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17 pages, 2528 KB  
Article
Physicochemical Characterization, Antimicrobial and Antibiofilm Activities of Thymus vulgaris and Rosmarinus officinalis Essential Oil Nanoemulsions with Potential Mouthwash Applications
by Cemre Irem Ayguler, Aleyna Ozveren, Timur Hakan Barak, Gamze Benli Yardimci, Ipek Tekin and Mujde Eryilmaz
Pharmaceuticals 2026, 19(8), 1229; https://doi.org/10.3390/ph19081229 - 4 Aug 2026
Viewed by 102
Abstract
Background/Objectives: In this study, nanoemulsions containing Thymus vulgaris (TV) and Rosmarinus officinalis (RO) essential oils (EOs) were prepared and characterized, and their antibacterial, antifungal, and antibiofilm activities were evaluated against some oral pathogens. Methods: Three nanoemulsions containing 5% TV-EO (F1), 5% [...] Read more.
Background/Objectives: In this study, nanoemulsions containing Thymus vulgaris (TV) and Rosmarinus officinalis (RO) essential oils (EOs) were prepared and characterized, and their antibacterial, antifungal, and antibiofilm activities were evaluated against some oral pathogens. Methods: Three nanoemulsions containing 5% TV-EO (F1), 5% RO-EO (F2), and a combination of 2.5% TV-EO and 2.5% RO-EO (F3) were prepared by emulsification followed by high-speed homogenization. Results: Gas chromatography–mass spectrometry coupled with flame ionization detection (GC-MS-FID) analysis revealed linalool (79.51%) as the major constituent of TV-EO and eucalyptol (58.13%), together with camphor (10.80%), as the predominant compounds of RO-EO. All nanoemulsions exhibited nanosized droplets, low polydispersity index (PDI) values (<0.3), and moderate to high colloidal stability. Transmission electron microscopy (TEM) analysis revealed predominantly spherical droplets in all formulations (F1–F3). All formulations exhibited antimicrobial activity against Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, and Candida albicans. Among the tested formulations, F2 showed the strongest antibacterial activity, whereas F3 exhibited the highest antibiofilm activity against mature S. mutans biofilms, achieving approximately 80% inhibition, comparable to chlorhexidine (CHX). Although F3 possessed the smallest droplet size, its antimicrobial activity was lower than that of F2, suggesting that biological activity was influenced not only by droplet size but also by EO composition. Conclusions: These findings demonstrate the potential of TV-EO and RO-EO nanoemulsions as promising natural alternatives for mouthwash formulations and highlight the importance of EO composition in determining antimicrobial and antibiofilm efficacy. Full article
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31 pages, 7908 KB  
Review
Therapeutic Potential of Cannabidiol in Dysbiosis-Related Oral Biofilm Diseases: Antibiofilm, Antivirulence and Host Response Evidence
by Jiaqi Zhu, Xinyan Huang, Shuangyue Wu, Xiaoran Xu, Siyuan Wu, Yuankun Zhai and Jianhang Bao
Pharmaceuticals 2026, 19(8), 1221; https://doi.org/10.3390/ph19081221 - 3 Aug 2026
Viewed by 137
Abstract
Dysbiosis-related oral biofilm diseases, particularly dental caries and periodontal diseases, pose major global health challenges because ecological shifts within oral microbial communities enhance biofilm virulence, resilience, and host inflammatory responses. Cannabidiol (CBD), a non-psychoactive phytocannabinoid with antimicrobial, antibiofilm, immunomodulatory, and antioxidant properties, has [...] Read more.
Dysbiosis-related oral biofilm diseases, particularly dental caries and periodontal diseases, pose major global health challenges because ecological shifts within oral microbial communities enhance biofilm virulence, resilience, and host inflammatory responses. Cannabidiol (CBD), a non-psychoactive phytocannabinoid with antimicrobial, antibiofilm, immunomodulatory, and antioxidant properties, has attracted increasing interest as an investigational, ecology-oriented adjunct for oral health applications. This narrative review evaluates current antibiofilm, antivirulence, and host response evidence for CBD in dysbiosis-related oral biofilm diseases, with emphasis on dental caries and periodontal diseases and selected supportive evidence from other oral biofilm-associated conditions. Current evidence suggests that CBD can inhibit biofilm formation, attenuate cariogenic and fungal virulence traits, modulate periodontal inflammation and immunity, and support tissue-protective responses. However, most evidence remains preclinical and model-dependent, particularly in caries research, and CBD’s hydrophobicity, limited stability, uncertain dose windows, and incomplete microbiome-level evidence remain major barriers to translation. Future studies should clarify CBD’s ecological effects on oral microbial communities, define clinically relevant dosing and exposure timing, and develop oral-retentive delivery systems. Full article
(This article belongs to the Special Issue The Therapeutic Potential of Cannabidiol)
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35 pages, 1091 KB  
Review
Bacterial Extracellular Vesicles at the Crossroads of Immune Regulation and Biofilm Dynamics: Biogenesis, Comparative Analysis, and Translational Challenges
by Qingyu Zhang, Beilei Zhang, Mohd Shafiq Aazmi, Lin Chen and Mohd Fakharul Zaman Raja Yahya
Biomolecules 2026, 16(8), 1132; https://doi.org/10.3390/biom16081132 - 3 Aug 2026
Viewed by 120
Abstract
Bacterial extracellular vesicles (BEVs) are nano-sized lipid bilayer particles secreted by bacteria, capable of carrying various proteins, lipids, nucleic acids, and pathogen-associated molecular patterns (PAMPs). The biosynthetic pathway of BEVs determines their load components, physicochemical properties, and different biological activities. Increasing evidence indicates [...] Read more.
