Search Results (1,035)

Background/Objectives: Conventional cancer treatments such as radiation therapy, chemotherapy, and surgery often severely reduce a patient’s quality of life due to their side effects. This underscores the urgent need to develop less toxic and more effective alternative therapies. Medicinal plants and their derivatives represent a valuable source for discovering such agents. In this study, we investigated the anticancer and antibacterial properties of ethanolic (EE), methanolic (ME), and aqueous (AE) leaf extracts from the Lebanese Eucalyptus camaldulensis. Methods: The cytotoxicity of the extracts and cis-diamminedichloroplatinum(II) (CDDP) was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against four human cancer cell lines: two breast cancer lines (MCF-7 and MDA-MB-231), colorectal carcinoma cells (HCT-116), and lung carcinoma epithelial cells (A-549). Morphological changes in the cells were observed via inverted phase-contrast microscopy. The antibacterial potential was determined through minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) broth microdilution assays, agar well diffusion, and time-kill tests. The antibiofilm activity of the extracts against Staphylococcus haemolyticus, Streptococcus intermedius, Escherichia coli, and Leclercia adecarboxylata was assessed using antibiofilm screening. Results: All three extracts demonstrated significant anticancer activity against the four cell lines, with particular potency against A-549 cells. The AE showed the greatest cytotoxic effect, with IC50 values ranging from 97.1 to 163.3 µg/mL after 24 h of treatment. Furthermore, all extracts induced notable morphological changes in the cancer cells. The MIC and MBC results indicated a bactericidal effect for the EE (MBC range: 1.5625–50%), ME (MBC range: 6.25–50%), and AE (MBC range: 3.125–100%). Agar well diffusion assays revealed inhibition zones of 8.1 to 20.1 mm, while time-kill tests showed bacterial inhibition within 1 to 24 h of incubation. The antibiofilm assays demonstrated that the extracts inhibited biofilm production and eradicated 50% of pre-formed biofilms. Conclusions: These findings suggest that Lebanese Eucalyptus camaldulensis leaf extracts are a promising source of bioactive compounds with both antibacterial and anticancer properties. Full article

Extensive clinical studies have convincingly demonstrated that fungal infections constitute a mounting global health concern. Candida albicans, C. auris, Aspergillus fumigatus, and Cryptococcus neoformans were classified by the WHO as critical priority fungi due to their widespread prevalence, high mortality rates associated with invasive mycoses, and the increasing spread of resistant strains. The currently available antifungal arsenal, comprising four major classes (azoles, polyenes, echinocandins, and flucytosine), is critically insufficient. Resistance to these agents is escalating worldwide, while the approval of new drugs remains alarmingly slow. This review provides a comprehensive overview of the molecular mechanisms underlying resistance to conventional and emerging antimycotics in critical priority fungi, including recently approved and investigational agents such as oteseconazole, rezafungin, ibrexafungerp, fosmanogepix, and olorofim. It examines current strategies to overcome resistance, including combination therapy, drug repurposing, targeted delivery, antibiofilm approaches, immunotherapy, and the application of natural antifungal compounds. Special emphasis is placed on antifungal peptides (AFPs) derived from plants, animals, bacteria, and fungi, along with their synthetic analogs, all of which have demonstrated efficacy against critical priority fungi. The review primarily focuses on the molecular targets of AFPs and on the development of fungal resistance under prolonged exposure, compared with conventional antifungals, in order to evaluate their potential as prototypes for next-generation antimycotics. Full article

Background/Objectives: Antibiotic resistance is a growing global health challenge, especially with tricky bacteria like Pseudomonas aeruginosa. Anthranilate, previously identified as a signaling molecule in P. aeruginosa, modulates various pathogenicity-related phenotypes by reducing virulence factor production, inhibiting biofilm formation, and increasing antibiotic susceptibility. Based on these good properties as an anti-virulence agent, this study evaluated the actual therapeutic potential of anthranilate using mouse skin infection and lung infection models. Specifically, the efficacy of anthranilate in a standalone treatment was investigated without reliance on conventional antibiotics. Methods: The therapeutic efficacy of anthranilate was evaluated in murine skin and lung infection models of P. aeruginosa. Anthranilate was administered as a standalone treatment, and its effects on bacterial burden, inflammatory responses, and tissue lesion progression were assessed and compared with those of gentamicin treatment. Results: Anthranilate effectively reduced bacterial suppuration, mitigated inflammatory responses, and suppressed lesion progression in infected skin and lung tissues. Notably, anthranilate alone produced therapeutic effects comparable to those of gentamicin. Conclusions: These findings highlight its potential of standalone treatment as an alternative to traditional antibiotics and offer a novel anti-virulence strategy that minimizes selective pressure for resistance development. Full article

