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Keywords = biofilms formation

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17 pages, 688 KiB  
Review
Mycoplasma Biofilms: Characteristics and Control Strategies
by Jingyi Liang, Baoyi Deng, Weihuo Li, Jingjing Qi, Yangshuo Li, Xueyan Wang, Ming Li, Hong Yang and Nan Zhang
Microorganisms 2025, 13(8), 1850; https://doi.org/10.3390/microorganisms13081850 (registering DOI) - 7 Aug 2025
Abstract
The Mycoplasmataceae are a family of bacteria that typically cause respiratory, arthritic, and genitourinary disease in humans. Mycoplasma spp. of animal origin are also the causative agents of porcine wheezing disease, chronic respiratory disease and arthritis in chickens and other conditions. These diseases [...] Read more.
The Mycoplasmataceae are a family of bacteria that typically cause respiratory, arthritic, and genitourinary disease in humans. Mycoplasma spp. of animal origin are also the causative agents of porcine wheezing disease, chronic respiratory disease and arthritis in chickens and other conditions. These diseases have a significant impact on public health and the economic development of livestock breeding. Clinical prevention and treatment of mycoplasma infections is primarily dependent on the use of antibiotics. However, inappropriate and excessive use of antimicrobials has enabled resistance development that has become a significant clinical concern. Mycoplasma are also robust biofilm producers, and this process is a major factor for the persistence of these infections, especially in conjunction with common antibiotic resistance mechanisms, including target gene mutations and the action of efflux pumps. A mycoplasma biofilm refers to a structured and stable microbial community formed by Mycoplasma spp. adhering to biological or non-biological surfaces under suitable conditions and secreting extracellular polymers (EPS) such as polysaccharides. This process allows the microorganisms to adapt to their surrounding environment and survive during the growth process. These biofilms render bacteria more resistant to antimicrobials than planktonic bacteria, resulting in biofilm-associated infections that are more challenging to eradicate and more likely to recur. The current study reviews progress from the fields of biofilm formation, structure and identification, correlations between biofilms and drug resistance and virulence as well as methods of biofilm prevention and control. Our aim was to provide a reference basis for the subsequent in-depth understanding of the research of mycoplasma biofilms. Full article
13 pages, 718 KiB  
Article
Evaluation and Verification of Starch Decomposition by Microbial Hydrolytic Enzymes
by Makoto Takaya, Manzo Uchigasaki, Koji Itonaga and Koichi Ara
Water 2025, 17(15), 2354; https://doi.org/10.3390/w17152354 (registering DOI) - 7 Aug 2025
Abstract
This study investigates the Enzyme Biofilm Method (EBM), a biological wastewater treatment technology previously developed by the authors. EBM employs microbial-derived hydrolytic enzyme groups in the initial treatment stage to break down high-molecular-weight organic matter—such as starch, proteins, and fats—into low-molecular-weight compounds. These [...] Read more.
This study investigates the Enzyme Biofilm Method (EBM), a biological wastewater treatment technology previously developed by the authors. EBM employs microbial-derived hydrolytic enzyme groups in the initial treatment stage to break down high-molecular-weight organic matter—such as starch, proteins, and fats—into low-molecular-weight compounds. These compounds enhance the growth of native microorganisms, promoting biofilm formation on carriers and improving treatment efficiency. Over the past decade, EBM has been practically applied in food factory wastewater facilities handling high organic loads. The enzyme groups used in EBM are derived from cultures of Bacillus mojavensis, Saccharomyces cariocanus, and Lacticaseibacillus paracasei. To clarify the system’s mechanism and ensure its practical viability, this study focused on starch—a prevalent and recalcitrant component of food wastewater—using two evaluation approaches. Verification 1: Field testing at a starch factory showed that adding enzyme groups to the equalization tank effectively reduced biological oxygen demand (BOD) through starch degradation. Verification 2: Laboratory experiments confirmed that the enzyme groups possess both amylase and maltase activities, sequentially breaking down starch into glucose. The resulting glucose supports microbial growth, facilitating biofilm formation and BOD reduction. These findings confirm EBM’s potential as a sustainable and effective solution for treating high-strength food industry wastewater. Full article
(This article belongs to the Special Issue Advanced Biological Wastewater Treatment and Nutrient Removal)
14 pages, 6774 KiB  
Article
Antimicrobial Activities of Propolis Nanoparticles in Combination with Ampicillin Sodium Against Methicillin-Resistant Staphylococcus aureus
by Kaiyue Feng, He Sang, Han Jin, Peng Song, Wei Xu, Hongzhuan Xuan and Fei Wang
Microorganisms 2025, 13(8), 1844; https://doi.org/10.3390/microorganisms13081844 - 7 Aug 2025
Abstract
Combining antibiotics with propolis is an effective method to combat bacterial drug resistance. Nanoparticles are of interest in the antimicrobial field because of their higher drug stability, solubility, penetration power, and treatment efficacy. In this study, propolis nanoparticles (PNPs) were synthesized, and their [...] Read more.