Bacterial extracellular vesicles (BEVs) are nano-sized lipid bilayer particles secreted by bacteria, capable of carrying various proteins, lipids, nucleic acids, and pathogen-associated molecular patterns (PAMPs). The biosynthetic pathway of BEVs determines their load components, physicochemical properties, and different biological activities. Increasing evidence indicates that BEVs play an important role in mediating host immune responses and the dynamic regulation of bacterial biofilms. Additionally, BEVs may serve as a molecular bridge between the two. BEVs derived from pathogens can trigger pro-inflammatory cascades, assist bacteria in immune evasion, and further accelerate the maturation of biofilms, forming a vicious cycle of persistent infection and inflammatory damage. In contrast, BEVs derived from probiotics can maintain host immune homeostasis and exert direct anti-biofilm and synergistic antibacterial effects, thereby breaking the pathological cycle. However, significant methodological research bottlenecks have greatly hindered the comparability and clinical translation of BEVs research. This article systematically summarizes the classification of BEVs and their biosynthetic mechanisms, compares the differential effects of BEVs from pathogenic bacteria and probiotic bacteria on immunity, clarifies the dual regulatory role of BEVs throughout the life cycle of biofilms, and highlights the bridging function of BEVs in the immune–biofilm interaction. Additionally, this article also discusses the current development of BEVs in clinical translation applications, such as vaccine development, antibiotic delivery, and mucosal inflammation intervention, and outlines the key industrial and clinical challenges faced in the future development of BEVs-based therapeutic approaches. Full article
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20 pages, 8492 KB  
Article
Antarctic Marine-Derived Fungi: Metabolomic Signatures and Antibiofilm-Driven Anti-Infective Potential Against Drug Resistant Pathogens
by İbrahim S. Uras, Pedro H. S. Candido, Catarina M. Luís, Vanda Marques, Cecília M. P. Rodrigues, Rita G. Sobral, Anelize Baurmeister, Belma Konuklugil and Susana P. Gaudêncio
Mar. Drugs 2026, 24(8), 267; https://doi.org/10.3390/md24080267 - 2 Aug 2026
Viewed by 228
Abstract
Antarctic marine-derived fungi represent an underexplored reservoir of bioactive secondary metabolites shaped by extreme environmental pressures. In this study, nine fungal isolates obtained from Antarctic macroalgae, lichens, sponge tissue, and sediments were evaluated for their antimicrobial, anticancer, antibiofilm, and metabolomic profiles. Untargeted LC–MS/MS [...] Read more.
Antarctic marine-derived fungi represent an underexplored reservoir of bioactive secondary metabolites shaped by extreme environmental pressures. In this study, nine fungal isolates obtained from Antarctic macroalgae, lichens, sponge tissue, and sediments were evaluated for their antimicrobial, anticancer, antibiofilm, and metabolomic profiles. Untargeted LC–MS/MS molecular networking (GNPS) revealed a chemically rich metabolome, dominated by alkaloids, followed by polyketides, meroterpenoids, and diketopiperazines, with Penicillium crustosum (A15A) emerging as a major biosynthetic contributor. The annotation of structurally diverse metabolites, including roquefortines, viridicatin derivatives, andrastins, and multiple diketopiperazines, highlights the metabolic plasticity of Antarctic fungi. Anticancer evaluation indicated cytotoxicity, with Aspergillus awamori (A30), Alternaria malorum (A36), and Cladosporium malorum (A38) displaying activity toward HCT 116 colorectal cancer cells at IC50 ≥ 30 µg/mL. Extracts were screened against methicillin-resistant Staphylococcus aureus (MRSA, COL), methicillin-susceptible S. aureus (MSSA, NCTC8325 4), and Escherichia coli K12, revealing low to no activity against these pathogens. Six of the nine isolates exhibited strong antibiofilm activity without inhibiting planktonic bacterial growth, indicating selective biofilm inhibition against MSSA and meeting the criteria for clinical developmental “hits”. Biofilm inhibition ranged from 81.10% to 98.50%, with A15A showing the highest activity (98.50% at 250 µg/mL), followed by A36 (91.16% at 31.35 µg/mL). Botrytis sp. (A22A), P. chrysogenum (A7), Ulocladium microsporum (A24B), and A30 also demonstrated strong antibiofilm activity (81.10–85.89%). To our knowledge, this is the first report describing antibiofilm activity of Antarctic fungal extracts. Full article
(This article belongs to the Section Marine Biotechnology Related to Drug Discovery or Production)
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27 pages, 1153 KB  
Review
Cold Atmospheric Plasma in Periodontitis: Mechanisms, Antimicrobial and Immunomodulatory Effects, and Therapeutic Potential—A Comprehensive Review
by Abbas Bumajdad, Ali Alnaser and Mohammad Qali
Dent. J. 2026, 14(8), 468; https://doi.org/10.3390/dj14080468 - 2 Aug 2026
Viewed by 298
Abstract
Background: Periodontitis is a chronic inflammatory disease characterized by a dysbiotic oral microbiome that leads to progressive periodontal tissue destruction. Current treatment primarily relies on scaling and root planing (SRP); however, this approach does not always completely eliminate pathogenic microorganisms or adequately modulate [...] Read more.