This study evaluated the anti-biofilm activity of the lipopeptide of Streptomyces lavendulae subsp. lavendulae 214 and aminoglycolipid biosurfactant of Streptomyces griseoflavus 2265 strains against biofilm formation by Xanthomonas strains. The biosurfactants of Streptomyces griseoflavus 2265 and Streptomyces lavendulae subsp. lavendulae 214 at a minimal concentration of 40 mg/L, characterized by antibiofilm activity against the development of Xanthomonas arboricola pv. juglandis 2417 strain and its mutant strains. The structure of the isolated biosurfactant was identified using Fourier Transform Infrared Spectroscopy (FTIR) and Nuclear Magnetic Resonance (NMR). The mutations in the rpfA gene of Xanthomonas arboricola pv. juglandis 8636 strain and deletions in the rpfGC genetic segment of Xanthomonas arboricola pv. juglandis 8737 strain did not ensure resistance to the effects of antimicrobial agents secreted by actinomycetes strains during the study. The concentration of aminoglycolipid biosurfactant of Streptomyces griseoflavus 2265 strain up to 20 mg/L and lipopeptide biosurfactant of Streptomyces lavendulae subsp. lavendulae 214 strain up to 10 mg/L stimulated cell motility in Xanthomonas arboricola pv. juglandis strains, while the increase in their value was accompanied by its inhibition, which was not affected by mutations in the rpfA gene of Xanthomonas arboricola pv. juglandis 8636 strain and deletions in the rpfGC genetic segment of Xanthomonas arboricola pv. juglandis 8737. These results may provide fundamental information for understanding the antagonistic effect of biological surfactants from soil microorganisms, which are dominated by actinomycetes, on the development and spread of Xanthomonas arboricola pv. juglandis strains of different genotypes on agricultural crops. Full article

Background/Objectives: Skin and soft tissue infections (SSTIs) remain a major clinical challenge due to the increasing prevalence of antimicrobial resistance and biofilm-associated pathogens. Conventional antibiotics are often limited by reduced efficacy, recurrent infections, and disruption of the resident skin microbiota. Consequently, bacteriocins have emerged as promising alternative or adjunctive antimicrobial agents for the treatment of skin and wound infections. Methods: A literature review was conducted using the PubMed, Scopus, and Web of Science databases. Experimental in vitro, ex vivo, and in vivo studies investigating bacteriocins in skin and wound infection models were analyzed, with a focus on antimicrobial activity, antibiofilm efficacy, activity against antimicrobial-resistant pathogens, and formulation strategies designed to improve therapeutic performance. Results: Available evidence demonstrates that numerous bacteriocins exhibit potent antimicrobial activity against clinically relevant skin-associated pathogens, particularly Staphylococcus aureus, including methicillin-resistant strains (MRSA). Several bacteriocins also showed significant antibiofilm properties and synergistic interactions with conventional antibiotics, resulting in enhanced bacterial eradication and reduced risk of resistance development. Experimental infection models further support their therapeutic potential in wound-associated infections. Additionally, advanced delivery platforms, including hydrogels, wound dressings, nanofibers, and lipid-based nanoparticles, improved peptide stability, sustained release, and local antimicrobial efficacy. Conclusions: Bacteriocins are promising candidates for the prevention and treatment of skin and wound infections due to their antimicrobial and antibiofilm activity, low propensity for resistance development, and suitability for topical administration. However, further studies addressing formulation optimization, safety, pharmacokinetics, and clinical validation are required before bacteriocin-based therapies can enter routine clinical practice. Full article

