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Bibliography on: Biofilm

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ESP: PubMed Auto Bibliography 25 Aug 2025 at 01:38 Created: 

Biofilm

Wikipedia: Biofilm A biofilm is any group of microorganisms in which cells stick to each other and often also to a surface. These adherent cells become embedded within a slimy extracellular matrix that is composed of extracellular polymeric substances (EPS). The EPS components are produced by the cells within the biofilm and are typically a polymeric conglomeration of extracellular DNA, proteins, and polysaccharides. Because they have three-dimensional structure and represent a community lifestyle for microorganisms, biofilms are frequently described metaphorically as cities for microbes. Biofilms may form on living or non-living surfaces and can be prevalent in natural, industrial and hospital settings. The microbial cells growing in a biofilm are physiologically distinct from planktonic cells of the same organism, which, by contrast, are single-cells that may float or swim in a liquid medium. Biofilms can be present on the teeth of most animals as dental plaque, where they may cause tooth decay and gum disease. Microbes form a biofilm in response to many factors, which may include cellular recognition of specific or non-specific attachment sites on a surface, nutritional cues, or in some cases, by exposure of planktonic cells to sub-inhibitory concentrations of antibiotics. When a cell switches to the biofilm mode of growth, it undergoes a phenotypic shift in behavior in which large suites of genes are differentially regulated.

Created with PubMed® Query: ( biofilm[title] NOT 28392838[PMID] NOT 31293528[PMID] NOT 29372251[PMID] ) NOT pmcbook NOT ispreviousversion

Citations The Papers (from PubMed®)

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RevDate: 2025-08-24

Lee YH, Hsu LH, Yu ZH, et al (2025)

Rad6 and Bre1 ubiquitin ligase negatively regulate biofilm formation and virulence in Candida glabrata.

The Journal of infection pii:S0163-4453(25)00195-1 [Epub ahead of print].

BACKGROUND: Candida glabrata is an opportunistic human fungal pathogen causing infections due to its innate antifungal drug resistance and ability to adhere to mucocutaneous surfaces. Epigenetic pathways may be important factors in the development of drug resistance. Our previous studies showed that deubiquitination of H2B, regulated by a module comprised of Ubp8, Sgf11, Sgf73, and Sus1, plays important roles in oxidative stress tolerance and biofilm formation of C. glabrata. However, the roles of the Rad6 and Bre1 ligase in regulating the ubiquitination of H2B in C. glabrata remain unclear.

METHODS: We characterized the functions of Rad6 and Bre1 in C. glabrata by generating deletion mutants (rad6, bre1, and rad6 bre1). We analyzed biofilm formation, gene expression of key adhesins (EPA1, EPA6, EPA20) and proteases (YPS4), antifungal drug susceptibility, stress responses, and virulence in a murine model of systemic candidiasis.

RESULTS: Deletion of RAD6 and BRE1 resulted in enhanced biofilm formation, correlating with upregulation of key adhesin genes and the protease gene YPS4. The mutants showed distinct patterns of antifungal drug susceptibility: rad6 and rad6 bre1 mutants exhibited increased sensitivity to azoles, while bre1 mutant showed enhanced resistance to azoles in solid YPD agar plates but no significant difference in liquid RPMI medium. All mutants demonstrated decreased resistance to echinocandins and amphotericin B, associated with altered expression of ergosterol biosynthesis genes (ERG11) and glucan synthase genes (FKS1, FKS2). The mutants also displayed decreased resistance to oxidative and cell wall stresses despite elevated basal expression of antioxidant genes (SOD1, GPX2, CTA1). In a murine model of systemic candidiasis, both rad6 and bre1 mutants exhibited enhanced virulence compared to the wild type.

CONCLUSION: Rad6 and Bre1 in C. glabrata function as negative regulators of biofilm formation and adhesion, and their related-genes expression, while RAD6 deletion also suppresses macrophage ROS production and enhances fungal survival. The enhanced virulence observed in the rad6 and bre1 mutants is primarily attributed to these combined effects of increased biofilm formation, enhanced adhesion capability, and macrophage immune evasion.

RevDate: 2025-08-24

Sakthivel I, Rangaswamy B, Rajagopal B, et al (2025)

Integrating kinetic models, gene circuits, and biofilm dynamics for enhanced exopolysaccharide production in nitrifying bacterial consortia.

Journal of microbiological methods pii:S0167-7012(25)00153-8 [Epub ahead of print].

Bacterial consortia enriched from domestic wastewater were studied through kinetic and genetic circuit modelling to optimize extracellular polysaccharide (EPS) production and nitrogen removal. This study integrates kinetic modelling and synthetic biology to optimize consortia performance. Growth kinetics were simulated using extended Monod and Verhulst models, under controlled nitrogen flux (10 ppm NH4Cl), yielding a maximum biomass concentration (OD590 = 5.39) and an EPS production of 2.63 g/L by day 45. The Monod model described the specific growth rate (μ) as a function of nitrogen concentration (Sn), while the Verhulst model estimated biomass accumulation over time. Scanning electron microscopy (SEM) showed the gradual development of biofilms, starting from scattered clusters, and progressing to dense structures. Nitrogen flux analysis revealed that 80 % of ammonia was oxidized by autotrophic bacteria (AOB/NOB). PCR amplification confirmed the presence of the exoY gene, which was used to build a BUFFER-gate logic gene circuit for controlling succinoglycan production. Through focused genetic and kinetic optimization, this study demonstrates effective nitrification, providing a strong framework for wastewater treatment and biofilm engineering.

RevDate: 2025-08-24

Wang L, Wang S, Zhang W, et al (2025)

ROS-induced allosteric regulation of NikR coordinates HP0910-mediated OMP2 methylation to modulate H. pylori biofilm dynamics and therapeutic targeting.

Microbiological research, 301:128319 pii:S0944-5013(25)00278-2 [Epub ahead of print].

Biofilm formation represents a critical survival strategy for Helicobacter pylori (H. pylori), facilitating antibiotic resistance and chronic colonization. In this study, we demonstrate that reactive oxygen species (ROS) released by macrophages enhance H. pylori biofilm formation through a novel epigenetic pathway. Transcriptomic and genetic analyses revealed that the nickel-responsive regulator NikR is allosterically activated by ROS, transitioning from its apo to holo conformation. This conformational shift markedly represses the expression of the DNA methyltransferase HP0910, resulting in hypomethylation of the omp2 gene, which encodes an outer membrane protein (OMP), and subsequent OMP2 overexpression, as validated by quantitative reverse transcription PCR (qRT-PCR) and reporter assays. Elevated OMP2 levels enhance extracellular polymeric substance (EPS) production, leading to a significant increase in biofilm biomass and thickness, as quantified by crystal violet staining and confocal laser scanning microscopy (CLSM). ROS scavenging reverses this phenotype, promoting biofilm dispersal. Furthermore, molecular docking and functional assays identified flopropione, a small-molecule compound targeting OMP2, led to an approximately 80 % reduction in biofilm biomass in vitro. When combined with standard triple therapy (omeprazole, amoxicillin, and clarithromycin), flopropione significantly improved bacterial clearance (>2-log10 reduction) in a murine infection model. Collectively, our findings elucidate the ROS-NikR-HP0910-OMP2 signaling axis that regulates H. pylori biofilm dynamics and identify flopropione as a promising anti-biofilm therapeutic candidate against multidrug-resistant infections.

RevDate: 2025-08-24

Yang D, Liang J, Yang Q, et al (2025)

Pyrite facet-dependent microbial oxidation and interfacial interaction mechanisms: An example of crystal facets variability with different modes of biofilm attachment.

Colloids and surfaces. B, Biointerfaces, 256(Pt 2):115063 pii:S0927-7765(25)00570-3 [Epub ahead of print].

Different crystal facets of pyrite are anisotropic, which affects the biogeochemical cycling of iron. However, the potential mechanisms of interfacial interactions between pyrite and microorganisms on different exposed crystal surfaces are unclear. Therefore, this study investigates the effect of the interaction of pyrite {100} and {210} facets with Sulfobacillus thermophilidoxidans YN22 on the dissolution of pyrite. The results showed that the {210} facets formed larger pearl-string corrosion pits and the biological oxidation rate was about 20 % higher than that of the {100} facets. This was attributed to the higher reactivity of the {210} facets with 4-coordinated iron atoms and electron transfer capacity, which promoted a wider and denser distribution of biofilms on their surfaces, thus accelerating the oxidative dissolution of the {210} facets and the formation of larger corrosion pits. In addition, the {100} facets showed a more dispersed biofilm distribution and their surfaces were prone to form more K-jarosite, which hindered the oxidation of the {100} facets. This study revealed that the surface properties of different crystal facets {100} and {210} of pyrite affect biocorrosion, providing a new perspective on the oxidation process of pyrite with microorganisms at different crystal facets.

RevDate: 2025-08-24
CmpDate: 2025-08-24

Motavaf F, Abbasi M, Asadalizadeh H, et al (2025)

Enhanced Antibacterial, Anti-Biofilm, and Anticancer Activities of Liposome-Encapsulated Selenium Nanoparticles: A Novel Therapeutic Approach.

Asian Pacific journal of cancer prevention : APJCP, 26(8):3005-3017 pii:91810.

BACKGROUND: This study investigates the green synthesis of selenium nanoparticles (SeNPs) and their encapsulation in liposomes as a novel drug delivery system to enhance the antibacterial and anticancer properties of SeNPs. Liposomes are well-known for their ability to improve the biological activity of encapsulated drugs, making them a promising candidate for targeted therapies, particularly in oral cancer treatment.

METHODS: Biosynthesised SeNPs were incorporated into liposomes via the thin-film hydration technique. Particle size and zeta potential were quantified by dynamic light scattering (DLS), whereas encapsulation efficiency (EE) was determined spectrophotometrically (UV-Vis).

RESULTS: The physicochemical properties of the liposome-loaded SeNPs were characterized, revealing an average size of 270 nm, spherical morphology, and an encapsulation efficiency of 50.5%. The release profile of SeNPs from the liposomes demonstrated a controlled release of 61% over 64 hours, while free SeNPs released 100% of their content during the same period. The antibacterial and anti-biofilm activities of both free and liposome-loaded SeNPs were tested against standard pathogenic bacterial strains, with the liposome formulation showing enhanced efficacy. The cytotoxicity assay revealed that liposome-loaded SeNPs exhibited significantly higher cytotoxic effects on oral cells compared to free SeNPs, indicating improved therapeutic potential.

CONCLUSION: The study demonstrates that liposome-loaded SeNPs are an effective and biocompatible drug delivery system with notable antibacterial, anti-biofilm, and anticancer properties, making them a promising candidate for targeted drug delivery in oral cancer therapy.

RevDate: 2025-08-23

Bai S, Wan S, Chen Y, et al (2025)

Design and synthesis of anti-biofilm derivatives from phospholipid amides.

Molecular diversity [Epub ahead of print].

Bacterial biofilms serve as a natural barrier, enabling bacteria residing within them to exist and potentially amplify bacterial resistance by shielding themselves from bactericide exposure. Despite considerable efforts directed toward inhibiting bacterial growth, research has overlooked bacterial biofilms to a significant extent, leading to the frequent deficiency of traditional antimicrobials in inhibiting such biofilms. This necessitates the development of antimicrobials capable of inhibiting biofilms for effective antibacterial intervention. Herein, we have developed a new bacteriostatic agent, A6, which has demonstrated the capability of inhibiting biofilm formation. It achieved a biofilm inhibition rate of 72.76% at a concentration of 47.94 μg/mL (2.0 EC50). Mechanistic studies revealed that A6 inhibits extracellular polymeric substances (EPS) production and bacterial motility, both critical for bacterial virulence, biofilm formation, maturation, or plant cell wall degradation. Additionally, the conductivity and protein leakage experiments demonstrated that compound A6 significantly affected various physiological processes of Xoc. In summary, A6 presents a promising antimicrobial solution by simultaneously inhibiting biofilms, addressing a crucial aspect of bacterial plant diseases.

RevDate: 2025-08-23
CmpDate: 2025-08-23

Mullin TJ, MacIsaac SA, Stoddart AK, et al (2025)

Impact of material properties for improved Pseudomonas aeruginosa biofilm inactivation with 280 nm UV LEDs.

Scientific reports, 15(1):31016.

Biofilm microbial communities encased in extracellular polymeric substances are a concern in drinking water premise plumbing and fixtures, and are challenging to remove and disinfect. Pseudomonas aeruginosa (P. aeruginosa), a commonly used surrogate organism, is employed in this study due to its widely documented occurrence in biofilms within drinking water systems. This study investigates 280 nm UV light emitting diodes (UV LEDs) for inactivating P. aeruginosa biofilms grown on common plumbing materials extruded Polytetrafluoroethylene, Acrylonitrile Butadiene Styrene, Viton®, Silicone, High Density Poly Ethylene, Stainless Steel, Porex (expanded PTFE), and Polycarbonate. Biofilms were cultivated in CDC biofilm reactors on 12.8 mm diameter coupons and then exposed to UV LED light at fluences ranging from 5 to 40 mJ/cm[2] with log reduction values between 0.851 and 2.05 CFU/cm[2] for Viton® (k = 0.133 ± 0.0625 cm[2]/mJ) and Silicone (k = 0.344 ± 0.145 cm[2]/mJ), respectively. This research demonstrates that material properties influence biofilm formation and the subsequent effectiveness of UV LED inactivation while illustrating that characteristics such as surface roughness and reflectivity significantly impact inactivation. This work advances the understanding of biofilm inactivation under UV LED exposure, thereby aiding in the development of more effective biofilm inactivation strategies.

RevDate: 2025-08-23

Ventura PVB, Chaves AC, Pereira MS, et al (2025)

Exploring Plant Compounds as Enhancers of Ciprofloxacin Activity Against Planktonic and Biofilm Cells of Poultry-Related Enterobacteriaceae.

Microbial pathogenesis pii:S0882-4010(25)00720-X [Epub ahead of print].

This study evaluated the antimicrobial and antibiofilm activities of ciprofloxacin (Cip) in combination with trans-cinnamaldehyde (Tc), geraniol (Ger), and eugenol (Eug) against multidrug-resistant Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae isolated from broiler feces. Disk diffusion and microdilution assays showed that Tc displayed the highest antibacterial activity in disk diffusion (IZD: 30 mm) and microdilution assays (MIC: 312.5 μg/mL), while Ger and Eug revealed smaller inhibition zones (∼9-12 mm) and higher MIC values (2,500 μg/mL). Checkerboard assays indicated that Ger and Eug synergized with Cip (FICi ≤ 0.5), decreasing Cip MIC by up to 130-fold. The combinations of Cip with Eug and Ger led to a 100% and 90% reduction in biofilms, respectively. These findings suggest that plant compounds, particularly Ger and Eug, enhance the efficacy of Cip against both planktonic and biofilm cells, presenting a promising strategy to combat antibiotic resistance in poultry farming.

RevDate: 2025-08-22

Rafi MO, Hasan MAE, Fahim NAI, et al (2025)

Environmental distribution of biofilm-forming antibiotic-resistant Escherichia coli associated with plastic surface materials.

Environmental science and pollution research international [Epub ahead of print].

Plastic pollution is now an emerging issue worldwide, and the amount of plastic debris is rapidly increasing day by day in this decade. The surface of plastic contains a wide variety of biofilm-forming microorganisms that can pose a risk to human health. Studies showed that Escherichia coli is resistant to numerous classes of antibiotics; however, the prevalence of the bacterium on the environmental plastic surface is still unknown. The current study aimed at identifying biofilm-forming E. coli from the plastic surface collected from various environmental origins and determining the antibiotic-resistant pattern. A total of 90 plastic samples were collected from wastewater and open surface environments of Mymensingh Medical College, Bangladesh Agricultural University, and BCIC industrial areas of Mymensingh. Among these, 65 samples were found to be positive for the presence of E. coli. The plastic samples collected from drainage sources displayed the highest E. coli prevalence. By targeting the malB gene of the cultured samples, 36 E. coli isolates were positive out of 65, and the prevalence rate was 55.4%. There was a considerable variation in terms of the antibiotic-resistant pattern of the isolates. Randomly, 29 isolates were subjected to an antibiogram study. All of the isolates were resistant to imipenam (100%) and ceftazidime (100%), 79.40% were resistant to ampicillin, and 44.82% resistant to gentamicin. The beta-lactamase-producing genes blaTEM were detected in 51% (14/29) isolates that showed resistance to ampicillin. The biofilm-forming study revealed that 91.16% strong biofilm-forming E. coli isolates were resistant to ampicillin. Additionally, 18.18% non-biofilm-forming tetracycline-resistant E. coli isolates have been found in this study. In summary, to the best of our knowledge, this is the first study in Bangladesh to isolate and identify biofilm-forming antibiotic-resistant E. coli collected from environmental plastic surfaces, but further pathogenicity tests and resistome analysis are required to know the exact genetic resistance pattern.

RevDate: 2025-08-22

Kwong PTH, Das T, Arnold JC, et al (2025)

The anti-biofilm activity of cannabinoids against methicillin-resistant Staphylococcus aureus.

Journal of applied microbiology pii:8239790 [Epub ahead of print].

AIMS: Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of hospital-acquired pneumonia with resistance against beta-lactam antibiotics. New, potent antibiotics against MRSA with other mechanisms of action are thus urgently needed. Recently, cannabinoids have been evaluated for antimicrobial activity in the ongoing search for new anti-infective agents, but their anti-biofilm effect has not been extensively studied. In this study, five main phytocannabinoids - canndibidiol (CBD), delta-9-tetrahydrocannabinol (THC), cannabinol (CBN), cannabigerol (CBG), and cannabichromene (CBC) were examined for their activity against a MRSA biofilm.

METHODS AND RESULTS: The anti-biofilm activity was assessed by crystal violet staining, resazurin metabolic assay, reactive oxygen species (ROS) assay, and propidium iodide membrane integrity test. The minimum inhibitory concentrations of all tested cannabinoids were between 1-2 µg/mL. CBN showed the most potent anti-MRSA biofilm activity, significantly reducing biofilm biomass and bacterial viability. It also induced the highest intracellular ROS levels. In contrast, CBD was the least effective among the tested cannabinoids in most of the anti-biofilm assays, yet it caused the greatest membrane damage to bacteria within the biofilm.

CONCLUSIONS: This study showed that despite being chemically similar, the cannabinoids demonstrated different potency and potentially different mechanisms of action against MRSA. More research is needed to investigate how they act on this pathogen and its biofilm.

RevDate: 2025-08-22
CmpDate: 2025-08-22

Kashi M, Varseh M, Askarinia M, et al (2025)

The interactions of natural compounds with Escherichia coli motility, attachment, communication systems, and mature biofilm.

Archives of microbiology, 207(10):236.

Bacterial biofilms play a significant role in increasing antibiotic resistance and the pathogenesis of Escherichia coli; their control is a major challenge in treating bacterial infections. In recent years, natural compounds have emerged as effective alternatives for inhibiting the formation and destruction of bacterial biofilms. Natural compounds such as curcumin, cinnamaldehyde, eugenol, carvacrol, quercetin, resveratrol, thymol, citral, and catechin are noteworthy in hindering and destroying E. coli biofilms. They inhibit bacterial motility (swarming and swimming), reducing attachment to surfaces, and downregulate genes related to attachment and motility (fimH, csgABC, sfaAS, papG, fliAC, flhCD, and motAB). Natural compounds can also disrupt the bacterial communication system and cause changes in the expression of luxS, sdiA, tnaA, qseBC, bssS, and lsrR genes. Studies have also shown that natural compounds can destroy mature biofilms' structure and decrease biofilm exopolysaccharides' production. This impact makes the bacteria more sensitive to the antimicrobial agents. However, one of the biggest challenges in using natural compounds is their low stability and restricted bioavailability in biological environments. For this reason, utilizing nanoparticles and novel drug delivery systems can improve these compounds' stability, penetrability, and efficacy. This article reviews the antibiofilm potential of natural compounds, their mechanisms of activity in hindering and destroying E. coli biofilms, and the role of nanotechnology in improving the performance of these compounds.

RevDate: 2025-08-22

Sankaran M, Kaliyamoorthy K, M Alagumuthu (2025)

Antimicrobial quinoline triazoles: synthesis, docking, and dynamic simulation studies against biofilm-associated infections.

Molecular diversity [Epub ahead of print].

The alarming rise of multidrug-resistant (MDR) bacterial pathogens poses a significant challenge to current antimicrobial therapy, challenging the development of novel, structurally diverse agents. In this study, a new series of phenylquinoline-triazoles (PQTs) 4a-l was rationally designed and synthesized using a copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) click chemistry approach. Synthesized PQTs were characterized by standard analytical techniques, including [1]H NMR, [13]C NMR, HRMS, and spectroscopic analyses. The antimicrobial efficacy of PQTs 4a-l was evaluated against a panel of clinically relevant biofilm-causing bacterial strains, including Streptococcus pneumoniae (MTCC 1936), Staphylococcus aureus (MTCC 737), Escherichia coli (MTCC 443), Pseudomonas aeruginosa (MTCC 424), and methicillin-resistant Staphylococcus aureus (MRSA). Candida albicans was the only fungal strain utilized, considering its role in biofilm formation in several infections, including UTI (Urinary Tract Infection). In the results, three PQTs exhibited potent broad-spectrum antibacterial activity, predominantly against Gram-positive strains and MRSA. Due to the activity selectivity, a molecular docking study was executed against the penicillin-binding protein 2a (PBP2a), a key resistance factor in MRSA (PDB ID: 6H5O), and the best compounds screened were subjected to test the PBP2a inhibition potential in vitro. The most active compounds exhibited strong binding affinities and favorable interaction forms within the active site of PBP2a, including hydrogen bonding and π-π stacking with key amino acid residues. Furthermore, the docked complexes were subjected to 100 ns molecular dynamics (MD) simulations, which confirmed their structural stability and robust interactions under physiological conditions. Furthermore, in silico ADME and drug-likeness profiling suggested good pharmacokinetic properties. In conclusion, we identified compounds 4d, 4i, and 4 k as are most effective PQTs among 4a-l with remarkable antimicrobial potentials. These findings determine that PQTs are promising scaffolds for combating resistant bacterial infections such as MRSA and warrant further preclinical investigation.

