Initial Biofilm Formation Research Articles

Antibacterial and Antibiofilm Potential of Chlorophyllin Against Streptococcus mutans In Vitro and In Silico.

Streptococcus mutans is a leading causative agent of dental caries and exerts pathogenicity by forming biofilms. Dental caries continues to be a significant public health issue worldwide, affecting an estimated 2.5 billion people, showing a 14.6% increase over the past decade. Herein, the antibacterial potential of Chlorophyllin extracted from Spinacia oleracea was evaluated against biofilm-forming S. mutans via in vitro and in silico studies. The antimicrobial activity of chlorophyllin extract against S. mutans isolates was tested using the agar well diffusion method. Chlorophyllin extract was also tested against biofilm-forming isolates of S. mutans. Chlorophyllin was docked with the antigen I/II (AgI/II) protein of S. mutans to evaluate its antimicrobial mechanism. The chemical structure and canonical SMILES format of Chlorophyllin were obtained from PubChem. Additionally, adsorption, distribution, metabolism, excretion, and toxicity (ADMET) analyses of Chlorophyllin were performed using ADMETlab 2.0 to assess its pharmacokinetic properties. An agar well diffusion assay revealed that all S. mutans isolates were susceptible to Chlorophyllin extract and showed a variety of inhibition zones ranging from 32 to 41 mm. Chlorophyllin reduces the biofilm strength of four isolates from strong to moderate and six from strong to weak. The antibiofilm potential of Chlorophyllin was measured by a reduction in the number of functional groups observed in the Fourier Transform Infrared Spectrometer (FTIR) spectra of the extracellular polymeric substance (EPS) samples. Chlorophyllin showed binding with AgI/II proteins of S. mutans, which are involved in adherence to the tooth surface and initiating biofilm formation. The ADMET analysis revealed that the safety of Chlorophyllin exhibited favorable pharmacokinetic properties. Chlorophyllin stands out as a promising antibacterial and antibiofilm agent against biofilm-forming S. mutans, and its safety profile highlights its potential suitability for further investigation as a therapeutic agent.

Metallo-β-lactamases producing Pseudomonas aeruginosa and the molecular mechanism of drug resistance variants

BackgroundMetallo-β-lactamases (MBLs) type carbapenemases are produced by pathogenic Pseudomonas spp. and exhibit carbapenemase activity. The study will look into drug resistance and the molecular mechanisms of drug-resistant Pseudomonas aeruginosa variants. MethodsA total of 74 P. aeruginosa strains were isolated from urine, pus, sputum, blood, throat swab, Foley’s catheter, and nasal swab. The isolates were screened for antibiotic susceptibility and the identified MDR strains were further tested for β-lactamase production. Multiplex PCR was used to identify the presence of mcr-1 and blaNDM-1 genes in MDR organisms. The minimum inhibitory concentration for ceftazidime and colistin was also determined, in addition to the biofilm inhibitor activity. Confocal microscopy was used to determine the production of biofilms. ResultsThe isolated P. aeruginosa strains exhibited antibiotic resistance to aminoglycosides (amikacin and gentamicin). Fifty three percent of the isolated P. aeruginosa strains produced metallo-β-lactamase and the remaining isolates were non-metallo-β-lactamase type (46.8 %) (p < 0.0001). The MIC value of β-lactamase producers against colistin ranges from 0.5 µg/mL to 6 µg/mL. The MDR bacteria exhibited mcr-1 and blaNDM−1 genes. The MDR P. aeruginosa strain treated with colistin and ceftazidime inhibited initial biofilm formation. These combination of antibiotics effectively prevented initial biofilm development than individual antibiotics (p < 0.001). ConclusionsThe current analysis detected MDR among P. aeruginosa isolates that carried drug-resistant genes.

Pseudomonas aeruginosa Activates Quorum Sensing, Antioxidant Enzymes and Type VI Secretion in Response to Oxidative Stress to Initiate Biofilm Formation and Wound Chronicity.

