sustainable and energy-efficient MABR systems for wastewater treatment.},
}

@article {pmid42321540,
year = {2026},
author = {Olorunshola, MM and Oladosu, VI},
title = {Evolved phage cocktails outsmart biofilm defences.},
journal = {Nature microbiology},
volume = {},
number = {},
pages = {},
pmid = {42321540},
issn = {2058-5276},
}

@article {pmid42312298,
year = {2026},
author = {Steffen, K and Bitter, K and Schlafer, S and Paris, S and Bauer, M},
title = {Combined treatment with bacteriophages and sodium hypochlorite against E. faecalis biofilm in an in vitro root canal model with evaluation of bacterial resistance.},
journal = {Journal of oral microbiology},
volume = {18},
number = {1},
pages = {2687225},
pmid = {42312298},
issn = {2000-2297},
abstract = {AIM: To assess the time-dependent antimicrobial activity of a newly isolated bacteriophage against Enterococcus faecalis biofilms in two in vitro models, aloneand combinedwith sodium hypochlorite, followed by the evaluation of potential resistance development.

METHODOLOGY: Bacteriophage CUB_EF02 was isolated from sewage water using E. faecalis ATCC 29212 as host, visualized by TEM, and sequenced. Its antimicrobial effect against biofilms grown on porous glass beads was assessed in a time-killing assay. Additionally, 10-day-old biofilms were established in 36 extracted single-rooted human teeth and treated with 0.9% NaCl, 1% NaOCl, phages, or their combination. Dentine-adherent biofilm was quantified by plate counts at baseline, after treatment, and after 48 h re-incubation. Logarithmic reduction factors were analyzed using one-way ANOVA and Tukey HSD. Phage resistance was evaluated by Top-Agar overlay spot assay.

RESULTS: Phage treatment reduced biofilm by 1.35 ± 0.2 SD-log after 6 h compared to control. Irrigation of the root canals using NaOCl achieved a significantly higher reduction (LRFred. = 2.98 ± 0.77 SD) than phages alone(LRFred. = 1.50 ± 0.23 SD). Combined treatment showed the strongest antimicrobial effect (LRFred. = 4.05 ± 1.17 SD). Bacterial regrowth occurred in all groups. Resistance developed after phage incubationfor ≥6 h.

CONCLUSION: Bacteriophages represent a promising adjunctive in root canal disinfection, but remain less effective than NaOCl and may inducebacterial resistance.},
}

@article {pmid42312514,
year = {2026},
author = {Singh, T and Singh, RP and Jain, M and Muthukumaran, J and Singh, AK},
title = {Targeting Biofilm Formation in Acinetobacter baumannii: In Silico Discovery of Novel Candidate Inhibitors for Acyl-Homoserine Lactone Synthase.},
journal = {Current drug discovery technologies},
volume = {},
number = {},
pages = {},
doi = {10.2174/0115701638450698260520051756},
pmid = {42312514},
issn = {1875-6220},
abstract = {INTRODUCTION: Acinetobacter baumannii, a Gram-negative member of the ESKAPE pathogens, has emerged as a pivotal cause of multidrug-resistant hospital-acquired or nosocomial infections worldwide. Its ability to regulate virulence and biofilm formation through quorum sensing (QS) significantly contributes to its virulence and pathogenicity. Acyl-homoserine lactone synthase (AHLS) from the A. baumannii AYE strain plays a key role in the QS pathway and represents a promising druggable target for the development of anti-bacterial strategies.

METHODOLOGY: A homology-modeled three-dimensional structure of AHLS (AYE strain) was predicted, optimized, and validated. High-throughput virtual screening of 975 natural antimicrobial compounds was performed, followed by Lipinski's and ADMET profiling to assess drug-likeness and safety. Promising drug candidates were further evaluated using 100ns molecular dynamics (MD) simulations to identify putative AHLS inhibitors.

RESULTS: MM/PBSA based binding free energy calculations revealed favorable interactions for CID_291096 (-14.74 ± 2.20 kcal/mol), CID_155586 (-15.26 ± 2.27 kcal/mol), and MSID_001127 (-28.44 ± 3.32 kcal/mol). Among these, MSID_001127 (Lovastatin) demonstrated superior structural stability and sustained intermolecular non-covalent interactions throughout the 100ns MD simulation. Structural stability was further supported by RMSD, RMSF, Rg, SASA, PCA, and hydrogen-bonding analyses.

DISCUSSION: Through virtual screening, three phytochemical lead compounds targeting AHLs with high negative binding free energies were identified. Stable protein-ligand interactions and favourable binding energetics were identified by molecular docking, 100 ns molecular dynamics simulations, and MM/PBSA analyses. Based on the results, ligand MSID_001127 was the most promising lead candidate compared with cipargamin. However, additional experimental validation is required to verify its therapeutic potential and biological activity.

CONCLUSION: These findings suggest that Lovastatin may be a promising drug candidate for AHLS targeting the QS pathway of A. baumannii. The results warrant further experimental validation to explore its potential as an anti-bacterial therapeutic agent.},
}

@article {pmid42313510,
year = {2026},
author = {Hatem, ZA and Kafe, FN and Musa, FH and Al-Taie, SF and Ateya, NH and Aziz, LM and Rajabi, E and Hasan, RN},
title = {Antibiotic Resistance and Biofilm Gene Distribution in Colistin-Resistant Acinetobacter baumannii.},
journal = {MicrobiologyOpen},
volume = {15},
number = {3},
pages = {e70332},
pmid = {42313510},
issn = {2045-8827},
mesh = {*Biofilms/growth & development ; *Acinetobacter baumannii/drug effects/genetics/isolation & purification/physiology/classification ; *Colistin/pharmacology ; *Anti-Bacterial Agents/pharmacology ; Microbial Sensitivity Tests ; Humans ; *Drug Resistance, Bacterial ; Acinetobacter Infections/microbiology ; Tigecycline ; Iran ; Multilocus Sequence Typing ; Bacterial Proteins/genetics ; Minocycline/analogs & derivatives/pharmacology ; },
abstract = {Acinetobacter baumannii, a multidrug-resistant opportunistic bacterium, poses a substantial hazard in hospital settings. The emergence of colistin- and tigecycline-resistant strains further limits treatment options and necessitates detailed investigation of resistance mechanisms. A total of 144 clinical A. baumannii isolates from multiple hospitals in Iran were identified using standard microbiological and molecular techniques. Antimicrobial susceptibility was assessed using both disk diffusion and broth microdilution techniques. Biofilm formation was quantified by crystal violet staining. Resistance and biofilm-related genes were detected by conventional polymerase chain reaction (PCR). The expression of key resistance genes (pmrA, pmrB, adeB, adeJ, and adeG) was evaluated by quantitative PCR (qPCR) in resistant isolates, and MLST was performed to determine the genetic relatedness among tigecycline- and colistin-resistant isolates. Resistance to colistin and tigecycline was observed in 3 (2.08%) and 2 (1.4%) isolates, respectively, and 90.9% of the isolates were biofilm producers, with higher odds of strong biofilm formation significantly correlating with the presence of blaPER1. All isolates carried pmrA and pmrB, but only colistin-resistant isolates showed overexpression of these genes compared to susceptible ones. MLST revealed diverse sequence types among resistant isolates, including ST188, ST138, ST387, ST2288, and ST3337. This study highlights the complex interplay between the presence of genes, their expression, and the resistance phenotype in A. baumannii and underscores the importance of monitoring chromosomal resistance determinants for effective control and treatment strategies.},
}

*Biofilms/growth & development
*Acinetobacter baumannii/drug effects/genetics/isolation & purification/physiology/classification
*Colistin/pharmacology
*Anti-Bacterial Agents/pharmacology
Microbial Sensitivity Tests
Humans
*Drug Resistance, Bacterial
Acinetobacter Infections/microbiology
Tigecycline
Iran
Multilocus Sequence Typing
Bacterial Proteins/genetics
Minocycline/analogs & derivatives/pharmacology

@article {pmid42314591,
year = {2026},
author = {Zhang, M and Xu, L and Du, X and Wan, J and Pei, W and Yun, J and Liu, X and Feng, M and Wei, T and Xu, K and Ge, F and Wang, X and Zhu, N},
title = {Mineral-biofilm interaction controls trophic transfer of PFAS along a biofilm-snail food chain.},
journal = {Aquatic toxicology (Amsterdam, Netherlands)},
volume = {298},
number = {},
pages = {107904},
doi = {10.1016/j.aquatox.2026.107904},
pmid = {42314591},
issn = {1879-1514},
abstract = {Natural biofilms developing on mineral substrates at the soil-water interface in freshwater ecosystems are important sinks of per- and polyfluoroalkyl substances (PFAS). However, the role of mineral-biofilm interactions in regulating their trophic transfer remains poorly understood. This study investigates how Fe2O3 is associated with changes in the distribution of PFAS within the biofilm matrix in a carbon chain length-dependent manner, which may further affect PFAS trophic transfer to snails. Fe2O3 preferentially localizes within the tightly bound extracellular polymeric substances (TB-EPS) layer, coincides with stimulated EPS production and altering PFAS partitioning within biofilms, and correlates with the redistribution of long-chain PFAS (e.g., PFNA) from loosely bound to tightly bound EPS fractions. In addition, Fe2O3 alleviates PFNA-induced toxicity and reshapes microbial community composition, increasing the relative abundance of Cyanobacteria and improving biofilm nutritional quality, which in turn may enhance snail grazing. Consequently, Fe2O3 is associated with a significant increase in the trophic transfer factor of PFNA from biofilms to snails (approximately 35%), while showing negligible effects on short-chain PFAS. Overall, these results suggest that Fe2O3 may enhance the trophic transfer of long-chain PFAS through combined effects on contaminant partitioning and biofilm quality, highlighting the important role of mineral-associated biofilms in PFAS bioaccumulation and providing new insights for ecological risk assessment in freshwater ecosystems.},
}