Combining antibiotics with propolis is an effective method to combat bacterial drug resistance. Nanoparticles are of interest in the antimicrobial field because of their higher drug stability, solubility, penetration power, and treatment efficacy. In this study, propolis nanoparticles (PNPs) were synthesized, and their antibacterial and anti-biofilm activities against methicillin-resistant Staphylococcus aureus (MRSA) in combination with ampicillin sodium (AS) were analyzed. The PNPs had an average particle diameter of 118.0 nm, a polydispersity index of 0.129, and a zeta potential of −28.2 mV. The fractional inhibitory concentration indices of PNPs and AS against tested MRSA strains highlighted this synergy, ranging between 0.375 and 0.5. Crystal violet staining showed that combined PNPs and AS significantly inhibited biofilm formation and reduced existing biofilm biomass. We then discovered that PNPs inhibited bacterial adhesion, extracellular polysaccharide synthesis, and mecR1, mecA, blaZ, and icaADBC gene expression. These results indicated that PNPs exerted a synergistic antibacterial effect with AS by inhibiting mecR1, mecA, and blaZ gene expressions to reduce the drug resistance of MRSA. Meanwhile, PNPs weakened bacterial adhesion and aggregation by suppressing icaADBC gene expression, allowing antibiotics to penetrate the biofilm, and exhibiting significant synergistic anti-biofilm activity. In summary, PNPs are promising candidates for combating MRSA-related diseases. Full article
(This article belongs to the Special Issue Bacterial Antibiotic Resistance, Second Edition)
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17 pages, 704 KiB  
Review
Marine Antimicrobial Peptides: Emerging Strategies Against Multidrug-Resistant and Biofilm-Forming Bacteria
by Rita Magalhães, Dalila Mil-Homens, Sónia Cruz and Manuela Oliveira
Antibiotics 2025, 14(8), 808; https://doi.org/10.3390/antibiotics14080808 - 7 Aug 2025
Abstract
The global rise in antimicrobial resistance poses a major threat to public health, with multidrug-resistant bacterial infections expected to surpass cancer in mortality by 2050. As traditional antibiotic pipelines stagnate, novel therapeutic alternatives are critically needed. Antimicrobial peptides (AMPs), particularly those derived from [...] Read more.