Pseudomonas aeruginosa (PA) is an opportunistic pathogen frequently isolated from cutaneous chronic wounds. How PA, in the presence of oxidative stress (OS), colonizes chronic wounds and forms a biofilm is still unknown. The purpose of this study is to investigate the changes in gene expression seen when PA is challenged with the high levels of OS present in chronic wounds. We used a biofilm-forming PA strain isolated from the chronic wounds of our murine model (RPA) and performed a qPCR to obtain gene expression patterns as RPA developed a biofilm in vitro in the presence of high levels of OS, and then compared the findings in vivo, in our mouse model of chronic wounds. We found that the planktonic bacteria under OS conditions overexpressed quorum sensing genes that are important for the bacteria to communicate with each other, antioxidant stress genes important to reduce OS in the microenvironment for survival, biofilm formation genes and virulence genes. Additionally, we performed RNAseq in vivo and identified the activation of novel genes/pathways of the Type VI Secretion System (T6SS) involved in RPA pathogenicity. In conclusion, RPA appears to survive the high OS microenvironment in chronic wounds and colonizes these wounds by turning on virulence, biofilm-forming and survival genes. These findings reveal pathways that may be promising targets for new therapies aimed at disrupting PA-containing biofilms immediately after debridement to facilitate the treatment of chronic human wounds.

Current Knowledge of Enterococcal Endocarditis: A Disease Lurking in Plain Sight of Health Providers.

Enterococcus faecalis is a bacterial pathogen that can cause opportunistic infections. Studies indicate that initial biofilm formation plays a crucial regulatory role in these infections, as well as in colonising and maintaining the gastrointestinal tract as a commensal member of the microbiome of most land animals. It has long been thought that vegetation of endocarditis resulting from bacterial attachment to the endocardial endothelium requires some pre-existing tissue damage, and in animal models of experimental endocarditis, mechanical valve damage is typically induced by cardiac catheterisation preceding infection. This section reviews historical and contemporary animal model studies that demonstrate the ability of E. faecalis to colonise the undamaged endovascular endothelial surface directly and produce robust microcolony biofilms encapsulated within a bacterially derived extracellular matrix. This report reviews both previous and current animal model studies demonstrating the resilient capacity of E. faecalis to colonise the undamaged endovascular endothelial surface directly and produce robust microcolony biofilms encapsulated in a bacterially derived extracellular matrix. The article also considers the morphological similarities when these biofilms develop on different host sites, such as when E. faecalis colonises the gastrointestinal epithelium as a commensal member of the common vertebrate microbiome, lurking in plain sight and transmitting systemic infection. These phenotypes may enable the organism to survive as an unrecognised infection in asymptomatic subjects, providing an infectious resource for subsequent clinical process of endocarditis.

Transcriptional analysis of oxidative and nitrosative stress on oral opportunistic Candida albicans.

The yeast Candida albicans is one of the most aggressive opportunistic pathogens in immunocompromised patients. The ability of the yeast to withstand stresses and radicals is of great concern. In the present study, four isolates of C. albicans were taken from patients with oral candidiasis and grown on RPMI for 24 hours at 37°C. Then, they were exposed to various concentrations of oxidative (H2O2) and nitrosative (HNO3) stress for two hours, and gene expression rates were measured through RT-PCR. After initial biofilm formation steps and growth validation, RNA extracted from the yeast and gene expression status were evaluated. Upon treatment with H2O2, the gene expression profile for ALS1, MLH1, and EXO1 showed approximately a fold increase in expression. While within HNO3 the yeast gene expression exhibited a dramatic increase in ALS1 up to 217 folds, while others such as MLH1, HWP1, and ERG11 showed a one-fold increase in the expression rate. The findings of this research indicate a considerable expression activity within the biofilm of Candida albicans, increased rate of DNA mismatch repair and break fixation may indicate the ability of the yeast to tolerate high concentrations of free radicals. It paves the way toward understanding the pathogenicity of the yeast and its survival capability inside macrophages. The study also revealed that the biofilm strategy of the yeast is more active within these stresses.