Antimicrobial peptides (AMPs) are increasingly regarded as next-generation antimicrobial agents because of their broad-spectrum activity, rapid killing, antibiofilm potential, immunomodulatory properties, and mechanisms of action that differ from those of many conventional antibiotics. Despite these advantages, their clinical translation remains limited by proteolytic instability, hemolysis or cytotoxicity, poor pharmacokinetics, salt and serum sensitivity, production costs, and delivery challenges. The AMP field is therefore shifting from natural peptide discovery toward integrated engineering pipelines that combine rational peptide modification, biomaterial-based delivery, high-throughput screening, and artificial intelligence (AI), particularly machine learning (ML) and deep learning approaches. Chemical and structural modifications, including D-amino acid substitution, N-glycine substitution, cyclization, lipidation, PEGylation, terminal amidation, hydrocarbon stapling, hybridization, sequence truncation, metal coordination, and biomaterial immobilization, are being used to improve stability, potency, selectivity, antibiofilm activity, and tissue localization. In parallel, AI-guided approaches, including ML, deep learning, and generative modeling, enable large-scale exploration of diverse peptide sources, including microbiomes and extinct proteomes, to entirely new sequences, while supporting optimization of potency, selectivity, stability, toxicity, and synthesizability. This focused review summarizes recent advances in AMP engineering, biomaterial-assisted delivery, and AI-guided discovery for biofilm-associated infections in the context of antimicrobial resistance. Full article

Vibriosis causes substantial losses in marine and brackish-water aquaculture, while increasing antimicrobial resistance limits the effectiveness of conventional antibiotic treatment. Biofilm formation and quorum-sensing-controlled virulence further contribute to the persistence of pathogenic Vibrio species. Prophages integrated into Vibrio genomes encode proteins and regulatory elements that may provide alternative approaches for controlling these pathogens. This review evaluates endolysins, polysaccharide depolymerases, nucleases, holins, spanins, tailocins, regulatory proteins, and small RNAs associated with prophages and related phages. The available evidence was classified according to molecular origin to distinguish validated prophage-derived molecules from those obtained from temperate or lytic phages and from molecules characterized in non-Vibrio bacteria. Endolysins and depolymerases can disrupt bacterial cells and biofilm matrices, whereas prophage regulatory elements may influence quorum sensing, adhesion, motility, toxin production, and secretion systems. However, the evidence directly supporting prophage-derived anti-Vibrio agents remains limited. Most experimentally demonstrated activity has been reported for endolysins and one validated depolymerase, and many of these molecules originated from lytic rather than temperate phages. No direct Vibrio-specific evidence is currently available for phage-derived nucleases, holins, spanins, or tailocins as isolated control agents. Prophage genomes nevertheless provide an extensive source of regulatory and antimicrobial candidates for further investigation. Progress toward aquaculture application will require experimental validation, effective delivery methods, safety assessment, scalable production, and clear regulatory standards. Full article

The increasing prevalence of biofilm-associated infections caused by Cutibacterium acnes has stimulated interest in food-derived natural products as alternative sources of anti-biofilm agents. This study investigated the effects of processing and extraction conditions on the phytochemical composition, antibacterial activity, and anti-biofilm properties of Dioscorea hispida Dennst. Reflux extraction of dried yam with 80% ethanol produced the crude extracts with the highest yields (1.39–1.80%), whereas fresh yam yielded 0.51–0.97% extract. Using Gas–liquid chromatography–mass spectrometry (GLC-MS) analysis, linoleic acid ethyl ester, n-hexadecanoic acid, 9,12-octadecadienoic acid (Z,Z)-, and stigmasterol were identified as the major constituents. Among the tested extracts, DH-W-F-H (D. hispida-water washing-fresh-hexane) and DH-W-F-E (D. hispida-water washing-fresh-ethanol) were extracted from fresh yam using hexane and ethanol, respectively, while DH-W-D-E (D. hispida-water washing-dry-ethanol) was isolated from dried yam using ethanol and exhibited the strongest antibacterial activity, with minimum inhibitory concentrations (MIC) ranging from 64 to 2048 µg/mL. These extracts demonstrated pronounced concentration-dependent inhibition of biofilm formation by Staphylococcus epidermidis, Staphylococcus aureus, and Cutibacterium acnes. The strongest anti-biofilm activity was observed against C. acnes, with biofilm formation nearly eliminated at MIC concentrations. Moreover, all three extracts significantly reduced established C. acnes biofilms, with DH-W-F-H exhibiting greater eradication efficacy than vancomycin under the tested conditions. To elucidate the underlying mechanism, major fatty acid derivatives were evaluated against C. acnes lipase (_CALipase), a virulence factor associated with biofilm development, using molecular docking, molecular dynamics simulations, and the Molecular Mechanics-Generalized Born Surface Area (MM-GBSA) binding free-energy calculations. The compounds exhibited favorable interactions with _CALipase, with linoleic acid ethyl ester (FA2) showing the strongest binding affinity, stable protein–ligand interactions throughout a 200 ns simulation, and the most favorable binding free energy. Collectively, the biological and computational findings suggest that fatty acid-rich extracts from processed D. hispida suppress biofilm formation through an antivirulence mechanism involving _CALipase inhibition. These results highlight the potential of D. hispida as a source of metabolites for the development of functional food ingredients and value-added cosmetic and dermatological applications. Full article