The global rise in antimicrobial resistance poses a major threat to public health, with multidrug-resistant bacterial infections expected to surpass cancer in mortality by 2050. As traditional antibiotic pipelines stagnate, novel therapeutic alternatives are critically needed. Antimicrobial peptides (AMPs), particularly those derived from marine organisms, have emerged as promising antimicrobial candidates due to their broad-spectrum activity, structural diversity, and distinctive mechanisms of action. Unlike conventional antibiotics, AMPs can disrupt microbial membranes, inhibit biofilm formation, and even modulate immune responses, making them highly effective against resistant bacteria. This review highlights the potential of marine AMPs as next-generation therapeutics, emphasizing their efficacy against multidrug-resistant pathogens and biofilm-associated infections. Furthermore, marine AMPs show promise in combating persister cells and disrupting quorum sensing pathways, offering new strategies for tackling chronic infections. Despite their potential, challenges such as production scalability and limited clinical validation remain; nevertheless, the use of new technologies and bioinformatic tools is accelerating the discovery and optimization of these peptides, paving the way for bypassing these challenges. This review consolidates current findings on marine AMPs, advocating for their continued exploration as viable tools in the fight against antimicrobial resistance. Full article
(This article belongs to the Section Antimicrobial Peptides)
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18 pages, 1500 KiB  
Article
Structure-Activity Relationships in Alkoxylated Resorcinarenes: Synthesis, Structural Features, and Bacterial Biofilm-Modulating Properties
by Mariusz Urbaniak, Łukasz Lechowicz, Barbara Gawdzik, Maciej Hodorowicz and Ewelina Wielgus
Molecules 2025, 30(15), 3304; https://doi.org/10.3390/molecules30153304 - 7 Aug 2025
Abstract
In this study, a series of novel alkoxylated resorcinarenes were synthesized using secondary and tertiary alcohols under mild catalytic conditions involving iminodiacetic acid. Structural characterization, including single-crystal X-ray diffraction, confirmed the successful incorporation of branched alkyl chains and highlighted the influence of substitution [...] Read more.
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. Full article
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18 pages, 7277 KiB  
Article
Comprehensive Analysis of the Molecular Epidemiological Characteristics of Duck-Derived Salmonella in Certain Regions of China
by Jiawen Chen, Xiangdi Li, Yanling Liu, Wenjia Rong, Laiyu Fu, Shuhua Wang, Yan Li, Xiaoxiao Duan, Yongda Zhao and Lili Guo
Microbiol. Res. 2025, 16(8), 184; https://doi.org/10.3390/microbiolres16080184 - 7 Aug 2025
Abstract
Salmonella is a major foodborne pathogen, yet real-time data on duck-derived strains in China remain scarce. This study investigated the epidemiology, antimicrobial resistance (AMR), gene profiles, and PFGE patterns of 114 Salmonella isolates recovered from 397 deceased ducks (2021–2024) across nine provinces (isolation [...] Read more.
Salmonella is a major foodborne pathogen, yet real-time data on duck-derived strains in China remain scarce. This study investigated the epidemiology, antimicrobial resistance (AMR), gene profiles, and PFGE patterns of 114 Salmonella isolates recovered from 397 deceased ducks (2021–2024) across nine provinces (isolation rate: 28.72%). Fourteen serotypes were identified, with S. Typhimurium (23.68%), S. Indiana (21.93%), S. Kentucky (18.42%), and S. Enteritidis (12.28%) being predominant. Most isolates showed high resistance to β-lactams, tetracyclines, quinolones, and sulfonamides, with extensive multidrug resistance (MDR) observed—especially in S. Indiana, S. Typhimurium, and S. Kentucky. Among the 23 detected resistance genes, tet(B) had the highest prevalence (75.44%), particularly in S. Indiana. Biofilm formation was observed in 99.12% of isolates, with 84.21% demonstrating moderate to strong capacity. Eighteen virulence genes were detected; S. Enteritidis carried more spvB/C, sipB, and sodC1, while S. Indiana had higher cdtB carriage. PFGE revealed substantial genetic diversity among strains. This comprehensive analysis highlights the high AMR and biofilm potential of duck-derived Salmonella in China, emphasizing the urgent need for enhanced surveillance and control measures to mitigate public health risks. Full article
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52 pages, 1574 KiB  
Review
Anti-QS Strategies Against Pseudomonas aeruginosa Infections
by Abdelaziz Touati, Nasir Adam Ibrahim, Lilia Tighilt and Takfarinas Idres
Microorganisms 2025, 13(8), 1838; https://doi.org/10.3390/microorganisms13081838 - 7 Aug 2025
Abstract
Pseudomonas aeruginosa poses significant health threats due to its multidrug-resistant profile, particularly affecting immunocompromised individuals. The pathogen’s ability to produce virulence factors and antibiotic-resistant biofilms, orchestrated through quorum-sensing (QS) mechanisms, complicates conventional therapeutic interventions. This review aims to critically assess the potential of [...] Read more.