Background: Periodontitis is a chronic inflammatory disease characterized by a dysbiotic oral microbiome that leads to progressive periodontal tissue destruction. Current treatment primarily relies on scaling and root planing (SRP); however, this approach does not always completely eliminate pathogenic microorganisms or adequately modulate the host inflammatory response. Cold atmospheric plasma (CAP) generates reactive oxygen and nitrogen species (RONS) with potential antimicrobial, antibiofilm, and immunomodulatory properties, making it a promising adjunctive therapeutic modality for the management of periodontitis. Literature Search Strategy: A comprehensive literature search was conducted using PubMed/MEDLINE, Scopus, Web of Science, and Embase to identify relevant studies published between January 2000 and June 2025. The search included in vitro, in vivo, and clinical studies evaluating the antimicrobial, immunomodulatory, and regenerative effects of CAP in periodontitis and related periodontal models. Reference lists of relevant articles were also manually screened to identify additional eligible studies. Objectives: This review aims to provide a comprehensive overview of the antimicrobial and immunomodulatory effects of CAP on periodontitis and to evaluate its potential as an adjunctive treatment modality. Furthermore, the review examines the regenerative potential and underlying mechanisms of CAP, while also addressing its clinical safety and biocompatibility. Key Findings: Preclinical evidence indicates that CAP reduces bacterial viability, disrupts biofilm architecture, and modulates host immune responses through RONS-mediated signaling pathways. In animal models of periodontitis, CAP has been shown to enhance markers of periodontal tissue repair, reduce the expression of pro-inflammatory cytokines, increase LC3-associated phagocytosis in macrophages, and accelerate wound healing. Clinical studies evaluating CAP as an adjunct to SRP have reported improvements in clinical attachment level (CAL) and reductions in the recolonization of periodontal pathogens. Nevertheless, substantial heterogeneity in CAP treatment parameters and a lack of long-term safety and efficacy data limit direct comparisons across studies and highlight the need for further well-designed clinical investigations. Conclusions: CAP appears to be a potentially useful adjunct to SRP in the management of periodontitis. Current preclinical evidence demonstrates significant antimicrobial, immunomodulatory, and regenerative effects; however, clinical evidence remains limited. Further high-quality clinical trials are required to validate these findings, establish standardized treatment protocols, optimize therapeutic parameters, and facilitate the development of regulatory frameworks for clinical implementation. Full article
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26 pages, 16165 KB  
Article
Enterococcus Phage Abitsa: Biological Characterization, Antibiofilm Activity and Evolutionary Insights into the Genus Efquatrovirus
by Konstantin S. Troshin, Lydia I. Ilyenko, Andrei V. Chaplin, Anastasiya A. Khritova, George A. Skvortsov, Anna A. Vasilyeva, Olga Y. Guseva, Igor S. Kopetskiy, Dmitriy A. Shagin, Lyudmila I. Kafarskaia, Boris A. Efimov, Artem A. Malkov, Maxim A. Sokolovskiy and Peter V. Evseev
Viruses 2026, 18(8), 842; https://doi.org/10.3390/v18080842 - 1 Aug 2026
Viewed by 207
Abstract
Enterococcus faecalis is an important opportunistic pathogen associated with persistent oral infections, biofilm formation, and antimicrobial tolerance, which makes phages targeting this species of both therapeutic and evolutionary interest. Here, we describe the isolation and characterization of Abitsa, a novel lytic Enterococcus phage [...] Read more.
