Enterococcus Faecalis Biofilm Formation Research Articles

Molecular docking and antimicrobial activities of photoexcited inhibitors in antimicrobial photodynamic therapy against Enterococcus faecalis biofilms in endodontic infections

Antimicrobial photodynamic therapy (aPDT) is a promising approach to combat antibiotic resistance in endodontic infections. It eliminates residual bacteria from the root canal space and reduces the need for antibiotics. To enhance its effectiveness, an in silico and in vitro study was performed to investigate the potential of targeted aPDT using natural photosensitizers, Kojic acid and Parietin. This approach aims to inhibit the biofilm formation of Enterococcus faecalis, a frequent cause of endodontic infections, by targeting the Ace and Esp proteins. After determining the physicochemical characteristics of Ace and Esp proteins and model quality assessment, the molecular dynamic simulation was performed to recognize the structural variations. The stability and physical movement of the protein-ligand complexes were evaluated. In silico molecular docking was conducted, followed by ADME/Tox profiling, pharmacokinetics characteristics, and assessment of drug-likeness properties of the natural photosensitizers. The study also investigated the changes in the expression of genes (esp and ace) involved in E. faecalis biofilm formation. The results showed that both Kojic acid and Parietin complied with Lipinski’s rule of five and exhibited drug-like properties. In silico analysis indicated stable complexes between Ace and Esp proteins and the natural photosensitizers. The molecular docking studies demonstrated good binding affinity. Additionally, the expression of the ace and esp genes was significantly downregulated in aPDT using Kojic acid and Parietin with blue light compared to the control group. This investigation concluded that Kojic acid and Parietin with drug-likeness could efficiently interact with Ace and Esp proteins with a strong binding affinity. Hence, natural photosensitizers-mediated aPDT can be considered a promising adjunctive treatment against endodontic infections.

Targeted metabolomics analysis approach to unravel the biofilm formation pathways of Enterococcus faecalis clinical isolates.

(i) To characterize Enterococcus faecalis biofilm formation pathways by semi-targeted metabolomics and targeted nitrogen panel analysis of strong (Ef63) and weak (Ef 64) biofilm forming E. faecalis clinical isolates and (ii) to validate the identified metabolic markers using targeted inhibitors. Previous proteomics profiling of E. faecalis clinical isolates with strong and weak biofilm formation revealed that differences in metabolic activity levels of small molecule, nucleotide and nitrogen compound metabolic processes and biosynthetic pathways, cofactor metabolic process, cellular amino acid and derivative metabolic process and lyase activity were associated with differences in biofilm formation. Hence, semi-targeted analysis of Ef 63, Ef 64 and ATC control strain Ef 29212 was performed by selecting metabolites that were part of both the previously identified pathways and a curated library with confirmed physical and chemical identity, followed by confirmatory targeted nitrogen panel analysis. Significantly regulated metabolites (p < .05) were selected based on fold change cut-offs of 1.2 and 0.8 for upregulation and downregulation, respectively, and subjected to pathway enrichment analysis. The identified metabolites and pathways were validated by minimum biofilm inhibitory concentration (MBIC) and colony forming unit (CFU) assays with targeted inhibitors. Metabolomics analysis showed upregulation of betaine, hypoxanthine, glycerophosphorylcholine, tyrosine, inosine, allantoin and citrulline in Ef 63 w.r.t Ef 64 and Ef 29212, and thesemetabolites mapped to purinemetabolism, urea cycle and aspartate metabolism pathways. MBIC and CFU assays using compounds against selected metabolites and metabolic pathways, namely glutathione against hypoxanthine and hydroxylamine against aspartate metabolism showed inhibitory effects against E. faecalis biofilm formation. The study demonstrated the importance of oxidative stress inducers such as hypoxanthine and aspartate metabolism pathway in E. faecalis biofilm formation. Targeted therapeutics against these metabolic markers can reduce the healthcare burden associated with E. faecalis infections.

Arginine impacts aggregation, biofilm formation, and antibiotic susceptibility in Enterococcus faecalis.