Objectives: The study determined antimicrobial resistance profiles of surgical wound multidrug-resistant (MDR) isolates belonging to the ESKAPE group and evaluated antibacterial and antibiofilm activities of geraniol (G), carvacrol (C) and eugenol (E), individually and in the selected mixtures. The cytotoxicity of monoterpenes and combinations was also assessed. Methods: The antibacterial and antibiofilm activity against tested isolates of Enterococcus faecium, Staphylococcus aureus MRSA, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter sp. was assessed in microdilution and crystal violet assay, respectively, while cytotoxicity was estimated in XTT assay on human MRC-5 fibroblasts. Results: C showed the strongest antibacterial activity (MIC 0.32 ± 0.24 mg mL⁻¹). Monoterpenes induced synergism in certain combinations (FICI 0.09–0.31). All individual monoterpenes inhibited biofilm formation, but G was the most active (38.78–84.72%, p < 0.05). The most pronounced biofilm eradication was observed for C (25.21–61.34%, p < 0.05). G-C mixtures showed notable inhibition of biofilm formation in all isolates except P. aeruginosa, but, on the contrary, induced biofilm eradication against P. aeruginosa only. Cytotoxicity was not detected with the applied concentrations of monoterpenes, while the G-C mixtures exhibited lower cytotoxicity than povidone-iodine (p < 0.05) used as the control. Conclusions: All monoterpenes and G-C mixtures proved significant antibacterial and antibiofilm potential against ESKAPE isolates and acceptable impact on cell viability. Practical significance: The results highlight the potential of plant monoterpenes and their binary combinations as complementary agents for controlling MDR ESKAPE pathogens and biofilm-associated infections following surgical procedures. Further in vivo and safety studies are needed to confirm their applicability in clinical settings. Full article

Background/Objectives: The increasing prevalence of antimicrobial resistance has reduced the effectiveness of conventional antibiotics, highlighting the need for alternative or adjunct antimicrobial agents. This study aimed to investigate the antibacterial, synergistic, and antibiofilm activities of Liquidambar orientalis Mill. essential oil against clinically relevant bacterial pathogens, using 24 bacterial strains representing 12 bacterial species. Methods: The volatile constituents of L. orientalis essential oil obtained by hydrodistillation of storax balsam were characterized by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID). Antibacterial activity was evaluated using minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), disk diffusion, and agar well diffusion assays against a panel of Gram-positive and Gram-negative bacterial pathogens. Synergistic interactions with erythromycin, amoxicillin, gentamicin, and enrofloxacin were assessed using fractional inhibitory concentration index (FICI) analysis. Antibiofilm activity was also evaluated against representative biofilm-forming strains. Results: A total of 90 volatile constituents were identified by GC-MS, of which styrene was the most abundant (68.6%), followed by (E)-ethyl cinnamate (6.8%), α-pinene (5.9%), and β-pinene (2.9%). The essential oil exhibited broad-spectrum antibacterial activity, with an overall inhibition rate of 60.4%. Activity was higher against Gram-positive (77.77%) than Gram-negative (73.33%) bacteria. MIC and MBC values ranged from 15.62 to 250 µg/mL and 31.25 to 250 µg/mL, respectively. FICI analysis revealed strain- and antibiotic-dependent interactions, ranging from synergistic to antagonistic effects. Variable antibiofilm activity was observed, with the strongest inhibition against Brucella melitensis and the weakest against Pseudomonas aeruginosa. No significant differences in MIC or MBC values were observed between bacterial groups (p > 0.05). Conclusions: L. orientalis essential oil demonstrated in vitro antibacterial activity against several clinically relevant pathogens, with particularly notable activity against B. melitensis. The essential oil also showed synergistic interactions with selected antibiotics and variable antibiofilm effects. These findings suggest that L. orientalis essential oil may have potential as an antibacterial agent or adjunct to antibiotic therapy; however, further in vivo and clinical studies are required to confirm its therapeutic applicability. Full article