Pseudomonas aeruginosa poses significant health threats due to its multidrug-resistant profile, particularly affecting immunocompromised individuals. The pathogen’s ability to produce virulence factors and antibiotic-resistant biofilms, orchestrated through quorum-sensing (QS) mechanisms, complicates conventional therapeutic interventions. This review aims to critically assess the potential of anti-QS strategies as alternatives to antibiotics against P. aeruginosa infections. Comprehensive literature searches were conducted using databases such as PubMed, Scopus, and Web of Science, focusing on studies addressing QS inhibition strategies published recently. Anti-QS strategies significantly attenuate bacterial virulence by disrupting QS-regulated genes involved in biofilm formation, motility, toxin secretion, and immune evasion. These interventions reduce the selective pressure for resistance and enhance antibiotic efficacy when used in combination therapies. Despite promising outcomes, practical application faces challenges, including specificity of inhibitors, pharmacokinetic limitations, potential cytotoxicity, and bacterial adaptability leading to resistance. Future perspectives should focus on multi-target QS inhibitors, advanced delivery systems, rigorous preclinical validations, and clinical translation frameworks. Addressing current limitations through multidisciplinary research can lead to clinically viable QS-targeted therapies, offering sustainable alternatives to traditional antibiotics and effectively managing antibiotic resistance. Full article
(This article belongs to the Collection Feature Papers in Medical Microbiology)
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11 pages, 910 KiB  
Article
Antimicrobial Effect of Gentamicin/Heparin and Gentamicin/Citrate Lock Solutions on Staphylococcus aureus and Pseudomonas aeruginosa Clinical Strains
by Daniel Salas-Treviño, Arantxa N. Rodríguez-Rodríguez, María T. Ramírez-Elizondo, Magaly Padilla-Orozco, Edeer I. Montoya-Hinojosa, Paola Bocanegra-Ibarias, Samantha Flores-Treviño and Adrián Camacho-Ortiz
Infect. Dis. Rep. 2025, 17(4), 98; https://doi.org/10.3390/idr17040098 - 6 Aug 2025
Abstract
Background/Objectives: Hemodialysis catheter-related bloodstream infection (HD-CRBSIs) is a main cause of morbidity in hemodialysis. New preventive strategies have emerged, such as using lock solutions with antiseptic or antibiotic capacity. In this study, the antimicrobial effect was analyzed in vitro and with a catheter [...] Read more.
Background/Objectives: Hemodialysis catheter-related bloodstream infection (HD-CRBSIs) is a main cause of morbidity in hemodialysis. New preventive strategies have emerged, such as using lock solutions with antiseptic or antibiotic capacity. In this study, the antimicrobial effect was analyzed in vitro and with a catheter model of lock solutions of gentamicin (LSG), gentamicin/heparin (LSG/H), and gentamicin/citrate (LSG/C) in clinical and ATCC strains of Pseudomonas aeruginosa and Staphylococcus aureus. Methods: The formation, minimum inhibitory concentration, and minimum inhibitory concentration of the biofilm and minimum biofilm eradication concentration of the lock solutions were determined. Additionally, colony-forming unit assays were performed to evaluate the antimicrobial efficacy of the lock solutions in a hemodialysis catheter inoculation model. Results: The minimum inhibitory concentration (MIC) of planktonic cells of both P. aeruginosa and S. aureus for LSG/H and LSG/C was 4 µg/mL. In the minimum biofilm inhibitory concentration (MBIC) tests, the LSG/H was less effective than LSG/C, requiring higher concentrations for inhibition, contrary to the minimum biofilm eradication concentration (MBEC), where LSG/H was more effective. All lock solutions eradicated P. aeruginosa biofilms in the HD catheter model under standard conditions. Nevertheless, under modified conditions, the lock solutions were not as effective versus ATCC and clinical strains of S. aureus. Conclusions: Our analysis shows that the lock solutions studied managed to eradicate intraluminal mature P. aeruginosa in non-tunneled HD catheters under standard conditions. Biofilm inhibition and eradication were observed at low gentamicin concentrations, which could optimize the gentamicin concentration in lock solutions used in HD catheters. Full article
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18 pages, 3014 KiB  
Article
Biocide Tolerance, Biofilm Formation, and Efflux Pump Activity in Clinical Isolates of Trichosporon asahii
by Yasmim Passos Lima, Jamile de Paiva Macedo, Alessandra Barbosa Ferreira Machado, Cláudio Galuppo Diniz, Vania Lucia da Silva and Vanessa Cordeiro Dias
Infect. Dis. Rep. 2025, 17(4), 97; https://doi.org/10.3390/idr17040097 - 6 Aug 2025
Abstract
Background: Trichosporon spp. are opportunistic fungi, capable of causing infection, especially in critically ill individuals who often use broad-spectrum antibiotics, invasive devices, and have comorbidities. Objectives The aim of this study was to analyze individuals’ clinical characteristics, evaluate tolerance to biocides, as well [...] Read more.