RevDate: 2025-08-22
CmpDate: 2025-08-22

Reem CSA, Chowdhury MAH, Ashrafudoulla M, et al (2025)

Leveraging Blockchain and AI for Biofilm Control in Food Processing Environments.

Comprehensive reviews in food science and food safety, 24(5):e70261.

Biofilm formation in food processing environments significantly threatens food safety and quality due to its resistance to conventional cleaning and disinfection methods. These resilient microbial communities contribute to contamination, spoilage and foodborne illnesses, highlighting the need for innovative and technology-driven control strategies. Emerging digital tools, particularly blockchain technology and artificial intelligence (AI), offer new opportunities for enhancing biofilm management. Blockchain ensures secure, real-time traceability of hygiene records, contamination events and compliance activities across the supply chain. Complementing this, AI technologies such as machine learning and sensor-based analytics support early detection of microbial growth, anomaly identification and predictive risk assessment. Together, these tools promote data-driven decision-making and more proactive contamination prevention. While pilot applications show promise in improving transparency and sanitation outcomes, challenges remain, including data integration, implementation costs and regulatory barriers. Addressing these issues will require interdisciplinary collaboration and supportive policy frameworks. This review summarizes the current and potential roles of blockchain and AI in biofilm control and outlines future directions for research and industrial application.

RevDate: 2025-08-22

Cintra TMF, Menezes RT, de Carvalho LS, et al (2025)

Rosemary Extract: Phytochemical Composition and Potential for Eliminating Polymicrobial Biofilm of Candida albicans and Multidrug-Resistant Bacteria.

Biotech (Basel (Switzerland)), 14(3): pii:biotech14030061.

Herbal medicines can be promising for the treatment of infections caused by multidrug-resistant microorganisms. This study aimed to evaluate Rosmarinus officinalis (Rosemary) hydroalcoholic extract (RHE) regarding its phytochemical composition and potential for eliminating polymicrobial biofilm of Candida albicans with multidrug-resistant bacteria (Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa). The extraction and quantification of the extract (flavonoids and phenols) were performed, and its antioxidant activity (DPPH) and the presence of bio-active compounds were investigated using high-performance liquid chromatography with Diode Array Detection (HPLC-DAD) and Gas Chromatography-Mass Spectrometry (GC-MS). The minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) were determined, and the extract's action on polymicrobial biofilms was evaluated using the MTT assay. Data were analyzed using one-way ANOVA and Tukey's tests, as well as Kruskal-Wallis and Dunn's tests, with a significance level of 5%. RHE showed compatible amounts of flavonoids and phenols, with an EC50 of 19.53 µg/mL. Through HPLC-DAD and GC-MS, biomolecules such as rosmarinic acid and α-Pinene were identified. The extract exhibited microbicidal activity and antibiofilm action, with reduction percentages of up to 69.6% (p < 0.05), showing superior performance compared to 0.12% chlorhexidine against C. albicans + A. baumannii. In conclusion, RHE may be a promising therapeutic agent against multidrug-resistant pathogens.

RevDate: 2025-08-22

Ismael M, Edwin M, K Juliah (2025)

Biogenic synthesis of silver nanoparticles using Sida cuneifolia leaf extract for enhanced antibacterial, cytotoxic, and anti-biofilm activities.

Biotechnology notes (Amsterdam, Netherlands), 6:196-208.

Antimicrobial resistance (AMR) is one of the global threats that needs to be addressed. Nanotechnology represents a promising way to address this issue due to its multifaceted mode of action. This study aimed to synthesize and evaluate the antimicrobial and anti-biofilm properties of silver nanoparticles using S. cuneifolia leaves extract. The formation and properties of AgNPs were characterized using a UV-Vis spectrophotometer, an FT-IR spectrophotometer, TEM, and XRD. Disc diffusion and MIC were used to evaluate the antibacterial activity of AgNPs towards E. coli, S. flexneri, and S. aureus. The antibacterial action of silver NPs was observed using SEM, and cytotoxicity was assessed using the hemolysis assay. The anti-biofilm was evaluated against E. coli and S. aureus. From the results obtained, a sharp peak in the UV-Vis spectra centered at 419 nm was associated with AgNPs, while the sharp, distinct peaks in the powder diffractograms were linked to the face-centered cubic (fcc) of crystalline AgNPs. TEM micrographs confirmed their spherical morphology, with dimensions varying from 4 to 31 nm. The nanoparticles showed significant antibacterial and anti-biofilm activities against the tested isolates. Additionally, SEM confirmed that they could destroy the cell membrane and cause death. The biocompatibility of the synthesized AgNPs was safe at 100 μg/mL. Therefore, S. cuneifolia leaf extract has the potential to be an environmentally friendly substitute for the fabrication of Ag nanoparticles. The findings reveal that the synthesized nanoparticles could serve as a secure and effective alternative for addressing AMR.

RevDate: 2025-08-22
CmpDate: 2025-08-22

Fernández-Calderón MC, Fernández-Babiano I, Navarro-Pérez ML, et al (2025)

Biofilm formation and role of other pathogenic factors in the virulence of Staphylococcus epidermidis clinical isolates.

Frontiers in cellular and infection microbiology, 15:1630341.

Medical device-associated infections represent a significant healthcare challenge, as sterilization of the biomaterial often necessitates device removal. The most frequently isolated microorganism in these infections is Staphylococcus epidermidis, a skin commensal capable of causing a wide range of nosocomial infections. The primary virulence factor of S. epidermidis is biofilm formation, which decreases antibiotic efficacy and host immune response. However, additional factors play crucial roles in infection establishment. Understanding the interplay between virulence factors is essential to developing preventive strategies that inhibit microbial adhesion and biofilm development. In this study, we analyzed the presence of genes associated with adhesion and biofilm formation (ica-dependent and ica-independent pathways) in 25 clinical isolates of S. epidermidis and four control strains: ATCC 12228, ATCC 35983, ATCC 35984, and the HAM 892 mutant. Resistance profile was determined, and biofilm quantification and composition of matrix was performed using multiple methodologies. Additionally, parameters associated with initial adherence as cell surface hydrophobicity (CSH) were investigated. A strong correlation was observed among different methods for measuring biofilm formation and matrix composition. The 14 icaADBC+ isolates exhibited higher prevalence of the aap, bhp, mecA, and IS256 genes, with polysaccharide intercellular adhesin (PIA) identified as the main matrix component. In contrast, icaADBC- biofilm-producing strains formed biofilms rich in other polysaccharides and proteins. The 15 non-biofilm-producing isolates showed significantly higher hydrophobicity levels, suggesting that this factor plays a critical role in initial adhesion and colonization. This study highlights the diverse mechanisms underlying biofilm formation in S. epidermidis and identifies hydrophobicity as a potential pathogenicity factor contributing to its virulence.

RevDate: 2025-08-22

Mandal AA, Upadhyay A, Mandal A, et al (2025)

Ru(II)-Based Photo-Antibiotics: Light-Driven Eradication of Bacterial Biofilm and Rapid Healing of Infective Wounds in Wistar Rat.

Chemistry (Weinheim an der Bergstrasse, Germany) [Epub ahead of print].

Three novel Ru(II)-based photoantibiotics, viz., [Ru(phtpy)(N,N,N)2](PF6)2, where N,N,N = 4'-phenyl-2,2':6',2″-terpyridine (phtpy, Ru1), 4-([2,2':6',2''-terpyridin]-4'-yl)-N,N-dimethylaniline (NMe2tpy, Ru2), trifluoromethylphenyl)-2,2':6',2''-terpyridine (CF3tpy, Ru3) were developed with excellent photostability for visible light-activated antibacterial activities and infective wound healing. Ru1-Ru3 exhibited an absorption in the 400-600 nm range, beneficial for antibacterial photodynamic therapy (aPDT) application. Upon visible light exposure (400-700 nm), Ru1-Ru3 inhibited bacterial growth of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Bacillus subtilis (B. subtilis), due to the effects of oxidative stress via ROS generation and photo-oxidation of NADH. Ru3 was identified as a lead antibiotic that further showed antibiofilm activities against E. coli under visible light exposure. Ru3 was found to be biocompatible against rat red blood cells and human embryonic kidney cells. Ru3 + light promoted rapid healing of infected wounds within 9 days in an E. coli-induced rat model, highlighting its potential future use in healthcare.

RevDate: 2025-08-21

Do E, McManus CJ, Zarnowski R, et al (2025)

Ume6 protein complexes connect morphogenesis, adherence and hypoxic genes to shape Candida albicans biofilm architecture.

Nature microbiology [Epub ahead of print].

Biofilms of the fungal pathogen Candida albicans can form on implanted medical devices and contribute to fungal virulence and are recalcitrant to antifungal therapy. The transcription factor Ume6 directs hyphal cell elongation and thus promotes biofilm formation in C. albicans. However, how exactly this key biofilm and virulence regulator functions has remained unclear. Here RNA sequencing and chromatin immunoprecipitation with sequencing data show that Ume6 binds to and activates multiple biofilm-relevant genes. Ume6-associated sequence motifs correspond to binding sites for biofilm master regulators Efg1 and Ndt80, and hypoxic response regulator Upc2. Co-immunoprecipitation assays show the existence of Ume6-Efg1, Ume6-Ndt80 and Ume6-Upc2 protein complexes. Promoter binding of Ume6 is partially dependent upon Efg1, Ndt80 or Upc2, as is Ume6 target gene activation, thus indicating that the protein complexes function to drive Ume6-target gene interaction. Ume6 therefore acts as a bridge that connects the hyphal morphogenesis and adherence genes that shape biofilm architecture and the hypoxic response genes required for growth in the low-oxygen biofilm environment. These findings are vital for our understanding of the pathobiology of C. albicans and could open the way to new treatment options.

RevDate: 2025-08-21

Nath R, Lahiri D, Nag M, et al (2025)

Retraction Note: Antibiofilm activity of exopolysaccharide-mediated ZnO nanoparticle against Pseudomonas aeruginosa biofilm.

Naunyn-Schmiedeberg's archives of pharmacology pii:10.1007/s00210-025-04569-y [Epub ahead of print].

RevDate: 2025-08-21

Østrup Jensen P, Rybtke M, T Tolker-Nielsen (2025)

Biofilm-associated molecular patterns: BAMPs.

Infection and immunity [Epub ahead of print].

Chronic infections involving bacterial biofilms are a major clinical challenge due to the ability of biofilm to resist antimicrobial treatments and host immune responses. The resulting persistent infections are often accompanied by collateral damage mainly executed by activated components of the innate immune system in response to the infectious biofilm. The innate immune system responds to the recognition of pathogen-associated molecular patterns (PAMPs), which are broadly expressed by both planktonic and biofilm-forming bacteria. However, the expression of special PAMPs in association with biofilms remains poorly defined. Here, we review prior studies that provide experimental evidence of the existence of immune-activating molecular patterns that are expressed at immunostimulatory levels in biofilms but not in planktonic bacteria. Accordingly, we introduce the concept of biofilm-associated molecular patterns (BAMPs) as a subset of PAMPs that are expressed in biofilms. Identifying BAMPs and elucidating their role in innate immune activation may inform the development of targeted therapies to reduce collateral tissue damage in biofilm-associated infections.

RevDate: 2025-08-21

Tuttobene MR, Bruna RE, Molino MV, et al (2025)

PrtA-mediated flagellar turnover is essential for robust biofilm development in Serratia marcescens.

Applied and environmental microbiology [Epub ahead of print].

Biofilm formation is crucial for bacterial persistence, requiring precise regulatory mechanisms to transition from motility to sessility. Here, we uncover the role of the metalloprotease PrtA in Serratia marcescens biofilm development and its interaction with flagellar components. Loss of PrtA leads to reduced biofilm biomass, thickness, and viable cell counts, as shown through high-resolution confocal microscopy. The biofilm-deficient phenotype is rescued by wild-type PrtA expression but not by a proteolytically inactive PrtAE177A mutant, underscoring the essential role of PrtA's enzymatic activity. Exogenous addition of purified PrtA restores biofilm formation, confirming its enzymatic necessity. Proteomic profiling identified flagellar proteins as primary PrtA targets, with an overrepresentation of flagellar components in prtA mutant biofilms. In addition, PrtA selectively degrades depolymerized flagellar filaments, facilitating biofilm progression by removing excess flagellar material. Transcriptional analysis reveals an inverse expression pattern of flagellar master regulator (flhDC) and prtA during biofilm establishment, suggesting a coordinated regulatory axis that suppresses flagellar function while promoting biofilm development. Confocal microscopy at the liquid-air interface shows increased flagellar content in prtA mutant biofilms, supporting PrtA's role in matrix organization and biofilm integrity. Collectively, these findings establish PrtA as a crucial mediator of flagellar turnover and extracellular proteolysis, linking motility suppression to robust biofilm formation. This work not only advances our understanding of biofilm regulation in S. marcescens but also identifies PrtA as a potential target for novel biofilm control strategies.IMPORTANCEBiofilms are central to the persistence and pathogenicity of Serratia marcescens, particularly in clinical settings where they contribute to chronic infections and antimicrobial resistance. This study identifies the metalloprotease PrtA as a critical regulator of biofilm development, acting through the selective degradation of flagellar components to mediate the transition from motility to sessility. By demonstrating that PrtA's proteolytic activity is essential for proper biofilm architecture and viability, and that it directly targets excess flagellar material, we provide mechanistic insight into how biofilm maturation is coordinated with motility suppression. The discovery of an inverse regulatory relationship between prtA and the flagellar master regulator flhDC further supports the existence of a finely tuned system controlling biofilm establishment. Together, these findings enhance our understanding of biofilm regulation in Serratia marcescens, an opportunistic human pathogen increasingly associated with antibiotic resistance, and highlight PrtA as a promising target for novel anti-biofilm strategies.

RevDate: 2025-08-21

O'Connor F, Lazanas A, B Prieto Simón (2025)

Carbonised porous silicon as scaffold and sensor for the electrochemical detection and characterisation of bacterial biofilm growth.

Journal of materials chemistry. B [Epub ahead of print].

The use of carbonised porous silicon (C-pSi) substrates as both scaffolds and electrodes for the electrochemical detection and characterisation of bacterial biofilm growth is reported for the first time. Three types of C-pSi are presented showcasing versatility in their hydrophilicity and surface chemistry. The tunability of these properties is taken as a springboard for the biological, morphological and electrochemical evaluation of the biofilms. The combined characterisation data obtained from environmental scanning electron microscopy, confocal scanning laser microscopy, cyclic voltammetry and electrochemical impedance spectroscopy, confirm C-pSi as a powerful tool for sensing bacterial biofilm growth. The collective assessment of biofilm growth at the C-pSi surface reveals changes related both to morphological and temporal parameters.

RevDate: 2025-08-21

Flumignan VK, Sircili MP, Germano LG, et al (2025)

Characterization of Bacterial Biofilm Composition in Occluded Plastic Biliary Stents.

Clinical and experimental gastroenterology, 18:179-189.

PURPOSE: Plastic biliary stents are an effective treatment for biliary obstruction. Despite being resolutive and accessible, they are known to have a low patency rate, estimated at 3 to 6 months. This can be attributed to the formation of bacterial biofilm, which leads to the luminal obstruction of the stent. The aim of this study is to identify the bacterial composition of biofilms from obstructed plastic biliary stents removed through ERCP.

METHODS: Obstructed plastic biliary stents were retrieved from patients undergoing ERCP. The stents were fragmented into three segments of 2.0 cm each: proximal, medial, and distal. Gram staining was performed on each fragment, followed by assessment using optical microscopy. Subsequently, 4 µm cross-sections were made of each fragment, with subsequent analysis by confocal microscopy. The material from the inside of the stents was also placed in culture medium and colony-forming units were counted.

RESULTS: Optical microscopy and analysis by confocal microscopy showed a seemingly higher number of bacterial colonies in the distal portion of the stents compared to the proximal and medial regions. A greater presence of bacteria in the distal segments of the stents was confirmed, with growth reaching up to 10[14], while growth in the proximal and medial segments was only observed up to 10[9] and 10[8], respectively. Biochemical identification using Gram staining identified both Gram-positive and Gram-negative species: Enterococcus faecium; Aeromonas hydrophila/caviae; Escherichia coli; Enterobacter cloacae; Citrobacter freundii; Klebsiella oxytoca; Proteus vulgaris; Proteus mirabilis; Pantoea sp; Morganella morganii.

CONCLUSION: The composition of the biofilm in biliary stents confirmed to be polymicrobial. The distal portion of the stents is likely the most frequent site of obstruction. New strategies, such as the development and improvement of plastic stents, should be considered to slow this growth and enhance permeability.

RevDate: 2025-08-21

Smutkeeree A, Rattanapakdeekul N, Lapirattanakul J, et al (2025)

Cariogenic Potential of Lactose-Free Infant Formulas: An In Vitro Analysis of Streptococcus mutans Biofilm and Acidogenicity.

Journal of International Society of Preventive & Community Dentistry, 15(3):265-274.

AIM: Lactose-free infant formulas (LFs) are commonly used for children with lactose intolerance, but their potential contribution to early childhood caries (ECC) remains poorly understood, particularly with varying sugar compositions. The present study aimed to assess the biofilm-forming ability, acidogenicity, and structural biofilm characteristics of LFs with and without sucrose using a Streptococcus mutans in vitro model.

MATERIALS AND METHODS: Two LFs (with and without 24% sucrose) were tested alongside brain heart infusion (BHI) (negative control) and BHI + 10% sucrose (positive control). S. mutans biofilm formation was quantified using crystal violet staining (A590nm). Biofilm pH was measured after 24 h of incubation. Structural characteristics were assessed via confocal laser scanning microscopy (CLSM). Statistical analysis included Kruskal-Wallis tests, Bonferroni post hoc tests, and linear regression modeling (P < 0.05).

RESULTS: Both LF samples showed significantly greater biofilm formation than the negative control (A590nm: 3.57 ± 0.06 vs. 0.08 ± 0.02; P < 0.05). Biofilm pH was significantly lower in both LFs (4.23 and 4.28) than in BHI alone (5.26; P < 0.05). However, there were no significant differences between the LFs with and without sucrose. CLSM revealed denser S. mutans aggregation in sucrose-supplemented LF, but without distinct biofilm boundaries.

CONCLUSION: LFs, even those without supplemental sucrose, promote S. mutans biofilm formation and acidogenicity, suggesting a potential cariogenic risk. These findings underscore the need for careful dietary recommendations and oral hygiene strategies for infants consuming LF formulas. Further studies involving in vivo and clinical trials are warranted to validate the results and assess the long-term implications for ECC development.

RevDate: 2025-08-21

Whiteley A, Nonglaton G, V Jousseaume (2025)

Poly(VP-co-V3D3), an Insoluble Hydrophilic Copolymer Coating Deposited via Initiated Chemical Vapor Deposition to Prevent Bacterial Adhesion and Biofilm Formation.

ACS applied bio materials [Epub ahead of print].

While bacterial biofilm remains one of the world's major health issues, many traditional strategies for combatting it are being questioned. Indeed, as antibioresistance is rising and other biocides are raising concerns due to their potential toxicity, these curative methods no longer seem to be the solution. Therefore, biomimetic preventive methods, based on impeding bacterial adhesion, are being developed. One strategy is creating superhydrophilic surfaces to which bacteria cannot adhere as a water barrier forms at the interface. Polymer thin films are an efficient way for crafting such surfaces, and more precisely, the vacuum-based method initiated chemical vapor deposition (iCVD) allows the formation of polymers through radical polymerization mechanisms, therefore giving good control over the chemical composition of the resulting coatings. Polyvinylpyrrolidone (poly(VP)) has previously been used for its antibiofouling properties due to its high wettability and can be easily deposited into thin films by iCVD. However, its strong affinity with water makes it soluble, and thus, it cannot be used in bacterial suspensions. By coinjecting the VP precursor with a cross-linker, in this case organosilicate 1,3,5-trivinyl-1,3,5-trimethylcyclotrisiloxane (V3D3), the iCVD technique allows the formation of cross-linked copolymer thin films. The conservation of the functional groups of both monomers, whose proportion can be finely tuned by adjusting the injected fractions of each precursor, was studied by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). The Fineman-Ross equation was used to evaluate the reactivity of both monomers, allowing us to describe the copolymer structure. Water contact angle (WCA) and atomic force microscopy (AFM) measurements were carried out to characterize the surface state of the coatings and correlate it with the copolymer composition. Finally, solubility tests were carried out in phosphate buffer saline (PBS), and microbiological adhesion assays were carried out with the model species Escherichia coli to highlight the potential of these copolymers for antifouling applications.

RevDate: 2025-08-20

Abdelaziz HTO, Seif Mohamed EM, Younis SKA, et al (2025)

Selenium nanoparticle loaded on PVA/chitosan biofilm synthesized from orange peels: antimicrobial and antioxidant properties for plum preservation.

BMC chemistry, 19(1):245.

RevDate: 2025-08-20

Jang S, Im H, A Jang (2025)

Comparative evaluation of nitrogen removal in moving bed biofilm reactors (MBBRs) treating anaerobic digestion effluent: effect of external carbon source and biocarrier autopsy.