Enterococcus faecalis is an important opportunistic pathogen associated with persistent oral infections, biofilm formation, and antimicrobial tolerance, which makes phages targeting this species of both therapeutic and evolutionary interest. Here, we describe the isolation and characterization of Abitsa, a novel lytic Enterococcus phage representing a new species within the genus Efquatrovirus. Abitsa formed small plaques and displayed siphovirus-like morphology. It showed an optimal multiplicity of infection of 0.01, a short latent period of 10 min and a burst size of 28 ± 5 virions per infected cell. The phage efficiently suppressed planktonic growth of E. faecalis over a broad multiplicity of infection (MOI) range and significantly disrupted pre-formed biofilms, with the strongest effect observed at MOI 0.1, resulting in an 82.14% reduction in biofilm biomass. Abitsa remained stable at pH 4–8, at 5–50 °C for 1 h, and in up to 75% chloroform, but exhibited a narrow host range, lysing only one additional clinical E. faecalis isolate among 34 tested enterococcal isolates. Genome analysis showed that Abitsa has a 41,581-bp linear genome lacking lysogeny-associated genes and represents a novel Efquatrovirus species. Comparative structural and phylogenetic analyses additionally supported mosaic evolution and extensive domain shuffling in receptor-binding and lysis-related proteins of Efquatrovirus-like phages. Together, these results identify Abitsa as a biologically unusual and evolutionarily informative lytic phage with antibiofilm activity. Full article
(This article belongs to the Section Bacterial Viruses)
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25 pages, 4399 KB  
Article
Biosynthesis of Silver Nanoparticles from Lotus sanguineus: Photocatalytic, Antimicrobial, and Antibiofilm Activities with Molecular Docking Against Quorum-Sensing Proteins
by Suzan Şahin Doğan
Biology 2026, 15(15), 1261; https://doi.org/10.3390/biology15151261 - 31 Jul 2026
Viewed by 266
Abstract
Industrial effluents containing toxic dyes, pathogenic microorganisms, and other hazardous contaminants pose a serious threat to environmental and public health, creating an urgent need for multifunctional materials capable of addressing both environmental pollution and microbial contamination. Silver nanoparticles (AgNPs) have attracted considerable attention [...] Read more.
Industrial effluents containing toxic dyes, pathogenic microorganisms, and other hazardous contaminants pose a serious threat to environmental and public health, creating an urgent need for multifunctional materials capable of addressing both environmental pollution and microbial contamination. Silver nanoparticles (AgNPs) have attracted considerable attention because of their broad-spectrum antimicrobial and photocatalytic properties. In this study, AgNPs were biosynthesized for the first time using Lotus sanguineus, an endemic plant rich in bioactive phytochemicals, through an environmentally friendly green synthesis approach. To comprehensively evaluate their multifunctional performance, the photocatalytic, antimicrobial, and antibiofilm activities of the biosynthesized AgNPs were investigated, while molecular docking analysis was performed to elucidate their interactions with quorum-sensing-related proteins. AgNPs were characterized using UV–Vis spectroscopy, FTIR, XRD, DLS, and SEM. Under sunlight, AgNPs showed degradation efficiencies of 60.6% for methylene blue and 29.5% for eriochrome black T, indicating higher photocatalytic activity toward the cationic dye. Antimicrobial evaluation demonstrated notable antimicrobial activities against Escherichia coli, Bacillus cereus, Pseudomonas aeruginosa, and Staphylococcus aureus, with minimum inhibitory concentration (MIC) values ranging from 31.25 to 62.5 μg/mL and minimum bactericidal concentration (MBC) values between 125 and 250 μg/mL. Additionally, AgNPs exhibited a maximum inhibition of 94.5% at 62.5 μg/mL against Pseudomonas aeruginosa. Furthermore, molecular docking results revealed favorable binding affinities between the ligand and quorum-sensing transcriptional activator (LasR), N-terminal domain of anthranilate-CoA ligase (PqsA), P. aeruginosa aminopeptidase (PaAP), and peptidoglycan N-acetylglucosamine deacetylase (Pgd) proteins. Overall, the results highlighted the potential of AgNPs for pharmaceutical applications, prevention of bacterial surface colonization, and environmental remediation. Full article
(This article belongs to the Section Microbiology)
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56 pages, 2904 KB  
Review
Functional Liposomal Nanocarriers for the Treatment of Antimicrobial-Resistant and Biofilm-Associated Ocular Infections
by Paula Stefana Pintilei, Roya Binaymotlagh, Farid Hajareh Haghighi, Laura Chronopoulou and Cleofe Palocci
Macromol 2026, 6(3), 56; https://doi.org/10.3390/macromol6030056 - 31 Jul 2026
Viewed by 221
Abstract
Nanomedicine-based liposomal delivery systems are gaining increasing attention as advanced therapeutic platforms for managing ocular infections associated with antimicrobial resistance (AMR) and biofilm formation. The primary objective of this review is to critically evaluate the potential of liposomal drug delivery systems for improving [...] Read more.