@article {pmid42316807,
year = {2026},
author = {Chowdhury, MAH and Reem, CSA and Ashrafudoulla, M and Yoon, HJ and Ha, SD},
title = {Biofilm Formation and Spore-Mediated Persistence of Clostridium perfringens in Meat and Poultry Processing Environments and Their Implications for Control Strategies.},
journal = {Journal of food science},
volume = {91},
number = {6},
pages = {e71193},
doi = {10.1111/1750-3841.71193},
pmid = {42316807},
issn = {1750-3841},
support = {//Chung-Ang University Young Scientist Scholarship in 2024/ ; //Chung-Ang University research grant in 2025/ ; },
mesh = {*Biofilms/growth & development ; *Clostridium perfringens/physiology/drug effects ; Animals ; *Spores, Bacterial/physiology ; *Meat/microbiology ; Poultry/microbiology ; *Food Handling ; Food Microbiology ; Food Contamination/prevention & control ; Food, Processed ; Anti-Bacterial Agents/pharmacology ; Quorum Sensing ; },
abstract = {Clostridium perfringens (C. perfringens) biofilms pose a persistent challenge in meat and poultry processing environments due to their structural resilience, spore-mediated survival and toxin-associated virulence. These biofilms readily develop on food-contact surfaces under typical processing conditions including organic residue accumulation, temperature fluctuations, and localized anaerobic niches, leading to increased tolerance to sanitation and thermal treatments. Mechanistically, biofilm resilience in C. perfringens is governed by the integration of sporulation processes, quorum sensing-regulated gene expression and extracellular polymeric substance (EPS) matrix formation, which collectively enhance stress tolerance, limit antimicrobial penetration, and facilitate persistence under fluctuating environmental conditions. The interaction between spore formation and EPS architecture further promotes survival during thermal processing and enables rapid re-establishment of biofilms following sanitation. This review synthesizes current knowledge on the formation and persistence of C. perfringens biofilms, key environmental drivers in meat and poultry processing systems and the mechanistic basis of their stress resistance and survival strategies. It also critically examines how these mechanisms influence the efficacy of existing intervention strategies. It further evaluates the limitations of conventional control strategies and highlights emerging approaches for biofilm prevention and control, including food-grade antimicrobials, surface engineering, enzymatic disruption, and microbiome-based interventions, with emphasis on their modes of action and applicability in industrial settings. Overall, this review provides a mechanistic and systems-level perspective to support the development of more effective biofilm control strategies in meat processing environments.},
}

*Biofilms/growth & development
*Clostridium perfringens/physiology/drug effects
Animals
*Spores, Bacterial/physiology
*Meat/microbiology
Poultry/microbiology
*Food Handling
Food Microbiology
Food Contamination/prevention & control
Food, Processed
Anti-Bacterial Agents/pharmacology
Quorum Sensing

@article {pmid42316914,
year = {2026},
author = {Liu, B and Wang, C and Tian, L and Li, R and Lv, S and Sun, Z and Xiao, M and Zheng, Q and Shi, L and Zhu, C},
title = {Acid-Triggered, Enzyme-Enabled EPS-Degrading Nanoplatform With Enhanced In Situ Retention for Intravenous Biofilm Therapy.},
journal = {Advanced materials (Deerfield Beach, Fla.)},
volume = {},
number = {},
pages = {e73749},
doi = {10.1002/adma.73749},
pmid = {42316914},
issn = {1521-4095},
support = {2024YFC2418700//National Key R&D Program of China/ ; 23JCZDJC00860//Natural Science Foundation of Tianjin/ ; BNLMS202308//Beijing National Laboratory for Molecular Sciences/ ; 63251169//Fundamental Research Funds for the Central Universities/ ; 63253194//Fundamental Research Funds for the Central Universities/ ; },
abstract = {Bacterial biofilms present a major challenge to antibacterial therapy due to their dense extracellular polymeric substance (EPS) matrix, which limits nanoparticle penetration and reduces drug efficacy. Here, we report a pH-responsive, surface charge-adaptive multifunctional nanosystem (DA-L@DTTB/Bro) for efficient in vivo treatment of biofilm-associated infections. The nanosystem integrates pH-triggered charge adaptation, in situ self-aggregation, photothermal responsiveness, enzymatic EPS degradation, and NIR-II imaging. Cationic phospholipid AGPDP, together with cholesterol and thermosensitive DPPC, self-assembled into liposomes encapsulating bromelain in the hydrophilic core and an NIR-II-emissive photothermal agent (DTTB) in the hydrophobic layer. Surface modification with DA-functionalized chitosan (CS-DA) generates negatively charged nanoparticles for prolonged circulation. At acidic infection sites, DA hydrolysis restores the cationic surface, enhancing biofilm penetration, while residual CS-DA induces self-aggregation to improve retention. NIR irradiation triggers DTTB-mediated hyperthermia, directly killing bacteria and disassembling liposomes to release bromelain, which degrades EPSs and facilitates biofilm dispersion. The nanosystem eradicates methicillin-resistant Staphylococcus aureus biofilms in vitro with 99.99% efficiency, enables high-contrast NIR-II imaging, persists at abscess sites in vivo, and accelerates wound healing. Furthermore, it demonstrates effective therapeutic activity against biofilm-associated infections in deep pulmonary tissues. This study presents a versatile intravenous strategy for targeted, synergistic therapy against biofilm-associated infections in vivo.},
}

@article {pmid42317001,
year = {2026},
author = {Yunus, J and Jemon, K and Mohamad, SE and Jonet, MA and Jamaluddin, H and Wan Dagang, WRZ},
title = {Protease-driven approaches for wound eschar debridement and biofilm disruption: current advances, future prospects, and limitations.},
journal = {Critical reviews in microbiology},
volume = {},
number = {},
pages = {1-17},
doi = {10.1080/1040841X.2026.2688093},
pmid = {42317001},
issn = {1549-7828},
abstract = {Chronic wounds frequently develop eschar, a dense layer of necrotic tissue that impedes healing and fosters bacterial biofilm formation. Biofilms, protected within an extracellular polymeric substance, show high resistance to antimicrobial agents, making wound management challenging. Conventional surgical debridement, though effective, is invasive, painful, and damages healthy tissue. Enzymatic debridement using proteolytic enzymes provides a less invasive, selective alternative. This review summarizes advances in protease-based therapies from plant, bacterial, and recombinant sources, focusing on their dual role in degrading eschar proteins and disrupting biofilm architecture. By elucidating the mechanisms through which proteases hydrolyze structural proteins and dismantle biofilms, we highlight their potential as minimally invasive interventions that enhance healing, reduce infection, and improve patient outcomes.},
}

@article {pmid42317929,
year = {2026},
author = {Singh, U and Jain, P and Singhai, A and Rashmi, and Verma, S and Kalyan, R and Venkatesh, V},
title = {Exploring the Genetic and Morphological Basis of Biofilm-Linked Drug Resistance in Clinical Isolates of Acinetobacter baumannii.},
journal = {Cureus},
volume = {18},
number = {5},
pages = {e108958},
pmid = {42317929},
issn = {2168-8184},
abstract = {Background Acinetobacter baumannii is a major nosocomial pathogen combining multidrug resistance (MDR) with strong biofilm-forming ability, enabling persistence on medical devices. Studying the correlation between biofilm-related genes and antibiotic resistance is critical for successful infection management. This study aimed to determine the prevalence of antibiotic resistance, phenotypic biofilm formation (by Congo red agar (CRA) and tissue culture plate (TCP) methods), and selected biofilm-associated genes (bap, csuD, ompA, blaper-1) in 153 clinical A. baumannii isolates from respiratory specimens, which were further characterized using scanning electron microscopy (SEM) analysis. In addition, the study aimed to evaluate the association between these genes, biofilm strength, and drug resistance patterns. Methodology In this study, an investigation was performed on 153 respiratory isolates from the laboratory of the Department of Microbiology at a tertiary care hospital (2022-2024). The Kirby-Bauer method was used for antibiotic susceptibility testing and minimum inhibitory concentration for colistin following the 2022 Clinical and Laboratory Standards Institute norms. Biofilm testing was performed using the CRA and TCP methods and validated by SEM and genetic analysis of biofilm association genes such as bap, csuD, ompA, and blaPER-1. Results The present study demonstrated an extremely high rate of antibiotic resistance, with meropenem resistance observed in 152 (99.3%) isolates and ceftriaxone resistance in 148 (96.7%) isolates. Regarding biofilm formation, the TCP method identified 85 (55.6%) isolates as biofilm producers, whereas the CRA method detected 57 (37.3%) isolates. Strong biofilm-producing isolates identified by the TCP method were confirmed by SEM to exhibit dense, mature biofilm structures. Genotypic analysis revealed csuD (99, 64.7%)and bap (82, 53.6%) as the most prevalent genes, both significantly associated with strong biofilm formation (p < 0.05). The combined presence of bap + csuD (51, 33.3%) showed a strong correlation with enhanced biofilm strength in the TCP assay (p < 0.05). Conclusions High MDR and strong biofilm formation were observed in A. baumannii, with a significant association between biofilm and antibiotic resistance. The csuD gene and bap + csuD combination correlated with increased resistance and biofilm production. SEM confirmed dense biofilm architecture, highlighting their role in virulence and persistence.},
}