Bioresource technology pii:S0960-8524(25)01142-3 [Epub ahead of print].

This study investigated nitrogen removal performance in an anoxic/oxic moving bed biofilm reactor (A/O MBBR) using two external carbon sources: food waste condensate (FWC) and methanol (MeOH). FWC supported effective denitrification (95.4 ± 4.2 %) due to the enrichment of diverse heterotrophic denitrifiers, including Thauera, Paracoccus, and Azospira, despite its chemical complexity. However, nitrification efficiency in the FWC-fed system was lower than in the MeOH-fed system (5.5 % lower), likely due to inhibitory organic constituents and fewer nitrifying bacteria (Nitrosomonas and Nitrolancea). In contrast, the MeOH-fed system achieved higher nitrification efficiency (97.7 ± 2.9 %) and stable biofilm formation. Despite the limitations of MeOH's substrate diversity, FWC proved to be a viable, sustainable carbon source for nitrogen removal in A/O MBBR systems. Future research should focus on enhancing nitrification through microbial enrichment and process optimization to achieve balanced nitrogen removal under complex substrate conditions.

RevDate: 2025-08-20

Upadhyay A, Pal D, A Kumar (2025)

Development of an innovative method of Salmonella Typhi biofilm quantification using Tetrahydrofuran and Response Surface Methodology.

Microbial pathogenesis pii:S0882-4010(25)00717-X [Epub ahead of print].

Salmonella Typhi (S. Typhi), the causative agent of typhoid fever, poses a severe global health threat, particularly due to its ability to form robust biofilms that confer high levels of antibiotic resistance. Biofilms act as a physical and chemical barrier, protecting the bacterial community from hostile environments and therapeutic interventions, thereby complicating infection management. Accurate and reproducible quantification of S. Typhi biofilms is essential for understanding pathogenesis, evaluating anti-biofilm agents, and improving clinical outcomes. However, conventional biofilm quantification methods, such as crystal violet staining, suffer from limitations, including biofilm loss during washing steps, variability in adherence to surfaces, and inconsistent readings due to biofilm accumulation at the air-liquid interface. In this study, we systematically investigated the efficiency of six chemicals (five solvents and one dye), dimethyl sulfoxide (DMSO), methanol, ethanol, acetone, tetrahydrofuran (THF), and crystal violet, for their interaction with pre-formed S. Typhi biofilms in 96-microtiter well plates. Our goal was to identify an optimal solvent that can effectively solubilize biofilm while preserving structural integrity, thus enhancing measurement precision. We developed a new solvent-assisted method that significantly reduces washing steps to minimize biofilm detachment and sample variability. The process was statistically optimized and validated using response surface methodology (RSM), enabling the identification of THF as a highly effective solvent that maximizes optical density signal without compromising reproducibility. Our findings demonstrate that THF-based biofilm quantification provides superior accuracy, consistency, and ease of use compared to traditional approaches. Notably, THF was found to be the most effective biofilm-dissolving solvent among the six chemicals. This optimized method not only streamlines biofilm measurement workflows but also holds great potential for high-throughput screening of anti-biofilm compounds.

RevDate: 2025-08-20

Li J, Liu X, Zuo J, et al (2025)

Domain driven mining of high activity anti-biofilm glycoside hydrolases in pathogenic bacteria.

International journal of biological macromolecules pii:S0141-8130(25)07530-0 [Epub ahead of print].

Biofilm formation represents a critical antimicrobial resistance mechanism contributing to persistent chronic infections. Glycoside hydrolases encoded within the exopolysaccharide synthesis gene clusters of pathogenic bacteria play pivotal roles in biofilm formation and dispersal processes, making the mining of anti-biofilm glycoside hydrolases from pathogen genomes a promising therapeutic strategy. In this study, we established a systematic pipeline for screening anti-biofilm glycoside hydrolases from Klebsiella pneumoniae and Acinetobacter baumannii genomes. Among 13 candidate enzymes screened, 7 were successfully expressed in soluble form, with PgaBAb1 and PgaBKp demonstrating potent anti-biofilm activity against Staphylococcus epidermidis biofilms. Among them, PgaBAb1 exhibited the highest anti-biofilm activity, with EC50 values for the disruption and inhibition of S. epidermidis biofilms being 2.83 ± 0.65 nM and 4.53 ± 1.19 nM, respectively. Enzymatic characterization revealed optimal pH 7.0 activity and remarkable stability at 37 °C for both enzymes. Mechanistic investigations confirmed that PgaBAb1 and PgaBKp mediate biofilm dispersal through enzymatic degradation of dPNAG. Notably, enzymatic pretreatment with these glycoside hydrolases significantly enhanced methicillin susceptibility in S. epidermidis. Our findings highlight the untapped potential of pathogenic bacteria as sources of therapeutic glycoside hydrolases and provide novel insights for addressing biofilm-associated chronic infections.

RevDate: 2025-08-20

Ren X, Wang F, Zhao H, et al (2025)

Synergistic effect of biochar and Phanerochaete chrysosporium regulating biofilm to promote microplastics degradation during composting.

Journal of environmental management, 393:127016 pii:S0301-4797(25)02992-5 [Epub ahead of print].

This study aimed to explore the impact of sawdust biochar (T2), Phanerochaete chrysosporium (PC, T3), and their mixtures (T4) on the polyethylene microplastics (PE-MPs) degradation and related mechanisms during composting, the treatment without addition was regarded as control (T1). Results showed that compared with the control, adding biochar, PC and their mixture could reduce the PE-MPs abundance from 27,833 to 17,267-22600 items/kg, and the minimum value was observed in T4. Meanwhile, the pronounced surface cracks, highest carbon loss and carbonyl index observed in T4 also confirmed this result. Additionally, adding the mixture of biochar and PC optimized composting environments and selectively enriched bacteria on PE-MPs biofilms, especially NS9_marine_group, main contributor to the PE-MPs degradation with 0-200 μm and 1000-3000 μm. Furthermore, the application of combined amendments enhanced connectivity between PE-MPs and microorganisms, thus benefiting PE-MPs degradation. These findings provide new insights into MPs reduction in compost and a theoretical basis for the safe recycling of organic wastes.

RevDate: 2025-08-20

Li MT, Cui YW, Mi YN, et al (2025)

A novel cascade A/O biofilm reactor coupled with an iron autotrophic denitrification biofilter process for treating real municipal wastewater: Deep nutrient removal and ultra-low sludge production.

Journal of environmental management, 393:127001 pii:S0301-4797(25)02977-9 [Epub ahead of print].

Currently, wastewater treatment processes are confronted with the challenge of high nutrient discharge standards and the need for low residual sludge production. In this study, a novel cascade A/O biofilm coupled with an iron autotrophic denitrification biofilter process was proposed to resolve these issues. The process showed high nitrogen removal with no apparent observed suspended sludge when treating real municipal wastewater with low C/N ratio. The process characteristics were investigated under various reflux ratios of iron ions liquid from the iron autotrophic denitrification biofilter to the cascade A/O biofilm reactor. Four reflux ratios experiments were set up successively, enhancing nutrient removal. The process showed the maximum TN removal efficiency of 90.94 % ± 2.02 %, phosphate removal efficiency of 66.38 % ± 3.36 % and the sludge reduction ratio of 86.59 ± 3.15 %. The sludge reduction ratio was affected by reflux ratio, due to the toxic effect of iron ions on microfauna. Following the increase of reflux ratios, the relative abundance of ciliates and nematodes significantly improved in the biofilm; Telotrochidium and Diplogasterida gradually became the dominant microfauna in the aerobic cells, playing key roles in sludge reduction. This study proposes a novel process which could deeply remove nitrogen from the real municipal wastewater and produce ultra-low residual sludge, solving the challenges faced by existing biological treatment technology.

RevDate: 2025-08-20

Avgoulas DI, Festa D, Petala M, et al (2025)

Flow geometry effect on Pseudomonas fluorescens SBW25 biofilm structure.

Colloids and surfaces. B, Biointerfaces, 256(Pt 2):115048 pii:S0927-7765(25)00555-7 [Epub ahead of print].

This study investigated the impact of flow path geometry on Pseudomonas fluorescens SBW25 biofilm structure, on different surface materials. A custom- experimental setup featuring vertically oriented millimeter-scale-flow channels was employed to grow biofilms on test coupons made of stainless steel electropolished (SSEP) and Teflon fluoroethylenepropylene (FEP), at 72 h after flow onset under constant flow conditions. Each flow channel accommodated an upstream and a downstream coupon placed in a row. Two channel types were employed: (i) one with a straight flow path throughout, and (ii) one with an upstream straight section and a downstream square-wave (zig-zag) flow path. Biofilm thickness and structure were quantified using Optical Coherence Tomography (OCT) images analyzed by a novel, in-house Matlab software. Results demonstrated that SSEP consistently supported thicker and more uniform biofilms compared to FEP. In straight channels (type i), biofilms on SSEP reached mean thicknesses of approximately 29 ± 9 μm (upstream) and 46 ± 17 μm (downstream), while FEP showed thinner biofilms (19-20 μm, COV ≈ 47 %). In square-wave channels (type ii), thicker biofilms developed on the upstream surfaces, with thicknesses of 86 ± 14 μm (COV ≈ 17 %) for SSEP and 81 ± 34 μm (COV ≈ 42 %) for FEP, respectively. Furthermore, biofilms on SSEP increased further along the downstream zig-zag path, indicating enhanced accumulation due to flow geometry. This effect was absent on FEP, where detachment occurred in certain zig-zag sections. Overall, the findings emphasize the critical interplay between surface properties and flow dynamics in shaping biofilm structure.

RevDate: 2025-08-20
CmpDate: 2025-08-20

Ghimire N, Rayamajhi M, Sun Y, et al (2025)

Chitosan-coated titanium screws: Enhancing biofilm resistance, mechanical stability, and osseointegration in orthopedic implants.

Journal of applied biomaterials & functional materials, 23:22808000251358057.

Orthopedic implant-associated infections, primarily caused by biofilm-forming Staphylococcus aureus, pose significant clinical challenges. These infections often lead to implant failure, prolonged antibiotic treatments, and an increased risk of revision surgeries, emphasizing the need for effective biofilm-resistant implant materials. In this study, we present a dual-functional titanium screw (Ti-S) grafted with chitosan (Cs), a biocompatible polymer known for its osteogenic and antimicrobial properties while maintaining mechanical integrity. The chitosan-modified titanium screw (Cs-Ti-S) was prepared via chemical immobilization to enhance resistance to biofilm formation while promoting osseointegration and preserving biomechanical integrity. Biomechanical testing confirmed that chitosan modification did not compromise mechanical performance, as Cs-Ti-S exhibited a torsional yield strength of 1.70 ± 0.00 Nm compared to 1.76 ± 0.05 Nm for unmodified titanium screws (Un-Ti-S), and an axial pullout force of 68.66 ± 14.36 N for Cs-Ti-S versus 70.33 ± 9.71 N for Un-Ti-S. Micro-scratch tests revealed similar hardness values (1.26 ± 0.03 GPa for Cs-Ti-S vs. 1.40 ± 0.07 GPa for Un-Ti-S) and scratch resistance, ensuring surface durability. Gene expression analysis showed upregulated β1-integrin on Cs-Ti-S at 24 h post-infection, indicating improved osteoblast adhesion. Scanning electron microscopy (SEM) analysis confirmed significantly reduced bacterial biofilm formation on Cs-Ti-S. Moreover, the combination of povidone-iodide (PI) treatment on Cs-Ti-S surfaces significantly inhibited biofilm formation over 7 days, unlike Un-Ti-S, which retained significant adhesion. These results suggest chitosan grafting as a scalable, non-antibiotic strategy to enhance antimicrobial resistance and osseointegration.

RevDate: 2025-08-20

Thakur D, L Kumar (2025)

Biofilm-associated Escherichia coli infections: pathogenesis, clinical implications, and treatment strategies.

Critical reviews in microbiology [Epub ahead of print].

Biofilm formation is a complex process in which bacteria adhere to surfaces and create a protective matrix. Biofilms shield bacteria, such as Escherichia coli, from antibiotics and the host immune system, greatly facilitating their pathogenesis by enabling immune evasion and antimicrobial resistance. This review examines the stages of E. coli biofilm formation and their role in infections across various body sites, including the central nervous system, eyes, ears, teeth, respiratory tract, cardiovascular system, gastrointestinal tract, urinary tract, and medical device-related infections. Each infection site is thoroughly analyzed in terms of clinical manifestations, diagnostic challenges, treatment resistance, and implications for patient management. Furthermore, this review discusses therapeutic advancements, which are crucial for combating biofilm-associated infections. By unraveling the complexities of biofilms and developing novel therapeutics, researchers and clinicians can enhance strategies for diagnosing, treating, and preventing persistent E. coli infections.

RevDate: 2025-08-20

Abdullah EM, Elariny EYT, Abdelaziz R, et al (2025)

Lytic bacteriophage disrupts biofilm and inhibits growth of pan-drug-resistant Listeria monocytogenes in dairy products.

Frontiers in microbiology, 16:1653368.

Listeria monocytogenes is a major foodborne pathogen whose presence presents a continuous challenge in the food industry. A key issue is the formation of biofilms, which are complex microbial communities that cling to surfaces. These biofilms are incredibly resilient, making them tough to eliminate and manage. Therefore, it is crucial to find new and innovative ways to prevent and remove them. This study investigated the prevalence of L. monocytogenes in raw milk and Kareish cheese samples, as well as its resistance to antimicrobials and its ability to form biofilms. We also isolated and characterized a lytic bacteriophage to explore its anti-biofilm potential. Listeria species prevalence was 20% (n = 24/120 samples), higher in raw milk (31.7%) than Kareish cheese (8.3%). Eighteen isolates (15%) were identified as L. monocytogenes. High resistance rates were observed, notably to cefotaxime and cotrimoxazole. One pan-drug resistant (PDR) isolate was found in Kareish cheese, and the other 17 isolates were multidrug resistant (MDR). All L. monocytogenes isolates formed biofilms, categorized as weak: n = 7, moderate: n = 9, and strong: n = 2. We isolated a lytic bacteriophage, vB_LmoP_M15, which demonstrated lytic activity against all L. monocytogenes isolates, including both MDR and PDR strains. This phage belongs to the Podoviridae family, characterized by a short, non-contractile tail and an icosahedral head. Its genome size was estimated to be approximately 48.5 kb based on agarose gel electrophoresis of undigested phage DNA using a high molecular weight marker, and its restriction pattern was analyzed using HinfI, HindIII, and HaeIII enzymes. It has a latent period of 15 min and a burst size of 172 phage particles per infected cell. Phage vB_LmoP_M15 demonstrated significant antibiofilm activity (p < 0.05 to p < 0.0001). It effectively disrupted preformed biofilms and inhibited biofilm formation by MDR/PDR isolates. Application of vB_LmoP_M15 in pasteurized milk resulted in a significant reduction of L. monocytogenes counts by 2.45 log10 CFU/ml over 7 days at 30°C. These findings underscore the significant potential of phage vB_LmoP_M15 for controlling L. monocytogenes contamination and biofilms in dairy products.

RevDate: 2025-08-20
CmpDate: 2025-08-20

Saffari Natanzi A, Poudineh M, Karimi E, et al (2025)

Innovative approaches to combat antibiotic resistance: integrating CRISPR/Cas9 and nanoparticles against biofilm-driven infections.

BMC medicine, 23(1):486.

The increasing prevalence of antibiotic-resistant bacterial infections is a major global health concern, with biofilms playing a key role in bacterial persistence and resistance. Biofilms provide a protective matrix that limits antibiotic penetration, enhances horizontal gene transfer, and enables bacterial survival in hostile environments. Conventional antimicrobial therapies are often ineffective against biofilm-associated infections, necessitating the development of novel therapeutic strategies. The CRISPR/Cas9 gene-editing system has emerged as a revolutionary tool for precision genome modification, offering targeted disruption of antibiotic resistance genes, quorum sensing pathways, and biofilm-regulating factors. However, the clinical application of CRISPR-based antibacterials faces significant challenges, particularly in efficient delivery and stability within bacterial populations. Nanoparticles (NPs) present an innovative solution, serving as effective carriers for CRISPR/Cas9 components while exhibiting intrinsic antibacterial properties. Nanoparticles can enhance CRISPR delivery by improving cellular uptake, increasing target specificity, and ensuring controlled release within biofilm environments. Recent advances have demonstrated that liposomal CRISPR-Cas9 formulations can reduce Pseudomonas aeruginosa biofilm biomass by over 90% in vitro, while gold nanoparticle carriers enhance editing efficiency up to 3.5-fold compared to non-carrier systems. These hybrid platforms also enable co-delivery with antibiotics, producing synergistic antibacterial effects and superior biofilm disruption. Additionally, they can facilitate co-delivery of antibiotics or antimicrobial peptides, further enhancing therapeutic efficacy. This review explores the synergistic integration of CRISPR/Cas9 and nanoparticles in combating biofilm-associated antibiotic resistance. We discuss the mechanisms of action, recent advancements, and current challenges in translating this approach into clinical practice. While CRISPR-nanoparticle hybrid systems hold immense potential for next-generation precision antimicrobial therapies, further research is required to optimize delivery platforms, minimize off-target effects, and assess long-term safety. Understanding and overcoming these challenges will be critical for developing effective biofilm-targeted antibacterial strategies.

RevDate: 2025-08-19

Yao L, Ma Y, Liu R, et al (2025)

LIPUS-responsive meropenem-loaded nanobubbles enable biofilm disruption and bone repair in orthopedic implant-associated infections.

Journal of controlled release : official journal of the Controlled Release Society pii:S0168-3659(25)00730-8 [Epub ahead of print].

Orthopedic implant-associated infections (OII), often driven by biofilm formation, pose persistent challenges to infection control and bone healing. This study introduces a dual-action therapeutic strategy employing low-intensity pulsed ultrasound (LIPUS)-responsive meropenem-loaded nanobubbles (NBs@MER) to simultaneously eradicate Escherichia coli biofilms and promote bone repair. In vitro, LIPUS activation of NBs@MER exhibited robust cavitation effects, which disrupted biofilm structure, enhanced antibiotic penetration, and improved antimicrobial efficacy. Concurrently, LIPUS stimulation modulated the immune microenvironment by polarizing macrophages toward the reparative M2 phenotype and enhancing osteogenic differentiation of bone marrow mesenchymal stem cells. In vivo studies using a rat OII model confirmed the dual functionality of the system, demonstrating marked reductions in bacterial burden and inflammation, alongside enhanced bone mineral density and trabecular architecture around infected implants. Collectively, this integrated strategy offers a promising platform for addressing the dual challenge of infection clearance and bone repair in OII management.

RevDate: 2025-08-19

Mendoza-Barrón DE, Hernández-Iturriaga M, A Godínez-Oviedo (2025)

Variability in biofilm formation dynamics by Salmonella enterica isolated from animal-origin foods, plant-based foods, environment, clinical, and unspecified food sources: A three-day in vitro study in tryptic soy broth at ambient temperature.

Canadian journal of microbiology [Epub ahead of print].

Bacterial biofilm production is linked to its adaptive capacity to environments throughout its lifecycle. This study aimed to assess the variability in biofilm formation (BF) dynamic by Salmonella enterica and to explore the potential impact of the cell's prior history, primarily shaped by strain and its isolation source. In vitro BF of 141 S. enterica strains isolated from animal-origin foods, plant-based foods, unspecified food sources, the environment, and clinical cases, was evaluated using the crystal violet assay at 25°C for up to 72 h. Kruskal-Wallis test were used to assesses the effect of time, source, and strain. The Aryani method was used to characterize microbial response variability. The BF capacity of S. enterica strains ranged from 0.07 to 2.3, 0.07 to 2.7, and 0.06 to 2.7OD595nm at 24, 48, and 72 h, respectively. At 24 hours (66.0%; 93/141) and 48 hours (56.0%; 79/141), most isolates were classified as non-biofilm producers, while at 72 hours, the majority were weak biofilm producers (39.7%; 56/141), Time, strain, and isolation source significantly influenced BF, with an overall increase in BF occurring over time, and clinical strains being the highest biofilm producers. ...

RevDate: 2025-08-19

Di Marco NI, Iriarte HJ, Colocho FA, et al (2025)

Biofilm formation of Yersinia enterocolitica and its response against GRAS compounds in meat juice.

Letters in applied microbiology pii:8238062 [Epub ahead of print].

Yersinia enterocolitica is a food-borne pathogen that causes yersiniosis, which primary sources are animal-originated foods. This work aimed to analyze the effect of fresh pork meat juice (MJ) on planktonic and biofilm growth of Y. enterocolitica strains and to investigate the activity of some GRAS food preservatives. Twenty-eight Y. enterocolitica strains were used to test growth and biofilm formation in MJ, trypticase soy broth supplemented with 0.25% of glucose (TSBG) alone and in combination with 50% MJ (TSBG: MJ). All strains grew in MJ but most of them to lesser extent than in TSBG. Although TSBG: MJ was the most favorable medium for biofilm formation, many strains were able to form biofilm in MJ. To determine GRAS compounds activity, one B1A and one B1B Y. enterocolitica strain were selected. In nutrient broth with MJ, acetic acid was the most effective compound with MIC of 0.78 mg ml-1, MBC of 3.14 mg ml-1 and BIC of 1.57 mg ml-1 for both strains; for B1A strain, BBC was 12.56 mg ml-1 and for B1B strain, it was 25.12 mg ml-1. Although not all strains exhibited the same ability to form biofilms in MJ, it reduces the susceptibility of Y. enterocolitica to GRAS compounds.