Biofilm volume and acidification within initial biofilms formed in situ on buccally and palatally exposed bracket material.

Acidification by bacterial biofilms at the bracket/tooth interface is one of the most common problems in fixed orthodontic treatments, which can lead to white spot lesions (WSL) and caries. As lingual brackets were shown to exhibit reduced WSL formation clinically, the aim of this in situ study was to compare initial intraoral biofilm formation and acidification on bracket-like specimens placed buccally and palatally in the upper jaw as apossible cause for this observation. Intraoral biofilm was collected from splints equipped with buccally and palatally exposed test specimens, which were worn by 12volunteers for atotal of 48 h. The test specimens consisted of standard bracket material cylinders on top of ahydroxyapatite disc to represent the bracket/tooth interface. They were analyzed for three-dimensional biofilm volume and live/dead distribution by fluorescence staining and confocal laser scanning microscopy as well as for acidification by fluorescence-based pH ratiometry. Similar general biofilm morphology with regard to volume and viability could be detected for buccally and palatally exposed specimens. For pH values, biofilms from both positions showed increased acidification at the bottom layer. Interestingly, the pH value at the top layers of the biofilms was slightly lower on palatally than on buccally exposed specimens, which may likely be due to anatomic conditions. Based on the results of this study, initial intraoral biofilm formation and acidification is almost similar on the bracket material/biomimetic tooth interface when placed buccally or palatally in the upper jaw. As lingual brackets were shown to exhibit reduced WSL formation clinically, future studies should investigate further factors like bracket geometry.

Time-lapse confocal microscopy to study in vitro Streptococcus mutans surface colonization.

The cariogenicity of Streptococcus mutans relates to its ability to form biofilms on dental surfaces. The aim of this work was to develop a flowcell system compatible with time-lapse confocal microscopy to compare the adhesion and accumulation of S. mutans cells on surfaces in unsupplemented media against media containing sucrose or sucralose (a non-metabolized sweetener) over a short period of time. Fluorescent S. mutans 3209/pVMCherry was suspended in unsupplemented media or media supplemented with 1% sucrose or 1% sucralose and passed through a 3D-printed flowcell system. Flowcells were imaged over 60 minutes using a confocal microscope. Image analysis was performed, including a newly developed object-movement-based method to measure biomass adhesion. Streptococcus mutans 3209/pVMCherry grown in 1% sucrose-supplemented media formed small, dense, relatively immobile clumps in the flowcell system measured by biovolume, surface area, and median object centroid movement. Sucralose-supplemented and un-supplemented media yielded large, loose, mobile aggregates. Architectural metrics and per-object movement were significantly different (P<0.05) when comparing sucrose-supplemented media to either unsupplemented or sucralose-supplemented media. These results demonstrate the utility of a flowcell system compatible with time-lapse confocal microscopy and image analysis when studying initial biofilm formation and adhesion under different nutritional conditions.

Enhanced chlorobenzene removal by internal magnetic field through initial cell adhesion and biofilm formation.

Magnetic fields (MF) have been proven efficient in bioaugmentation, and the internal MFs have become competitive because they require no configuration, despite their application in waste gas treatment remaining largely unexplored. In this study, we firstly developed an intensity-regulable bioaugmentation with internal MF for gaseous chlorobenzene (CB) treatment with modified packing in batch bioreactors, and the elimination capacity increased by up to 26%, surpassing that of the external MF. Additionally, the microbial affinity to CB and the packing surface was enhanced, which was correlated with the ninefold increased secreted ratio of proteins/polysaccharides, 43% promoted cell surface hydrophobicity, and half reduced zeta potential. Furthermore, the dehydrogenase content was promoted over 3 times, and CB removal steadily increased with the rising intensity indicating enhanced biofilm activity and reduced CB bioimpedance; this was further supported by kinetic analysis, which resulted in improved cell adhesive ability and biological utilisation of CB. The results introduced a novel concept of adjustable magnetic bioaugmentation and provided technical support for industrial waste gas treatments. KEY POINTS: • Regulable magnetic bioaugmentation was developed to promote 26% chlorobenzene removal • Chlorobenzene mineralisation was enhanced under the magnetic field • Microbial adhesion was promoted through weakening repulsive forces.