Enterococcus faecalis is a commensal bacterium in the gastrointestinal tract (GIT) of humans and other organisms. E. faecalis also causes infections in root canals, wounds, the urinary tract, and on heart valves. E. faecalis metabolizes arginine through the arginine deiminase (ADI) pathway, which converts arginine to ornithine and releases ATP, ammonia, and CO 2 . E. faecalis arginine metabolism also affects virulence of other pathogens during co-culture. E. faecalis may encounter elevated levels of arginine in the GIT or the oral cavity, where arginine is used as a dental therapeutic. Little is known about how E. faecalis responds to growth in arginine in the absence of other bacteria. To address this, we used RNAseq and additional assays to measure growth, gene expression, and biofilm formation in E. faecalis OG1RF grown in arginine. We demonstrate that arginine decreases E. faecalis biofilm production and causes widespread differential expression of genes related to metabolism, quorum sensing, and polysaccharide synthesis. Growth in arginine also increases aggregation of E. faecalis and promotes decreased susceptibility to the antibiotics ampicillin and ceftriaxone. This work provides a platform for understanding of how the presence of arginine in biological niches affects E. faecalis physiology and virulence of surrounding microbes.

Enhancing Antimicrobial Efficacy and Synergistic Effects of Nano-Silica-Based Combinations With Doxycycline, Metronidazole, and Ciprofloxacin Against Enterococcus faecalis Biofilms.

Enterococcus faecalisbiofilm formation within root canals poses a challenging problem in endodontics, often leading to treatment failure. To combat this issue, nanotechnology offers a promising avenue for enhancing antimicrobial efficacy. This study explores the potential synergistic effects of combining nanoscale silica particles with conventional antibiotics, including doxycycline, metronidazole, and ciprofloxacin, against E. faecalis biofilms. The unique characteristics of silica nanoparticles, such as their increased reactivity and ability to be functionalized with other compounds, make them ideal candidates for augmenting antibiotic efficacy. This research investigates the antimicrobial properties of these silica-based combinations and their potential to eliminate or inhibit E. faecalis biofilms more effectively than conventional treatments. Methodology: The methods involved the preparation of nanostructured silica particles and their combination with doxycycline, Flagyl, and ciprofloxacin at subinhibitory concentrations. These combinations were then tested against E. faecalis biofilms using the agar well diffusion technique. Preliminary results suggested that the synergistic interactions between silica nanoparticles and antibiotics can significantly enhance antimicrobial efficacy. The combined treatment exhibited superior inhibitory effects on E. faecalis compared to antibiotics or silica nanoparticles alone (P< 0.05). Conclusions: This study sheds light on the potential of nanoscale silica-based combinations to address the challenges posed by E. faecalis biofilms in endodontics. Understanding the mechanisms of synergy between nanoparticles and antibiotics can pave the way for the development of more effective and targeted strategies for root canal disinfection, ultimately improving the success rates of endodontic treatments.

Nitrogen and phosphorus eutrophication enhance biofilm-related drug resistance in Enterococcus faecalis isolated from Water Sources

Antibiotic resistance is a critical topic worldwide with important consequences for public health. So considering the rising issue of antibiotic-resistance in bacteria, we explored the impact of nitrogen and phosphorus eutrophication on drug resistance mechanisms in Enterococcus faecalis, especially ciprofloxacin, oxytetracycline, and ampicillin. For this purpose we examined the antibiotic-resistance genes and biofilm formation of Enterococcus faecalis under different concentration of nitrogen and phosphorus along with mentioned antibiotics. Mesocosms were designed to evaluate the impact of influence of eutrophication on the underlying mechanism of drugn resistence in Enterococcus faecalis. For this purpose, we explored the potential relation to biofilm formation, adhesion ability, and the expression levels of the regulatory gene fsrA and the downstream gene gelEI. Our results demonstrated that the isolates of all treatments displayed high biofilm forming potential, and fsrA and gelE genes expression. Additionally, the experimental group demonstrated substantially elevated Enterococcus faecalis gelE expression. Crystal violet staining was applied to observe biofilm formation during bacterial development phase and found higher biofilm formation. In conclusion, our data suggest that E. faecalis resistance to ciprofloxacin, oxytetracycline, and ampicillin is related to biofilm development. Also, the high level of resistance in Enterococcus faecalis is linked to the expression of the fsrA and gelE genes. Understanding these pathways is vital in tackling the rising problem of bacterial resistance and its potential effect on human health.