@article {pmid42318050,
year = {2026},
author = {Marimuthu, AK and Gopi, RR and Sridhar, JP and Ravi, K and Roy, A and Ramaiah, S and Anbarasu, A},
title = {Copper-coated carbon nanotube surfaces for inhibiting biofilm formation by Staphylococcus aureus and Pseudomonas aeruginosa.},
journal = {Cell surface (Amsterdam, Netherlands)},
volume = {16},
number = {},
pages = {100176},
pmid = {42318050},
issn = {2468-2330},
abstract = {Biofilm-associated infections caused by clinically important pathogens such as Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) remain a major challenge in healthcare settings due to enhanced antimicrobial resistance and persistence. In recent years, copper-coated carbon nanotubes (Cu-CNTs) have gained considerable attention as promising antimicrobial nanomaterials for preventing biofilm formation on medical devices and hospital-associated surfaces. This review summarizes recent advances in the development and application of Cu-CNT-based antimicrobial coatings, with an emphasis on their antibacterial and antibiofilm activities against Gram-positive and Gram-negative pathogens. The synergistic combination of the high surface-area-to-volume ratio of CNTs and the potent antimicrobial properties of copper (Cu) ions enhances microbial inhibition. Previous studies suggest that Cu-CNTs interfere with initial bacterial adhesion, inhibit biofilm maturation, and disrupt established biofilms through mechanisms involving oxidative stress generation, membrane destabilization, and cellular damage. Furthermore, the review discusses the physicochemical characteristics, antimicrobial mechanisms, biomedical applications, and potential challenges associated with Cu-CNT coatings, including toxicity and biocompatibility concerns. Overall, Cu-CNT-based coatings represent a promising strategy for developing durable antimicrobial surfaces to control chronic biofilm-associated infections and reduce healthcare-associated contamination.},
}

@article {pmid42318084,
year = {2026},
author = {Williamson, KS and Franklin, MJ},
title = {An optimized mung bean seedling model for characterizing virulence of Pseudomonas aeruginosa biofilm infections.},
journal = {Biofilm},
volume = {11},
number = {},
pages = {100363},
pmid = {42318084},
issn = {2590-2075},
abstract = {Plant-based infection models provide cost effective and biologically relevant systems for investigating bacterial pathogenesis and virulence in living hosts. The mung bean seedling model enables the study of bacterial biofilms on living surfaces by allowing attachment and biofilm development on plants, but its broader use has been limited by methodological complexity and variability in experimental outcomes. Here, we present a modified mung bean seedling biofilm infection model for assessing Pseudomonas aeruginosa virulence that improves both consistency and practicality. The assay incorporates a bleach-based seed sterilization protocol that effectively reduces surface associated contaminants while maintaining high seed germination percentages. Additional refinements, including dehulling germinated seedlings, a shortened bacterial inoculation period, and plate-based incubation of seedlings at 37 °C, minimize variability in plant health outcomes while supporting development of gnotobiotic plants. Plant mortality, cotyledon emergence, and root branching were identified as rapid and quantitative measures of biofilm associated disease. Using this modified assay, reproducible differences in virulence were detected among P. aeruginosa strains, including reduced pathogenicity in a pqsR quorum sensing mutant. This simplified mung bean seedling model provides an accessible platform for studying biofilm associated virulence and screening genes involved in biofilm-mediated pathogenicity on a biotic surface.},
}

@article {pmid42319152,
year = {2026},
author = {Attia, RG and Rizk, SA and Maamoun, SAM},
title = {Polyvinyl alcohol/chitosan biofilm incorporated with moringa oil nanoparticles: a sustainable and ecofriendly packaging strategy for managing Tribolium confusum infestation.},
journal = {Journal of economic entomology},
volume = {},
number = {},
pages = {},
doi = {10.1093/jee/toag174},
pmid = {42319152},
issn = {1938-291X},
abstract = {The development of a novel anti-stored grain pest membrane film of polyvinyl alcohol/chitosan-moringa nanoemulsion (PVA/CS-MNE) biofilm offers a promising alternative to conventional cereal packaging materials. The efficacy of different packaging films and thicknesses: polyethelene, cellophane, polypropylene, paper bags, and a prepared PVA/chitosan biofilm was evaluated for the control of Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) adults. The estimated LC50 values for surface-treated PVA/chitosan biofilm, cellophane, polyethelene, paper bag, and polypropylene with moringa oil against T. confusum were 38.48%, 48.94%, 57.59%, 79.94%, and 125.06%, respectively. The most effective materials (PVA/chitosan biofilm, cellophane, polyethelene) were selected, and MNE was subsequently applied to these selected material types, resulting in LC50 values of 0.15%, 0.28%, and 0.47%, respectively. The results revealed that packaging materials treated with MNE exhibited higher toxicity compared to those exposed to moringa oil. Particle size and morphology of MNE, as measured by transmission electron microscope, confirmed the formation of spherical shape and uniform size with an average particle size around 90 nm. The zeta potential of -18 mV confirmed the formation of moderate stability colloidal system. Morphological changes were observed in a PVA/chitosan biofilm after moringa oil incorporation, as revealed by field emission scanning electron microscopy. A series of penetration tests conducted over 2 mo indicated that PVA/chitosan-MNE biofilm exhibited the highest protective efficacy. The numbers of exuviae, larvae, pupae, adults, and the percentage weight loss in packed wheat flour were 0.33%, 0.33%, 0.00%, 0.00%, and 0%, respectively.},
}

@article {pmid42319510,
year = {2026},
author = {Marin-Dett, FH and Grejo, MP and Valente, V and Palaçon, MP and Bufalino, A and Peters, BM and Barbugli, PA},
title = {Candida albicans hyphae modulate Staphylococcus aureus cell-free supernatant during dual biofilm growth to drive molecular signatures of oral dysplasia.},
journal = {Medical microbiology and immunology},
volume = {215},
number = {1},
pages = {},
pmid = {42319510},
issn = {1432-1831},
mesh = {Humans ; *Candida albicans/physiology/growth & development/genetics ; *Biofilms/growth & development ; *Staphylococcus aureus/physiology/growth & development ; *Hyphae/growth & development ; Cytokines/metabolism ; Monocytes/immunology/drug effects ; Fungal Proteins/genetics/metabolism ; *Microbial Interactions ; Epithelial Cells/microbiology ; Virulence Factors/genetics/metabolism ; Coculture Techniques ; THP-1 Cells ; },
abstract = {Microbes frequently exist as biofilm-embedded multi-species communities where their interactions may establish or exacerbate chronic infection. Recently, fungi and bacteria have been associated with various human tumor microenvironments, suggesting that dynamic cross-kingdom interactions may directly or indirectly contribute to tumor-associated processes. Here, we aimed to investigate whether cell-free supernatants from mono- and dual-species biofilms of the commonly associated fungus Candida albicans and bacterium Staphylococcus aureus could alter human monocyte responses that promote a tumor-related genetic signature in dysplastic oral epithelial (DOK) cells. Treatment of THP-1 monocytes with S. aureus cell-free supernatant increased the production of proinflammatory cytokines (IL-8, IL-1β, and TNF) and CD86 expression. However, exposure to cell-free supernatants from dual-species biofilm suppressed these responses. To determine the fungal virulence factors responsible, C. albicans mutants deleted for genes involved in adhesion (als3Δ/Δ), hyphal growth (efg1Δ/Δ cph1Δ/Δ), or candidalysin production (ece1Δ/Δ) were assessed during co-culture. While candidalysin was dispensable, loss of hyphal growth or the adhesin Als3p phenocopied effects of S. aureus mono-culture treatment. Conditioned medium from THP-1 cells initially challenged with mono- or dual-biofilm cell-free supernatants was applied to DOK cells to assess TP53 and BCL2 gene expression. Conditioned medium from S. aureus treated THP-1 cells led to decreased epithelial TP53, but increased BCL2 expression, which was reversed by the presence of wild-type C. albicans. These phenotypes were similarly dependent on C. albicans hyphal growth during dual-biofilm co-culture. Collectively, our results reveal that fungal-bacterial interactions may shape the monocyte-epithelial axis by orchestrating immune responses that enhance tumor-associated gene expression in dysplastic oral epithelial cells.},
}

Humans
*Candida albicans/physiology/growth & development/genetics
*Biofilms/growth & development
*Staphylococcus aureus/physiology/growth & development
*Hyphae/growth & development
Cytokines/metabolism
Monocytes/immunology/drug effects
Fungal Proteins/genetics/metabolism
*Microbial Interactions
Epithelial Cells/microbiology
Virulence Factors/genetics/metabolism
Coculture Techniques
THP-1 Cells