Nanomedicine-based liposomal delivery systems are gaining increasing attention as advanced therapeutic platforms for managing ocular infections associated with antimicrobial resistance (AMR) and biofilm formation. The primary objective of this review is to critically evaluate the potential of liposomal drug delivery systems for improving the treatment of antimicrobial-resistant and biofilm-associated ocular infections by integrating current knowledge on antimicrobial resistance mechanisms, biofilm-targeted therapeutic strategies, and advances in liposomal formulations, while also identifying the major limitations, translational challenges, and knowledge gaps in this rapidly evolving field. Traditional ocular antimicrobial treatments are frequently limited by poor drug penetration, short precorneal residence time, low bioavailability, systemic side effects, and inadequate activity against resistant microorganisms and biofilm-embedded pathogens. This review provides a comprehensive overview of different liposomal systems, including conventional, cationic, polyethylene glycol (PEG)-modified, deformable, and stimulus-responsive liposomes, and discusses their advantages in ophthalmic drug delivery, such as enhanced corneal permeation, prolonged drug retention, controlled release, improved biocompatibility, and reduced ocular toxicity. The review further examines the mechanisms through which liposomes help overcome AMR, including improved epithelial transport, membrane disruption, intracellular drug delivery, efflux pump evasion, and enhanced antimicrobial efficacy. In addition, liposomal approaches targeting ocular biofilms are explored, focusing on improved biofilm penetration and the delivery of anti-biofilm agents such as antibiotics, enzymes, quorum-sensing inhibitors, and antimicrobial peptides. Current evidence from in vitro and in vivo ocular infection models is summarized together with disease-specific applications in keratitis, endophthalmitis, and contact lens-related infections. The article also compares liposomes with other ocular nanocarriers and addresses important considerations related to safety, stability, sterilization, large-scale production, and regulatory translation. In addition to highlighting recent advances, this review critically discusses the current limitations of liposomal formulations, the major barriers to clinical translation, and the key knowledge gaps that should be addressed to facilitate the future development and successful clinical application of these systems. Finally, emerging directions including ligand-targeted and stimulus-responsive liposomes, AI-driven formulation development, personalized nanotherapy, and gene therapy combinations are discussed as promising future strategies for combating resistant ocular infections. Full article
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23 pages, 5142 KB  
Article
Synthesis and Biological Evaluation of Novel C-28 Chloroacetamide-Modified Ursolic Acid Derivatives as Antibacterial Agents
by Nan Cai, Tian Luan, Junchao Zhang, Ning Li, Xiu Zhang, Peng Gao, Jiaxuan Li, Hongyu Zhan, Fanhao Meng and Dajun Zhang
Microorganisms 2026, 14(8), 1685; https://doi.org/10.3390/microorganisms14081685 - 31 Jul 2026
Viewed by 240
Abstract
The escalating crisis of bacterial antimicrobial resistance necessitates the discovery of novel antibacterial agents with distinct mechanisms of action. Ursolic acid (UA), a naturally abundant pentacyclic triterpenoid, serves as a promising scaffold for structural modification. In this study, a series of 26 novel [...] Read more.
The escalating crisis of bacterial antimicrobial resistance necessitates the discovery of novel antibacterial agents with distinct mechanisms of action. Ursolic acid (UA), a naturally abundant pentacyclic triterpenoid, serves as a promising scaffold for structural modification. In this study, a series of 26 novel UA derivatives (A1A7 and B1B19) were designed and synthesized by introducing various substituents at the C-28 carboxyl group via a chloroacetyl chloride linker. Their antibacterial activities were evaluated against S. aureus, S. epidermidis, E. coli, and P. aeruginosa using the microbroth dilution method. Among them, compound B1 exhibited the most potent activity, with a minimum inhibitory concentration (MIC) of 37.5 μg/mL against S. aureus and 75 μg/mL against E. coli. Antibacterial kinetic and time-kill curve assays confirmed the sustained bactericidal effect of B1. Furthermore, B1 significantly inhibited biofilm formation in both S. aureus and E. coli in a time-dependent manner. Molecular docking studies revealed that B1 binds spontaneously to the S. aureus SarA protein through three hydrogen bonds and π-π stacking interactions, providing a structural basis for its antibacterial and anti-biofilm activities. This study demonstrates that piperazine-modified UA derivative B1 is a promising antibacterial candidate and offers a new strategy for the structural optimization of ursolic acid. Full article
(This article belongs to the Section Antimicrobial Agents and Resistance)
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25 pages, 2165 KB  
Article
Evaluation of the Antioxidant, Anti-Inflammatory, and Antimicrobial Activity of a Cannabis sativa-Infused African Product Used for Wound Healing
by Siboniso Sithole, Xolani Shezi, Mlondolo Mavundla, Carl Mateta, Sphamandla Hlatshwayo, Sanele Mhlungu, Nokukhanya Thembane, Phumzile Afrika, Sibusiso Senzani, Exnevia Gomo, Nceba Gqaleni and Mlungisi Ngcobo
Int. J. Mol. Sci. 2026, 27(15), 6815; https://doi.org/10.3390/ijms27156815 - 29 Jul 2026
Viewed by 400
Abstract
Product Shezi (PS) is a polyherbal African traditional medicine (ATM) formulated from six known South African medicinal plants and is used for treating cutaneous wounds. However, its ethnopharmacological properties have not been scientifically validated. This study aimed to evaluate the antioxidant, anti-inflammatory, and [...] Read more.