Background: Trichosporon spp. are opportunistic fungi, capable of causing infection, especially in critically ill individuals who often use broad-spectrum antibiotics, invasive devices, and have comorbidities. Objectives The aim of this study was to analyze individuals’ clinical characteristics, evaluate tolerance to biocides, as well as biofilm formation and efflux pump activity in isolates of Trichosporon asahii. Methods: Clinical isolates of T. asahii collected between 2020 and 2023 from both hospitalized and non-hospitalized individuals, of both sexes, regardless of age, were tested for tolerance to sodium hypochlorite, hydrogen peroxide, benzalkonium chloride, and ethyl alcohol. Efflux pump activity was also assessed using ethidium bromide, and biofilm formation was measured with the Safranin test. Clinical parameters such as outcomes, source, and length of hospitalization were analyzed through electronic medical records. Results: A total of 37 clinical isolates of T. asahii were identified. Thirty-three (83.8%) isolates were from hospitalized individuals, with 81.82% collected in ICUs, an average hospital stay of 35 days, and a mortality rate of 51.6%. The tested strains displayed the largest mean inhibition zone for 2% sodium hypochlorite, indicating lower tolerance. A high level of efflux pump expression was detected among clinical isolates. Biofilm formation was detected in 25/67.5% of the isolates. Conclusions: These findings highlight the clinical relevance of T. asahii, particularly in critically ill individuals, and underscore the pathogen’s ability to tolerate biocides, express efflux pumps, and form biofilms, all of which may contribute to its persistence and pathogenicity in hospital environments. Enhanced surveillance and effective microbial control measures are essential to mitigate the risks associated with T. asahii infections. Full article
(This article belongs to the Section Fungal Infections)
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33 pages, 4268 KiB  
Review
Targeting Bacterial Biofilms on Medical Implants: Current and Emerging Approaches
by Alessandro Calogero Scalia and Ziba Najmi
Antibiotics 2025, 14(8), 802; https://doi.org/10.3390/antibiotics14080802 - 6 Aug 2025
Abstract
Biofilms are structured communities of microorganisms encased in a self-produced extracellular matrix, and they represent one of the most widespread forms of microbial life on Earth. Their presence poses serious challenges in both environmental and clinical settings. In natural and industrial systems, biofilms [...] Read more.