@article {pmid42319514,
year = {2026},
author = {Ståhle, M and Johnson, A and Turner, S and Mårtensson, P and Kalinowski, B and Dopson, M},
title = {Multi-Year Biofilm Formation on Granitic Surfaces Reveals Dynamic Microbial Communities in Fennoscandian Shield Deep Groundwaters.},
journal = {Microbial ecology},
volume = {},
number = {},
pages = {},
doi = {10.1007/s00248-026-02812-4},
pmid = {42319514},
issn = {1432-184X},
abstract = {The deep terrestrial biosphere is the vast biome beneath the soil layer that contains the majority of the Earth's prokaryotic biomass, yet it is one of the least investigated communities. Although, estimates of deep biosphere biomass suggest biofilm cells outnumber the planktonic biomass by several orders of magnitude, most investigations target planktonic communities captured from groundwaters. This multi-year study used 16S rRNA gene sequencing to compare planktonic and biofilm communities attached to natural granitic rock, demonstrating that biofilm formation selected for taxa with distinct relative abundances and exhibited temporal development. The biofilm communities also showed a decreasing influence of introduced populations on the natural rock surfaces (macadam) present at the onset of the incubations. After two- and four-years of biofilm incubation, a community developed that was dominated by sulfur/sulfate reducing Desulfocapsaceae, Desulfobacteraceae, and BM004 along with the families UBA5619, Rhodocyclaceae, Profunditerraquicolaceae, and UBA2206. This long-term community included populations predicted to be host-associated ultra-small cells. This contrasted with previous studies of early biofilm development in deep Fennoscandian Shield groundwaters that suggested biofilm initiation was mediated by lithotrophic carbon and nitrogen fixing populations. However, metabolic predictions based upon the 16S rRNA gene-based communities also showed an autotrophic and diazotrophic community including sulfur cycling in line with the previous studies. In conclusion, this study showed long-term biofilm composition to be dissimilar to the planktonic communities with a consistent strategy for energy conservation similar to previous studies of early biofilm formation from these groundwaters.},
}

@article {pmid42308838,
year = {2026},
author = {Ma, H and Liang, X and Li, B and Dong, X and Li, H and Ma, Y and Li, J and Shilin, MB and Leonteva, EO},
title = {Effects of biofilm-coated microplastics on the biological functions of RNA viruses in Mytilus coruscus.},
journal = {Aquatic toxicology (Amsterdam, Netherlands)},
volume = {298},
number = {},
pages = {107873},
doi = {10.1016/j.aquatox.2026.107873},
pmid = {42308838},
issn = {1879-1514},
abstract = {Microplastics (MPs) pollution poses emerging ecological risks through physical stress and its potential role in altering microbial and viral communities. Here, we investigated the effects of biofilm-associated polyethylene (PE) MPs exposure on RNA viral communities in the digestive glands of Mytilus coruscus (M. coruscus) during a 21-day in situ experiment. Metatranscriptomic analyzes revealed that lysogenic viruses were predominant in all samples (> 99%), while biofilm-associated MPs exposure was associated with increased viral richness and diversity in the digestive glands of M. coruscus (P < 0.05). Viral composition in MPs-exposed M. coruscus became more similar to that of MPs biofilms, with relative enrichment of Uroviricota and decreased relative abundance of Kitrinoviricota and Picornavirales. Functional annotation analysis showed higher abundances of annotations related to antibiotic resistance genes (ARGs), virulence factor genes (VFGs), and metal resistance genes (MRGs) in MPs-exposed digestive glands, while virus-bacteria co-occurrence networks exhibited reduced connectivity. Histopathological analysis further showed tissue-level alterations in digestive gland, gill, and mantle tissues following MPs exposure. These findings suggest that MPs exposure may alter host-associated microenvironments and contribute to changes in RNA virome composition, functional profiles, and virus-bacteria interactions. Overall, this study highlights the potential influence of MPs exposure on host-associated RNA viromes and its ecological implications in marine ecosystems.},
}

@article {pmid42310079,
year = {2026},
author = {Nouraei, H and Amirzadeh, N and Shabanzadeh, S and Zareshahrabadi, Z and Shamsdin, N and Nouraei, M and Naeimi, B and Pakshir, K},
title = {Effect of subsequent passages on biofilm formation intensity, ALS genes expression, and cell surface hydrophobicity variability in clinical Candida albicans isolates.},
journal = {Scientific reports},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41598-026-58680-y},
pmid = {42310079},
issn = {2045-2322},
support = {32536//Vice-Chancellor for Research, Shiraz University of Medical Sciences/ ; },
abstract = {Candida albicans is an opportunistic yeast pathogen that have several virulence factors included biofilm formation, cell surface hydrophobicity (CSH), and the expression of adhesion genes. Concerns exist that serial laboratory subculturing may diminish these traits, leading to inaccurate research findings. Aim of this study was evaluated the effect of subsequently subcultures on biofilm formation intensity, ALS gene expression, and surface hydrophobicity properties in clinical C. albicans isolates. Ten clinical C. albicans isolates were serially subcultured up to 20 passages (P). We used qPCR to quantify ALS1 and ALS3 gene expression, the Crystal Violet assay to measure biofilm formation intensity (P1, P5, P10, P15, P20), and a water-octane partitioning assay for CSH variability at different passages. Serial subculturing caused gradual downregulation of gene expression for both ALS1 and ALS3 (p < 0.001). This condition was accompanied by a biofilm-forming capacity that became progressively reduced in 90% of isolates, whereas 60% at P20 were already biofilm-negative (vs. 10% at P1). Cell surface hydrophobicity also decreased progressively, with 100% of isolates displaying low CSH at P15, compared with 40% in the initial P1 state. Serial subculturing leads to a rapid reduction of C. albicans pathogenic fitness with decreased expression of certain key adhesion genes, diminished biofilm formation, and lower CSH. These results highlight the plasticity of the organism and thus strongly suggest that low-passage clinical isolates should be used in studies to reflect true pathogenicity in vivo accurately.},
}

@article {pmid42310175,
year = {2026},
author = {Komatsu, K and Kim, J and Her, N and Alpers, R and Saito, N and Shibata, R and Fedorowicz, I and Kim, SJ and Kim, N and Lu, T and Tran, A and Lim, J and Sakaguchi, W and Sato, T and Haga, S and Matsuura, T and Park, W and Ogawa, T},
title = {The surface-agnostic advantage for peri-implant health: UV photofunctionalization as a positive-sum strategy for biofilm suppression and soft-tissue barrier-a systematic review with qualitative synthesis.},
journal = {International journal of implant dentistry},
volume = {},
number = {},
pages = {},
doi = {10.1186/s40729-026-00695-1},
pmid = {42310175},
issn = {2198-4034},
abstract = {PURPOSE: Long-term dental implant success depends on a biologic "race to the surface," in which osteogenic cells, peri-implant soft-tissue cells, and bacterial pathogens compete for early dominance at the implant-tissue interface. Because implant surface design is often optimized for one objective at the expense of another (e.g., micro-roughness to accelerate osteoconductivity but with increased plaque-retention risk; relatively smooth transmucosal surfaces to discourage bacterial attachment despite uncertainty regarding optimal soft-tissue integration), strategies that enhance peri-implant health without forcing topographical trade-offs are needed. Ultraviolet (UV) photofunctionalization-by removing storage-acquired hydrocarbons ("biological aging") and converting surfaces to a high-energy, superhydrophilic state-has been proposed as a chairside, topography-preserving approach to improve interfacial biology. This systematic review evaluates whether UV photofunctionalization of titanium and zirconia surfaces provides clinically relevant advantages for (1) reduction of bacterial attachment and biofilm formation, (2) peri-implant soft-tissue responses relevant to mucosal sealing, and (3) human clinical outcomes.

METHODS: After systematic literature search, screening and full-text evaluation, a total of 34 articles, including 9 bacterial/biofilm, 13 soft-tissue (1 overlapping between bacterial and soft-tissue), and 13 clinical studies were selected. Findings were synthesized qualitatively with attention to protocol heterogeneity (UV wavelength band, exposure duration, device configuration, and material and surface types).

RESULTS: Across experimental models, UV photofunctionalization most consistently reduced early bacterial attachment and/or early biofilm accumulation across several titanium surface topographies, supporting an early anti-adhesive and biofilm-suppressive phenotype. Soft-tissue studies generally demonstrated enhanced fibroblast/epithelial attachment, spreading, and functional behaviors relevant to sealing on both titanium and zirconia, although the optimal underlying topography for soft-tissue integration remains unresolved. Clinically, the most consistent signal was accelerated and enhanced implant stability development, while selected studies also suggested favorable trends in peri-implant soft-tissue parameters and/or crestal bone maintenance. However, clinical outcomes remained variable and were limited by heterogeneity in UV protocols, surface systems, endpoints, and follow-up duration.