Product Shezi (PS) is a polyherbal African traditional medicine (ATM) formulated from six known South African medicinal plants and is used for treating cutaneous wounds. However, its ethnopharmacological properties have not been scientifically validated. This study aimed to evaluate the antioxidant, anti-inflammatory, and antimicrobial activities of PS in vitro. Aqueous and methanolic extracts were prepared and qualitatively screened for phytochemical constituents. Cytotoxicity in fibroblasts and macrophages was assessed using an ATP-based viability assay. Antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and a hydrogen peroxide (H2O2)-induced oxidative stress model. Anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated macrophages were measured using the Griess reagent system, a human prostaglandin E2 (PGE2) ELISA, and a bovine serum albumin (BSA) anti-denaturation assay. Antimicrobial activity was assessed by twofold serial broth microdilution, agar well diffusion, and a crystal violet biofilm assay. Phytochemical screening confirmed the presence of saponins, alkaloids, tannins, glycosides, terpenoids, flavonoids, and steroids. PS exhibited IC10 values of 3 and 10 μg/mL in fibroblasts and macrophages, respectively. PS (1–5 μg/mL) showed 70% DPPH scavenging potential and potent H2O2 cytoprotection (p < 0.001). Nitric oxide inhibition was non-significant (p > 0.05) whilst PGE2 decreased significantly (p < 0.05) compared to LPS-stimulated cells. BSA denaturation was inhibited in a dose-dependent manner (p < 0.05). Staphylococcus aureus and S. epidermidis were the only microorganisms susceptible to PS (MIC 650–5000 μg/mL) with low SI (SI ≈ 0.05–0.06), while others were resistant. PS showed no antibiofilm activity (crystal-violet assay). Overall, PS demonstrated antioxidant and anti-inflammatory activities and selective antibacterial effects against planktonic bacteria in vitro. However, further optimization of extraction methods, detailed mechanistic studies, and in vivo investigations are warranted to substantiate therapeutic relevance. Full article
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19 pages, 2071 KB  
Article
Comparison of the Effect of Silver Nanoparticles Biosynthesized with Lavandula angustifolia Extract and Lavender Essential Oil Against Multidrug-Resistant and Biofilm-Forming Staphylococcus Species
by Patrícia Hudecová, Silvia Ondrašovičová, Vanda Hajdučková, Nikola Dančová, Gabriela Gregová, Lívia Mačák, Oksana Velgosová, Jana Ondrašovičová and Ján Király
Pharmaceutics 2026, 18(8), 930; https://doi.org/10.3390/pharmaceutics18080930 - 29 Jul 2026
Viewed by 235
Abstract
Background: Multidrug-resistant and biofilm-forming staphylococci pose a threat to the sustainability of public health, livestock health and the ecosystem. Pathogenic potential with a worsening prognosis of therapy is mainly due to methicillin-resistant Staphylococcus aureus (MRSA) or multidrug-resistant Non-aureus staphylococci and mammaliicocci [...] Read more.
Background: Multidrug-resistant and biofilm-forming staphylococci pose a threat to the sustainability of public health, livestock health and the ecosystem. Pathogenic potential with a worsening prognosis of therapy is mainly due to methicillin-resistant Staphylococcus aureus (MRSA) or multidrug-resistant Non-aureus staphylococci and mammaliicocci (NASM). Alternative approaches based on the use of biosynthesized nanoparticles or substances of natural origin appear to be promising solutions to the problem of ineffective suppression of infections caused by pathogenic microorganisms. Methods: The aim of this study was to monitor and compare the biological effects of silver nanoparticles prepared by green synthesis using Lavandula angustifolia and lavender essential oil. In particular, the antibacterial, antibiofilm, and biofilm-eradicating effects against biofilm-forming and multidrug-resistant reference strains and field isolates of staphylococci were monitored. Results: AgNPs inhibited the growth and formation of biofilms of S. aureus strains at a concentration of 0.05 μg/μL, but clinical NASM at 0.025 μg/μL. Sensitivity to Lavender essential oil (LEO) was the same against all tested staphylococcal strains, with an antibacterial MIC of 0.901 μg/μL. The essential oil also had an effect on biofilm formation against all tested strains, but its effect was recorded at a tenfold lower concentration (antibiofilm MIC = 0.0901 μg/μL). No eradication activity was recorded for either tested substance. Their activity against the formed biofilms was not recorded. Conclusions: The results demonstrate promising antibacterial and antibiofilm activities of biosynthesized AgNPs and lavender essential oil under in vitro conditions. These findings support further investigation of these materials as potential alternative antimicrobial approaches, particularly in combination with conventional antimicrobial agents. Full article
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34 pages, 27393 KB  
Review
Prevention and Treatment of Staphylococcus aureus Biofilms Using Promising Agr-QS-Targeting Anti-Biofilm Agents
by Salma Waheed Sheikh, Ahmad Ali, Asma Ahsan, Fei Shang, Ting Xue and Lauren Gollahon
Pathogens 2026, 15(8), 795; https://doi.org/10.3390/pathogens15080795 - 27 Jul 2026
Viewed by 189
Abstract
Staphylococcus aureus (S. aureus), a leading cause of nosocomial infections, contributes significantly to increased morbidity and mortality, especially when it forms biofilms on medical devices. This pathogen, specifically methicillin-resistant S. aureus (MRSA), remains a challenge to treat due to its ability [...] Read more.