Antimicrobial Resistance, Biofilm Formation, and Virulence Determinants in Enterococcus faecalis Isolated from Cultured and Wild Fish.

Fish has always been an integral part of Bengali cuisine and economy. Fish could also be a potential reservoir of pathogens. This study aimed to inquisite the distribution of virulence, biofilm formation, and antimicrobial resistance of Enterococcus faecalis isolated from wild and cultivated fish in Bangladesh. A total of 132 koi fish (Anabas scandens) and catfish (Heteropneustes fossilis) were collected from different markets in the Mymensingh district and analyzed to detect E. faecalis. E. faecalis was detected by conventional culture and polymerase chain reaction (PCR), followed by the detection of virulence genes by PCR. Antibiotic susceptibility was determined using the disk diffusion method, and biofilm-forming ability was investigated by crystal violet microtiter plate (CVMP) methods. A total of 47 wild and 40 cultured fish samples were confirmed positive for E. faecalis by PCR. The CVMP method revealed four per cent of isolates from cultured fish as strong biofilm formers, but no strong producers were found from the wild fish. In the PCR test, 45% of the isolates from the wild and cultivated fish samples were found to be positive for at least one biofilm-producing virulence gene, where agg, ace, gelE, pil, and fsrC genes were detected in 80, 95, 100, 93, and 100% of the isolates, respectively. Many of the isolates from both types of samples were multidrug resistant (MDR) (73% in local fish and 100% in cultured fish), with 100% resistance to erythromycin, linezolid, penicillin, and rifampicin in E. faecalis from cultured fish and 73.08, 69.23, 69.23, and 76.92%, respectively, in E. faecalis from wild fish. This study shows that E. faecalis from wild fish have a higher frequency of virulence genes and biofilm-forming genes than cultivated fish. However, compared to wild fish, cultured fish were found to carry E. faecalis that was more highly multidrug resistant. Present findings suggest that both wild and cultured fish could be potential sources for MDR E. faecalis, having potential public health implications.

Cinnamaldehyde inhibits Enterococcus faecalis biofilm formation and promotes clearance of its colonization by modulation of phagocytes in vitro

The nosocomial pathogen, Enterococcus faecalis plays a crucial role in the pathogenesis of variety of infections including endocarditis, urinary tract, and recurrent root canal infections. Primary virulence factors of E. faecalis such as biofilm formation, gelatinase production and suppression of host innate immune response can severely harm host tissue. Thus, novel treatments are needed to prevent E. faecalis biofilm development and pathogenicity due to the worrisome rise in enterococcal resistance to antibiotics. The primary phytochemical in cinnamon essential oils, cinnamaldehyde, has shown promising efficacy against a variety of infections. Here, we looked into how cinnamaldehyde affected the growth of biofilms, the activity of the enzyme gelatinase, and gene expression in E. faecalis. In addition, we looked at the influence of cinnamaldehyde on RAW264.7 macrophages' interaction with biofilm and planktonic E. faecalis in terms of intracellular bacterial clearance, NO generation, and macrophage migration in vitro. According to our research, cinnamaldehyde attenuated the biofilm formation potential of planktonic E. faecalis and gelatinase activity of the biofilm at non-lethal concentrations. The expression of the quorum sensing fsr locus and its downstream gene gelE in biofilms were also found to be significantly downregulated by cinnamaldehyde. Results also demonstrated that cinnamaldehyde treatment increased NO production, intracellular bacterial clearance, and migration of RAW264.7 macrophages in presence of both biofilm and planktonic E. faecalis. Overall these results suggest that cinnamaldehyde has the ability to inhibit E. faecalis biofilm formation and modulate host innate immune response for better clearance of bacterial colonization.