CONCLUSIONS: UV photofunctionalization can be conceptualized as a surface-agnostic physicochemical reactivation technology: a topography-preserving enhancement that restores high surface energy and favorable surface chemistry without altering the underlying surface architecture. Current evidence for this concept is strongest for titanium, whereas supportive evidence for zirconia is emerging primarily from soft-tissue and interface-focused models. This interface-first, positive-sum strategy may allow clinicians to select zone-specific topographies (e.g., smooth transmucosal regions and rough endosteal regions) while maximizing soft-tissue affinity and suppressing early colonization. Although current clinical evidence most strongly supports accelerated osseointegration/stability development, further longitudinal studies with standardized peri-implant health, microbiologic, and mucosal inflammatory endpoints are needed to clarify the long-term translational impact of UV photofunctionalization on peri-implant disease prevention.},
}

@article {pmid42310482,
year = {2026},
author = {Zaylaa, M and Farha, R and Taleb, J and Saad, A and Gharbawy, OE and Sarraf, R and Kassam, I and Hamdach, C and Bissar, N},
title = {Interplay Between Virulence Genes, Antimicrobial Resistance Profiles, and Biofilm Formation in Klebsiella pneumoniae Causing Urinary Tract Infections in North Lebanon.},
journal = {MicrobiologyOpen},
volume = {15},
number = {3},
pages = {e70336},
doi = {10.1002/mbo3.70336},
pmid = {42310482},
issn = {2045-8827},
mesh = {*Biofilms/growth & development ; Lebanon/epidemiology ; *Klebsiella pneumoniae/genetics/drug effects/isolation & purification/physiology/pathogenicity ; *Urinary Tract Infections/microbiology/epidemiology ; Humans ; *Klebsiella Infections/microbiology/epidemiology ; *Virulence Factors/genetics ; Anti-Bacterial Agents/pharmacology ; Virulence/genetics ; beta-Lactamases/genetics ; Microbial Sensitivity Tests ; Drug Resistance, Multiple, Bacterial ; Bacterial Proteins/genetics ; *Drug Resistance, Bacterial ; Community-Acquired Infections/microbiology ; Cross Infection/microbiology ; },
abstract = {Klebsiella pneumoniae (K. pneumoniae) is a major cause of urinary tract infections (UTIs) and poses a growing public health concern due to multidrug resistance and virulence potential. This study aimed to characterize antimicrobial resistance, biofilm formation, and virulence gene distribution among urinary K. pneumoniae isolates from North Lebanon and to explore correlations between these factors. A total of 153 non-duplicate isolates from hospital and community settings were analyzed for antibiotic susceptibility, hypermucoviscosity, biofilm formation, and presence of key virulence (fimH, mrkD, magA, rmpA, entB, iucA, iroN, kfu) and β-lactamase (blaTEM, blaSHV, blaCTX-M) genes. Adhesion-associated genes fimH and mrkD were highly prevalent, particularly in hospital-acquired and strong biofilm-producing isolates. Capsular and regulatory genes (magA) was more frequent in putative hypervirulent and community-acquired strains, while siderophore genes entB and iucA were strongly associated with biofilm formation and hospital-acquired infections. A significantly higher resistance to cephalosporins, along with an increased frequency of multidrug-resistant phenotypes, was detected in hospital-acquired, classical, and strong biofilm-forming isolates. ESBL production was significantly more common in hospital-acquired and biofilm-forming isolates. Strong biofilm formation was largely associated with classical K. pneumoniae and hospital-acquired infections, whereas putative hypervirulent strains were primarily weak biofilm producers and community-acquired. These findings highlight the interplay between virulence determinants, biofilm formation, and antibiotic resistance, emphasizing the need for targeted infection control and treatment strategies in North Lebanon.},
}

*Biofilms/growth & development
Lebanon/epidemiology
*Klebsiella pneumoniae/genetics/drug effects/isolation & purification/physiology/pathogenicity
*Urinary Tract Infections/microbiology/epidemiology
Humans
*Klebsiella Infections/microbiology/epidemiology
*Virulence Factors/genetics
Anti-Bacterial Agents/pharmacology
Virulence/genetics
beta-Lactamases/genetics
Microbial Sensitivity Tests
Drug Resistance, Multiple, Bacterial
Bacterial Proteins/genetics
*Drug Resistance, Bacterial
Community-Acquired Infections/microbiology
Cross Infection/microbiology

@article {pmid42311309,
year = {2026},
author = {Loaiza-Toscuento, DI and Perez-Corona, CE and Spezzia-Mazzocco, T and Peregrina-Barreto, H and Ramos-Garcia, R and Padilla-Martinez, JP and Ramirez-San-Juan, JC},
title = {Laser speckle imaging for biofilm viability assessment: a noninvasive alternative to the MTT assay.},
journal = {Biomedical optics express},
volume = {17},
number = {6},
pages = {3167-3179},
pmid = {42311309},
issn = {2156-7085},
abstract = {Accurate measurement of cell viability in biofilms is critical in biology and medicine. We propose two laser speckle-based methods to assess the viability of Candida tropicalis biofilms. Time-varying speckle images were processed using temporal correlation analysis and frequent-motion imaging, and the resulting metrics were compared against the standard MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. A high Pearson correlation coefficient (>0.99) was observed between both speckle-based metrics and MTT. For C. tropicalis biofilms in microplates, the proposed approach provides a rapid, low-cost, and non-destructive proxy for cell viability and is expected to be transferable to other fungal and bacterial biofilms and other cell-based systems.},
}

@article {pmid42311376,
year = {2026},
author = {Bachir, A and Altaie, AM and Bendardaf, R and Talaat, IM and Hamoudi, R},
title = {Microbial dysbiosis drives colorectal carcinogenesis via integrated inflammatory, metabolic, and biofilm pathways.},
journal = {Frontiers in microbiology},
volume = {17},
number = {},
pages = {1795882},
pmid = {42311376},
issn = {1664-302X},
abstract = {Colorectal cancer (CRC) arises from a multifaceted interplay among the intestinal microbiota, chronic inflammation, and host genomic instability, with microbial dysbiosis serving as an active driver rather than a by-product of malignant transformation. Genotoxic Escherichia coli (colibactin-positive), enterotoxigenic Bacteroides fragilis, and Fusobacterium nucleatum contribute to distinct stages of CRC progression by engaging the DNA-damage response and activating β-catenin-dependent Wnt signaling and NF-κB/STAT3 transcriptional programs controlling pro-inflammatory (IL-6, IL-8), pro-survival (BCL-2, BCL-XL), and proliferative (MYC, CCND1) gene expression.. Here, we propose a tri-axial pathogenic framework in which (i) cyclic dinucleotide-mediated activation of the cGAS-STING pathway engages TBK1-IRF3 and NF-κB signaling, driving type I interferons (IFN-β) and pro-inflammatory cytokines (IL-6, TNF-α) that couple microbial genotoxic stress to innate inflammation; (ii) altered microbial metabolites, including indoles and bile acids, reprogram AhR and FXR/TGR5 signaling; and (iii) crypt-anchored biofilms spatially amplify IL-6 leading to activation of STAT3, epigenetic silencing of tumor suppressors, and immune evasion. This review critically synthesizes current evidence supporting these axes and maps them onto CRC molecular subsets and tumor location. Recognition of these integrated microbial-host circuits identifies mechanistically grounded candidates for biomarker development, microbiome-based diagnostics, and targeted interventions to restore microbial and immune equilibrium, thereby providing a refined framework for the molecular classification and precision management of CRC.},
}

@article {pmid42311500,
year = {2026},
author = {Shambhavi, K and Singh, DV},
title = {Outer membrane vesicles in Vibrio species: Roles in biofilm formation and pathogenesis.},
journal = {Microbial cell (Graz, Austria)},
volume = {13},
number = {},
pages = {198-217},
pmid = {42311500},
issn = {2311-2638},
abstract = {Outer membrane vesicles (OMVs) have been increasingly recognized as common mediators of bacterial physiology in Gram-negative bacteria, including Vibrio species. The degree and function of OMV production can differ among strains and even within a single species. The secretion of OMVs is a prevalent trait among many Vibrio species, particularly in pathogenic organisms such as Vibrio cholerae, Vibrio vulnificus, and Vibrio parahaemolyticus. The OMVs released by these organisms are often associated with infection, transport of virulence factors into host cells, defense against stress, biofilm formation, flagella rotation, transportation of active enzymes, signaling molecules in the surrounding environment, and facilitating bacterial translocation. All of these are advantageous to the bacteria. These OMVs also possess immunogenic properties that regulate the innate and adaptive immune responses, which are beneficial to host cells. Few species, such as Vibrio ordalii, Vibrio coralliilyticus, Vibrio natriegens Vibrio alginolyticus, and Vibrio europaeus, have been recently studied for the first time that secrete OMVs; future research is necessary to determine any other activities that these vesicles may possess beyond those that are now documented.},
}