Biofilms are structured communities of microorganisms encased in a self-produced extracellular matrix, and they represent one of the most widespread forms of microbial life on Earth. Their presence poses serious challenges in both environmental and clinical settings. In natural and industrial systems, biofilms contribute to water contamination, pipeline corrosion, and biofouling. Clinically, biofilm-associated infections are responsible for approximately 80% of all microbial infections, including endocarditis, osteomyelitis, cystic fibrosis, and chronic sinusitis. A particularly critical concern is their colonization of medical devices, where biofilms can lead to chronic infections, implant failure, and increased mortality. Implantable devices, such as orthopedic implants, cardiac pacemakers, cochlear implants, urinary catheters, and hernia meshes, are highly susceptible to microbial attachment and biofilm development. These infections are often recalcitrant to conventional antibiotics and frequently necessitate surgical revision. In the United States, over 500,000 biofilm-related implant infections occur annually, with prosthetic joint infections alone projected to incur revision surgery costs exceeding USD 500 million per year—a figure expected to rise to USD 1.62 billion by 2030. To address these challenges, surface modification of medical devices has emerged as a promising strategy to prevent bacterial adhesion and biofilm formation. This review focuses on recent advances in chemical surface functionalization using non-antibiotic agents, such as enzymes, chelating agents, quorum sensing quenching factors, biosurfactants, oxidizing compounds and nanoparticles, designed to enhance antifouling and mature biofilm eradication properties. These approaches aim not only to prevent device-associated infections but also to reduce dependence on antibiotics and mitigate the development of antimicrobial resistance. Full article
(This article belongs to the Special Issue Antibacterial and Antibiofilm Properties of Biomaterial)
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24 pages, 957 KiB  
Review
Biofilm and Antimicrobial Resistance: Mechanisms, Implications, and Emerging Solutions
by Bharmjeet Singh, Manju Dahiya, Vikram Kumar, Archana Ayyagari, Deepti N. Chaudhari and Jayesh J. Ahire
Microbiol. Res. 2025, 16(8), 183; https://doi.org/10.3390/microbiolres16080183 - 6 Aug 2025
Abstract
Biofilms are a spontaneously formed slimy matrix of extracellular polymeric substances (EPS) enveloping miniature bacterial colonies, which aid in pathogen colonization, shielding the bacteria from antibiotics, as well as imparting them resistance towards the same. Biofilms employ a robust communication mechanism called quorum [...] Read more.
Biofilms are a spontaneously formed slimy matrix of extracellular polymeric substances (EPS) enveloping miniature bacterial colonies, which aid in pathogen colonization, shielding the bacteria from antibiotics, as well as imparting them resistance towards the same. Biofilms employ a robust communication mechanism called quorum sensing that serves to keep their population density constant. What is most significant about biofilms is that they contribute to the development of bacterial virulence by providing protection to pathogenic species, allowing them to colonize the host, and also inhibiting the activities of antimicrobials on them. They grow on animate surfaces (such as on teeth and intestinal mucosa, etc.) and inanimate objects (like catheters, contact lenses, pacemakers, endotracheal devices, intrauterine devices, and stents, etc.) alike. It has been reported that as much as 80% of human infections involve biofilms. Serious implications of biofilms include the necessity of greater concentrations of antibiotics to treat common human infections, even contributing to antimicrobial resistance (AMR), since bacteria embedded within biofilms are protected from the action of potential antibiotics. This review explores various contemporary strategies for controlling biofilms, focusing on their modes of action, mechanisms of drug resistance, and innovative approaches to find a solution in this regard. This review interestingly targets the extracellular polymeric matrix as a highly effective strategy to counteract the potential harm of biofilms since it plays a critical role in biofilm formation and significantly contributes to antimicrobial resistance. Full article
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19 pages, 1579 KiB  
Article
Plasma-Treated Water Effect on Sporulating Bacillus cereus vs. Non-Sporulating Listeria monocytogenes Biofilm Cell Vitality
by Samantha Nestel, Robert Wagner, Mareike Meister, Thomas Weihe and Uta Schnabel
Appl. Microbiol. 2025, 5(3), 80; https://doi.org/10.3390/applmicrobiol5030080 - 5 Aug 2025
Abstract
Foodborne illness caused by bacterial pathogens is a global health concern and results in millions of infections annually. Therefore, food products typically undergo several processing stages, including sanitation steps, before being distributed in an attempt to remove pathogens. However, many sanitation methods have [...] Read more.