Gram-negative quorum sensing signalling enhances biofilm formation and virulence traits in gram-positive pathogen Enterococcus faecalis

ABSTRACT Acyl-homoserine lactones (AHLs) are typical quorum-sensing molecules of gram-negative bacteria. Recent evidence suggests that AHLs may also affect gram-positives, although knowledge of these interactions remains scarce. Here, we assessed the effect of AHLs on biofilm formation and transcriptional regulations in the gram-positive Enterococcus faecalis. Five E. faecalis strains were investigated herein. Crystal violet was employed to quantify the biomass formed, and confocal microscopy in combination with SYTO9/PI allowed the visualisation of biofilms’ structure. The differential expression of 10 genes involved in quorum-sensing, biofilm formation and stress responses was evaluated using reverse-transcription-qPCR. The AHL exposure significantly increased biofilm production in strain ATCC 29212 and two isolates from infected dental roots, UmID4 and UmID5. In strains ATCC 29212 and UmID7, AHLs up-regulated the quorum-sensing genes (fsrC, cylA), the adhesins ace, efaA and asa1, together with the glycosyltransferase epaQ. In strain UmID7, AHL exposure additionally up-regulated two membrane-stress response genes (σV, groEL) associated with increased stress-tolerance and virulence. Altogether, our results demonstrate that AHLs promote biofilm formation and up-regulate a transcriptional network involved in virulence and stress tolerance in several E. faecalis strains. These data provide yet-unreported insights into E. faecalis biofilm responses to AHLs, a family of molecules long-considered the monopole of gram-negative signalling.

Antibiofilm activity of Cyanobacteria spirulina as an irrigation solution against Enterococcus faecalis

Sodium hypochlorite (NaOCl) is currently the golden standard for root canal irrigation. NaOCl at a concentration of 5.25% to 6% can eliminate E. faecalis, but this concentration can increase the risk of toxic effects. Cyanobacteria spirulina is known to produce several secondary metabolites that have antimicrobial activity against gram-positive and gram-negative bacteria. The aim of this study was to determine the antibiofilm power of Cyanobacteria spirulina against the biofilms of Enterococcus faecalis at concentrations of 60 mg/ml, 70 mg/ml, 80 mg/ml, and 90 mg/ml. This research was a true experiment with a post-test only group design. The object of the research was divided into 6 groups. Group 1 was a negative control group, group 2 was a positive control group with 5% NaOCl, group 3, 4, 5, 6 were treatment groups given Cyanobacteria spirulina solution at a concentration of 60 mg/ml, 70 mg/ml, 80 mg/ml, and 90 mg/ml, respectively. Optical density (OD) of bacteria was bound by staining and analyzed by ELISA auto reader with a wave length of 595 nm (OD 595 nm). The results of the LSD test showed that the significance between K+ and P1, P2, P3 (p&lt; 0.005), K+ and P4 (p= 0.129), P1 and P2 (p=0.449), P3 and P4 (p=0.178). Significance of p&lt;0.0005 showed a significant difference between the groups. The data were analyzed using the one-way ANOVA test followed by a double comparison test with the Least Significance Different (LSD) Post Hoc test method. Cyanobacteria spirulina solution at a concentration of 90 mg/ml had the greatest inhibitory effect on the biofilm formation of Enterococcus faecalis.

Effect of nisin and p-coumaric acid on autoinducer-2 activity, biofilm formation, and sprE expression of Enterococcus faecalis.