RevDate: 2025-08-19

Wimmer A, Jernej L, Liu J, et al (2025)

Chlorophyllin-based Photodynamic Inactivation against Candidozyma auris planktonic cells and dynamic biofilm.

Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology [Epub ahead of print].

Candidozyma auris likely gained thermotolerance as a result of climate change and emerged from its origin in Indian wetlands to a severe healthcare threat because of its resistance to common antifungals and antiseptics in only a few decades. This development identifies the yeast as a perfect example for the relevance of the One Health concept for human well-being. Here, we compare the effectiveness of Photodynamic Inactivation (PDI) based on the economic and ecofriendly natural photosensitizer sodium-magnesium-chlorophyllin (Chl) to that of a synthetic chlorin e6 derivative carrying cationic moieties, B17-0024, suggesting both as alternative to common disinfectants against C. auris. Experiments were conducted on planktonic cells and-for the very first time-on dynamic biofilms using the CDC bioreactor. Treatment of planktonic cells with 50 µM Chl and blue light (395 nm, 7.5 J cm[-2], 15 min drug to light interval) achieved a 7 log10 step reduction of viable C. auris. B17-0024 induced a more than 5 log10 step photokilling at 10 µM. Illumination of the same concentrations with red light (600-700 nm, 30 J cm[-2]) resulted in a relative inactivation of 7 log10 steps for Chl and 6 log10 steps for B17-0024. Dynamic biofilm samples were illuminated with 3.33-times higher radiant exposure (25 J cm[-2] at 395 nm or 100 J cm[-2] at 600-700 nm). The antimicrobial effect of a 99.9% reduction of C. auris was exceeded with 10 µM B17-0024 and blue light illumination and with 50 µM Chl and B17-0024 activated by red light. Biofilms were completely eradicated when doubling the photosensitizer concentrations. Our results demonstrate that PDI based on Chl represents a rapid and effective tool to eliminate emerging pathogens even if resistant to conventional treatment. Due to its low costs and eco-friendliness PDI based on Chl may be applicable for disinfection of larger areas in hospitals.

RevDate: 2025-08-19

Põhako-Palu K, Preem L, Randmäe K, et al (2025)

Development of Rapid and Economic In Vitro Assay and Biorelevant Ex Vivo Biofilm Inhibition Wound Model to Test the Antibacterial Efficacy of Wound Dressings.

Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society, 33(4):e70080.

Chronic wounds are a major healthcare problem, consuming resources globally and necessitating innovative wound dressing development. All antimicrobial wound dressings must be tested for safety and antibacterial effectiveness prior to patient use. This study aimed to develop a rapid, economical in vitro assay and biorelevant ex vivo wound biofilm model on porcine skin to test the antibacterial efficacy of antimicrobial wound dressings. The methods were validated using five commercially available wound dressings and experimental electrospun (ES) wound dressing containing chloramphenicol in polycaprolactone and polyethylene oxide fibres (PCL/PEO/CAM). An in vitro assay was used to assess the growth inhibition, killing efficacy, and dressing sterility against multiple bacterial strains and inoculum sizes. Ex vivo models using porcine skin were used to evaluate biofilm inhibition with dressings on top of or inside infected wounds. The in vitro assay allowed rapid initial screening, whilst ex vivo models provided more biorelevant conditions for understanding the efficacy in wound-mimicking environments. The assay and model are suitable for rapid evaluation of antimicrobial efficacy before animal studies and clinical trials. Using various commercially available wound dressings alongside novel dressings for validation ensures that the method is broadly applicable. The antibacterial efficacy of commercial antimicrobial wound dressings and experimental ES PCL/PEO/CAM fibre mat was confirmed. This study highlights the importance of using multiple complementary assays and models to comprehensively assess antimicrobial wound dressing materials.

RevDate: 2025-08-19

Namba-Koide N, Yoshida Y, Nagaoka N, et al (2025)

Phosphorylated pullulan as a local drug delivery matrix for cationic antibacterial chemicals to prevent oral biofilm.

BMC oral health, 25(1):1333.

BACKGROUND: Preventing oral infections, such as oral caries and periodontal disease, helps reduce the risks of various systemic diseases. In this study, the polysaccharide pullulan produced by the black yeast Aureobasidium pullulans was modified in combination with the cationic surfactant cetylpyridinium chloride (CPC) to create a local drug delivery system, and its antibacterial potential on oral bacteria was examined in vitro.

METHODS: Pullulan was phosphorylated at the CH2OH residue of α6 in the maltotriose structure and mixed with CPC. Bacterial attachment of cariogenic Streptococcus mutans on hydroxyapatite plates (HAPs) treated with the phosphorylated pullulan (PP) and CPC compound (0.01% PP and 0.001- 0.03% CPC, and vice versa) was assessed by observing bacteria using a field emission scanning electron microscope (FE-SEM) and quantified through 16 S rRNA amplification via real-time polymerase chain reaction (PCR). Additionally, the quartz crystal microbalance (QCM) method was employed to evaluate the sustained release of CPC.

RESULTS: PP-CPC compound maintained significant bactericidal activity even at 0.01%, which is one-fifth of the conventional applicable concentration of CPC. Additionally, a residual mixture was detected by the hydroxyapatite sensor of the crystal oscillator microbalance detector, suggesting an unknown molecular interaction that enables the sustained release of CPC after attachment to hydroxyapatite.

CONCLUSIONS: The combination of PP and CPC may contribute to the low concentration and effective prevention of oral infections, such as dental caries.

RevDate: 2025-08-17

Valdivia-Tapia AC, Lippert F, Castelluccio PF, et al (2025)

In vitro effect of fluoride-free mouthwashes on Streptococcus mutans biofilm.

Clinical oral investigations, 29(9):413.

OBJECTIVE: To evaluate the efficacy of commercially available, fluoride-free mouthwashes sold in Indianapolis, IN, on Streptococcus mutans biofilm.

MATERIALS AND METHODS: Eighty-one different mouthwashes were purchased. A 16-h culture of S. mutans UA159 was treated with the mouthwashes in three dilutions (1:3, 1:6, and 1:12), prepared in Tryptic Soy broth supplemented with 1% sucrose. The minimum inhibitory concentrations (MIC), planktonic, and biofilm growth were evaluated using a spectrophotometer. In addition, the growth for minimum bactericidal concentration (MBC) was evaluated using five μL of the dilution and incubated on blood agar. For the analysis of the results, the mouthwashes were separated into six groups according to their active ingredients (cetylpyridinium chloride/CPC, n = 25; essential oils/EO n = 10; whitening/W (hydrogen peroxide/ sodium hexametaphosphate), n = 12; Natural-Derived Actives / NDA, n = 15; zinc chloride/ZC, n = 3; others/O, n = 16). ANOVA followed by the Tukey test was performed (p < 0.05).

RESULTS: Regarding MIC, planktonic, and biofilm growth of S. mutans, there was a significant decrease for the W and CPC groups (p < 0.001). The EO and W groups had more inhibition on S. mutans biofilm compared to the CPC group (p < 0.05). For ZC, NDA, and O groups, there were different effects within the same group, presenting a large variability. About MBC, W and CPC groups presented the higher inhibition (W > CPC > EO > NDA/ZC/O).

CONCLUSION: The mouthwashes demonstrated significant effect on S. mutans biofilm, especially in the 1:3 dilution. W and CPC groups had a more significant effect on S. mutans biofilm.

CLINICAL RELEVANCE: S. mutans is an important bacterium in dental caries and periodontal diseases. Our study showed that non-fluoridated mouthwashes affect the initial stages of biofilm formation.

RevDate: 2025-07-18

Pudipeddi A, Bijle MN, C Yiu (2025)

Effect of arginine-based synbiotics on multispecies biofilm.

Journal of dentistry, 161:105974 pii:S0300-5712(25)00418-X [Epub ahead of print].

OBJECTIVE: To examine the effect of arginine (Arg)-based synbiotics on multispecies biofilm.

METHODS: In vitro biofilms (Streptococcus mutans UA159, S. gordonii DL1, S. sanguinis DSS-10) were grown on HA discs under anaerobic conditions (37°C, 5 % CO₂, 24 h) and treated with: (1) Arg (0.25 %, 0.5 % w/v.), (2) Lacticaseibacillus rhamnosus GG (LRG) at 10⁷ CFU/mL, or (3) their combinations, 2× daily for 3 days. At 96 h, biofilm matrix components (exoproteins, eDNA, and carbohydrates) and microbial viability (confocal microscopy and PMA-qPCR) were quantified. Relative gene expression analysis was also conducted with species-specific genes (gtfB, sagP, arcA, argG, argH).

RESULTS: For carbohydrates, no significant difference was identified among treatment groups (p>0.05). Protein content for 0.5 % Arg+LRG was significantly lower than LRG (p<0.05). The lowest eDNA content was observed in LRG (p<0.05); while eDNA content of 0.5 % Arg+LRG, was similar to 0.5 % Arg and control (p>0.05). Using confocal imaging, the highest proportion of live cell was found in 0.5 % Arg+LRG (p<0.05), followed by 0.25 % Arg+LRG, 3-D biofilm imaging demonstrated increased biomass with smoother architecture in biofilms treated with 0.5 % Arg+LRG (p<0.05). 0.5 % Arg+LRG significantly enhanced growth of commensal streptococci (S. gordonii, S. sanguinis) compared to monotherapies and control, while also reducing viability of S. mutans compared to control (p<0.05). Gene expression analysis revealed downregulation of S. mutans virulence (gtfB) and upregulation of commensal metabolism (sagP, arcA) for 0.5 % Arg+LRG (p<0.05).

CONCLUSION: The 0.5 % Arg+LRG synbiotics uniquely integrates ecological modulation by regulating biochemical matrix components, promoting commensal enrichment and suppressing cariogenic pathogens.

CLINICAL SIGNIFICANCE: A deliverable Arg-LRG synbiotics for caries prevention addresses a global public health priority. This strategy aligns with microbial homeostasis principles, presenting a novel paradigm for caries prevention. The Arg-LRG synbiotics can counter the limitations of fluorides to establish a diverse oral microbiome, imparting an ecologically driven approach to caries prevention.

RevDate: 2025-08-12
CmpDate: 2025-08-12

Dinelli RG, Shibli JA, Tolentino PHMP, et al (2025)

5-Aminolevulinic Acid Gel Associated with Light-Emitting Diode Modulates the in Vitro Subgingival Multispecies Oral Biofilm.

Photobiomodulation, photomedicine, and laser surgery, 43(8):373-378.

Objective: This study assessed the impact of a 5% combination of 5-aminolevulinic acid (5-ALA) (Aladent) in a multispecies in vitro biofilm model subgingival pathogens. Methods: The 33-species biofilm model was established in the Calgary Biofilm Device during a duration of 7 days. The biofilm treatments comprised various groups: control, light-emitting diode (LED), Aladent (ALADA), and Aladent with LED (ALAD+L), administered on day 6. The Aladent was in contact with the biofilm for 45 min before to the 7-min LED treatment. The LED (λ = 630 nm, power ≈ 380 mW/cm[2]) was placed 2 mm from the biofilm. Subsequently, during 7 days of biofilm formation, the metabolic activity of the biofilms was assessed utilizing triphenyltetrazolium chloride, and the presence of 33 bacterial species was evaluated through DNA-DNA hybridization. Results: The findings indicated that the ALAD+L treatment was the sole intervention demonstrating a statistically significant reduction (∼70%) in the metabolic activity of the biofilms relative to the control group. Moreover, ALAD+L markedly diminished the overall biofilm count and the average counts of five bacterial species: S. intermedius, V. parvula, A. israelii, P. gingivalis, and E. saburreum. Conclusion: The integration of the photosensitizer Aladent with LED application significantly diminished metabolic activity and bacterial species count in the multispecies subgingival biofilm model, indicating substantial promise for the treatment of peri-implantitis.

RevDate: 2024-09-14
CmpDate: 2024-09-10

de Sousa DV, Maia PVS, Eltink E, et al (2024)

Biomolecules in Pleistocene fossils from tropical cave indicate fossil biofilm.

Scientific reports, 14(1):21071.

Finding biomolecules in fossils is a challenging task due to their degradation over time from physical, chemical, and biological factors. The primary hypothesis for explaining the presence of biomolecules in fossilized bones tissues suggests their survival in the fossilization process. In contrast, some of these biomolecules could either derive from bacteria biofilm, thus without a direct relationship with the fossil record or could be an artifact from measurement procedures. Raman spectroscopy studies across various fossil ages and environments have detected multiple bands ranging from 1200 to 1800 cm[-1] associative of organic compounds. However, the significance of these bands remains elusive. Our research aims to address this issue through a deep Raman spectroscopy investigation on Pleistocene teeth from Tayassu and Smilodon populator. These fossils were obtained from a well-preserved stratigraphic succession in Toca de Cima do Pilão cave, near the National Park of Serra da Capivara in semiarid Brazil. We propose two hypotheses to explain the presence of organic compounds related to 1200 to 1800 cm[-1] Raman spectral range in fossil tissues: (i) these bands are biological signatures of preserved fossil biomolecules, or (ii) they are exogenous biological signatures associated with the bacterial biofilm formation during post-depositional processes. Our results align with the latter hypothesis, followed by biofilm degradation. However, the specific mechanisms involved in the natural biofilm degradation in fossil records remain unexplored in this study. In our case, the formation of biofilm on fossil bones is attributed to the oligotrophic conditions of the cave sediment matrix. We present a comprehensive model to elucidate the existence of biofilm on fossilized tissues, emphasizing the pivotal role of post-depositional processes, especially water action, in the cave environment. As the fossils were discovered in a cave setting, post-depositional processes significantly contribute to the formation of the biofilm matrix. Although our study provides insights into biofilm formation, further research is needed to delve into the specific mechanisms driving natural biofilm degradation in fossils.

RevDate: 2024-04-26
CmpDate: 2024-04-24

Li C, Teng F, Wu F, et al (2024)

Enhanced cavitation dose and reactive oxygen species production in microbubble-mediated sonodynamic therapy for inhibition of Escherichia coli and biofilm.

Ultrasonics sonochemistry, 105:106853.

Sonodynamic therapy (SDT) is an emerging antibacterial therapy. This work selected hematoporphyrin monomethyl ether (HMME) as the sonosensitizer, and studied the enhanced inhibition effect of Escherichia coli and biofilm by microbubble-mediated cavitation in SDT. Firstly, the influence of microbubble-mediated cavitation effect on different concentrations of HMME (10 µg/ml, 30 µg/ml, 50 µg/ml) was studied. Using 1,3-diphenylisobenzofuran (DPBF) as an indicator, the effect of microbubble-mediated cavitation on the production of reactive oxygen species (ROS) was studied by absorption spectroscopy. Secondly, using agar medium, laser confocal microscopy and scanning electron microscopy, the effect of microbubble-mediated cavitation on the activity and morphology of bacteria was studied. Finally, the inhibitory effect of cavitation combined with SDT on biofilm was evaluated by laser confocal microscopy. The research results indicate that: (1) Microbubble-mediated ultrasound cavitation can significantly increase cavitation intensity and production of ROS. (2) Microbubble-mediated acoustic cavitation can alter the morphological structure of bacteria. (3) It can significantly enhance the inhibition of SDT on the activity of Escherichia coli and its biofilm. Compared with the control group, the addition of microbubbles resulted in an increase in the number of dead bacteria by 61.7 %, 71.6 %, and 76.2 %, respectively. The fluorescence intensity of the biofilm decreased by 27.1 %, 80.3 %, and 98.2 %, respectively. On the basis of adding microbubbles to ensure antibacterial and biofilm inhibition effects, this work studied the influence of cavitation effect in SDT on bacterial structure, providing a foundation for further revealing the intrinsic mechanism of SDT.

RevDate: 2025-04-28
CmpDate: 2025-04-28

Aragão MGB, Tedesco AC, Borges HS, et al (2025)

Chitosan nanoparticles loaded with epigallocatechin-3-gallate: synthesis, characterisation, and effects against Streptococcus mutans biofilm.

Natural product research, 39(9):2550-2557.

This study evaluated the effects of chitosan nanoparticles loaded with epigallocatechin-3-gallate (EGCG) against Streptococcus mutans biofilm. EGCG-loaded chitosan (Nchi + EGCG) nanoparticles and Chitosan (Nchi) nanoparticles were prepared by ion gelation process and characterised regarding particle size, polydispersion index, zeta potential, and accelerated stability. S mutans biofilms were treated twice daily with NaCl 0.9% (negative control), Nchi, Nchi + EGCG, and chlorhexidine (CHX) 0.12% (positive control). After 67 h, the biofilms were evaluated for acidogenesis, bacterial viability and dry weight. Biofilm morphology and structure were analysed by scanning electron microscopy. The nanoformulations presented medium to short-term stability, size of 500 nm, and polydispersion index around 0.400. Treatments affected cell morphology and biofilm structure. However, no effects on microbial viability, biofilm dry weight, and acidogenesis were observed. Thus, the nanoformulations disassembled the biofilm matrix without affecting microbial viability, which makes them promising candidates for the development of dental caries preventive and therapeutic agents.

RevDate: 2023-11-08
CmpDate: 2022-02-28

Baliarda A, Winkler M, Tournier L, et al (2021)

Dynamic interspecies interactions and robustness in a four-species model biofilm.

MicrobiologyOpen, 10(6):e1254.

Interspecific interactions within biofilms determine relative species abundance, growth dynamics, community resilience, and success or failure of invasion by an extraneous organism. However, deciphering interspecific interactions and assessing their contribution to biofilm properties and function remain a challenge. Here, we describe the constitution of a model biofilm composed of four bacterial species belonging to four different genera (Rhodocyclus sp., Pseudomonas fluorescens, Kocuria varians, and Bacillus cereus), derived from a biofilm isolated from an industrial milk pasteurization unit. We demonstrate that the growth dynamics and equilibrium composition of this biofilm are highly reproducible. Based on its equilibrium composition, we show that the establishment of this four-species biofilm is highly robust against initial, transient perturbations but less so towards continuous perturbations. By comparing biofilms formed from different numbers and combinations of the constituent species and by fitting a growth model to the experimental data, we reveal a network of dynamic, positive, and negative interactions that determine the final composition of the biofilm. Furthermore, we reveal that the molecular determinant of one negative interaction is the thiocillin I synthesized by the B. cereus strain, and demonstrate its importance for species distribution and its impact on robustness by mutational analysis of the biofilm ecosystem.

RevDate: 2024-04-04
CmpDate: 2019-02-08

Melo MAS, Weir MD, Passos VF, et al (2018)

Human In Situ Study of the effect of Bis(2-Methacryloyloxyethyl) Dimethylammonium Bromide Immobilized in Dental Composite on Controlling Mature Cariogenic Biofilm.

International journal of molecular sciences, 19(11):.

Cariogenic oral biofilms cause recurrent dental caries around composite restorations, resulting in unprosperous oral health and expensive restorative treatment. Quaternary ammonium monomers that can be copolymerized with dental resin systems have been explored for the modulation of dental plaque biofilm growth over dental composite surfaces. Here, for the first time, we investigated the effect of bis(2-methacryloyloxyethyl) dimethylammonium bromide (QADM) on human overlying mature oral biofilms grown intra-orally in human participants for 7[-]14 days. Seventeen volunteers wore palatal devices containing composite specimens containing 10% by mass of QADM or a control composite without QADM. After 7 and 14 days, the adherent biofilms were collected to determine bacterial counts via colony-forming unit (CFU) counts. Biofilm viability, chronological changes, and percentage coverage were also determined through live/dead staining. QADM composites caused a significant inhibition of Streptococcus mutans biofilm formation for up to seven days. No difference in the CFU values were found for the 14-day period. Our findings suggest that: (1) QADM composites were successful in inhibiting 1[-]3-day biofilms in the oral environment in vivo; (2) QADM significantly reduced the portion of the S. mutans group; and (3) stronger antibiofilm activity is required for the control of mature long-term cariogenic biofilms. Contact-killing strategies using dental materials aimed at preventing or at least reducing high numbers of cariogenic bacteria seem to be a promising approach in patients at high risk of the recurrence of dental caries around composites.

RevDate: 2018-11-13
CmpDate: 2016-12-13

Charnock C, AL Nordlie (2016)

Proteobacteria, extremophiles and unassigned species dominate in a tape-like showerhead biofilm.

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology], 47(2):345-351.

The development of showerhead biofilms exposes the user to repeated contact with potentially pathogenic microbes, yet we know relatively little about the content of these aggregates. The aim of the present study was to examine the microbial content of tape-like films found protruding from a domestic showerhead. Culturing showed that the films were dominated by aerobic α- and β-proteobacteria. Three isolates made up almost the entire plate count. These were a Brevundimonas species, a metalophilic Cupriavidus species and a thermophile, Geobacillus species. Furthermore, it was shown that the Cupriavidus isolate alone had a high capacity for biofilm formation and thus might be the initiator of biofilm production. A clone library revealed the same general composition. However, half of the 70 clones analyzed could not be assigned to a particular bacterial phylum and of these 29 differed from one another by only 1-2 base pairs, indicating a single species. Thus both the culture dependent and culture independent characterizations suggest a simple yet novel composition. The work is important as the biofilm is fundamentally different in form (tape-like) and content from that of all previously reported ones, where variously Mycobacterium, Methylobacterium and Xanthomonas species have dominated, and extremophiles were not reported.

RevDate: 2022-03-16
CmpDate: 2015-07-20

Prada-López I, Quintas V, Casares-De-Cal MA, et al (2015)

Ex vivo vs. in vivo antibacterial activity of two antiseptics on oral biofilm.

Frontiers in microbiology, 6:655.

AIM: To compare the immediate antibacterial effect of two application methods (passive immersion and active mouthwash) of two antiseptic solutions on the in situ oral biofilm.