A guide to Stenotrophomonas maltophilia virulence capabilities, as we currently understand them.

The Gram-negative pathogen Stenotrophomonas maltophilia causes a wide range of human infections. It causes particularly serious lung infections in individuals with cystic fibrosis, leading to high mortality rates. This pathogen is resistant to most known antibiotics and harbors a plethora of virulence factors, including lytic enzymes and serine proteases, that cause acute infection in host organisms. S. maltophilia also establishes chronic infections through biofilm formation. The biofilm environment protects the bacteria from external threats and harsh conditions and is therefore vital for the long-term pathogenesis of the microbe. While studies have identified several genes that mediate S. maltophilia's initial colonization and biofilm formation, the cascade of events initiated by these factors is poorly understood. Consequently, understanding these and other virulence factors can yield exciting new targets for novel therapeutics.

RpoN (sigma factor 54) contributes to bacterial fitness during tracheal colonization of Bordetella bronchiseptica.

The Gram-negative pathogenic bacterium Bordetella bronchiseptica is a respiratory pathogen closely related to Bordetella pertussis, the causative agent of whooping cough. Despite sharing homologous virulence factors, B. bronchiseptica infects a broad range of mammalian hosts, including some experimental animals, whereas B. pertussis is strictly adapted to humans. Therefore, B. bronchiseptica is often used as a representative model to explore the pathogenicity of Bordetella in infection experiments with laboratory animals. Although Bordetella virulence factors, including toxins and adhesins have been studied well, our recent study implied that unknown virulence factors are involved in tracheal colonization and infection. Here, we investigated bacterial genes contributing to tracheal colonization by high-throughput transposon sequencing (Tn-seq). After the screening, we picked up 151 candidate genes of various functions and found that a rpoN-deficient mutant strain was defective in tracheal colonization when co-inoculated with the wild-type strain. rpoN encodes σ54 , a sigma factor that regulates the transcription of various genes, implying its contribution to various bacterial activities. In fact, we found RpoN of B. bronchiseptica is involved in bacterial motility and initial biofilm formation. From these results, we propose that RpoN supports bacterial colonization by regulating various bacteriological functions.

Biofilm dynamic changes in drip irrigation emitter flow channels using reclaimed water

Reusing reclaimed water through drip irrigation offers an effective way to overcome freshwater scarcity. However, biofilm accumulation in the flow channel of drip emitters is the primary obstacle. So far, biofilm development in the emitters remains largely unknown. Here, industrial computed tomography was used to quantify the emitter biofilm developments. Results showed that biofilm typically accumulated in the inlet of the emitter flow channel and decreased toward the flow direction. Biofilm was also typically accumulated in dead areas having low flow velocities, such as edge, corner, and adjacent surface zones in flow channels. The dynamics of the emitter biofilm variation mechanisms were also elucidated. Specifically, particle and nutrient transport appeared to be dominant in shaping the initial biofilm formation (i.e., 5% emitter clogging degree), resulting in biofilm abundant in the inlet of the flow channel. However, it changed the hydraulic conditions in emitter flow channels, increased local water shear stress, which made the biofilm easier to be washed away in the inlet, and caused biofilm to grow faster in the middle and outlet regions of the flow channel at 20% emitter clogging degree. At 50% clogging degree, biofilm in inlet regions grew faster again. Finally, some biofilm control approaches such as water quality management and emitter flow channel structure optimization were proposed. This study provided a better understanding of biofilm formation mechanisms in the emitter flow channels, which is critical for designing appropriate biofilm control technologies and will facilitate the reuse of reclaimed water in agricultural irrigation.