Quorum sensing (QS) is an inter- and intracellular communication mechanism that regulates gene expression in response to population size. Autoinducer-2 (AI-2) signaling is a QS signaling molecule common to both Gram-negative and Gram-positive bacteria. Enterococcus faecalis is one of the leading causes of nosocomial infections worldwide. There has been an increasing interest in controlling infectious diseases through targeting the QS mechanism using natural compounds. This study aimed to investigate the effect of nisin and p-coumaric acid (pCA), on biofilm formation and AI-2 signaling in E. faecalis. Their effect on the expression of the QS-regulated virulence encoding gene sprE was also investigated. Nisin exhibited a MIC ranging from 0.25 to 0.5mg/mL, while the MIC of pCA was 1mg/mL. The luminescence-based response of the reporter strain Vibrio harveyi BB170 was used to determine AI-2 activity in E. faecalis strains. Nisin was not effective in inhibiting AI-2 activity, while pCA reduced AI-2 activity by ≥ 60%. Moreover, pCA and nisin combination showed higher inhibitory effect on biofilm formation of E. faecalis, compared to the treatment of pCA or nisin alone. qRT-PCR analysis showed that nisin alone and the combination of nisin and pCA, at their MIC values, led to a 32.78- and 40.22-fold decrease in sprE gene expression, respectively, while pCA alone did not have a significant effect. Considering the demand to explore new therapeutic avenues for infectious bacteria, this study was the first to report that pCA can act like a quorum sensing inhibitor (QSI) against AI-2 signaling in E. faecalis.

Antibiofilm activity of neem leaf (Azadirachta indica A. Juss) ethanolic extracts against Enterococcus faecalis in vitro

Background: Enterococcus faecalis commonly infects root canals by forming a biofilm. Extracts from neem leaves (Azadirachta indica A. Juss) have been shown to have antibacterial properties, indicating their potential in preventing or treating biofilm formation caused by bacteria. Purpose: This study aims to investigate the phytochemical compounds present in neem leaves (Azadirachta indica A. Juss) and establish the concentration of ethanol-based neem leaf extract that can effectively inhibit the in vitro growth of Enterococcus faecalis biofilm. Methods: This study employed the maceration technique for extraction, gas chromatography mass spectroscopy for the analysis of plant chemicals, and a microtiter plate assay for measuring biofilm formation with treatment concentrations of 6.25%, 12.5%, 25%, 50%, and 75%, with a positive control of 0.2% chlorhexidine. Results: A phytochemical analysis revealed that the ethanol extract of neem leaves contained 22 different metabolites, mainly terpenoids and fatty acids. The extract demonstrated antibiofilm activity only at a concentration of 12.5% with an average biofilm inhibition of 36.85%. However, lower concentrations of 6.25%, 25%, 50%, and 75% had the opposite effect, promoting biofilm formation in Enterococcus faecalis. Conclusion: Phytochemical metabolite contained in the ethanolic extracts of neem leaves might contribute a promising agent in treating a biofilm-mediated root canal infection of Enterococcus faecalis.

An in vitro study on the efficacy of hydrogen peroxide mediated high-power photodynamic therapy affecting Enterococcus faecalis biofilm formation and dispersal.

Biofilms are involved in failure of root canal treatment due to their high resistance to antimicrobial agents, which make their removal as a big challenge. The present study aims at utilizing hydrogen peroxide (HP) plus high frequency laser reinforced antimicrobial photodynamic therapy (a-PDT) as a complementary therapy against Enterococcus faecalis (E. faecalis) at planktonic and biofilm stages. E. faecalis at planktonic and biofilm stages was treated with the photosensitizer HP, followed by no irradiation or irradiation with a power of 2.5W (ʎ=980nm). The cell viability, anti-biofilm, anti-metabolic potential, and temperature changes were evaluated. The combination of HP and 980nm diode laser intensely boosted antibacterial and anti-biofilm efficacy compared with either component alone, affirming HP reinforcement as a bacteriostatic agent. The maximum effect on biofilm occurs in 5.25% sodium hypochlorite (NaOCl) group. During laser irradiations, the mean of temperature changes remains below 5.6°C. It could be concluded that the HP could improve anti-biofilm efficacy as a photosensitizer in a-PDT.