MATERIAL AND METHODS: A randomized observer-masked crossover study was conducted. Fifteen healthy volunteers wore a specific intraoral device for 48 h to form a biofilm in three glass disks. One of these disks was used as a baseline; another one was immersed in a solution of 0.2% Chlorhexidine (0.2% CHX), remaining the third in the device, placed in the oral cavity, during the 0.2% CHX mouthwash application. After a 2-weeks washout period, the protocol was repeated using a solution of Essential Oils (EO). Samples were analyzed for bacterial viability with the confocal laser scanning microscope after previous staining with LIVE/DEAD® BacLight™.

RESULTS: The EO showed a better antibacterial effect compared to the 0.2% CHX after the mouthwash application (% of bacterial viability = 1.16 ± 1.00% vs. 5.08 ± 5.79%, respectively), and was more effective in all layers (p < 0.05). In the immersion, both antiseptics were significantly less effective (% of bacterial viability = 26.93 ± 13.11%, EO vs. 15.17 ± 6.14%, 0.2% CHX); in the case of EO immersion, there were no significant changes in the bacterial viability of the deepest layer in comparison with the baseline.

CONCLUSIONS: The method of application conditioned the antibacterial activity of the 0.2% CHX and EO solutions on the in situ oral biofilm. The in vivo active mouthwash was more effective than the ex vivo passive immersion in both antiseptic solutions. There was more penetration of the antiseptic inside the biofilm with an active mouthwash, especially with the EO. Trial registered in clinicaltrials.gov with the number NCT02267239. URL: https://clinicaltrials.gov/ct2/show/NCT02267239.

RevDate: 2019-11-10
CmpDate: 2008-04-09

Bercy P, J Lasserre (2007)

Susceptibility to various oral antiseptics of Porphyromonas gingivalis W83 within a biofilm.

Advances in therapy, 24(6):1181-1191.

The origin of chronic periodontal disease is strongly related to the nature and physiology of the subgingival bacterial biofilm, of which Porphyromonas gingivalis is a main protagonist. This study was conducted in vitro, to test the susceptibility of the W83 strain of P gingivalis to several oral antiseptics, bearing in mind its mode of growth as biofilm. To this end, the investigators inoculated a brain-heart infusion broth with Streptococcus gordonii, to which P gingivalis was added before perfusion for 7 d via a closed circuit containing a modified Robbins device. Then, various antiseptics were perfused through the circuit over 30 min, and their bactericidal effects were evaluated after culture by comparison of the mean proportion of bacteria killed. The average proportion of P gingivalis W83 killed after 15 min of contact with antiseptics was 90.51% (+/-4.78) for chlorhexidine 0.2%, 89.87% (+/-6.58) for povidone-iodine 1%, and 98.6% (+/-0.86) for Listerine (McNeil-PPC, Inc., Morris Plains, NJ). After 30 min of contact, survival of P gingivalis was nil, irrespective of the antimicrobial agent used. Preliminary results show that antiseptic mouth rinses, when used in pure concentrations in the traditional way, are effective in killing P gingivalis W83 within a biofilm. Furthermore, in light of these experiments, it appears that the best results are obtained when Listerine is used; however, new trials should be conducted to confirm this observation.

RevDate: 2025-08-18

Gandra RM, Giovanini L, Branquinha MH, et al (2025)

Secretion of serine proteases by planktonic- and biofilm-growing cells of Candida parapsilosis.

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] [Epub ahead of print].

Candida parapsilosis is a common cause of candidiasis worldwide, with biofilm formation and secretion of aspartic proteases (Saps) as key virulence factors. Conversely, serine protease secretion by this fungus is poorly understood. In this study, we investigated the secretion of serine-type proteases by planktonic- and biofilm-forming cells of C. parapsilosis cultured in brain heart infusion (BHI) medium. Cell-free supernatant from the reference strain (ATCC 22019) was screened against various serine protease substrates, revealing pronounced activity toward N-benzoyl-Phe-Val-Arg-pNa (0.74 nmol pNA.mg[- 1].min[- 1]), with optimal activity at pH 9.0 and temperatures between 32 °C and 40 °C. Proteolytic activity was significantly reduced by serine protease inhibitors PMSF (32.8%), TLCK (40.2%) and benzamidine (50.7%), while inhibitors of other protease classes had no effect, confirming its serine-type specificity. Notably, serine protease activity was detected in supernatants from cells grown in BHI but absent in those cultured in albumin-supplemented yeast carbon base medium, a known inducer of Saps, suggesting culture-dependent regulation of protease expression. Serine protease activity also increased over time, rising from 0.36 pNA.mg[- 1].min[- 1] at 24-hour to 1.14 pNA.mg[- 1].min[- 1] at 72-hour. Clinical isolates of C. parapsilosis exhibited significantly higher serine protease activity than the reference strain under optimal conditions. Serine-type protease activity was also detected in the supernatant of mature biofilms, showing a correlation with metabolic activity and biomass. Infection of Galleria mellonella larvae with C. parapsilosis isolates revealed no correlation between larval mortality and serine protease production. These findings suggest that C. parapsilosis serine proteases contribute to fungal growth and biofilm development, representing potential targets for antifungal intervention.

RevDate: 2025-08-18

Al-Monofy KB, Abdelaziz AA, Abo-Kamar AM, et al (2025)

Coating silicon catheters with the optimized and stable carotenoid bioproduct from Micrococcus luteus inhibited the biofilm formation by multidrug-resistant Enterococcus faecalis via downregulation of GelE gene expression.

Microbial cell factories, 24(1):186.

BACKGROUND: Microbial carotenoids have gained industry interest due to their safety and diverse biological activities; however, the low yield of carotenoids hinders their applications. Hence, this study focused on optimizing carotenoid pigment production from Micrococcus luteus strains by studying 54 physical and chemical independent conditions. The chronic infections by Enterococcus faecalis are related to its ability to form biofilms on the surface of several implanted medical devices, such as urinary catheters. Therefore, the potential antibacterial and antibiofilm activities of the purified pigment against E. faecalis were investigated in our study.

RESULTS: Using one-factor-at-a-time experiments, the top-examined parameters were tryptic soya broth (TSB), agitation, temperature, pH, incubation time, inoculum size (IS), sodium chloride, tryptophan, glycerol, tryptone, glutaric acid, toluene, ferric sulphate, and disodium hydrogen phosphate. The data from the Plackett-Burman design showed that temperature, sodium chloride, tryptophan, and toluene were fundamental factors controlling carotenoid production. The conditions for the upstream process were determined via response surface methodology design, which included TSB medium, agitation speed of 120 rpm, temperature of 32.5 °C, pH = 7, incubation time of 96 h, 2% IS, sodium chloride (12.5 g/l), tryptophan (12.5 g/l), glutaric acid (5 g/l), toluene (12.5%), and disodium hydrogen phosphate (5 g/l). Submerged fermentation model validation using the M6 isolate (accession number of PP197163) revealed an increase in carotenoid production up to 6-fold (1.2 g/l). The produced pigment was purified and characterized as β-carotene, and the stability study showed that the extracted β-carotene was stable for a year in dimethyl sulfoxide at 4 °C. The MTT test data proved that the pigment was safe on human dermal fibroblasts with an IC50 equal to 542.7 µg/ml. For the first time, it was reported that the stable purified β-carotene exhibited powerful antibacterial activity against multidrug-resistant (MDR) E. faecalis, with inhibition zones ranging from 13 to 32 mm and minimum inhibitory concentrations (MICs) ranging from 3.75 to 30 µg/ml at safe concentrations. In addition, it was found that our stable purified β-carotene showed up to 94% inhibition in biofilm formation by strong biofilm-forming E. faecalis. In addition, the β-carotene-coated catheter manifested a lower biofilm formation by E. faecalis by up to 75.3%. Moreover, crystal violet staining, dual staining, and fluorescence staining techniques displayed immature biofilms of E. faecalis when treated with 0.25 and 0.5 MICs of β-carotene. The mechanistic pathway for the purified β-carotene's antibiofilm activity was strongly linked to the inhibition of gelatinase enzyme production (up to 100% inhibition) as manifested phenotypically, genotypically, and by molecular docking.

CONCLUSION: This work provided a deeper insight into optimizing carotenoid production from M. luteus by investigating the influence of 54 diverse conditions. Also, this is the first time to report the antibacterial and antibiofilm actions of the stable purified microbial β-carotene against strong biofilm-forming MDR E. faecalis colonizing urinary catheters.

RevDate: 2025-08-18

Bludau D, Volkenandt S, Wagenhofer J, et al (2025)

Tire Wear Particles Drive Size-Dependent Loss of Freshwater Bacterial Biofilm Diversity.

Environmental pollution (Barking, Essex : 1987) pii:S0269-7491(25)01377-6 [Epub ahead of print].

Tire wear particles (TWPs) represent a significant, yet understudied, source of anthropogenic pollution in aquatic environments, contributing to microplastic contamination and potentially altering microbial ecosystems. As TWPs are continuously released through abrasion during vehicle use, they accumulate in rivers and sediments, where they can act as surfaces for microbial biofilm formation. In this study, we investigated how TWP characteristics - particle size (small & large), wear condition (unused & used), and vehicle type (car & truck) - affect microbial biofilm composition after four weeks of exposure in the River Rhine, compared to natural sediment. Using 16S rRNA amplicon sequencing, we found that TWP-associated biofilms harboured significantly lower bacterial diversity than natural sediment biofilms. While a substantial number of OTUs were shared between both habitats, TWPs exhibited increased richness but decreased overall diversity, indicating selective enrichment of specialised taxa. Larger particles further reduced diversity, and specific genera such as Aquabacterium and Ketobacter were highly enriched on TWPs, indicating selective microbial colonization. These findings reveal the effects of TWPs on microbial biofilm communities, emphasizing their potential role in altering freshwater microbial ecosystems. Given the widespread release of TWPs, understanding their ecological impact is crucial for assessing their role in aquatic pollution and ecosystem functioning.

RevDate: 2025-08-18

Jothi R, Malligarjunan N, Vidhya K, et al (2025)

Lupeol mitigates biofilm formation and attenuates virulence dimorphism in Candida albicans.

Microbial pathogenesis pii:S0882-4010(25)00714-4 [Epub ahead of print].

BACKGROUND: The global burden of Candida infections, especially in immunocompromised individuals, underscores the need for novel therapies. Candida's virulence is driven by traits like hyphal elongation and biofilm formation, making these key targets for managing candidiasis and preventing its progression to life-threatening systemic infections.

PURPOSE: This study investigates the anti-candidal efficacy of lupeol at sub-MIC levels against Candida albicans, Candida tropicalis, and Candida glabrata, focusing on its effects on biofilm formation, hyphal progression, and the expression of virulence genes.

METHODS: The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of lupeol were determined against the Candida species. Sub-MIC concentrations (25-200 μg/mL) inhibited biofilm formation, as assessed by confocal and scanning electron microscopy. Lupeol's effects on hyphal progression and germ tube formation in C. albicans were evaluated, and qPCR analyzed key virulence gene expression.

RESULTS: The MIC and MFC of lupeol were found to be 256 and 512 μg/mL, respectively, against C. albicans, C. tropicalis, and C. glabrata. At sub-MIC levels, lupeol dose-dependently inhibited biofilm formation without affecting metabolic activity. It altered biofilm structure, inhibited hyphal growth, reversed hyphae to yeast morphology, and modified colony appearance. qPCR revealed downregulation of virulence genes (RAS1, UME6, HWP1, EFG1, CPH1) and upregulation of TUP1.

CONCLUSION: These findings highlight lupeol as a promising therapeutic candidate for combating Candida infections by targeting critical virulence factors and disrupting biofilm integrity. This study paves the way for further exploration of lupeol's clinical applications in antifungal therapy.

RevDate: 2025-08-18

Comlekcioglu U, Aydogan MY, Aygan A, et al (2025)

Characterization of biofilm and exopolysaccharide production in Bacillus strains isolated from the bovine uterus.

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] [Epub ahead of print].

The bovine uterus hosts a diverse microbiome whose role in reproductive physiology and pathology is increasingly recognized. While Bacillus species have been occasionally isolated from the uterus, their biofilm and exopolysaccharide (EPS) forming capabilities have not been systematically characterized. In this study, four Bacillus strains (BU13, BU14, BU15, and BU16) were isolated from the bovine uteri and examined for their taxonomic affiliation, phenotypic characteristics, EPS production, biofilm formation, and antibiotic susceptibility. Phylogenetic analyses based on nearly full-length 16S rRNA gene sequences revealed that all isolates belonged to the B. subtilis group, with BU13, BU14, and BU16 closely related to B. licheniformis, and BU15 related to B. amyloliquefaciens and B. siamensis. BU13 and BU16 demonstrated high levels of EPS and biofilm production, especially in sucrose-supplemented media and under nutrient-rich conditions. Notably, these strains also exhibited relatively smaller inhibition zones against β-lactam antibiotics, which may be associated with their robust EPS-biofilm phenotypes. In contrast, larger inhibition zones were observed with gentamicin, enrofloxacin, and trimethoprim. These findings underscore the importance of characterizing commensal Bacillus spp. in the uterus and highlight that certain strains may possess traits that facilitate persistence and reduce antimicrobial responsiveness. This is the first study to comprehensively evaluate the biofilm-forming potential of uterine Bacillus isolates and provides a foundation for future investigations into their role in reproductive health and disease.

RevDate: 2025-08-18

Hussain MA, Zafar M, Khan YS, et al (2025)

Correction: Molecular identification, antibiotic susceptibility, and biofilm formation of airborne bacteria.

AMB Express, 15(1):118.

RevDate: 2025-08-18

Glowacki RW, Engelhart MJ, Till JM, et al (2025)

Identification of strain-specific cues that regulate biofilm formation in Bacteroides thetaiotaomicron.

Microbiology spectrum [Epub ahead of print].

Members of the gut microbiome encounter a barrage of host- and microbe-derived microbiocidal factors that must be overcome to maintain fitness in the intestine. The long-term stability of many gut microbiome strains within the microbiome suggests the existence of strain-specific strategies that have evolved to foster resilience to such insults. Despite this, little is known about the mechanisms that mediate this resistance. Biofilm formation represents one commonly employed defense strategy against stressors like those found in the intestine. Here, we demonstrate strain-level variation in the capacity of the gut symbiont Bacteroides thetaiotaomicron to form biofilms. Despite the potent induction of biofilm formation by bile in most strains, we show that the specific bile acid species driving biofilm formation differs among strains and uncover that a secondary bile acid, lithocholic acid, and its conjugated forms potently induce biofilm formation in a strain-specific manner. Additionally, we found that the short-chain fatty acid, acetic acid, could suppress biofilm formation. Thus, our data define molecular components of bile that can promote biofilm formation in B. thetaiotaomicron and reveal that distinct molecular cues trigger the induction or inhibition of this process. Moreover, we uncover strain-level variation in these responses, thus identifying that both shared and strain-specific determinants govern biofilm formation in this species.IMPORTANCEIn order to thrive within the intestine, it is imperative that gut microbes resist the multitude of insults derived from the host immune system and other microbiome members. As such, they have evolved strategies that ensure their survival within the intestine. We investigated one such strategy, biofilm formation, in Bacteroides thetaiotaomicron, a common member of the human microbiome. We uncovered significant variation in natural biofilm formation in the absence of an overt stimulus among different B. thetaiotaomicron strains and revealed that different strains adopted a biofilm lifestyle in response to distinct molecular stimuli. Thus, our studies provide novel insights into factors mediating gut symbiont resiliency, revealing strain-specific and shared strategies in these responses. Collectively, our findings underscore the prevalence of strain-level differences that should be factored into our understanding of gut microbiome functions.

RevDate: 2025-08-18

Chen F, Lei M, Luo J, et al (2025)

Ultrathin DNA-copper nanosheets with antibacterial and anti-biofilm activity for treatment of infected wounds.

Nanoscale horizons [Epub ahead of print].

The development of innovative antibacterial materials is crucial for addressing wounds infected with bacterial biofilms. Advanced nanomaterials that enable non-antibiotic antibacterial strategies offer new possibilities for treating bacterial infections by eliminating pathogens without relying on antibiotics. Herein, we introduce non-toxic and biocompatible DNA-copper cluster nanosheets (DNS/CuNCs) as effective antibacterial agents. DNS/CuNCs can reduce bacterial surface motility and the secretion of virulence factors by interfering with quorum sensing, and thereby inhibit biofilm formation and enhance their potential as prophylactic antibacterial agents. Notably, DNS/CuNCs exhibit significant in vitro bactericidal activity against Staphylococcus aureus and Pseudomonas aeruginosa and disrupt established surface biofilms in the presence of hydrogen peroxide (H2O2). This is attributed to the synergistic effects of their physical ultrathin properties and peroxidase-like activity, which lead to an increase in intracellular ROS levels in bacteria, thereby achieving antibacterial and biofilm-disrupting effects. In vivo, DNS/CuNCs effectively eradicate bacterial infections, promote wound healing, and restore normal tissue morphology without toxicity to mammalian cells. With their combined abilities to inhibit biofilm formation, exhibit antibacterial activity, and disrupt biofilms, along with excellent biocompatibility, DNA-templated CuNCs emerge as highly promising candidates for preventive and clinical antibacterial therapies.

RevDate: 2025-08-18

Thabit AG, Sediek MN, MSE Mohamed (2025)

The presence of exotoxin genes and biofilm production in carbapenem-resistant Pseudomonas aeruginosa clinical isolates.

Germs, 15(1):26-36 pii:germs.2025.1452.

INTRODUCTION: Pseudomonas aeruginosa produces many exotoxins which are essential for the bacterial pathogenesis. The aim of this study was to identify Pseudomonas aeruginosa from clinical specimens, detect the sensitivity pattern, biofilm production, and the frequency of exogenes.

METHODS: Pseudomonas aeruginosa clinical isolates were identified by conventional and genotypic methods. Antibiotic susceptibility patterns and biofilm production were performed. Molecular detection of exotoxin genes exoS, exoT, exoU, and exoY in Pseudomonas aeruginosa isolates was performed by PCR.

RESULTS: Seventy-five Pseudomonas aeruginosa were identified in 400 clinical specimens. Sixty-six (88%) isolates were carbapenem-resistant. A total of 25 (33.3%) isolates were extensively drug resistant, 18 (24%) were multidrug resistant, and 11 (14.7%) were pandrug resistant. Sixty-three (84%) isolates were biofilm producers. Biofilm formation was detected in 56 (85%) of carbapenem-resistant isolates. Totally, 70 (93.3%) isolates carried exoS, 68 (90.7%) carried exoY, 65 (86.7%) carried exoT, and 28 (37.3%) carried exoU. Exogenes were highly expressed in carbapenem-resistant isolates. Coexistence of more than one gene was detected in nearly all isolates.

CONCLUSIONS: Pseudomonas aeruginosa clinical isolates were resistant to many anti-pseudomonal antibiotics. Most of isolates were biofilm-producers. The genes exoT, exoS and exoY were identified in almost all P. aeruginosa strains and are considered an inevitable component of P. aeruginosa virulence.

RevDate: 2025-08-18

van der Ploeg K, de Vogel CP, Klaassen CHW, et al (2025)

Proteomic identification of PA2146 as a biofilm marker of Pseudomonas aeruginosa on endoscope channel material.

Biofilm, 10:100310 pii:S2590-2075(25)00058-9.

STUDY BACKGROUND AND AIMS: Pseudomonas aeruginosa can persistently contaminate endoscopes by forming biofilms within internal channels, complicating both detection and eradication. Current microbiological surveillance methods have limited efficacy and may yield false-negative results. This study aimed to identify proteomic markers of P. aeruginosa biofilms on endoscope channel material.

METHODS: Three genetically unrelated P. aeruginosa isolates from contaminated duodenoscopes and two reference strains (ATCC 27853 and PAO1) were used. Biofilms were grown on disinfected endoscope biopsy channel rings and incubated for 24, 48, and 72 h. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was employed to analyze temporal changes in protein spectra. Peaks of interest were further characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and whole-genome sequencing to identify associated proteins. To further confirm the origin of these peaks, strains naturally lacking the corresponding genes were analyzed.

RESULTS: MALDI-TOF MS revealed distinct time- and strain-specific spectral profiles, with two notable peaks at approximately 2723 m/z and 5450 m/z. LC-MS/MS identified the 5450 m/z peak as PA2146, corresponding to a 5449.1 Da protein after in vivo methionine cleavage. The 2723 m/z peak was confirmed as its doubly charged ion. Both peaks were absent in strains naturally lacking PA2146, confirming it as the source.

CONCLUSION: PA2146 expression increases during P. aeruginosa biofilm development on endoscope channel surfaces, indicating its potential as a biomarker for contamination. MALDI-TOF MS could enhance biofilm detection in endoscope surveillance. Further research should assess the clinical utility of proteomic approaches for improving endoscopic microbiological safety.

RevDate: 2025-08-18

Hong L, Hjort K, Andersson DI, et al (2025)

Linoleic acid addition prevents Staphylococcus aureus biofilm formation on PMMA bone cement.

Biofilm, 10:100311 pii:S2590-2075(25)00059-0.