Accelerating the environmental biodegradation of poly-3-hydroxybutyrate (PHB) via plasma surface treatment

The surface of poly-3-hydroxybutyrate (PHB) was modified using a low-pressure plasma system with air as the process gas to accelerate its biodegradation rate in soil. The water contact angle of PHB was reduced from 98° to 57° after plasma treatment, rendering the surface hydrophilic and also induced an increase in the surface free energy. Etching on the surface was observed after the plasma treatment without a significant change in the surface crystallinity. AFM imaging showed that the plasma treatment increased the surface roughness by about 10 folds and created diverse surface structures. The soil burial test showed an approximately 1.5-fold increase in the biodegradation rate for the plasma-treated sample. Initial microbial attachment and biofilm formation were higher on the modified surface. This study demonstrated that the surface morphology created by plasma treatment promoted initial colonization and subsequent biofilm formation on the PHB surface, facilitating and accelerating its biodegradation in soil.

Insights into the effect of ethyl xanthate on the adhesion and biofilm formation by Acidianus manzaensis on chalcopyrite

Adhesion and biofilm formation play important roles in the bio-dissolution of metal sulfides, which can initiate dissolution and enhance bioleaching. Ethyl xanthate, a commonly used flotation reagent, remained in flotation concentrates of metal sulfides and may have an impact on the bioleaching of metal sulfides. Therefore, it is important to analyze the effect of ethyl xanthate on microbial initial adhesion and biofilm formation. The results showed that the adhesion percentage of Acidianus manzaensis on chalcopyrite decreased with increasing ethyl xanthate concentration after adsorption equilibrium, due to the toxicity of ethyl xanthate to A. manzaensis. However, a low concentration (0.01 mg/L) of ethyl xanthate had little effect on the growth of A. manzaensis. The hydrophobicity of chalcopyrite increased after the addition of ethyl xanthate, leading to an increase in total interaction energy and adhesion force. The presence of ethyl xanthate inhibited the early growth of biofilm and delayed the formation of microcolonies on chalcopyrite. While biofilm coverage was larger with the addition of ethyl xanthate than that of the control when biofilm was mature. Furthermore, in the presence of ethyl xanthate, more and larger microcolonies were observed on the chalcopyrite surface when biofilm was matured, resulting in an increased corrosion degree and more jarosite produced on the chalcopyrite surface. This study suggested that a low concentration (0.01 mg/L) of ethyl xanthate could promote the dissolution of chalcopyrite, which could be significant for bioleaching of flotation reagent-bearing flotation concentrates.

PlzD modifies Vibrio vulnificus foraging behavior and virulence in response to elevated c-di-GMP.

Many free-swimming bacteria propel themselves through liquid using rotary flagella, and mounting evidence suggests that the inhibition of flagellar rotation initiates biofilm formation, a sessile lifestyle that is a nearly universal surface colonization paradigm in bacteria. In general, motility and biofilm formation are inversely regulated by the intracellular second messenger bis-(3´-5´)-cyclic dimeric guanosine monophosphate (c-di-GMP). Here, we identify a protein, PlzD, bearing a conserved c-di-GMP binding PilZ domain that localizes to the flagellar pole in a c-di-GMP-dependent manner and alters the foraging behavior, biofilm, and virulence characteristics of the opportunistic human pathogen, Vibrio vulnificus. Our data suggest that PlzD interacts with components of the flagellar stator to decrease bacterial swimming speed and changes in swimming direction, and these activities are enhanced when cellular c-di-GMP levels are elevated. These results reveal a physical link between a second messenger (c-di-GMP) and an effector (PlzD) that promotes transition from a motile to a sessile state in V. vulnificus.

The bioaccessibility of adsorped heavy metals on biofilm-coated microplastics and their implication for the progression of neurodegenerative diseases.