Staphylococcus aureus (S. aureus), a leading cause of nosocomial infections, contributes significantly to increased morbidity and mortality, especially when it forms biofilms on medical devices. This pathogen, specifically methicillin-resistant S. aureus (MRSA), remains a challenge to treat due to its ability to form biofilms and rapidly develop resistance against antibiotics. Biofilm formation allows bacteria to adhere to biotic and abiotic surfaces, creating a protective matrix that shields them from immune responses and antibiotic therapies. The widespread prevalence of multidrug-resistant S. aureus biofilms poses a significant therapeutic challenge in clinical settings. Several novel therapeutic strategies have been developed to combat S. aureus biofilm-associated infections. Accumulating evidence suggests that natural plants and their derivatives possess antimicrobial and chemo preventive properties that can disrupt established biofilms. Several plant-derived compounds with anti-biofilm activities have been reported to target the regulatory proteins involved in the Agr quorum sensing (Agr-QS) system, underscoring their potential as therapeutic candidates for the prevention and treatment of biofilm-associated infections. However, despite these encouraging findings, clinical validation of these plant-based agents is essential to ensure their efficacy, safety, and optimal application in treating S. aureus biofilm infections. The continued exploration of natural biofilm inhibitors anticipates the urgent need for new treatments to combat biofilm-associated infections and multidrug-resistant pathogens like MRSA. This review provides a detailed overview of preventive and therapeutic interventions to eradicate biofilm-forming S. aureus infections. Full article
(This article belongs to the Section Bacterial Pathogens)
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31 pages, 3173 KB  
Article
Genome-Guided Discovery of an Antimicrobial Peptide Scaffold from an Undescribed Antarctic Rhodococcus Lineage: A Proof-of-Concept Study
by Dayaimi González, Andrés Santos, Eulàlia Sans-Serramitjana, Fanny Guzmán, Carla Gallardo-Benavente, Dayani Hernández, Mercyleidi Díaz Reyes and Francisca Acevedo
Int. J. Mol. Sci. 2026, 27(15), 6655; https://doi.org/10.3390/ijms27156655 - 25 Jul 2026
Viewed by 200
Abstract
Rhodococcus species display remarkable metabolic versatility and adaptation to extreme environments, yet their antimicrobial potential remains largely unexplored, particularly in strains representing undescribed genomic lineages. This study investigated whether the genome of Rhodococcus sp. GB-02, a strain isolated from Antarctic soil, encodes antimicrobial [...] Read more.
Rhodococcus species display remarkable metabolic versatility and adaptation to extreme environments, yet their antimicrobial potential remains largely unexplored, particularly in strains representing undescribed genomic lineages. This study investigated whether the genome of Rhodococcus sp. GB-02, a strain isolated from Antarctic soil, encodes antimicrobial peptides with demonstrable activity against clinically relevant pathogens. Hybrid genome assembly revealed a 6.58 Mb high-quality draft genome belonging to a previously undescribed genomic lineage within the genus Rhodococcus, based on average nucleotide identity and digital DNA–DNA hybridization analyses. Genome mining identified 20 biosynthetic gene clusters, predominantly non-ribosomal peptide synthetases, several of which showed low or no similarity to known pathways. Eight putative antimicrobial peptides (AMPs) were computationally predicted; among these, three lacked detectable homology to sequences in current databases, and experimental validation of one predicted peptide, designated Peptide 5764, demonstrated antibacterial activity at high micromolar concentrations against Gram-positive and Gram-negative pathogens and concentration-dependent inhibition of biofilm formation, particularly in Pseudomonas aeruginosa. However, Peptide 5764 exhibited considerable hemolytic activity at antibacterial concentrations, indicating that optimization for membrane selectivity will be required before therapeutic application. These findings establish Rhodococcus sp. GB-02 as an underexplored Antarctic lineage with a diverse biosynthetic repertoire and provide proof-of-concept that genome-guided AMP prediction can identify experimentally active peptide scaffolds from this lineage. Full article
(This article belongs to the Special Issue Computational Studies of Natural Products)
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24 pages, 2186 KB  
Article
LyeTx I mnΔKL, a New Synthetic Peptide Derived from a Lycosa erythrognatha Toxin, with Potent In Vitro and In Vivo Activity Against Methicillin-Resistant Staphylococcus aureus
by Waleska Stephanie da Cruz Nizer, Giulliana Altaf dos Santos, William Gustavo Lima, Felipe Henrique de Souza Silva, Wanderson Aparecido Brandão Candido, Amanda Neves de Souza, Giovanna Paula Araújo, Rodrigo Moreira Verly and Maria Elena de Lima
Toxins 2026, 18(8), 323; https://doi.org/10.3390/toxins18080323 - 25 Jul 2026
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Abstract
The emergence of multidrug-resistant (MDR) bacteria, particularly methicillin-resistant Staphylococcus aureus (MRSA), represents a major global health challenge by limiting the current therapeutic options. In this context, antimicrobial peptides (AMPs) have been widely studied for their potent antimicrobial properties. In this study, we evaluated [...] Read more.