Background: Fresh leafy vegetables are widely consumed because of their high nutritional value but are also recognized as important vehicles for foodborne pathogens. Escherichia coli readily attaches to lettuce surfaces and forms biofilms, enhancing bacterial persistence and reducing the effectiveness of conventional washing procedures. This study evaluated the antibiofilm activity of dill, basil, winter savory, and peppermint essential oils (EOs) against E. coli during initial bacterial attachment and against preformed biofilms on polystyrene surface and lettuce leaves. Methods: Biofilm formation was assessed using Congo Red agar, the crystal violet microtiter plate assay, and a bacterial attachment assay on lettuce leaves, while biofilm architecture and the effects of EO treatment were examined by scanning electron microscopy (SEM). Antibacterial activity was determined by the broth microdilution method, and antibiofilm activity on initial bacterial attachment and preformed biofilms was evaluated at 0.5 × MIC, MIC, and 2 × MIC using the crystal violet assay and bacterial attachment assay on lettuce leaves, respectively. Results: The examined E. coli isolates predominantly exhibited the rdar morphotype and produced stronger biofilms at 25 °C than at 37 °C. All tested EOs significantly reduced initial bacterial attachment and disrupted preformed biofilms in a concentration-dependent manner. Dill EO showed the strongest antibacterial and antibiofilm effects, followed by basil, winter savory, and peppermint EOs. At 2 × MIC, the tested EOs reduced initial bacterial attachment by 74.17–84.63%, while dill EO reduced preformed biofilm on polystyrene by 59.90% after 60 min. SEM analysis confirmed extensive disruption of biofilm architecture and pronounced morphological damage to bacterial cells following EO treatment. Conclusions: These findings demonstrate that essential oils, particularly dill EO, effectively inhibit both the establishment and persistence of E. coli biofilms on polystyrene and lettuce leaves, highlighting their potential as natural antibiofilm agents for improving the microbiological safety of fresh produce. Full article

N-acetylcysteine (NAC) is a synthetic derivative of L-cysteine, known since the mid-20th century as a mucolytic agent and, in recent decades, has attracted attention for its antioxidant and antibiofilm properties. Bacterial biofilms are structured communities of microorganisms enclosed in an extracellular polymeric matrix, which accounts for their markedly increased resistance to antibiotics (up to 1000-fold higher than in planktonic forms) and to the host immune response. According to the literature, up to 65% of infectious agents are associated with biofilm formation, making them a challenging therapeutic target. This review systematizes current data on the molecular mechanisms of the antibiofilm action of NAC, including disruption of matrix proteins and polysaccharides, degradation of extracellular DNA, suppression of the quorum sensing system, and disturbance of bacterial redox homeostasis. Particular attention is given to synergistic combinations of NAC with antibiotics of five major classes; effective concentrations are provided, and the types of interaction are characterized. The results of clinical studies from the last decade are reviewed, demonstrating the potential of NAC as an adjuvant in urinary tract infections, chronic rhinosinusitis, diabetic osteomyelitis, and cystic fibrosis. The main limitations (pH dependence, instability, low oral bioavailability) are critically evaluated, and approaches to overcoming them using nanoparticles, hydrogels, and combinations with propolis or chitosan are proposed. The review is intended for researchers in antimicrobial chemotherapy and developers of new drug delivery systems. Full article

Periprosthetic joint infection (PJI) remains one of the most serious complications of arthroplasty, largely due to the formation of microbial biofilms on implant surfaces. Biofilm-associated infections exhibit increased tolerance to antimicrobial therapy and host immune responses, making eradication difficult and often requiring repeated surgical interventions. Consequently, there is a growing need for effective local therapeutic strategies capable of delivering high concentrations of antimicrobial agents directly to the site of infection while minimizing systemic toxicity. Hydrogels have emerged as promising drug delivery platforms for the management of biofilm-associated PJI. Their biocompatibility, injectability, high water content, and tunable physicochemical properties enable controlled and localized release of therapeutic agents within the infected peri-implant environment. This narrative review summarizes recent advances in hydrogel-based approaches, including antibiotic-loaded hydrogels, systems incorporating anti-biofilm enzymes, bacteriophage-loaded formulations, and nanoparticle-enhanced platforms. It also highlights future research directions, with particular emphasis on the need for expanded clinical studies to facilitate the translation of emerging hydrogel-based therapies into clinical practice. Further development of these systems should focus on the incorporation of novel therapeutic agents into hydrogel platforms, aiming to enhance biofilm eradication and improve treatment outcomes in patients with PJI. Particular attention is given to stimuli-responsive (“smart”) hydrogels that release therapeutic payloads in response to infection-related triggers such as pH changes, with emphasis on the need for expanded clinical studies to facilitate the translation of emerging hydrogel-based therapies into clinical practice. Further development of these systems should focus on the incorporation of novel therapeutic agents into hydrogel platforms, aiming to enhance biofilm eradication and improve treatment outcomes in patients with PJI. Full article