@article {pmid42311627,
year = {2026},
author = {Northage, N and Gomilšek, M and Modic, M and Vengust, D and Zorko, A and Cvelbar, U and Walsh, JL},
title = {Physicochemical and Antimicrobial Characterization of Nanobubbles Reveals Physical Disruption is the Primary Mode of Biofilm Inactivation.},
journal = {ACS ES&T water},
volume = {6},
number = {6},
pages = {3852-3863},
pmid = {42311627},
issn = {2690-0637},
abstract = {Biofilm-associated contamination represents a persistent and costly challenge across environmental systems, causing reduced efficacy of disinfectants. Recently, nanobubbles (NBs) have shown promise for biofilm decontamination; yet, their underpinning mode of action remains a topic of debate. In this study, the interaction of air-generated NBs with Escherichia coli and Staphylococcus aureus biofilms was investigated. NBs were generated using a venturi nozzle and characterized using Nanoparticle Tracking Analysis, revealing a NB density of 5.66 × 10[8] particles/mL and a mean diameter of 84 nm. Application of NB solution to microbial biofilms resulted in a 2.16 log reduction for E. coli and 1.52 log reduction for S. aureus, along with visible morphological changes such as cell collapse, wrinkling, and matrix disruption. ESR spin trapping confirmed hydroxyl radical formation, but intracellular ROS and lipid peroxidation levels were minimal and, in some cases, not significantly different from Milli-Q water controls. After 28 days, NBs remained present and continued to demonstrate antimicrobial activity, biofilm disruption, and some ROS activity. These findings indicate that although hydroxyl radicals are generated, oxidative stress is not the dominant antimicrobial mechanism under the examined conditions, suggesting physical biofilm disruption is the primary mode of action.},
}

@article {pmid42312039,
year = {2026},
author = {Li, J and Gao, P and Kao, RY and Li, H and Sun, H},
title = {Bismuth drug as an antibiotic adjuvant to inhibit biofilm formation via a dual mechanism.},
journal = {RSC medicinal chemistry},
volume = {},
number = {},
pages = {},
pmid = {42312039},
issn = {2632-8682},
abstract = {Burkholderia cepacia (B. cepacia) exhibits intrinsic resistance to many conventional antibiotics. A key factor contributing to this resistance is its ability to form biofilms, which hinder antibiotic penetration and make this bacterial infection very difficult to treat. Developing novel biofilm inhibitors to restore antibiotic efficacy is a promising strategy to combat antimicrobial resistance (AMR). Here, we show the combination of a bismuth drug, e.g., colloidal bismuth subcitrate (CBS), with antibiotics potently inhibits biofilm formation in B. cepacia. Mechanistic studies reveal that CBS promotes the degradation of cyclic diguanylate (c-di-GMP) by inducing accumulation of the biofilm signalling molecule, nitric oxide (NO), driven by bindings of bismuth(iii) to cytochrome bo 3 ubiquinol oxidase (CyoC) and cytochrome bd-I ubiquinol oxidase subunit 1 (CydA). Furthermore, CBS reduces the cAMP level, downregulates crp, (encoding the cAMP receptor protein), and disrupts the formation of cAMP-CRP complex, leading to upregulation of rpoS, which further suppresses biofilm formation. These two bismuth-mediated pathways, i.e., NO-mediated c-di-GMP degradation and cAMP-CRP-regulated rpoS expression, work cooperatively to inhibit biofilm formation. Additionally, bismuth-antibiotic combinations effectively inhibit biofilm formation across a diverse range of bacteria. These findings emphasise the great potential of bismuth drugs as antibiotic adjuvants to tackle antimicrobial resistance.},
}

@article {pmid42302871,
year = {2026},
author = {Huang, J and Dang, J and Zhang, H and Zhu, J and Hu, J and Xu, J and Zhang, X and Zhang, H and Zeng, W},
title = {Quantifying the combined effects of substrate microstructure and environmental factors on microalgal biofilm growth: a novel kinetic modeling approach.},
journal = {Bioresource technology},
volume = {},
number = {},
pages = {135187},
doi = {10.1016/j.biortech.2026.135187},
pmid = {42302871},
issn = {1873-2976},
abstract = {Microalgal biofilm cultivation is a promising strategy for achieving efficient carbon sequestration and biomass production. However, existing growth kinetics models predominantly focus on environmental stimuli, neglecting the decisive roles of substrate microstructure in mass transfer and cell attachment. In this study, a multi-factor growth kinetics model was developed for non-submerged microalgal biofilm systems. A substrate structural term of cotton fabric substrate (porosity, tortuosity, thickness, and average pore diameter) was formulated and multiplicatively coupled with light intensity, CO2, and nitrate concentrations. The model showed good agreement with experimental μ under the tested conditions (R[2] > 0.9389) for Chlorella vulgaris biofilms cultivated on cotton fabrics, forecasting a maximum μ of 1.33 d[-1] in the integrated model. Sobol global sensitivity analysis showed that nitrogen supply and light irradiance were the dominant contributors to the specific growth rate (μ). Notably, within the investigated parameter range, the model suggested an attachment response associated with pore size, with a fitted pore-suitability diameter of approximately 2.04 μm. This may indicate a potential trade-off between cell attachment stability and substrate-mediated nitrate diffusion in the tested non-submerged cotton-based biofilm system. This framework describes the combined effects of substrate structural factors and environmental parameters on microalgal biofilm growth and may be useful for substrate design under similar non-submerged biofilm cultivation configurations.},
}

@article {pmid42303231,
year = {2026},
author = {Slate, AJ and Lawler, CRE and Clarke, OE and Jones, BV},
title = {Drug Repurposing as an approach to control biofilm formation and encrustation of urinary catheters: Preclinical evidence and future challenges.},
journal = {Journal of applied microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1093/jambio/lxag139},
pmid = {42303231},
issn = {1365-2672},
abstract = {Indwelling urethral catheters are the most widely used medical devices across the world, and catheter-associated urinary tract infections (CAUTIs) are the most common type of healthcare acquired infection. For many patients, urinary catheter blockage is a common and recurring problem, which can have considerable negative impact on patient health and well-being. Blockage primarily stems from the formation of crystalline bacterial biofilms on catheter surfaces, which can lead to upper urinary tract infection (UTI) and the onset of serious clinical complications. Potential solutions to this important clinical problem include the development of novel antibiofilm agents that can prevent formation of these communities on urinary catheters. However, traditional de novo methods of drug discovery are laborious, expensive, have long lead times and carry a high risk of failure in the clinical trial stages. One potential approach to mitigate this risk and cost, is the evaluation of pre-existing licensed drugs for those with useful antibiofilm or antimicrobial activity. Here we review current preclinical evidence for antibiofilm and antimicrobial activities in licensed drugs from a range of classes, such as urease inhibitors, Selective Serotonin Reuptake Inhibitors (SSRIs), phenothiazines, oncology therapeutics and non-steroidal anti-inflammatory drugs (NSAIDs). In doing so, we consider the application of the repurposing approach to control CAUTI and catheter blockage, and identify key challenges and opportunities related to delivery of repurposed drugs to the catheterised urinary tract.},
}

@article {pmid42304081,
year = {2026},
author = {Shabiya, MA and Ranjani, S and Hemalatha, S},
title = {Microfluidics-based engineered silver nanoparticles to control growth and biofilm formation in bacterial pathogens causing dental infection.},
journal = {Scientific reports},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41598-026-58510-1},
pmid = {42304081},
issn = {2045-2322},
abstract = {Dental infections caused by Streptococcus mutans and Enterococcus faecalis are major contributors to dental caries and root canal infections due to their strong biofilm forming ability across all age groups. This present study aimed to develop a microfluidics-assisted green synthesis approach for the fabrication of Clitoria ternatea-derived silver nanoparticles (CtAgNPs) and to evaluate their antibacterial and antibiofilm potential in dental pathogens. The bioactive compounds present in Clitoria ternatea floral extract act as a natural reducing and stabilizing agent within a microfluidics system. The synthesized CtAgNPs were characterized to confirm their physicochemical properties. UV-Visible spectroscopy reveals the surface plasmon resonance peak at 423 nm, FESEM imaging showed the spherical morphology with an average particle size of 151.6 nm, and a zeta potential of - 26.1 mV, indicating good colloidal stability. FT-IR analysis confirmed the presence of phytochemical-derived functional groups on the surface of CtAgNPs. Toxicity assessment using zebrafish (Danio rerio) embryos confirmed the biocompatibility and non-toxic nature of CtAgNPs. CtAgNPs demonstrated strong antibacterial activity with MIC values ranging from 3.125 to 6.25 µg/mL and exhibited more than 75% inhibition of biofilm formation in both S. mutans and E. faecalis. Mechanistic investigations revealed that the nanoparticles induced oxidative stress, characterized by increased lipid peroxidation (MDA), reduced antioxidant enzyme activities (CAT and SOD), and leakage of intracellular proteins and sugars, indicating membrane damage. Overall findings demonstrate that microfluidics-assisted green-synthesized CtAgNPs effectively inhibit the growth and biofilm formation of dental pathogens, highlighting their potential as an eco-friendly nanotherapeutic strategy for the prevention and management of dental biofilm-associated infections.},
}