The emergence of multidrug-resistant (MDR) bacteria, particularly methicillin-resistant Staphylococcus aureus (MRSA), represents a major global health challenge by limiting the current therapeutic options. In this context, antimicrobial peptides (AMPs) have been widely studied for their potent antimicrobial properties. In this study, we evaluated the anti-MRSA effect of a novel AMP, LyeTx I mnΔKL, derived from a toxin of Lycosa erythrognatha. Its activity was evaluated in vitro by minimal inhibitory and bactericidal concentrations (MIC and MBC), antibiofilm effect, membrane interaction, cytotoxicity, synergistic interaction with vancomycin, and in vivo in an MRSA murine wound/abscess infection model. LyeTx I mnΔKL showed enhanced antimicrobial activity against clinical MRSA isolates compared to its prototype (LyeTx I mnΔK), with MIC50 and MBC50 of 2 and 8 µM and 16 and 32 µM, respectively. Furthermore, LyeTx I mnΔKL exhibited a rapid bactericidal effect and a pronounced ability to inhibit biofilm formation and disrupt mature biofilms. LyeTx I mnΔKL interacts with bacterial membranes, adopts an α-helical structure, and induces membrane disruption and leakage of intracellular material. In vivo, topical treatment with LyeTx I mnΔKL reduced MRSA burden compared with LyeTx I mnΔK and untreated controls (log10 CFU/g of wound of 2.4, 4.4, and 6.5 for LyeTx I mnΔKL, LyeTx I mnΔK, and the saline group, respectively). However, increased cytotoxicity remains a significant limitation. Overall, LyeTx I mnΔKL is a promising anti-MRSA candidate for topical use with potent antibiofilm and in vivo activity. Full article
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19 pages, 5446 KB  
Article
Potentiating Gentamicin Efficacy Against Biofilms of Clinically Relevant Gram-Negative Bacteria Using Biosynthesized ZnO Nanoparticles
by Akshit Malhotra, Kwthar Debbarma, Sangita Jana, Irusan Dhinakaran, Surisetty Jaya Prasanthi, Elvira Rozhina, Ram Karan and Ashwini Chauhan
Pharmaceutics 2026, 18(8), 913; https://doi.org/10.3390/pharmaceutics18080913 - 24 Jul 2026
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Abstract
Background: Gram-negative bacteria resistant to multiple drugs are a major cause of illness and death worldwide. Their remarkable capacity to develop resistance to antibiotics makes them a serious concern in medical practice. Methods: A simple, green, novel method is used to [...] Read more.
Background: Gram-negative bacteria resistant to multiple drugs are a major cause of illness and death worldwide. Their remarkable capacity to develop resistance to antibiotics makes them a serious concern in medical practice. Methods: A simple, green, novel method is used to synthesize ZnO nanoparticles (ZnO NPs) using ethanolic extracts of Diplazium esculentum via precipitation. Results: ZnO NPs exhibit a hexagonal structure with a particle size of ~30 nm and a band gap of 3.24 eV. The defect sites formed in ZnO NPs were estimated using prominent peaks in the photoluminescence spectra. ZnO NPs displayed a more than 4-log reduction in multi-drug-resistant E. coli and K. pneumoniae clinical isolates at a 500 μg/mL concentration. Moreover, ZnO NPs significantly reduced the biofilm bacterial cell viability of clinical isolates of Gram-negative bacteria. Complete eradication of biofilms was achieved for drug-resistant E. coli clinical isolates using a combination of sub-MIC of gentamicin and 500 μg/mL ZnO NPs. Green-synthesized ZnO NPs did not induce oxidative stress in mice, as indicated by unchanged GST, GSH, and thiol levels across all the tested organs. ZnO NPs showed both antibacterial and antibiofilm efficacy against drug-resistant strains of E. coli, K. pneumoniae, and S. aureus and completely eradicated E. coli biofilm in combination with gentamicin. Conclusions: Our study focuses on the sustainable synthesis of biocompatible ZnO NPs for the treatment of infections caused by pathogens belonging to the high-priority ESKAPE group. Full article
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