In Vitro Study of Irrigation solution of Chitosan Nanoparticles to Inhibit the Adhesion and Biofilm Formation of Enterococcus faecalis in the Root Canal

Enterococcus faecalis (E. faecalis) plays a role in the pathogenesis of dental root canal infections. Chitosan has antibacterial properties and a chelating agent in the tooth root canal and is biodegradable and non-toxic. They analyzed the irrigation response of Chitosan nanoparticles to the virulence properties of E. faecalis in the dental root canal. Examination of E. faecalis virulence properties was carried out with violet crystals to obtain biofilm inhibition strength, Gram staining to determine lysis and coagulation of bacterial cells, and Atomic Force Microscopy (AFM) to analyze the surface roughness of the tooth root canal. Chitosan nanoparticles combined with 2.5% NaOCl has a strong inhibition of the formation of E. faecalis biofilm, both based on the control group (p&gt; 0.05; 0.088) and incubation time of 48 h and 72 h, also inhibitory power, which was better at 24 h (p&gt; 0.05; 0.185) than the other groups. Irrigation solution of Chitosan nanoparticles combined with 2.5% NaOCl has better lysis and cell agglutination of E. faecalis bacteria compared to other groups, especially at all incubation times, based on the control group (p&gt; 0.05; 0.104) and incubation time (p&gt; 0.05; 0.580) can reduce the surface roughness of the dental root canal, but the impact of giving irrigation materials to each treatment group with the incubation time showed significant differences (p&lt;0.05). The Chitosan nanoparticles solution as an irrigation material has a strong ability to suppress the formation of biofilms, coagulation, and lysis of E. faecalis cells and better reduce the walls' surface roughness dental root canal at 24 hours incubation time.

Association between virulence factors and biofilm formation in Enterococcus faecalis isolated from semen of infertile men.

Enterococcus faecalis is a common microbial semen contaminant. Although virulence factors and biofilm formation have often been analyzed in Enterococcus spp., there is little information about these features in isolates obtained from the genitourinary tract. This study was intended to characterize and determine the relationship between biofilm-forming ability and the presence of E. faecalis virulence factors isolated from human semen. A total of 32 patients diagnosed with primary infertility and 28 healthy men were included in the study. Semen analyses were performed according to the WHO guidelines. PCR reactions were applied for the detection of ace, esp, efeA, gelE, asa1, and cylA genes. Microtiter plate assay, via measurement of OD560, was used to measure the biofilm-forming ability of the isolates. Sixty E. faecalis isolates from semen of infertile and fertile men were characterized by phenotypic and genotypic methods. The prevalence of ace, esp, efeA, gelE, asa1 and cylA were reported to be 81.3%/100.0%, 81.3%/89.3%, 81.3%/85.7%, 71.9%/53.6%, 8.8%/75.0%, and 62.5%/67.9% in infertile/fertile groups; respectively. Strong, weak, and non-biofilm reactions were reported to be 50.0%/21.4%, 40.6%/64.3%, and 9.4%/14.3% in infertile and fertile groups; respectively. There was a significant relationship between fertility and weak biofilm reaction and also between biofilm formation and possession of the esp gene (P < .05). It could be speculated that colonization with E. faecalis with a strong ability for biofilm formation could become a potential threat to men's fertility.

Bacterial Adhesion and Biofilm Formation of Enterococcus faecalis on Zwitterionic Methylmethacrylat and Polysulfones.

Biofilm-associated implant infections represent a major challenge for healthcare systems around the world due to high patient burden and enormous costs incurred. Enterococcus faecalis (E. faecalis) is the most prevalent enterococcal species identified in biofilm-associated infections. The steadily growing areas of application of implants demand a solution for the control of bacterial infections. Therefore, the development of modified anti-microbial implant materials and the testing of the behavior of different relevant bacterial strains towards them display an indispensable task. Recently, we demonstrated an anti-microbial effect of zwitterionic modified silicone rubber (LSR) against Staphylococcus aureus. The aim of this study was to evaluate bacterial colonization and biofilm formation of another clinically relevant strain, E. faecalis, on this material in comparison to two of the most commonly used thermoplastic polyurethanes (TPUs) and other modified LSR surfaces. By generating growth curves, crystal violet, and fluorescence staining, as well as analyzing the expression of biofilm-associated genes, we demonstrated no anti-microbial activity of the investigated materials against E. faecalis. These results point to the fact that anti-microbial effects of novel implant materials do not always apply across the board to all bacterial strains.