Microplastic (MP) tiny fragments (< 5mm) of conventional and specialized industrial polymers are persistent and ubiquitous in both aquatic and terrestrial ecosystem. Breathing, ingestion, consumption of food stuffs, potable water, and skin are possible routes of MP exposure that pose potential human health risk. Various microorganisms including bacteria, cyanobacteria, and microalgae rapidly colonized on MP surfaces which initiate biofilm formation. It gradually changed the MP surface chemistry and polymer properties that attract environmental metals. Physicochemical and environmental parameters like polymer type, dissolved organic matter (DOM), pH, salinity, ion concentrations, and microbial community compositions regulate metal adsorption on MP biofilm surface. A set of highly conserved proteins tightly regulates metal uptake, subcellular distribution, storage, and transport to maintain cellular homeostasis. Exposure of metal-MP biofilm can disrupt that cellular homeostasis to induce toxicities. Imbalances in metal concentrations therefore led to neuronal network dysfunction, ROS, mitochondrial damage in diseases like Alzheimer's disease (AD), Parkinson's disease (PD), and Prion disorder. This review focuses on the biofilm development on MP surfaces, factors controlling the growth of MP biofilm which triggered metal accumulation to induce neurotoxicological consequences in human body and stategies to reestablish the homeostasis. Thus, the present study gives a new approach on the health risks of heavy metals associated with MP biofilm in which biofilms trigger metal accumulation and MPs serve as a vector for those accumulated metals causing metal dysbiosis in human body.

Microalgae cell adhesions on hydrophobic membrane substrates using quartz crystal microbalance with dissipation.

Microalgal cell adhesion and biofilm formation are affected by interactions between microalgae strains and membrane materials. Variations of surface properties of microalgae and membrane materials are expected to affect cell-membranes and cell-cell interactions and thus initial microalgal cell adhesion and biofilm formation rates. Hence, it should be possible to identify the dominant mechanisms controlling microalgal cell adhesion and biofilm formation. The effects of surface properties of three different microalgal strains and three different types of membrane materials on microalgal cell adhesion and biofilm formation were systematically investigated in real time by monitoring changes in the oscillation frequency and dissipation of the quartz crystal resonator (QCM-D). The results revealed that in general a higher surface free energy, more negative zeta potential, and higher surface roughness of membrane materials positively correlated with a larger quantity of microalgae cell deposition, while a more hydrophilic microalgae with a larger negative zeta potential preferred to attach to a more hydrophobic membrane material. The adhered microalgal layers exhibited viscoelastic properties. The relative importance of these mechanisms in controlling microalgae cell attachment and biofilm formation might vary, depending on the properties of specific microalgae species and hydrophobic membrane materials used.

Surface properties of membrane materials and their role in cell adhesion and biofilm formation of microalgae

In this study, the effects of surface properties of membrane materials on microalgae cell adhesion and biofilm formation were investigated using Chlorella vulgaris and five different types of membrane materials under hydrodynamic conditions. The results suggest that the contact angle (hydrophobicity), surface free energy, and free energy of cohesion of membrane materials alone could not sufficiently elucidate the selectivity of microalgae cell adhesion and biofilm formation on membrane materials surfaces, and membrane surface roughness played a dominant role in controlling biofilm formation rate, under tested hydrodynamic conditions. A lower level of biofilm EPS production was generally associated with a larger amount of biofilm formation. The zeta potential of membrane materials could enhance initial microalgae cell adhesion and biofilm formation through salt bridging or charge neutralization mechanisms.

Supramolecular Self-Healing Antifouling Coating for Dental Materials.

In orthodontic treatment, orthodontic appliances are prone to bacterial infections, which pose a risk to oral health. Surface modification of orthodontic appliances has been explored to improve their antifouling properties and impart antibacterial capabilities, inhibiting initial bacterial adhesion and biofilm formation. However, coatings are susceptible to damage in the complex oral environment, leading to a loss of functionality. Here, we have prepared an antifouling self-healing coating based on supramolecular bonding by employing a simple spin coating method. The presence of the hydrophilic zwitterionic trimethylamine N-oxide (TMAO) and the hydrophobic antimicrobial moieties triclosan acrylate (TCSA) imparts to the polymers an amphiphilic structure and enhances the interaction with bacteria, resulting in excellent antimicrobial activity and surface antifouling properties. The multiple hydrogen bonds of ureido-pyrimidinone methacrylate (UPyMA) and ionic interactions contained in the polymers not only increased the adhesion of the coating to the material substrate (approximately 3 times) but also endowed the coating with the intrinsic self-healing ability to restore the antibiofouling properties at oral temperature and humidity. Finally, the polymer coating is biologically safe both in vitro and in vivo, showing no cytotoxic effects on cells and tissues. This research offers a promising avenue for improving the performance of orthodontic appliances and contributes to the maintenance and treatment of oral health.