Acrylic bone cement is widely used in vertebroplasty to treat osteoporosis-induced vertebral compression fractures. However, infection after vertebroplasty is problematic and previous work has suggested loading the bone cement with an antibiotic for prophylaxis. Linoleic acid (LA) has been investigated as a promising additive to improve the mechanical properties of bone cement for vertebroplasty, but LA could potentially also have an antibacterial effect. In this study, we evaluated the antibacterial properties of LA-loaded bone cement by comparing its antibiofilm properties with that of original bone cement through quantification of bacterial growth using viable cell count and scanning electron microscopy. The released monomer (MMA) concentration and the monomer minimum inhibitory concentration were determined to clarify the monomer's potential role in inhibiting bacterial growth. The LA release profile was measured, and a checkerboard assay was done to determine any synergistic effects of LA and the commonly used antibiotic gentamicin. Results show that LA-loaded bone cement could significantly inhibit Staphylococcus aureus biofilm formation, including gentamicin-resistant strains, but with limited effect on Escherichia coli. Furthermore, the released MMA did not have a significant influence on bacterial growth. The checkerboard assay results show that the LA and gentamicin combination could broaden the antibacterial spectrum and increase gentamicin efficacy. In conclusion, LA merits further investigation as an antibacterial agent in bone cement, alone or in combination with antibiotics.

RevDate: 2025-08-18

Mahale RP, K A, Princy A, et al (2025)

Comparative evaluation of biofilm-forming capacity in uropathogenic and commensal Escherichia coli.

Frontiers in cellular and infection microbiology, 15:1570422.

INTRODUCTION: Escherichia coli (E. coli) causes most cases of the urinary tract infections (UTIs) via virulence factors like biofilms. This study identifies key phenotypic and genotypic virulence attributes of Uropathogenic Escherichia coli.

METHODOLOGY: A total of 180 uropathogenic E. coli (UPEC) isolated from patients with different categories (cystitis, pyelonephritis, recurrent UTI, catheter-associated UTI, and asymptomatic bacteriuria) of UTI and 30 commensal E. coli isolated from healthy individuals were evaluated for biofilm production by phenotypic methods using tissue culture plate, tube adherence, and Congo red method, and RT-PCR was used to genetically characterize them.

RESULTS: This study analyzed 1,600 urine samples from UTI patients, with 498 showing significant bacterial growth and 180 identifying E. coli as the pathogen. The female-to-male ratio of UTI cases was 0.74. Antibiotic susceptibility testing revealed 100% sensitivity to tigecycline and fosfomycin as well as 89.44%, 86.11%, 81.66%, and 72.22% sensitivity to nitrofurantoin, amikacin, imipenem, and meropenem, respectively. Only 64.44% were sensitive to ciprofloxacin, with 10% being multidrug-resistant (MDR). Moreover, 18.33% of the UPEC isolates produced mettalo-beta-lactamases (MBL), and 13.33% produced AmpC beta-lactamases. Biofilm production was observed in 72.22% of UPEC isolates compared to 16.66% in commensal isolates. The biofilm-forming UPEC, compared to commensal E. coli, has significantly higher antibiotic resistance, with a 128-fold reduction in ciprofloxacin susceptibility. Additionally, the fimH gene was detected in 98.33% of the UPEC isolates.

CONCLUSION: This study shows that UPEC strains produce specific virulence determinants like adhesion to uroepithelial cells. Screening for virulence factors should be integrated into microbiology laboratories. Specific virulence genes linked to UPEC may serve as potential targets for prophylactic strategies to prevent recurrent infections and improve management.

RevDate: 2025-08-18

Tang Y, Li Y, Xiao N, et al (2025)

Antibiotic Resistance, Biofilm Formation, and Molecular Epidemiology of Foodborne Staphylococcus aureus Isolated in Northwest Hubei Province, China.

Food science & nutrition, 13(8):e70791 pii:FSN370791.

Staphylococcus aureus is a common pathogen responsible for foodborne infections worldwide. This study investigated the antibiotic resistance profiles, biofilm formation capacity, and molecular epidemiological characteristics of foodborne S. aureus isolates from northwest Hubei Province, China, as well as the correlation among these factors. Among the 303 food samples collected from Xiangyang, Suizhou, and Shiyan cities, 41 yielded non-duplicate S. aureus strains. Of the 41 S. aureus isolates, 8 (19.51%) were identified as MRSA, while 33 (80.49%) were methicillin-susceptible S. aureus (MSSA). High resistance was observed to penicillin (78.05%), tetracycline (43.90%), and erythromycin (31.71%), with MRSA strains demonstrating significantly stronger resistance profiles than MSSA strains. Among MRSA isolates, 50% (4/8) demonstrated strong biofilm-forming capacity, compared to only 9.09% (3/33) of MSSA isolates. Strong biofilm formation was observed more frequently in isolates from frozen meat (66.67%, 4/6) than in those from vegetarian salads (0%, 0/9) or fresh meat (12.50%, 2/16). The prevalence of Panton-Valentine leukocidin (PVL) gene was higher in MRSA strains, whereas enterotoxin genes were more commonly found in MSSA strains, though differences between groups were not statistically significant. The primary epidemic clones identified were CC88-ST88-t1376/t437, CC7-ST7-t091/t3884, and CC5-ST6/ST462-t701/t165, constituting 63.41% (26/41) of isolates. The CC59-ST59/338 strain exhibited a pronounced capacity for strong biofilm formation. SCCmecII-CC15-ST15-t085 and SCCmecIII-CC7-ST7-t3884 strains exhibited the highest antibiotic resistance, with resistance to 9 and 7 antibiotics, whereas CC88-ST88-t1376, CC7-ST7-t091, and CC5-ST6-t701 showed resistance to fewer than three antibiotics. The findings enhance the understanding of the drug resistance profiles and molecular epidemiology of foodborne S. aureus, providing a foundation for more effective control measures.

RevDate: 2025-08-17

Nofouzi K, Shakeri S, Nikkhah S, et al (2025)

Gallibacterium anatis as an emerging pathogen in pet birds: biofilm formation contributes to treatment challenges and persistence.

BMC microbiology, 25(1):518.

BACKGROUND: Gallibacterium anatis (G. anatis), a microorganism of the Pasteurellaceae family, is an emerging avian pathogen associated with reproductive and respiratory diseases in poultry. However, its role in ornamental birds is still poorly understood. The aim of the present study was to conduct the first comprehensive survey of the prevalence of G. anatis in pet birds, to investigate its antimicrobial resistance (AMR) profile and to assess its ability to form biofilms using cultural, biochemical, molecular and histopathological methods.

METHODS: In this study, 191 fecal and tissue samples were collected from various companion birds. Clinical samples were cultured on 5% sheep blood agar and MacConkey agar plates to isolate bacterial pathogens. After incubation, colonies were evaluated based on their macroscopic characteristics such as size, color, and hemolytic properties on blood agar-and a Gram stain was performed as an essential preliminary step for bacterial identification. The 16-23 S rRNA gene region of G. anatis was amplified by PCR method. The disc diffusion method was used to assess microbial susceptibility and resistance. Biofilm formation was analyzed using a microtiter plate assay. Tissue samples were routinely processed, embedded in paraffin, sectioned and stained with common haematoxylin-eosin (H&E).

RESULTS: In this study, 20 G. anatis strains were isolated from 191 clinical samples of pet birds, representing a prevalence of 10.5%. Isolates were identified by colony morphology, Gram staining, biochemical testing and PCR for the -intergenic spacer region of 16-23 S rRNA, which yielded diagnostic amplicons of 790 bp and 1080 bp. Antimicrobial susceptibility testing showed complete susceptibility to ciprofloxacin (100%) and remarkable resistance to erythromycin (80%). β-lactam resistance was prevalent, with 70% and 75% of isolates resistant to ampicillin and amoxicillin, respectively. Biofilm formation tests showed that 80% of isolates had moderate biofilm formation. Gross and histopathological examinations of infected birds revealed severe respiratory and systemic lesions, including tracheitis, bronchopneumonia with focal necrosis, multifocal hepatic necrosis, and vascular congestion in multiple organs.

CONCLUSIONS: These results support the idea that G. anatis is a potentially important pathogen, with a biofilm-forming ability that contributes to treatment failure and environmental persistence. The 45% prevalence of multidrug resistance (MDR) highlights the pressing need for antimicrobial stewardship in avian veterinary medicine. Given the zoonotic potential of G. anatis, our study underscores the importance of One Health surveillance efforts in mitigating risk to both poultry and humans.

RevDate: 2025-08-17

Le DD, Wang WH, CY Lan (2025)

Exploring the role of the CCAAT-binding complex in cell wall maintenance and biofilm formation in Candida albicans.

Journal of microbiology, immunology, and infection = Wei mian yu gan ran za zhi pii:S1684-1182(25)00156-2 [Epub ahead of print].

BACKGROUND: The conserved CCAAT-binding complex (CBC) specifically recognizes and binds to the CCAAT motif present in eukaryotic promoters, thereby controlling gene transcription. In Candida albicans, the CBC cooperates with another transcription factor, Hap43, to regulate iron homeostasis. Moreover, several Hap43-independent functions have also been uncovered. However, the functions of CBC have not been extensively characterized.

METHODS: Deletion mutants lacking each component of the CBC were independently compared to the wild-type strain with regard to cell wall properties, composition, and structure. The effects of CBC deletion on biofilm formation were also investigated. Finally, RNA-seq analysis was performed to reveal functional divergence between the two Hap3 paralogs, Hap31 and Hap32.

RESULTS: CBC deletion significantly impacts cell wall properties, composition, exposure of glucan and chitin, as well as cell wall remodeling. These effects appear to be associated with the small GTPase Rhb1 and the Mkc1 signaling pathway. Moreover, we showed that CBC deletion affects biofilm formation, which appears to be independent of Rhb1. RNA-seq analysis further revealed the broad roles of the Hap3 paralogs within the CBC.

CONCLUSION: Notably, this work provides new insights into the relationship among CBC, cell wall maintenance, and biofilm formation in C. albicans.

RevDate: 2025-08-17

Saxena P, Samanta D, Gnimpieba EZ, et al (2025)

Sulfate-reducing bacteria: Unraveling biofilm complexity, stress adaptation, and strategies for corrosion control.

The Science of the total environment, 998:180226 pii:S0048-9697(25)01866-2 [Epub ahead of print].

Sulfate reducing bacteria (SRB) are anaerobes that play a pivotal role in the global sulfur cycle, influencing both environmental and industrial processes. SRB-mediated sulfate reduction facilitates the bioprecipitation of heavy metals, offering potential applications in bioremediation; however, the metabolic byproducts (e.g., H2S) contribute to microbiologically influenced corrosion (MIC) in industrial infrastructure. Over the decades, MIC has been a significant contributor to global corrosion-related economic losses, notably in oil pipelines, marine vessels, and gas storage facilities. MIC is primarily driven by SRB biofilms, which accelerate metal degradation through extracellular electron transfer mechanisms and sulfide-induced pitting corrosion. The ability of SRB to form structured biofilms enhances their resilience under environmental stressors (e.g., oxidative stress, metal toxicity, and pH fluctuations). However, the regulatory mechanisms underlying SRB biofilm development and stress adaptation remain poorly understood. Recent advances in omics strategies- transcriptomics, proteomics, and metabolomics, have provided deeper insights into SRB physiology, revealing key genetic determinants of biofilm formation and quorum sensing-mediated regulation. Nevertheless, knowledge gaps persist regarding the molecular basis of SRB community interactions, extracellular electron transfer pathways, and the impact of multi-stressor environments on biofilm resilience. This review critically examines the dual role of SRB in both detrimental and beneficial processes, highlighting recent molecular developments in SRB biofilm research and MIC mitigation strategies. We also discuss emerging approaches, including systems biology interventions, quorum sensing inhibitors, and computational modeling, to develop targeted solutions for controlling MIC while harnessing SRB for sustainable environmental applications.

RevDate: 2025-08-17

Rocha Garcia MA, Bastos Dos Santos M, de Cássia Orlandi Sardi J, et al (2025)

Synthesis, anti-biofilm and molecular docking of amino-substituted chalcones targeting Staphylococcus aureus sortase A.

Future medicinal chemistry [Epub ahead of print].

Antibiotic resistance is an urgent global health challenge that requires the development of new antibacterial agents. In this study, 14 aminochalcones bearing electron-withdrawing groups were synthesized and evaluated for antibacterial activity. Chalcone C5, with an ortho-chlorine on ring B, demonstrated the most potent effect, notably against methicillin-susceptible and methicillin-resistant Staphylococcus aureus (MICs of 1.9 and 3.9 µg/mL, respectively), comparable to vancomycin. C5 showed synergistic interaction with vancomycin, reducing its MIC tenfold. Time-kill assays confirmed C5's bactericidal action within 8 h, with no bacterial regrowth up to 12 h. C5 also significantly inhibited bacterial adhesion to keratinocytes (HaCaT) and reduced biofilm formation and survival at both MIC and 10× MIC, showing effects comparable to vancomycin. In silico, ADMET predictions indicated favorable pharmacokinetic and safety profiles, including high intestinal absorption and lack of hERG inhibition or cytotoxicity. Molecular docking against S. aureus sortase A (SrtA) suggests strong interactions with key residues (Arg197, Glu105, Asn114), supporting the anti-adhesion activity. Furthermore, in vivo toxicity assessment using Galleria mellonella larvae showed minimal toxicity at 100× MIC. These findings support chalcone C5 as a promising lead compound for the development of new antibacterial agents, particularly for combating S. aureus infections and biofilm-associated pathologies.

RevDate: 2025-08-16

Reigada I, Togbe E, Sovegnon T, et al (2025)

Potent antimicrobial and Anti-Biofilm Effects of Uvaria chamae on Staphylococcus aureus strains via Potential Cell Wall Interference and Lack of Resistance Induction.

Journal of ethnopharmacology pii:S0378-8741(25)01126-2 [Epub ahead of print].

Antimicrobial resistance (AMR) is a critical global health issue, particularly in sub-Saharan Africa. Traditional remedies, such as Uvaria chamae, are commonly used in West Africa to treat bacterial infections.

AIM OF THE STUDY: To comprehensively evaluate the antimicrobial, anti-biofilm, and resistance induction profiles of U. chamae.

MATERIALS AND METHODS: Ethanolic, hydroethanolic, and aqueous root and leaves extracts were assayed against Gram-positive and -negative bacteria. Biofilm inhibition, time-kill kinetics, and bacterial morphology were studied. In vivo efficacy was assessed using a Caenorhabditis elegans infection model. Chemical profiling was conducted via UPLC-PDA-QTOF/MS, and cytotoxicity evaluated in A549 and THP-1 cells.

RESULTS: The ethanolic extract of the roots was the most potent, showing selective activity against S. aureus and MRSA (MIC = 4 μg/mL, MBC = 8 μg/mL). Time-kill assays confirmed bacteriostatic activity at MIC and bactericidal effects at higher concentrations. The extract effectively disrupted biofilms and did not induce resistance, unlike ciprofloxacin. It did not damage bacterial membranes but altered cell wall morphology. Low cytotoxicity was observed in human cells, and no toxicity was detected in C. elegans. In vivo, the extract significantly reduced bacterial burden in infected nematodes at 14 and 32 μg/mL. LC-MS analysis on active fractions revealed several known antimicrobial compounds, aligning with previously reported metabolite profile of U. chamae roots.

CONCLUSION: U. chamae root extract exhibits strong, selective antibacterial activity with low toxicity, no resistance induction, and in vivo efficacy. These findings support its potential as a locally relevant option to combat AMR in Africa.

RevDate: 2025-08-16

Sun Y, Chu M, Zhuhuang C, et al (2025)

Vibrio parahaemolyticus outer membrane porin Vpa0810 regulates its biofilm formation and plays a vital role in its stresses tolerance.

International journal of food microbiology, 442:111390 pii:S0168-1605(25)00335-6 [Epub ahead of print].

Vibrio parahaemolyticus (V. parahaemolyticus), a Gram-negative halophilic bacterium, is a leading seafood-borne pathogen that can cause acute gastroenteritis. Outer membrane (OM) porins are involved in exporting extracellular polymeric substances, which is essential for biofilm formation. However, the contribution of porins to the biofilm formation of V. parahaemolyticus is still obscure. We recently found that a mutation in the DHH/DHHA1 family gene vp2835 enhances biofilm formation in V. parahaemolyticus but inhibits its motility. Here, we identified an OM porin gene, vpa0810, from Δvp2835 strain. The deletion of vpa0810 in the wild type strain resulted in the inhibition of both swarming and swimming motility, while reverted the motility defect of Δvp2835. The Δvpa0810 decreased its biofilm formation capability in both the wild type and the Δvp2835 strain. Additionally, the surface hydrophobicity, auto-aggregation, OM permeability, and exopolysaccharides (EPS) content were decreased in the Δvpa0810 strain. Interestingly, qRT-PCR results demonstrated that the genes involved in EPS synthesis (cpsA, scvE, and cpsF) and the porin gene ompW were significantly up-regulated in the Δvpa0810 strain, but down-regulated in the Δvp2835-Δvpa0810 double mutant. Furthermore, the deletion of vpa0810 compromised its ability to form biofilm on silicon wafers, glass, stainless-steel plates, as well as on shrimp and crab surfaces. The vpa0810 deletion mutation also reduced its stress tolerance to bile salt, low pH, low temperature, and the antibiotics colistin. In summary, our data suggest that Vpa0810 may regulate biofilm formation through EPS biosynthesis and exportation, which might provide a new target for developing control strategies against V. parahaemolyticus.

RevDate: 2025-08-16

Habib MB, Shah NA, Amir A, et al (2025)

Antimicrobial resistance and biofilm formation in implants related infections: Pathogens profiling and implants susceptibility.

Diagnostic microbiology and infectious disease, 113(4):117061 pii:S0732-8893(25)00384-0 [Epub ahead of print].

Medical implant-associated infections are becoming increasingly hazardous because of the development of antimicrobial resistance (AMR). This study examined the burden of infectious medical implants (IMI), the possibility of biofilm generation, and the relationship between the type of implant material. A total of 135 infectious samples from medical implants were collected for this study. Matrix-assisted laser desorption Ionization time-of-flight (MALDI-TOF) was used to identify bacterial isolates. Disk diffusion and broth microdilution were used to test antimicrobial susceptibility, and biofilm potential was determined using a microtitre plate assay. The most prevalent pathogen was Acinetobacter baumannii (n=50, 37%), followed by Pseudomonas aeruginosa (n=42, 31.1%) and Escherichia coli (n=21, 15.6%). Antimicrobial susceptibility profiling showed MDR (n=89, 66%), XDR (n=13, 7%), PDR (n=2, 4%), and sensitive isolates (n=31,23%). The biofilm assay showed 66 (49%) strong, 46 (33%) moderate, and 23 (18%) weak biofilm producers, respectively. The strongest biofilms were found on the interlocking nails of the tibia and orthopaedic staples. A higher biofilm potential has been reported for stainless-steel implants. Binary logistic regression revealed that A. baumannii and stainless steel implants were significant predictors of strong biofilm formation. Similarly, E. coli and orthopedic staples were independently associated with multidrug resistance. Enterobacter hormaechie subsp. Oharae and Gemella haemolysans were reported first time in Pakistan among implants related infections. This study highlights the need for health professionals and policymakers to address concerns regarding implant-associated infections and alternative therapeutic strategies.

RevDate: 2025-08-16

Kannan KP, Smiline Girija AS, V Priyadharsini J (2025)

Deciphering the twinning act of antimicrobial resistance and biofilm-associated virulence in multidrug-resistant Acinetobacterbaumannii.

Biochemical and biophysical research communications, 780:152487 pii:S0006-291X(25)01202-1 [Epub ahead of print].

Acinetobacter baumannii has emerged as a predominant healthcare-associated pathogen, exhibiting high levels of antibiotic resistance and biofilm-forming capability, contributing to its persistence in hospital environments. This study evaluated the association between antibiotic resistance and biofilm-associated genes in A. baumannii clinical isolates. A total of 247 A. baumannii isolates were collected from a tertiary care hospital in Chennai, India, over three years (March 2023-January 2025). Antibiotic susceptibility testing revealed multi-drug resistance (MDR), particularly to carbapenems and β-lactam/β-lactamase inhibitors, while colistin and tigecycline remained effective against all isolates. Biofilm formation assays demonstrated that 82.8 % of isolates exhibited biofilm-producing capabilities, with 46.2 % categorized as strong biofilm producers. Molecular characterization confirmed the widespread presence of antibiotic resistance genes, including blaOXA-51 and ISAba1 (100 %) and blaOXA-23 (97.05 %), along with genes associated with quinolone, aminoglycoside, and tetracycline resistance. Virulence gene analysis revealed the high prevalence of biofilm-associated genes (pgaA-D, csuA-E, ompA and bap), indicating their role in bacterial adherence and persistence. Statistical analysis revealed a significant association between MDR and biofilm-forming ability in the isolates. These findings offer region-specific molecular epidemiological insights and underscore the need for ongoing surveillance and enhanced infection control strategies.

RevDate: 2025-08-16

Liu M, Wu C, Zhang X, et al (2025)

Discovery of a dual-action compound for metallo-β-lactamase inhibition and biofilm clearance to reverse CRE resistance.

Journal of inorganic biochemistry, 273:113028 pii:S0162-0134(25)00208-9 [Epub ahead of print].

The emergence of carbapenem-resistant Enterobacteriaceae (CRE) poses a severe threat to global public health due to its multidrug resistance and limited treatment options. In this study, QA, a zinc ion chelator with potent inhibitory activity against metallo-β-lactamases (MBLs), was identified through high-throughput screening and biochemical analysis. QA effectively inhibits MBLs by chelating zinc ions and demonstrates potent biofilm clearance capabilities, ultimately reversing carbapenem-resistant Enterobacteriaceae (CRE) resistance. Synergistic antibacterial effects were observed when QA was combined with meropenem and other β-lactam antibiotics, significantly enhancing their efficacy. These findings highlight the dual-action potential of QA as a zinc ion chelator and biofilm disruptor, offering a promising strategy for combating CRE infections and multidrug-resistant bacteria.