@article {pmid42304178,
year = {2026},
author = {Poddar, K and Tripathi, G and Khairnar, SV and Srivastav, S and Anand, A},
title = {Enhanced Respiratory Electron Dissipation by Immunometabolites Promotes Mycobacterial Biofilm Longevity.},
journal = {ACS infectious diseases},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsinfecdis.6c00483},
pmid = {42304178},
issn = {2373-8227},
abstract = {Mycobacterial species inhabit diverse ecological niches and frequently adopt a biofilm lifestyle, including within host environments, where this organization critically influences the persistence and pathophysiological outcomes. Here, using Mycobacterium smegmatis, we demonstrate that nitrate, a host-derived immunometabolite, markedly extends the biofilm lifespan. Mechanistically, nitrate sustains respiratory activity and suppresses the induction of the dormancy response. This effect is accompanied by the maintenance of intracellular redox balance, consistent with enhanced electron dissipation. Importantly, fumarate, a distinct host-relevant metabolite capable of facilitating electron dissipation, recapitulates this phenotype, indicating that the observed biofilm longevity is not limited to nitrate but instead is linked to the availability of alternative electron sinks. Together, our findings establish that the electron dissipation capacity is a key determinant of respiratory homeostasis and biofilm persistence in mycobacteria. We report a similar phenotype for M. abscessus, a nontuberculous opportunistic pathogen. These results highlight respiratory flexibility as a central adaptive axis through which mycobacteria exploit host metabolic cues to prolong survival.},
}

@article {pmid42304237,
year = {2026},
author = {Nasher, AA and Assayaghi, RM and Al-Shamahy, HA and Al-Jaufy, AY and Othman, AM and Al-Shawkany, AM and Al-Shamahi, EH},
title = {Vulvovaginal candidiasis among Yemeni women: prevalence of Candida species, biofilm formation rates, antifungal susceptibility patterns, and biofilm-associated genes ALS1 and HWP1.},
journal = {BMC microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1186/s12866-026-05241-y},
pmid = {42304237},
issn = {1471-2180},
abstract = {BACKGROUND: Vulvovaginal candidiasis (VVC) is a common fungal infection in women, primarily caused by Candida species. Biofilm formation is a key virulence factor contributing to pathogenicity and antifungal resistance. This study aimed to identify Candida species, evaluate antifungal susceptibility, quantify biofilm formation, and detect virulence gene markers (ALS1 and HWP1) in vaginal isolates from women with VVC.

METHODS: A cross-sectional study was conducted from December 2021 to June 2024 among 400 women attending obstetrics and gynecology clinics in Sana'a City. Vaginal swabs were collected and cultured in a microbiology laboratory. Antifungal susceptibility of isolates to nystatin, voriconazole, fluconazole, ketoconazole, clotrimazole, miconazole, itraconazole, and amphotericin B was assessed using the disk diffusion method. Biofilm formation was measured using the microtiter plate assay. The presence of ALS1 and HWP1 genes was determined by PCR. Data were analyzed using SPSS version 20, and associations were evaluated with the Chi-square test; P < .05 was considered statistically significant.

RESULTS: C. albicans was identified in 367 (91.8%) isolates, while non-albicans species accounted for 33 (8.2%). Of the 367 C. albicans isolates, 174 (47.4%) formed biofilms: 32 (8%) strong, 72 (18%) moderate, and 70 (17.5%) weak. Biofilm- forming isolates were associated with higher antifungal resistance, with amphotericin B (113; 64.9%) and itraconazole (108; 62.1%) showing the highest resistance, and nystatin the lowest (23; 13.2%). The ALS1 gene was detected in all biofilm- forming C. albicans isolates (174; 100%), while HWP1 was present in 82 (47.1%). Non-biofilm-forming isolates demonstrated lower resistance rates across all drugs.

CONCLUSIONS: VVC remains highly prevalent in Sana'a, Yemen, with C. albicans as the dominant pathogen. These findings suggest a potential association between biofilm formation and increased antifungal resistance also the presence of key virulence genes-particularly ALS1 and HWP1. However, due to the study's cross-sectional design, a definitive causal relationship cannot be inferred. This study emphasizes the importance of integrating phenotypic and molecular characterization into routine diagnostics and surveillance to improve the management of resistant Candida infections. Continuous monitoring of species distribution is also warranted to detect emerging non-albicans species.},
}

@article {pmid42304313,
year = {2026},
author = {Arafa, AM and Yahya, G and Abdel-Halim, MS and Osman, A and Nazeih, SI and Abbas, HA and Mostafa, I},
title = {Chemical profile, anti-biofilm and antioxidant activities of Cymbopogon citratus (DC.) Stapf essential oil.},
journal = {BMC complementary medicine and therapies},
volume = {},
number = {},
pages = {},
doi = {10.1186/s12906-026-05431-1},
pmid = {42304313},
issn = {2662-7671},
abstract = {BACKGROUND: The urgent need for novel antimicrobial agents arises from the escalating threat of antibiotic-resistant pathogens. Tackling the resistance mechanisms through the application of resistance modifying agents in combination to antimicrobials represents an effective strategy to combat antimicrobial resistance and to enhance the antimicrobial activity. Screening of plant extracts, essential oils and their active compounds for potential resistance modifying properties has proven effective on both a small and large scale. In this study, we define the metabolomic profile of essential oils extracted from different parts of Cymbopogon citratus (DC.) Stapf (lemongrass), and characterize their antioxidant and virulence attenuating activities.

METHODS: The major active ingredients of essential oils extracted from different parts of Cymbopogon citratus (lemongrass) were identified by GC-MS analysis. The minimum inhibitory concentration (MIC) of the essential oils against P. aeruginosa PAO1, Staphylococcus aureus ATCC 6538, and Candida albicans ATCC 10,261 was determined using the broth microdilution assay. Antibiofilm and antiprotease activities were phenotypically evaluated for the extracted lemongrass essential oils for the same standard strains and clinical isolates for the same microbes. Also, the effects on microbial virulence were validated by RT-qPCR against a subset of genes regulating biofilm, quorum sensing, and stress response in P. aeruginosa, and S. aureus. Furthermore, the antioxidant activities of the different essential oils extracted were evaluated using DPPH, β-carotene/linoleic acid and FRAP methods.

RESULTS: GC-MS analysis of the essential oils of Cymbopogon citratus revealed the presence of 47 different compounds distributed in the different plant organs. Active constituents such as geranial, neral, myrcene, nerolic acid, linalool, iso-citral, trans iso-citral, and neryl acetate were more abundant in the essential oil extracted from leaves (89.77%), followed by stems (82.92%) and finally roots (52.02%). S. aureus, and C. albicans were more sensitive to C. citratus essential oils than Gram negative P. aeruginosa. Incorporation of sub-MIC doses of essential oils into the culture media was sufficient to disrupt the formation of microbial biofilms in P. aeruginosa, S. aureus, and C. albicans, and to inactivate the proteolytic activities of microbial proteases. The essential oils of C. citratus showed promising and broad-spectrum biofilm eradicating activity. Sub-MIC doses of lemongrass oil dramatically reduced the expression of relA, pslA, and spoT in P. aeruginosa, and agrA, icaA, and sigB in S. aureus indicating a broad-spectrum anti-virulence activity.

CONCLUSION: Our study identifies C. citratus essential oils as potential virulence-attenuating agents. Essential oils extracted from the leaves, stems, and roots of C. citratus exhibited significant antimicrobial, anti-biofilm, anti-virulence, and antioxidant activities, effectively combating microbial resistance. These findings suggest that C. citratus essential oils could be a valuable natural alternative in the fight against resistant pathogens.},
}

@article {pmid42305672,
year = {2026},
author = {Liang, J and Chen, W and Wang, C and Wang, M and Wei, P and Xu, Z},
title = {The EnvZ/OmpR two-component regulatory system regulates biofilm formation in Salmonella pullorum via interaction with the LuxS/AI-2 quorum sensing system and activation of the SoxR-AcrAB-TolC pathway.},
journal = {Frontiers in microbiology},
volume = {17},
number = {},
pages = {1817019},
pmid = {42305672},
issn = {1664-302X},
abstract = {To investigate the mechanisms of biofilm (BF) formation in Salmonella pullorum (SP), a strong BF-forming strain designated 1904D10 was selected. Differential proteomics based on tandem mass tag (TMT) labeling was performed to compare the whole bacterial proteome between planktonic and biofilm states. A total of 219 differentially expressed proteins were identified (96 up-regulated, 123 down-regulated in biofilm state). Key up-regulated proteins included those involved in the EnvZ/OmpR two-component system (TCS), LuxS/AI-2 quorum sensing (QS) system, and AcrAB-TolC efflux pump. An ompR deletion mutant was constructed using Red homologous recombination. Deletion of ompR significantly reduced biofilm formation and significantly increased susceptibility to all tested antibiotics. Electrophoretic mobility shift assay (EMSA) demonstrated that OmpR protein directly binds to the promoter region of soxR (a transcriptional activator of AcrAB-TolC), but not to soxS. Molecular docking predicted a potential interaction between the AI-2 QS signaling molecule and EnvZ protein (binding energy: -23.99 kJ/mol), with hydrogen bonds forming at five amino acid residues. This study provides evidence that OmpR directly binds to the soxR promoter and that AI-2 potentially interacts with EnvZ. These findings offer new mechanistic insights specific to Salmonella pullorum-biofilm (SP-BF) regulation and provide potential targets for biofilm control strategies.},
}