Plant Extract-Based Fabrication of Silver Nanoparticles and Their Effective Role in Antibacterial, Anticancer, and Water Treatment Applications.

Ammi visnaga is a biennial or annual herbaceous plant belonging to the family Apiaceae. For the first time, silver nanoparticles were synthesized using an extract of this plant. Biofilms are a rich source of many pathogenic organisms and, thus, can be the genesis of various disease outbreaks. In addition, the treatment of cancer is still a critical drawback for mankind. The primary purpose of this research work was to comparatively analyze antibiofilms against Staphylococcus aureus, photocatalytic activity against Eosin Y, and in vitro anticancer activity against the HeLa cell line of silver nanoparticles and Ammi visnaga plant extract. The systematic characterization of synthesized nanoparticles was carried out using UV-Visible spectroscopy (UV-Vis), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), atomic force microscopy (AFM), dynamic light scattering (DLS), zeta potential, and X-ray diffraction microscopy (XRD). The initial characterization was performed with UV-Vis spectroscopy, where a peak appeared at 435 nm, which indicated the SPR band of the silver nanoparticles. AFM and SEM were performed to determine the morphology and shape of the nanoparticles, while EDX confirmed the presence of Ag in the spectra. The crystalline character of the silver nanoparticles was concluded with XRD. The synthesized nanoparticles were then subjected to biological activities. The antibacterial activity was evaluated by determining the inhibition of the initial biofilm formation with Staphylococcus aureus using a crystal violet assay. The response of the AgNPs against cellular growth and biofilm formation was found to be dose dependent. Green-synthesized nanoparticles showed 99% inhibition against biofilm and bacteria, performed excellent anticancer assay with an IC50 concentration of 17.1 ± 0.6 µg/mL and 100% inhibition, and photodegradation of the toxic organic dye Eosin Y up to 50%. Moreover, the effect of the pH and dosage of the photocatalyst was also measured to optimize the reaction conditions and maximum photocatalytic potential. Therefore, synthesized silver nanoparticles can be used in the treatment of wastewater contaminated with toxic dyes, pathogenic biofilms, and the treatment of cancer cell lines.

Novel Lactotransferrin-Derived Antimicrobial Peptide LF-1 Inhibits the Cariogenic Virulence Factors of Streptococcus mutans

We previously developed a novel lactotransferrin-derived antimicrobial peptide, LF-1, with selective antibacterial activity against the characteristic cariogenic bacterium Streptococcus mutans. This study further investigated the effects of LF-1 on the cariogenic virulence factors of S. mutans and evaluated the changes in virulence-associated enzymes and genes; the viability, acidogenicity, and aciduricity of planktonic S. mutans; and initial colonisation and biofilm formation after treatment with LF-1. The method of qRT-PCR was used to evaluate S. mutans virulence-associated gene expression. LF-1 interfered with the cell viability of S. mutans within 6 h. LF-1 inhibited the acidogenicity and aciduricity of S. mutans, with reduced lactic acid production and survival in a lethal acidic environment, and inactivated lactate dehydrogenase and F1F0-ATPase activity. LF-1 decreased surface-adherent S. mutans within 60 min and inhibited S. mutans biofilm formation, where scanning electron microscopy and confocal laser scanning microscopy showed reduced extracellular matrix and bacteria. LF-1 downregulates S. mutans virulence-associated gene expression. LF-1 inhibited the growth and cariogenic virulence factors of S. mutans in vitro with a reduction in key enzymatic activity and downregulation of virulence-associated gene expression. LF-1 has promising application prospects in the fight against S. mutans and dental caries.