RevDate: 2025-08-18

Hao M, J Wang (2025)

Molecular epidemiology, antimicrobial resistance, and virulence characteristics of predominant methicillin-resistant Staphylococcus aureus clones with strong biofilm-producing capability from a tertiary teaching hospital in China.

BMC microbiology, 25(1):510.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most prevalent bacterial pathogens leading to various kinds of infections, but the characteristics of this superbug with both strong biofilm-producing and intracellular invasive capabilities is rarely reported. This study aimed to investigate the genotypic and phenotypic features of this superbug with above two properties.

METHODS: Phenotypic resistance profiling of MRSA clinical isolates was performed via the VITEK 2 AST-GP67 Test Kit. Biofilm production was assessed via crystal violet staining and the Congo red agar (CRA) method. The biofilm-degrading activity was tested using Proteinase K, Dispersin B, and DNase I. The intracellular invasive capability was evaluated via dilution plate count and immunofluorescence assay. Genotyping was performed using multilocus sequence typing and staphylococcal protein A typing methods, and virulence genes were detected via polymerase chain reaction. Flow cytometry was performed to assess the cytotoxicity of the dominant MRSA clones.

RESULTS: A high prevalence (21.6%) of MRSA isolates exhibiting strong biofilm-forming capability was observed in this study, including 70 strains with the highest level of biofilm production (optical density > 0.4). DNase I exhibited the most effective biofilm-degrading activity, with the biofilm-degrading percentage of 78.6% of the strains exceeding 50%. Simultaneously, 71.4% of the isolates exhibited strong invasive capability into A549 cells. ST5-t2460 (48.6%), ST59-t437 (20%), and ST239-t030 (11.4%) were identified as the predominant clones. In particular, ST5-t2460 and ST239-t030 clones exhibited broader antibiotic resistance to gentamicin, ciprofloxacin, levofloxacin, moxifloxacin, and tetracycline compared with ST59-t437 clone. In addition, a higher percentage of the isolates belonging to ST5-t2460 (91.2%) and ST239-t030 (100%) clones demonstrated stronger intracellular invasive capability relative to those belonging to ST59-t437 clone (14.3%). Furthermore, ST5-t2460 and ST239-t030 clones displayed stronger cytotoxicity and carried higher proportions of adhesion-related genes (fnbA, sdrD, sasC) and other virulence genes (sea, seb, sec, isdB, lukE-D, tsst-1).

CONCLUSIONS: This is the first report of the phenotypic-genotypic characteristics of MRSA with both strong biofilm-producing and virulence potential, with ST5-t2460, ST59-t437, and ST239-t030 clones accounting for the major genotypes. Further exploration of specific virulence genes correlating to the pathogenesis of this superbug is deemed essential for developing targeted infection control and treatment strategies in the future.

RevDate: 2025-08-16

Gao Z, Wang Y, Chen H, et al (2025)

Enhanced pollutant removal in moving bed biofilm reactor under high-salinity condition via specialized quorum sensing bacteria.

Environmental research, 285(Pt 4):122610 pii:S0013-9351(25)01862-6 [Epub ahead of print].

The study aims to accelerate biofilm formation and operational performance of moving bed biofilm reactor (MBBR) in a high salt environment by adding specific quorum sensing (QS) strain (Vibrio sp. LV-Q1). Results showed that the addition of QS strain significantly promoted the pollutant removal process of the bioreactor. At the end of the reaction, the NH+ 4-N and COD removal rates of the bioaugmented MBBR (R1) were about 13.20 % and 10.15 % higher than the unbioaugmented MBBR (R0), respectively. The bioaugmentation of LV-Q1 enhanced the biofilm activity and adhesion of the system, which was related to the increase of polysaccharides and proteins in the biofilm. On day 70, the polysaccharides and proteins of the biofilm in R1 were increased by 41.66 % and 87.66 % compared with that in R0, respectively. And throughout the entire operation process, the biological system of R1 maintained a higher concentration of signal molecules, while the bioaugmentation of LV-Q1 made the biofilm distribution more uniform. Analysis of the microbial community structure further reflected that the strain Vibrio sp. LV-Q1 successfully proliferated (the relative abundance increased to 59.81 %). The bioaugmentation of exogenous addition of QS strain enhanced the expression of functional microorganisms and promoted the enrichment of Nitrincola, Marinobacterium, and Vibrio in the biofilm in a high salt environment. This work provides a new option for improving the MBBR for treating nitrogen-containing wastewater in a high salt environment.

RevDate: 2025-08-15

Alamrani A, JG Eldiasty (2025)

Bacterial-extract mediated gold nanoparticles: Enhanced uptake, oxidative stress, and cytotoxicity in HepG2 cells with biofilm inhibition and antibiotic synergy.

Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy, 345:126763 pii:S1386-1425(25)01070-4 [Epub ahead of print].

Gold nanoparticles (AuNPs) are widely studied for cancer therapy due to their biocompatibility and unique physicochemical properties. However, limitations in cellular uptake and biological activity remain. Green synthesis using bacterial extracts offers a sustainable alternative that may improve therapeutic outcomes. This study compares the biological and physicochemical performance of chemically synthesized AuNPs (Chem-AuNPs) with those produced via bacterial biosynthesis (Bacterial-AuNPs) in HepG2 liver cancer cells and pathogenic bacteria. AuNPs were synthesized via classical citrate reduction or bacterial extract-mediated biosynthesis. Characterization included UV-Vis, TEM, DLS, XRD, and EDX. Cytotoxicity, ROS generation, mitochondrial membrane potential (ΔΨm), and gene expression were assessed in HepG2 cells. Biofilm inhibition and antibiotic synergy (checkerboard assay) were evaluated against multi-drug resistant bacterial strains. Bacterial-AuNPs were smaller and more monodisperse (PDI = 0.18) compared to Chem-AuNPs (PDI = 0.32)., with enhanced cellular uptake (95 % vs. 70 %) and ROS induction (3.0- vs. 2.0-fold). NAC co-treatment confirmed ROS-mediated cytotoxicity. Bacterial-AuNPs significantly inhibited biofilm formation and exhibited strong synergy with antibiotics (FICI ≤0.5) in key pathogens. The primary uptake mechanism was identified as clathrin-mediated endocytosis, with caveolae-mediated uptake contributing at higher concentrations. Bacterial-AuNPs demonstrate superior cellular uptake, ROS generation, and mitochondrial dysfunction, key mechanisms in cancer cell apoptosis. Biogenically synthesized AuNPs demonstrate superior anticancer and antimicrobial activity compared to chemically synthesized counterparts. These findings highlight the potential of bacterial-extract-mediated AuNPs as multifunctional therapeutic agents in oncology and infectious disease contexts.

RevDate: 2025-08-15

Bhattacharya S, Venkatasubramanian A, Chatterjee A, et al (2025)

Effects of various physicochemical parameters on the Biofilm formation and Pyocyanin production in Pseudomonas aeruginosa PA14.

Biotechnology letters, 47(5):91.

BACKGROUND: Biofilm formation in Pseudomonas aeruginosa provides protection against multiple stressors and contributes to its pathogenicity. Pyocyanin, a virulence factor regulated by quorum sensing, is crucial for infections. This study aimed to evaluate how various physicochemical conditions impact biofilm formation and pyocyanin production in P. aeruginosa PA14.

METHODS: Biofilm formation and pyocyanin production were assessed under varying conditions, including nutrient availability, NaCl concentrations, pH, temperature, heavy metal salts, light exposure, and microbial competition. Biofilm levels were quantified using a crystal violet assay, while pyocyanin levels were measured spectrophotometrically. Statistical analyses were performed to identify significant trends and correlations.

RESULTS: Key findings revealed that biofilm formation and pyocyanin production were reduced under most stress conditions examined in this study, compared to controls, with few exceptions. FeCl3 enhanced biofilm formation, while NaCl concentrations above 3% and extreme pH values inhibited it. NiCl2 was the most effective at reducing biofilm amount among the salts which we examined. Pyocyanin production followed similar trends, peaking under neutral pH and nutrient-enriched conditions. Positive correlations between biofilm and pyocyanin production were observed, particularly in nutrient-limited media. Additionally, light exposure and inter-microbial competition significantly reduced biofilm levels.

CONCLUSION: This study highlights the differential responses of P. aeruginosa to various stress conditions, underscoring the importance of environmental factors in modulating biofilm formation and virulence. These findings provide insights into bacterial adaptive strategies and offer potential avenues for developing targeted interventions against biofilm-associated infections.

RevDate: 2025-08-15

Mukherjee R, Klempt F, Fuchs F, et al (2025)

Biofilm development of Porphyromonas gingivalis on titanium surfaces in response to 1,4-dihydroxy-2-naphthoic acid-a hybrid in vitro-in silico approach.

Microbiology spectrum [Epub ahead of print].

Colonization of titanium dental implants by the oral pathogen Porphyromonas gingivalis can lead to peri-implant diseases and, ultimately, implant failure. P. gingivalis growth can be stimulated by 1,4-dihydroxy-2-naphthoic acid (DHNA), a menaquinone precursor from various oral bacteria; however, its impact on biofilm formation remains unclear. The aim of the study was to evaluate P. gingivalis growth and metabolic activity over 6 days in response to DHNA on two titanium grade IV surfaces with different roughness using a hybrid in vitro-in silico approach. P. gingivalis growth was modestly stimulated by DHNA and exhibited an inverse correlation with ammonia concentration in culture medium. Notably, this growth pattern transitioned from an initial linear phase to a later exponential phase, with DHNA-treated biofilms reaching this exponential shift at an earlier stage than untreated controls. Confocal microscopy revealed that DHNA-treated biofilms exhibited surface-dependent growth patterns, with larger biofilm volumes observed on rougher surfaces in later biofilm stages, compared with smoother surfaces. Regardless of surface characteristics, the area occupied by biofilms and the size of the aggregates exhibited a consistent and progressive increase over time and were larger in late DHNA-treated biofilms. The experimental data were used to calibrate a coupled finite element method (FEM)-based model that simulated P. gingivalis biofilm dynamics and nutrient utilization. Summarizing, DHNA moderately stimulated P. gingivalis growth, accelerated its transition to ammonia-independent growth, and promoted an increase in biofilm area and aggregate size. Our coupled approach offers significant potential for advancing in vitro biofilm research.IMPORTANCEThe results of our hybrid in vitro-in silico experiments advance the research on P. gingivalis physiology and its DHNA-dependent colonization of implant surfaces. Our findings reveal that DHNA accelerates P. gingivalis growth, induces aggregation, and promotes colonization of titanium surfaces. For the first time, DHNA-induced P. gingivalis growth acceleration and an earlier shift away from ammonia dependency were observed fluorometrically, highlighting ammonia assimilation as a promising marker of P. gingivalis physiology during early biofilm expansion. Understanding how growth factors together with surface properties influence P. gingivalis colonization offers a basis for future preventive strategies. Our study's stringent characterization of 3D surface texture parameters is expected to improve the reproducibility of biofilm-surface interactions experiments. The findings were validated using a continuum-based in silico model, initiating a hybrid approach where computational models complement in vitro research. Our interdisciplinary approach offers a versatile framework for investigating additional aspects of oral biofilms on titanium.

RevDate: 2025-08-15

Abd Allah RH, Samir S, Nasr SM, et al (2025)

Effect of Kaempferol against Biofilm Formation by Klebsiella pneumoniae Clinical Isolates.

Recent patents on biotechnology pii:BIOT-EPUB-150049 [Epub ahead of print].

BACKGROUND: Biofilm production is a key factor in the development of antibiotic resistance in multidrug-resistant Klebsiella pneumoniae (K. pneumoniae), a significant contributor to healthcare-associated infections (HAIs). Kaempferol, a flavonoid, is widely recognized for its ability to combat various microorganisms.

AIM: Our goal is to assess the impact of kaempferol on K. pneumoniae biofilms by determining the level of gene expression for the biofilm-forming genes.

METHODS: Fifty K. pneumoniae isolates were studied. Different doses of kaempferol with a concentration range of 0.04 to 100% in Luria Bertani broth (LB) medium were incubated at 37℃ for 24 h with forty-three K. pneumoniae strong and intermediate biofilm producers. The minimum inhibitory concentration (MIC) of kaempferol was determined. Molecular detection of the biofilm-forming genes (mrkA, pgaA, wbbM, and wzm) was performed on all isolates before and after kaempferol treatment at 0.5 x MIC.

RESULTS: Seven isolates out of 50 (14%) exhibited weak biofilm formation ability, 6 out of 50 (12%) were moderate producers, and 37 out of 50 (74%) were strong producers. The MIC values of kaempferol for K. pneumoniae ranged from 50% to 6.25% (p = 0.0003). The levels of expression of the studied genes were slightly decreased after treatment compared with their corresponding values before treatment.

CONCLUSION: Based on current knowledge, few research studies have investigated the impact of kaempferol on K. pneumoniae biofilms. Our results show that its effect on the biofilms of this bacterium is moderate to weak. Further research is necessary to determine potential synergies with other treatments.

RevDate: 2025-08-15

Moraes SM, Araújo TT, Abuna GF, et al (2025)

Impact of TiF4 Varnish on the Oral Biofilm Microbiome in High-Caries-Risk Patients.

JDR clinical and translational research [Epub ahead of print].

INTRODUCTION: Dental caries is a prevalent global disease, influenced by biofilm formation, dietary sugars, and host factors. Fixed orthodontic appliances increase the risk of noncavitated lesions, highlighting the need for effective prevention. Fluoride varnishes reduce demineralization and promote remineralization; however, their impact on the oral biofilm microbiome in high-caries-risk patients remains underexplored.

OBJECTIVE: To profile microbial biofilms involved in caries lesions after treatment with sodium fluoride (NaF) and titanium tetrafluoride (TiF4) varnishes using 16S rRNA sequencing of the oral biofilm microbiome.

METHODS: A randomized crossover study was conducted with 13 participants (12-18 y) with fixed orthodontic appliances and at least 1 active noncavitated lesion. Participants underwent 4 stages: G1 (nontreatment), G2 (professional prophylaxis; PP), G3 (PP + NaF varnish), and G4 (PP + TiF4 varnish). Clinical analyses (Nyvad and plaque indices) and supragingival biofilm sampling were performed. Bacterial DNA was extracted and amplified for 16S rRNA sequencing. Repeated-measures analysis of variance, Friedman/Wilcoxon with Bonferroni correction, Pearson chi-squared, and permutational multivariate analysis of variance tests were performed ( P < 0.05).

RESULTS: Shannon diversity (median, 25%-75%) values were as follows: G1 (6.25, 6.21-6.27), G2 (5.81, 5.77-5.83), G3 (5.63, 5.64-5.71), and G4 (5.76, 5.72-5.78). G2, G3, and G4 differed significantly from G1, with no difference among them ( P < 0.05). The most abundant genera were Veillonella (G1: 7.6%, G2: 10.6%, G3: 9.4%, G4: 5.7%), Corynebacterium (G1: 8.2%, G2: 7.3%, G3: 6.8%, G4: 10.4%), and Neisseria (G1: 4.0%, G2: 9.2%, G3: 9.6%, G4: 9.6%). Significant reductions were observed in the Prevotella/Haemophilus, Prevotella/Neisseria, and Prevotella/Rothia log-ratios compared with G1 ( P = 0.001). G2 reduced Prevotella/Haemophilus. G3 reduced Prevotella/Haemophilus and Prevotella/Neisseria. G4 reduced Prevotella relative to all 3 genera, indicating broader microbiome modulation.

CONCLUSION: PP, whether or not combined with fluoride varnishes, modified the biofilm microbiota. PP + TiF4 varnish affected a greater number of bacterial log-ratios associated with commensal-dysbiotic balance, although no significant differences were found between treatment groups.Knowledge Transfer Statement:The findings from this study can guide clinicians in selecting the most effective fluoride varnish for high-caries-risk patients. By understanding how sodium fluoride (NaF) and titanium tetrafluoride (TiF4) varnishes modulate the oral microbiome, clinicians can develop more targeted and effective prevention strategies. This knowledge has the potential to enhance patient outcomes by optimizing caries prevention during orthodontic treatment, allowing for more personalized and microbiome-focused approaches.

RevDate: 2025-08-16

Yang X, Shi H, Zimba BL, et al (2025)

Smart bacteriophage release from bovine β-lactoglobulin fibrils based hydrogel with anti-MRSA, biofilm degradation and hair follicle regeneration functions for infected burn wound.

International journal of biological macromolecules, 322(Pt 2):146855 pii:S0141-8130(25)07412-4 [Epub ahead of print].

The emergence and spread of antibiotic-resistant bacteria negatively impact the effectiveness of antibiotics in treating burn wound infections, which significantly hinders the healing process. To address this, a β-lactoglobulin fibrils/oxidized dextran/phage (BLGFs/ODEX/Phage) hydrogel dressing has been developed to treat burn wounds that are infected with methicillin-resistant Staphylococcus aureus (MRSA). First, a highly efficacious MRSA phage is isolated and purified, and it exhibits excellent bactericidal efficiency, storage stability, biofilm degradation ability, and biocompatibility. Then, a pH-responsive BLGFs/ODEX hydrogel is developed to load MRSA phage by utilizing a Schiff base reaction between BLGFs and ODEX. The hydrogel possesses excellent mechanical properties, self-healing ability, cell adhesion promotion, and biocompatibility. The results of burn wound model experiments indicate that the MRSA phages encapsulated within the hydrogel can be released in a controlled manner dependent on bacterial growth, effectively mitigating the accumulation of excessive bacterial toxins. Therefore, the BLGFs/ODEX/Phage hydrogel dressing would display superior abilities in regulating inflammation, promoting collagen deposition, facilitating angiogenesis, and enhancing hair follicle regeneration, thereby accelerating wound healing. Overall, this work demonstrates that the combination of bacteriophage therapy and hydrogel dressing would be a promising and innovative strategy for the treatment of infected burn wounds, presenting expansive prospects for clinical application.

RevDate: 2025-08-16

Patel RR, Mishra A, Singh SK, et al (2025)

Mechanistic evaluation of Clerodendrum serratum anti-biofilm potency against Mycobacterium species.

Microbial pathogenesis, 208:107985 pii:S0882-4010(25)00710-7 [Epub ahead of print].

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

RevDate: 2025-08-14

Larsson Y, Nikolausz M, Kisielius V, et al (2025)

Metabolic pathways for biotransformation of benzalkonium compounds in fungal- and bacteria-based biofilm reactors.

Journal of hazardous materials, 496:139494 pii:S0304-3894(25)02410-0 [Epub ahead of print].

Benzalkonium compounds (BACs) are quaternary ammonium biocides widely used in healthcare and industrial animal production, resulting in high concentrations in manure and wastewater. While BAC biodegradation has been demonstrated in wastewater and soil, little is known about the relevant biodegradation pathways in multi-species biofilm systems. This study investigated BAC degradation pathways in three aerobic moving bed biofilm reactor (MBBR) systems: (1) a fungi (Trametes versicolor) inoculated reactor, and heterotrophic biofilm in (2) water, and (3) manure. We characterised transformation products and elucidated their fragmentation mechanisms to explain the spectra. ω-oxidation and β/α-oxidation dominated in the water-MBBR, whereas formation of benzyldimethylamine (BDMA), was observed only in the manure-MBBR, which was the only system hosting Pseudomonas and Aeromonas species. The water-MBBR microbial community showed a significant increase in abundance of Rhodococcus, known for ω- and β-oxidation, suggesting a role in BAC degradation. None of the five ARGs (sul1, sul2, qnrD, tetM, tetA) tested in this study were elevated in the systems by exposure to BAC. These results demonstrate that BACs are degradable in different aerobic biofilm systems via diverse pathways, influenced by microbial composition and matrix complexity. However, the different metabolites indicate towards complex risk assessments and bioremediation strategies.

RevDate: 2025-08-17

Liu B, Bai M, Tu W, et al (2025)

The Virulence Factor LLO of Listeria monocytogenes Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect.

Foods (Basel, Switzerland), 14(15):.

Listeria monocytogenes is a life-threatening bacterial foodborne pathogen that can persist in food-processing facilities for years. Although phages can control L. monocytogenes during food production, phage-resistant bacterial subpopulations can regrow in phage-treated environments. In this study, an L. monocytogenes hly defective strain, NJ05-Δhly, was produced, which considerably regulated the interactions between L. monocytogenes and phages. Specifically, we observed a 76.92-fold decrease in the efficiency of plating of the defective strain following infection with the Listeria phage vB-LmoM-NJ05. The lytic effect was notably diminished at multiplicities of infection of 1 and 10. Furthermore, the inactivation of LLO impaired biofilm formation, which was completely suppressed and eliminated following treatment with 10[8] PFU/mL of phage. Additionally, phages protected cells from mitochondrial membrane damage and the accumulation of mitochondrial reactive oxygen species induced by L. monocytogenes invasion. Transcriptomic analysis confirmed these findings, revealing the significant downregulation of genes associated with phage sensitivity, pathogenicity, biofilm formation, and motility in L. monocytogenes. These results underscore the vital role of LLO in regulating the pathogenicity, phage susceptibility, and biofilm formation of L. monocytogenes. These observations highlight the important role of virulence factors in phage applications and provide insights into the potential use of phages for developing biosanitizers.

RevDate: 2025-08-17

Urbaniak M, Lechowicz Ł, Gawdzik B, et al (2025)

Structure-Activity Relationships in Alkoxylated Resorcinarenes: Synthesis, Structural Features, and Bacterial Biofilm-Modulating Properties.

Molecules (Basel, Switzerland), 30(15):.