Candida albicans biofilms are a major cause of persistent infections and contribute to antifungal resistance as well as limitations in drug delivery. Targeting virulence traits such as biofilm formation and hyphal transition represents an effective strategy for controlling fungal pathogenicity without exerting strong selective pressure on planktonic growth. In this study, a library of structurally diverse benzophenone derivatives was screened to identify compounds with antibiofilm and anti-hyphal activities against azole-resistant C. albicans. Most benzophenone derivatives exhibited weak antifungal activity (MIC ≥ 200 µg/mL). However, several halogenated benzophenones markedly suppressed biofilm formation. Among them, decafluorobenzophenone at 10 µg/mL displayed the strongest inhibition, reducing biofilm formation to approximately 1–2% of control levels while maintaining substantial planktonic cell viability. Microscopy confirmed hyphal suppression, while qRT-PCR showed a 36-fold reduction in ALS3 expression. These findings indicate that multi-halogenated benzophenones act primarily as anti-virulence agents targeting biofilm formation and hyphal development. Molecular docking suggested a possible interaction of decafluorobenzophenone with the Als3 binding pocket. Decafluorobenzophenone showed low toxicity, with unaffected Caenorhabditis elegans viability at 10 µg/mL, plant germination at 100 µg/mL, and only slight hemolysis at 100 µg/mL. The results highlight halogen substitution as a key structural determinant and identify benzophenone scaffolds as promising leads for developing novel antibiofilm strategies against azole-resistant Candida infections. Full article

Background/Objective: Mature oral biofilms resist conventional antiseptics via dense extracellular polymeric substance (EPS) matrices. Consequently, the prolonged use of broad-spectrum agents like chlorhexidine (CHX) raises profound ecological concerns. Using a mature oral microcosm biofilm model, we evaluated the multifaceted efficacy of 0.5% black cumin seed oil (BCSO) in disrupting structural biomass, suppressing aciduric bacterial viability, and attenuating porphyrin-associated anaerobic biofilm maturation. Methods: Saliva-derived microcosm biofilms were cultivated on hydroxyapatite discs in a mucin-containing medium (0.5% sucrose) for 6 days. Mature biofilms were treated daily for 1 min for 6 days (n = 19/group) with 0.5% BCSO, 0.12% CHX (positive control), or 0.5% dimethyl sulfoxide (negative control). EPS and bacterial biovolumes were quantified using confocal microscopy. Viability was strictly evaluated as aciduric bacterial colony-forming units. Pathogenic potential was specifically assessed as porphyrin-associated maturation by the red-to-green fluorescence intensity ratio (${\mathrm{Ratio}}_{\mathrm{R}/\mathrm{G}}$) using quantitative light-induced fluorescence-digital technology. Results: Compared with the negative control, 0.5% BCSO significantly degraded the EPS matrix (p < 0.001) and bacterial biovolumes (p = 0.045), and reduced aciduric bacterial counts (p = 0.028). Importantly, 0.5% BCSO significantly reduced the ${\mathrm{Ratio}}_{\mathrm{R}/\mathrm{G}}$ (p = 0.015), compared with the negative control, indicating severely attenuated porphyrin-associated anaerobic maturation. In the CHX group, the reduction did not reach statistical significance compared with the negative control (p = 0.399). Conclusions: In vitro, 0.5% BCSO and 0.12% CHX effectively reduced structural biomass and aciduric bacterial viability. Furthermore, BCSO significantly reduced late-stage, porphyrin-associated anaerobic maturation compared with the negative control, whereas the reduction observed with CHX did not reach statistical significance. These findings highlight the potential of BCSO as a targeted natural antibiofilm adjunct, warranting rigorous in vivo and mechanistic validation before clinical translation. Full article