Foodborne illness caused by bacterial pathogens is a global health concern and results in millions of infections annually. Therefore, food products typically undergo several processing stages, including sanitation steps, before being distributed in an attempt to remove pathogens. However, many sanitation methods have compounding effects on the color, texture, flavor, and nutritional quality of the product or do not effectively reduce the pathogens that food can be exposed to. Some bacterial pathogens particularly possess traits and tactics that make them even more difficult to mitigate such as biofilm formation. Non-thermal plasma sanitation techniques, including plasma-treated water (PTW), have proven to be promising methods that significantly reduce pathogenic bacteria that food is exposed to. Published work reveals that PTW can effectively mitigate both gram-positive and gram-negative bacterial biofilms. This study presents a novel analysis of the differences in antimicrobial effects of PTW treatment between biofilm-forming gram-positive bacteria, commonly associated with foodborne illness, that are sporulating (Bacillus cereus) and non-sporulating (Listeria monocytogenes). After treatment with PTW, the results suggest the following hypotheses: (1) that the non-sporulating species experiences less membrane damage but a greater reduction in metabolic activity, leading to a possible viable but non-culturable (VBNC) state, and (2) that the sporulating species undergoes spore formation, which may subsequently convert into vegetative cells over time. PTW treatment on gram-positive bacterial biofilms that persist in food processing environments proves to be effective in reducing the proliferating abilities of the bacteria. However, the variance in PTW’s effects on metabolic activity and cell vitality between sporulating and non-sporulating species suggest that other survival tactics might be induced. This analysis further informs the application of PTW in food processing as an effective sanitation method. Full article
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17 pages, 2170 KiB  
Article
RcsB and H-NS Both Contribute to the Repression the Expression of the csgDEFG Operon
by Hiroshi Ogasawara, Azusa Tomioka and Yuki Kato
Microorganisms 2025, 13(8), 1829; https://doi.org/10.3390/microorganisms13081829 - 5 Aug 2025
Abstract
Curli fimbriae are a major component of biofilm formation in Escherichia coli, and their expression is regulated by numerous transcription factors and small regulatory RNAs (sRNAs). The RcsD-RcsC-RcsB phosphorelay system, which is involved in the envelope stress response, plays a role in [...] Read more.
Curli fimbriae are a major component of biofilm formation in Escherichia coli, and their expression is regulated by numerous transcription factors and small regulatory RNAs (sRNAs). The RcsD-RcsC-RcsB phosphorelay system, which is involved in the envelope stress response, plays a role in this regulation. In this study, we report that DNase-I footprinting analysis revealed that the response regulator RcsB interacts with the −31 to +53 region of the promoter region of csgD, which encodes a major regulator of biofilm formation, and thus contributes to its transcriptional repression. Additionally, overexpression of RcsB or RcsB D56A that could not be phosphorylated by the histidine kinases RcsC and D both significantly reduced csgD expression and suppressed Curli formation. This indicates that the phosphorylation of RcsB has an insignificant impact on its affinity for its operator sites. Furthermore, we confirm that RcsB binds cooperatively to the csgD promoter region in the presence of the nucleoid-associated protein H-NS. Our study also confirms that RcsB positively regulates the expression of an sRNA, RprA, which is known to reduce mRNA csgD mRNA translation RprA via its binding to the 5′-untranslated region (UTR) of csgD. These findings indicate that, in E. coli, the RcsBCD system suppresses csgD expression through both direct transcriptional repression by the regulator RcsB and translational repression by the sRNA RprA. Full article
(This article belongs to the Special Issue Transcriptional Regulation in Bacteria, 2nd Edition)
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18 pages, 4470 KiB  
Article
Cloning, Heterologous Expression, and Antifungal Activity Evaluation of a Novel Truncated TasA Protein from Bacillus amyloliquefaciens BS-3
by Li-Ming Dai, Li-Li He, Lan-Lan Li, Yi-Xian Liu, Yu-Ping Shi, Hai-Peng Su and Zhi-Ying Cai
Int. J. Mol. Sci. 2025, 26(15), 7529; https://doi.org/10.3390/ijms26157529 - 4 Aug 2025
Viewed by 166
Abstract
TasA gene, encoding a functional amyloid protein critical for biofilm formation and antimicrobial activity, was cloned from the endophytic strain Bacillus amyloliquefaciens BS-3, isolated from rubber tree roots. This study identified the shortest functional TasA variant (483 bp, 160 aa) reported to date, [...] Read more.