In this study, a series of novel alkoxylated resorcinarenes were synthesized using secondary and tertiary alcohols under mild catalytic conditions involving iminodiacetic acid. Structural characterization, including single-crystal X-ray diffraction, confirmed the successful incorporation of branched alkyl chains and highlighted the influence of substitution patterns on molecular packing. Notably, detailed mass spectrometric analysis revealed that, under specific conditions, the reaction pathway may shift toward the formation of defined oligomeric species with supramolecular characteristics-an observation that adds a new dimension to the synthetic potential of this system. To complement the chemical analysis, selected derivatives were evaluated for biological activity, focusing on bacterial growth and biofilm formation. Using four clinically relevant strains (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Bacillus subtilis), we assessed both planktonic proliferation (OD600) and biofilm biomass (crystal violet assay). Compound 2c (2-pentanol derivative) consistently promoted biofilm formation, particularly in S. aureus and B. subtilis, while having limited cytotoxic effects. In contrast, compound 2e and the DMSO control exhibited minimal impact on biofilm development. The results suggest that specific structural features of the alkoxy chains may modulate microbial responses, potentially via membrane stress or quorum sensing interference. This work highlights the dual relevance of alkoxylated resorcinarenes as both supramolecular building blocks and modulators of microbial behavior.

RevDate: 2025-08-13

Frantar A, Seme K, Gašperšič R, et al (2025)

Isolation and characterization of two novel oral bacteriophages with anti-biofilm activity against Cutibacterium acnes.

International journal of medical microbiology : IJMM, 320:151668 pii:S1438-4221(25)00024-4 [Epub ahead of print].

Bacteriophage therapy offers a promising solution to combat antibiotic-resistant infections, yet its potential against biofilm-associated pathogens in oral diseases remains underexplored. This study investigates the opportunistic bacterium Cutibacterium acnes, an overlooked contributor to dental implant and prosthetic joint infections. Biofilms formed by C. acnes are highly resilient and resistant to antibiotics, complicating treatment. Two novel lytic bacteriophages, Ristretto and Corretto, targeting C. acnes, were isolated from human saliva, with morphological analysis confirming their classification as siphoviruses. Their genome sequencing revealed no harmful antimicrobial resistance or virulence genes, making them suitable for therapeutic use. Remarkably, phage Corretto demonstrated a broad host range and achieved near-complete eradication of mature biofilms across multiple C. acnes strains, outperforming Ristretto in efficacy and strain coverage. The activity of these phages was dosage-dependent and varied across bacterial strains, revealing potential strain-specific resistance mechanisms within biofilms. These findings highlight bacteriophage therapy's potential to disrupt persistent biofilms where antibiotics fail, offering a new approach for treating biofilm-driven infections in dental and medical implantology. This study underscores the need for further research into phage-based strategies to address the growing global challenge of antimicrobial resistance and improve outcomes in biofilm-related diseases.

RevDate: 2025-08-13

Shen Y, Zhang X, Feng X, et al (2025)

Unraveling tetracycline-induced biofilm stability and resistance mechanisms in aerobic granular sludge via proteomics.

Water research, 287(Pt A):124385 pii:S0043-1354(25)01291-6 [Epub ahead of print].

Tetracycline (TC) stress disrupts microbial communities, yet its impact on aerobic granular sludge (AGS) formation and stability remain poorly understood. This study employs quantitative proteomics to unravel the AGS adaptive mechanisms under continuous TC exposure (1 mg/L). The results demonstrated that TC accelerated AGS granulation, achieving stable structure within 20 days, accompanied by improved pollutants removal (COD: 87.4%; TN: 84.1%; TP: 66.5%; TC: 83.0%) and settling performance (SVI: 50 mL/g). A critical granule size threshold of 3-4 mm was identified, beyond which AGS exhibited destabilization tendencies. Extracellular polymeric substances (EPS), particularly proteins, play pivotal role in maintaining AGS stability, with the α-helix / (β-sheet + random coil) ratio correlated with granule integrity. Proteomic analysis revealed the upregulation of outer membrane protein A (OmpA), facilitating biofilm formation, while TC-targeted ribosomes and bacterial chemotaxis were identified as central mechanisms for TC resistance and stress adaptation, respectively. These findings uncover molecular adaptations driving AGS stability under antibiotic pressure and offer insights for optimizing high-antibiotic wastewater treatment.

RevDate: 2025-08-13

Yang W, Zheng X, Jia D, et al (2025)

Engineered nanoplatform with DNase-mimetic catalysis and photothermal ablation for synergistic biofilm eradication.

Colloids and surfaces. B, Biointerfaces, 256(Pt 2):115019 pii:S0927-7765(25)00526-0 [Epub ahead of print].

Bacterial infections represent an increasing global health threat, exacerbated by the decline in antibiotic effectiveness due to widespread resistance. Biofilms, structured microbial communities embedded in extracellular polymeric substances (EPS), significantly hinder treatment by limiting antibiotic penetration and promoting bacterial persistence. With over 80 % of bacterial infections involving biofilms, there is an urgent need for antibiotic-free approaches that can disrupt these protective matrices. This study presents a dual-functional nanoplatform (Au-Ce NR) combining gold nanorods (Au NRs) as photothermal agents with synthetic deoxyribonuclease (DNase) mimics (Ce[4][+]/nitrilotriacetic acid (NTA) complexes) for synergistic biofilm eradication. The system was developed through covalent conjugation of Ce[4+]/NTA complexes onto polyethylene glycol-functionalized Au NRs. Within biofilm microenvironments, Ce[4+]/NTA complexes selectively degrade extracellular DNA (eDNA), a vital EPS component, thereby destabilizing the biofilm and facilitating nanoplatform penetration. Near-infrared irradiation subsequently induces localized hyperthermia via Au NRs, effectively eliminating dispersed bacteria while minimizing the risk of resistance development. In vitro experiments demonstrated efficient eDNA degradation in methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa biofilms, significantly enhancing bactericidal activity compared to unmodified Au NRs. In vivo assessments using an MRSA-infected wound model confirmed the therapeutic efficacy through reduced inflammation and accelerated wound healing. By combining enzymatic matrix disruption with photothermal ablation, this strategy addresses key limitations of conventional treatments for biofilm-associated infections in the post-antibiotic era.

RevDate: 2025-08-13

Huo S, Chang L, Liu Y, et al (2025)

Polyamine-Activated Carbonyl Stress Nanoplatform Synergistically Reverses Biofilm-Driven Immunosuppressive Microenvironment.

ACS nano [Epub ahead of print].

Polyamine metabolic dysregulation induced by implant-associated infections (IAIs) is a pivotal contributor to the formation of an immunosuppressive microenvironment. Excessive polyamines facilitate pathogen persistence by suppressing bacterial membrane lipid peroxidation (LPO) and enhancing DNA repair mechanisms. Simultaneously, polyamines promote biofilm formation via quorum sensing (QS) modulation and inhibit host immunity to facilitate immune escape. Herein, we developed a polyamine-responsive carbonyl stress nanoplatform MIL-100@PAO@PVP (MPP), which synergistically integrates metabolic intervention and chemodynamic therapy (CDT), addressing the limitations inherent to conventional oxidative damage-based therapies. Specifically, plasma amine oxidase (PAO) within MPP catalyzes polyamine degradation at infection sites, generating highly toxic acrolein and hydrogen peroxide (H2O2). The produced H2O2 markedly enhances MIL-100-mediated CDT, triggering a burst of hydroxyl radicals ([•]OH) that induces severe bacterial membrane LPO and DNA damage. Importantly, the generated acrolein further amplifies bacterial DNA damage via the induction of carbonyl stress. Additionally, bacterial debris resulting from MPP-induced cell death acts as endogenous antigens, effectively activating the antigen-presenting functions of macrophages and dendritic cells (DCs), thus reshaping the local immune response and reversing immunosuppression. Experimental results demonstrated robust antibiofilm efficacy and immunostimulatory effects of MPP in both in vitro and in vivo models, highlighting a promising therapeutic strategy for treating IAIs.

RevDate: 2025-08-13

Conceição K, de Andrade VM, de Oliveira VDM, et al (2025)

Unleashing IbKTP-NH2 a kyotorphin derivative, against bacterial and fungal multispecies biofilm adhesion and viability on materials.

Journal of applied microbiology pii:8233669 [Epub ahead of print].

AIMS: Healthcare-Associated Infection (HAI) are a major global health concern, contributing to increased mortality and substantial economic burden on healthcare systems. This study aims to evaluate the efficacy of the peptide kyotorphin conjugated with ibuprofen (IbKTP-NH2) in inhibiting multispecies biofilms formed by Candida albicans, Pseudomonas aeruginosa, and Streptococcus pneumoniae, particularly in the context of biofilm-associated infections linked to implanted medical devices.

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

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

RevDate: 2025-08-15

Soon KL, Rashid SA, Nazakat R, et al (2025)

Hidden risks of biofilm-forming and multidrug-resistant Gram-negative bacteria in water vending machines.

Antonie van Leeuwenhoek, 118(9):128.

Water vending machines are widely utilised in various locations, including workplaces, universities and urban areas due to their convenience and accessibility. However, the microbial quality of the water dispensed from these machines, particularly in residential colleges, has not been thoroughly studied. This study aims to analyse the physicochemical and microbiological quality of drinking water collected from water vending machines located in 14 residential colleges around Universiti Malaya, Kuala Lumpur. Our study revealed that all drinking water samples met WHO acceptable limits for pH, TDS and electrical conductivity. However, dissolved oxygen and free chlorine levels were slightly below the acceptable range. No faecal contamination was detected in the drinking water samples, as indicated by the absence of enterococci and coliform bacteria, including faecal coliforms. Nevertheless, Gram-negative non-coliform bacteria were identified in water samples from 11 residential colleges. The identified bacteria included Pseudomonas spp., Acinetobacter spp., Bosea sp., Stenotrophomonas sp., Achromobacter sp., Roseatels sp. and Cupriavidus sp. Using CLSI guidelines as the standard, antibiotic sensitivity tests showed that all bacterial isolates were resistant to penicillin G (100%), while most were susceptible to ofloxacin (> 80%). Notably, Pseudomonas spp., Acinetobacter spp., Stenotrophomonas sp., Achromobacter sp. and Cupriavidus sp. were identified as multidrug-resistant bacteria, showing resistance to three or more categories of antimicrobial agents tested in this study. The biofilm assay confirmed that Bosea sp., Roseateles sp. and Acinetobacter sp. possess biofilm formation capacity. Overall, this study highlights that while the water from vending machines generally meets physicochemical standards and is free of faecal contamination, multidrug-resistant Gram-negative bacteria are still prevalent. Although these bacteria may not pose immediate health risks upon consumption, their presence poses long-term risks due to biofilm accumulation, resistance gene transfer and poor maintenance. This underscores the need for proper maintenance measures of water vending machines, including regular cleaning and effective disinfection, to ensure the microbial safety of drinking water.

RevDate: 2025-08-16

Leiva-Sabadini C, Berríos P, Saavedra P, et al (2025)

Biofilm formation on collagen substrates modulates Streptococcus mutans bacterial extracellular nanovesicle production and cargo.

Nanoscale advances [Epub ahead of print].

Streptococcus mutans is the major microbial etiological agent of dental caries and can adhere to surfaces such as type-I collagen, which is present in dentin and periodontal tissues. Recent studies have characterized planktonic S. mutans bacterial extracellular vesicles (bEVs) at the nanoscale range and demonstrated environmental-induced changes due to sugar presence or pH alterations. However, to date, no studies have explored whether surface-derived changes can modulate bEV production in the context of oral biofilm formation in the elderly. Therefore, this work aimed to determine the role of biofilm formation and collagen glycation on the nanoscale morphology and proteomic composition of S. mutans bEVs. For this, bEVs from S. mutans biofilms on native and glycated collagen surfaces were isolated, characterized, and compared to bEVs from planktonic cells. Nanoparticle tracking analysis (NTA), atomic force microscopy (AFM), and electron microscopy confirmed bEV production and showed that bEVs from biofilms are smaller in size and less abundant than those from planktonic cells. Furthermore, proteome analysis revealed that S. mutans biofilm formation on native and glycated collagen led to the enrichment of several key virulence proteins. Also, a shift towards proteins involved in metabolic processes was found in bEVs following biofilm formation on collagen surfaces, whereas glucan metabolism proteins were overexpressed in vesicles from the planktonic state. These results demonstrate that biofilm formation, as well as the glycation of collagen associated with aging and hyperglycaemia, can modulate bEV characteristics and cargo and could play a central role in S. mutans virulence and the development of diseases such as dental caries and periodontal disease.

RevDate: 2025-08-16

Doll-Nikutta K, Mikolai C, Heine N, et al (2025)

Biocompatible liquid-infused titanium minimizes oral biofilm adhesion in flow chamber and 3D implant-tissue-biofilm in vitro models.

Bioactive materials, 53:706-717.

Biomedical implants are susceptible to bacterial colonization, which can lead to challenging implant-associated infections. In particular, dental implant abutments - which are continuously exposed to bacteria within the oral cavity - stand to greatly benefit from strategies which inhibit the development of bacterial biofilms. Liquid-infused titanium surfaces have demonstrated excellent biofilm repellency, but to date have not been analyzed with substances suitable for medical device approval in terms of biocompatibility under conditions mimicking the environment of dental implant abutments. In this study, different medical-grade lubricants coated onto laser-structured titanium were screened for stability and water-repellency - with the results suggesting that unmodified structured titanium coated with silicone oil was the most promising combination of materials. When analyzing biofilm formation, the coated surfaces showed a statistically significant reduction in oral commensal Streptococcus oralis biofilms grown under static conditions as well as oral multispecies biofilms grown under salivation-resembling flow conditions. This biofilm-reducing effect was also observed when the coated surface interfaced with a 3D implant-tissue-oral-bacterial-biofilm (INTERbACT) in vitro model, which allows for the direct interaction of human tissue and oral multispecies biofilm at the implant interface. Importantly, this biofilm reduction was not due to toxicity of the coated surfaces, but is most likely attributable to inhibition of bacterial attachment. Additionally, the surfaces were not cytotoxic, without altering adjacent soft tissue or causing elevated pro-inflammatory cytokine secretion. These findings highlight the promise of biocompatible liquid-infused titanium surfaces as biofilm-repellent implant abutment modifications and provide the basis for further investigations in targeted pre-clinical studies.

RevDate: 2025-08-16

Miranda A, Brandquist ND, Johnson K, et al (2025)

Quaternized chitosan derivatives inhibit growth and affect biofilm formation of Staphylococcus aureus.

Scientific reports, 15(1):29606.

Antimicrobial resistance (AMR) poses a global health threat, severely impeding the effective treatment of bacterial infections and jeopardizing the safety of routine medical procedures. Methicillin-resistant Staphylococcus aureus (MRSA) is particularly problematic because of its resistance to beta-lactams and the ability to form resilient biofilms. Conventional antibiotics, including last-resort options, have serious side effects and may contribute to further resistance. Chitosan, a natural biopolymer, offers a promising alternative due to its biocompatibility and antimicrobial properties, though its effectiveness against biofilms is limited. Recent studies suggest that increasing the positive charge density and adding hydrophobic moieties to chitosan, can enhance its antimicrobial properties. In this work, the antibacterial activity of quaternized chitosan derivatives against AMR S. aureus strains was assessed. Quaternization of chitosan's amino group and introduction of hydrophobic side chains was found to significantly inhibit bacterial growth in both methicillin-sensitive (MSSA) and MRSA strains. Notably, nanofibrous materials composed of polyethylene oxide and hexyl-modified chitosan demonstrate alterations in S. aureus biofilm development, leading to significant accumulation of dead cells. Combined, these results highlight the potential of modified chitosan derivatives as effective antimicrobial agents for surface treatments and medical device coatings, particularly in applications where antibiotics are traditionally used, such as biofilm-prone environments.

RevDate: 2025-08-12

Zhang Y, Xu R, Zhu X, et al (2025)

UV-Induced VBNC state formation and resuscitation in E. coli: ATP-based metabolic activation and biofilm-mediated recovery.

Water research, 287(Pt A):124380 pii:S0043-1354(25)01286-2 [Epub ahead of print].

In this study, we systematically investigated the induction mechanism, metabolic characterization, and resuscitation behavior of viable but non-culturable bacteria (VBNC) by ultraviolet (UV) disinfection under different conditions, using E. coli as our example. Flow cytometry (FCM), ATP metabolism analyses and genetic assays revealed that low-dose UV (4.5 mJ/cm[2]) significantly inhibited bacterial culturability while maintaining cell membrane integrity and activating metabolic activity (total ATP levels were elevated to 182 % of initial values). Higher doses of UV (14.1 mJ/cm[2]), on the other hand, resulted in metabolic inhibition (ATP decreased to 58 %), but did not completely inactivate the bacteria. Gene expression analysis showed that the DNA repair gene recA and the antioxidant gene katG were significantly up-regulated with increasing UV money dose, while the stress response gene rpoS decreased, suggesting that the SOS repair system dominated the maintenance of the VBNC state. Resuscitation tests showed that biofilm greatly sped up the recovery of VBNC bacteria by releasing tyrosine-like extracellular polymers and quorum sensing signaling molecules, with resuscitation time shortened to 4 h in LB medium and 17 h in sterile water. This study reveals that UV disinfection-induced VBNC bacteria are characterized by enhanced metabolic activity and elucidates the mechanism by which biofilm promotes their resuscitation by secreting tyrosine-like EPS and C14 signaling molecules.

RevDate: 2025-08-12

Xu H, Chen X, Jia Y, et al (2025)

A trifunctional coating for biofilm prevention: integrating antifouling, photothermal killing, and quorum sensing interference.

Journal of colloid and interface science, 701:138628 pii:S0021-9797(25)02019-3 [Epub ahead of print].

Bacterial biofilm-associated infections pose a significant challenge in biomedical fields, especially in the context of medical devices and implants, where conventional treatments often fail to combat mature biofilms. To overcome this limitation, an antibiofilm coating was developed, incorporating three functional components: carbon nanoparticles derived from candle soot (CS), hydrophilic and pH-responsive copolymer brushes of 2-hydroxyethyl methacrylate (HEMA) and 3-(acrylamido)-phenylboronic acid (APBA), and natural quorum sensing inhibitors (QSIs). The carbon nanostructure of the CS substrate provides inherent photothermal conversion properties, while its high surface area facilitates efficient chemical functionalization. The copolymer design strategically incorporates hydrophilic poly(HEMA) domains that form bacteria-repellent hydration layers, alongside APBA moieties that covalently bind plant-derived QSIs (quercetin or baicalein) through pH-sensitive boronate ester linkages. This combination of components enables the coating to suppress biofilm formation via three synergistic mechanisms: bacterial adhesion is initially inhibited by hydration barriers, surface-bound pathogens are subsequently eradicated through photothermal conversion upon near-infrared activation, and the release of QSIs triggered by the acidic microenvironment disrupts biofilm maturation. The coating demonstrated sustained antibiofilm efficacy against both Pseudomonas aeruginosa and Staphylococcus aureus, while preserving mammalian cell viability and substrate compatibility across various biomedical materials. By integrating physical barriers, thermal ablation, and quorum sensing interference through precise material engineering, this coating offers a promising solution for preventing biofilm-associated infections in clinical settings.

RevDate: 2025-08-12

De Marque MB, Silva BG, E Foresti (2025)

Ammonia diffusivity in biofilm reactors: impact of polyurethane foam thickness and pore clogging on simultaneous nitrification and denitrification.

Bioprocess and biosystems engineering [Epub ahead of print].

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

RevDate: 2025-08-12

Wen X, Tang T, Bao T, et al (2025)

gltB encoding glutamate synthase is involved in persister and biofilm formation and virulence in Staphylococcus aureus.

Microbiology spectrum [Epub ahead of print].

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

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In the early 1990's, Robert Robbins was a faculty member at Johns Hopkins, where he directed the informatics core of GDB — the human gene-mapping database of the international human genome project. To share papers with colleagues around the world, he set up a small paper-sharing section on his personal web page. This small project evolved into The Electronic Scholarly Publishing Project.

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In 1995, Robbins became the VP/IT of the Fred Hutchinson Cancer Research Center in Seattle, WA. Soon after arriving in Seattle, Robbins secured funding, through the ELSI component of the US Human Genome Project, to create the original ESP.ORG web site, with the formal goal of providing free, world-wide access to the literature of classical genetics.

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Although the methods of molecular biology can seem almost magical to the uninitiated, the original techniques of classical genetics are readily appreciated by one and all: cross individuals that differ in some inherited trait, collect all of the progeny, score their attributes, and propose mechanisms to explain the patterns of inheritance observed.

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This is a must read book for anyone with an interest in invasion biology. The full title of the book lays out the author's premise — The New Wild: Why Invasive Species Will Be Nature's Salvation. Not only is species movement not bad for ecosystems, it is the way that ecosystems respond to perturbation — it is the way ecosystems heal. Even if you are one of those who is absolutely convinced that invasive species are actually "a blight, pollution, an epidemic, or a cancer on nature", you should read this book to clarify your own thinking. True scientific understanding never comes from just interacting with those with whom you already agree. R. Robbins

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Papers in Classical Genetics

The ESP began as an effort to share a handful of key papers from the early days of classical genetics. Now the collection has grown to include hundreds of papers, in full-text format.

Digital Books

Along with papers on classical genetics, ESP offers a collection of full-text digital books, including many works by Darwin and even a collection of poetry — Chicago Poems by Carl Sandburg.

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ESP now offers a large collection of user-selected side-by-side timelines (e.g., all science vs. all other categories, or arts and culture vs. world history), designed to provide a comparative context for appreciating world events.

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Bibliographies on several topics of potential interest to the ESP community are automatically maintained and generated on the ESP site.

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