@article {pmid42305676,
year = {2026},
author = {Bai, Y and Zhang, Z and Xu, J and Hu, R and Shang, Z and Wei, X and Wang, W and Li, B and Zhu, Z and Zhang, J},
title = {Isoferulic acid suppresses Escherichia coli biofilm formation via LuxS/AI-2 quorum sensing inhibition and synergizes with fosfomycin.},
journal = {Frontiers in microbiology},
volume = {17},
number = {},
pages = {1837128},
pmid = {42305676},
issn = {1664-302X},
abstract = {Escherichia coli (E. coli) O157:H7, a highly virulent foodborne pathogen, poses a substantial threat to public and veterinary health. Its ability to form robust biofilms significantly amplifies virulence and confers resistance to conventional antibiotics, often leading to therapeutic failure. In this study, we employed a genetically engineered LuxS-eGFP reporter strain to screen for compounds targeting the LuxS/AI-2 quorum sensing (QS) system-a key regulator of biofilm formation. Our investigation identified Isoferulic Acid (IFA) as a potent inhibitor of this pathway. IFA effectively suppressed de novo biofilm formation in E. coli O157:H7 without exerting bactericidal effects or impairing general metabolic activity, and it also demonstrated efficacy in dispersing pre-established mature biofilms. Furthermore, we established a novel combinatorial therapeutic strategy by integrating IFA with the antibiotic sodium fosfomycin. This combination exhibited a marked synergistic effect, significantly enhancing antibacterial efficacy against E. coli both in vitro and in vivo. These results substantiate that IFA possesses significant antibiofilm activity and a unique capacity to potentiate antibiotic action, thereby offering a promising new avenue for combating recalcitrant E. coli infections.},
}

@article {pmid42306528,
year = {2026},
author = {Zhao, P and Wang, Z and Jia, N and Wang, X and Liu, J and Zhu, Y},
title = {Mapping the knowledge landscape of Pseudomonas aeruginosa biofilm-mediated drug resistance: a bibliometric analysis and clinical trial landscape overview.},
journal = {Frontiers in cellular and infection microbiology},
volume = {16},
number = {},
pages = {1830404},
pmid = {42306528},
issn = {2235-2988},
mesh = {*Biofilms/drug effects/growth & development ; *Pseudomonas aeruginosa/drug effects/physiology ; Humans ; Bibliometrics ; *Anti-Bacterial Agents/pharmacology/therapeutic use ; *Pseudomonas Infections/microbiology/drug therapy ; *Drug Resistance, Bacterial ; Clinical Trials as Topic ; *Drug Resistance, Multiple, Bacterial ; },
abstract = {BACKGROUND: Pseudomonas aeruginosa is a major opportunistic pathogen whose ability to form biofilms greatly enhances antimicrobial tolerance and contributes to persistent infection. Although increasing attention has been paid to biofilm-mediated drug resistance, the overall knowledge structure and translational development of this field remain unclear.

METHODS: A bibliometric analysis was performed using publications retrieved from the Web of Science Core Collection and Scopus on December 20, 2025. The search covered the period 2014-2025 and focused on P. aeruginosa, antimicrobial resistance, and biofilms, resulting in 6,537 publications for bibliometric analysis. To complement the bibliometric findings, a supplementary narrative review of published clinical studies and a separate registered trial landscape overview were conducted. After screening, 6 published clinical studies and 18 registered interventional trials were included.

RESULTS: Global research output on P. aeruginosa biofilm-mediated resistance increased steadily from 2014 to 2025. China, the United States, and India were the most productive countries, while the United States showed the leading role in the international collaboration network. Keyword clustering and temporal analyses indicated three major research directions: multidrug resistance evolution and pathogenic synergy, novel antibacterial interventions and functional materials, and clinical translation and efficacy evaluation. The supplementary clinical component showed growing interest in adjunctive and mechanistically targeted strategies, particularly in chronic airway and wound-associated infections, although mature efficacy data remain limited.

CONCLUSIONS: Research on P. aeruginosa biofilm-mediated drug resistance is shifting from mechanistic exploration toward translational application. This study provides a data-driven overview of the field's intellectual structure, research hotspots, and emerging trends, and may help guide future anti-biofilm and anti-resistance research.},
}

*Biofilms/drug effects/growth & development
*Pseudomonas aeruginosa/drug effects/physiology
Humans
Bibliometrics
*Anti-Bacterial Agents/pharmacology/therapeutic use
*Pseudomonas Infections/microbiology/drug therapy
*Drug Resistance, Bacterial
Clinical Trials as Topic
*Drug Resistance, Multiple, Bacterial

@article {pmid42307328,
year = {2026},
author = {Moehammad, KS and Al Zamzami, IM and Pramudia, Z and Susanti, YAD and Faqih, AR and Yahya, and Kurniawan, A},
title = {Probiotic Bacillus spp. modulate biofilm microbial assembly in Litopenaeus vannamei culture water.},
journal = {Brazilian journal of biology = Revista brasleira de biologia},
volume = {86},
number = {},
pages = {e303812},
doi = {10.1590/1519-6984.303812},
pmid = {42307328},
issn = {1678-4375},
mesh = {*Biofilms/growth & development/drug effects ; Animals ; *Penaeidae/microbiology ; *Bacillus/physiology ; *Probiotics/pharmacology ; Microscopy, Electron, Scanning ; RNA, Ribosomal, 16S/genetics ; Aquaculture ; *Water Microbiology ; },
abstract = {Shrimp aquaculture faces persistent challenges arising from microbial imbalances and pathogenic dominance, often linked to biofilm-forming bacterial communities. Probiotics, particularly Bacillus spp., offer a sustainable alternative to antibiotic and chemical management; however, their structural and ecological roles in biofilm modulation remain insufficiently understood. This study applied an integrative approach combining Scanning Electron Microscopy (SEM), 16S rRNA amplicon sequencing, and nutrient monitoring to evaluate the influence of Bacillus-based probiotic supplementation on biofilm communities in Litopenaeus vannamei culture water. Amplicon-based analysis revealed a distinct reconfiguration of bacterial assemblages: untreated biofilms were dominated by Vibrio spp., whereas probiotic supplementation reduced their relative abundance and transiently increased marine-associated genera such as Marivita, Pseudoalteromonas, and Marinobacter during early succession. Bacillus exhibited increased relative abundance under probiotic treatment, coinciding with a shift toward a more compositionally balanced community structure. SEM observations corroborated these findings, showing that probiotic application was associated with less compact and more heterogeneous biofilm architecture. Nutrient analyses demonstrated moderately lower concentrations of ammonia, nitrite, and nitrate in probiotic-treated systems, whereas phosphate concentrations remained consistently low across treatments. Although Bacillus abundance was negatively associated with inorganic nitrogen parameters, functional mechanisms were not directly measured. These findings therefore represent ecological associations rather than mechanistic confirmation of enhanced nutrient turnover. Overall, the results highlight the potential of Bacillus-based probiotics as environmentally compatible biofilm modulators in shrimp aquaculture systems.},
}

*Biofilms/growth & development/drug effects
Animals
*Penaeidae/microbiology
*Bacillus/physiology
*Probiotics/pharmacology
Microscopy, Electron, Scanning
RNA, Ribosomal, 16S/genetics
Aquaculture
*Water Microbiology

@article {pmid42308220,
year = {2026},
author = {Arens, DK and Jensen, M and Rose, MA and Zamora, HM and Huh, EY and Hwang, YY},
title = {Phage mediated growth inhibition and biofilm disruption of the endodontic pathogen Enterococcus faecalis.},
journal = {PloS one},
volume = {21},
number = {6},
pages = {e0350657},
doi = {10.1371/journal.pone.0350657},
pmid = {42308220},
issn = {1932-6203},
mesh = {*Biofilms/growth & development ; *Enterococcus faecalis/virology/growth & development/physiology ; *Bacteriophages/physiology/isolation & purification ; Humans ; Host Specificity ; },
abstract = {The failure of endodontic procedures such as root canal therapy is primarily indicated by persistent microbial infections. Root canal therapy involves the removal of decaying dental pulp (internal blood vessels and nerves of teeth), sanitizing the canal, and filling the space with biocompatible materials. Improper cleaning or the breakdown of these materials can lead to secondary infections. These infections, if left untreated, can lead to severe pain, bone and tooth loss, and potentially systemic infection. The bacterium Enterococcus faecalis is one of the most commonly associated organisms with failed root canal therapy, in addition to its prevalence in urinary tract infections, endocarditis, wounds, and sepsis. E. faecalis is known to survive in low nutrient environments and produce extensive biofilms, making it difficult to eradicate. In addition to antibiotic treatment, bacteriophages (phages), which are bacteria-specific viruses that kill their host are an interesting companion or alternative to antibiotics. In this study we isolated and characterized 14 E. faecalis phages from wastewater samples by testing their host range, growth inhibition, and biofilm eradication capabilities against several E. faecalis strains including two that were orally derived. Several phages showed broad host ranges (up to 16 strains), strong bacterial growth inhibition even when applied at low concentrations, and significant eradication of mature biofilms (97% reduction). The phages presented here represent a unique repertoire of antibacterial agents for use in treating endodontic infections and add to the growing library of E. faecalis phages to treat diverse infections.},
}

*Biofilms/growth & development
*Enterococcus faecalis/virology/growth & development/physiology
*Bacteriophages/physiology/isolation & purification
Humans
Host Specificity

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

COVARIATES: Not applicable.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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