TasA gene, encoding a functional amyloid protein critical for biofilm formation and antimicrobial activity, was cloned from the endophytic strain Bacillus amyloliquefaciens BS-3, isolated from rubber tree roots. This study identified the shortest functional TasA variant (483 bp, 160 aa) reported to date, featuring unique amino acid substitutions in conserved domains. Bioinformatics analysis predicted a signal peptide (1–27 aa) and transmembrane domain (7–29 aa), which were truncated to optimize heterologous expression. Two prokaryotic vectors (pET28a and pCZN1) were constructed, with pCZN1-TasA expressed solubly in Escherichia coli Arctic Express at 15 °C, while pET28a-TasA formed inclusion bodies at 37 °C. Purified recombinant TasA exhibited potent antifungal activity, achieving 98.6% ± 1.09 inhibition against Colletotrichum acutatum, 64.77% ± 1.34 against Alternaria heveae. Notably, TasA completely suppressed spore germination in C. acutatum and Oidium heveae Steinmannat 60 μg/mL. Structural analysis via AlphaFold3 revealed that truncation enhanced protein stability. These findings highlight BS-3-derived TasA as a promising biocontrol agent, providing molecular insights for developing protein-based biopesticides against rubber tree pathogens. Full article
(This article belongs to the Section Biochemistry)
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31 pages, 698 KiB  
Review
Mechanistic Role of Heavy Metals in Driving Antimicrobial Resistance: From Rhizosphere to Phyllosphere
by Rahul Kumar, Tanja P. Vasić, Sanja P. Živković, Periyasamy Panneerselvam, Gustavo Santoyo, Sergio de los Santos Villalobos, Adeyemi Nurudeen Olatunbosun, Aditi Pandit, Leonard Koolman, Debasis Mitra and Pankaj Gautam
Appl. Microbiol. 2025, 5(3), 79; https://doi.org/10.3390/applmicrobiol5030079 - 4 Aug 2025
Viewed by 71
Abstract
Heavy metal pollution represents a pervasive environmental challenge that significantly exacerbates the ever-increasing crisis of antimicrobial resistance and the capacity of microorganisms to endure and proliferate despite antibiotic interventions. This review examines the intricate relationship between heavy metals and AMR, with an emphasis [...] Read more.
Heavy metal pollution represents a pervasive environmental challenge that significantly exacerbates the ever-increasing crisis of antimicrobial resistance and the capacity of microorganisms to endure and proliferate despite antibiotic interventions. This review examines the intricate relationship between heavy metals and AMR, with an emphasis on the underlying molecular mechanisms and ecological ramifications. Common environmental metals, including arsenic, mercury, cadmium, and lead, exert substantial selective pressures on microbial communities. These induce oxidative stress and DNA damage, potentially leading to mutations that enhance antibiotic resistance. Key microbial responses include the overexpression of efflux pumps that expel both metals and antibiotics, production of detoxifying enzymes, and formation of protective biofilms, all of which contribute to the emergence of multidrug-resistant strains. In the soil environment, particularly the rhizosphere, heavy metals disrupt plant–microbe interactions by inhibiting beneficial organisms, such as rhizobacteria, mycorrhizal fungi, and actinomycetes, thereby impairing nutrient cycling and plant health. Nonetheless, certain microbial consortia can tolerate and detoxify heavy metals through sequestration and biotransformation, rendering them valuable for bioremediation. Advances in biotechnology, including gene editing and the development of engineered metal-resistant microbes, offer promising solutions for mitigating the spread of metal-driven AMR and restoring ecological balance. By understanding the interplay between metal pollution and microbial resistance, we can more effectively devise strategies for environmental protection and public health. Full article
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