Pectolinarin Inhibits the Bacterial Biofilm Formation and Thereby Reduces Bacterial Pathogenicity.

Bacterial biofilms are a growing problem as it is a major cause of nosocomial infection from urinary catheters to chronic tissue infections and provide resistance to a variety of antibiotics and the host’s immune system. The effect of pectolinarin on the biofilm formation in Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Streptococcus mutans, Streptococcus sobrinus, Staphylococcus aureus, Pseudomonas aeruginosa, Cutibacterium acnes, and Porphyromonas gingivalis was studied in TSBg (tryptic soy broth supplemented with 1% glucose). Pectolinarin inhibited biofilm formation of E. faecalis (IC50 = 0.39 μg/mL), E. faecium (IC50 = 0.19 μg/mL), E. coli (IC50 = 0.25 μg/mL), S. mutans (IC50 = 1.2 μg/mL), S. sobrinus (IC50 = 1.4 μg/mL), S. aureus (IC50 = 0.39 μg/mL), P. aeruginosa (IC50 = 0.9 μg/mL), P. acnes (IC50 = 12.5 μg/mL), and P. gingivalis (IC50 = 9.0 μg/mL) without inhibiting the bacterial growth. Pectolinarin also showed increased susceptibility of antibacterial activity with commercially available antibiotics including ampicillin, vancomycin, streptomycin, and oxytetracyclin against E. faecalis and E. faecium. Finally, pectolinarin dose-dependently reduced the expression of genes including cytolysin genes (cylLS, cylR2 and cylM), quorum sensing (QS) genes (fsrB, fsrC, gelE, ebpA, ebpB, acm, scm and bps), and biofilm virulence genes (esp) of E. faecalis and E. faecium. Pectolinarin reduced the bacterial biofilm formation, activated the antibacterial susceptibility, and reduced the bacterial adherence. These results suggest that bacterial biofilm formation is a good target to develop the antibacterial agents against biofilm-related infections.

Identification of Protein Drug Targets of Biofilm Formation and Quorum Sensing in Multidrug Resistant Enterococcus faecalis.

Enterococcus faecalis (E. faecalis) is an opportunistic multidrug-resistant (MDR) pathogen found in the guts of humans and farmed animals. Due to the occurrence of (MDR) strain there is an urgent need to look for an alternative treatment approach. E. faecalis is a Gram-positive bacterium, which is among the most prevalent multidrug resistant hospital pathogens. Its ability to develop quorum sensing (QS) mediated biofilm formation further exacerbates the pathogenicity and triggers lifethreatening infections. Therefore, developing a suitable remedy for curing E. faecalis mediated enterococcal infections is an arduous task. Several putative virulence factors and proteins are involved in the development of biofilms in E. faecalis. Such proteins often play important roles in virulence, disease, and colonization by pathogens. The elucidation of the structure-function relationship of such protein drug targets and the interacting compounds could provide an attractive paradigm towards developing structure-based drugs against E. faecalis. This review provides a comprehensive overview of the current status, enigmas that warrant further studies, and the prospects toward alleviating the antibiotic resistance in E. faecalis. Specifically, the role of biofilm and quorum sensing (QS) in the emergence of MDR strains had been elaborated along with the importance of the protein drug targets involved in both the processes.

Antimicrobial evaluation of polytetrafluoroethylene used as part of temporary restorations: An ex vivo study.

The use of polytetrafluoroethylene (PTFE) tape as the base layer of temporary restorations had gained popularity mainly due to the ease of manipulation. The aim of this study was to assess whether this method changes the potential for bacterial growth and leakage of temporary restorations. The direct contact test and live/dead fluorescent staining were used for comparing Enterococcus faecalis growth and biofilm formation on PTFE, composite, intermediate restorative material (IRM) and Coltosol F. Confocal laser scanning microscopy was employed to evaluate E.faecalis penetration through 2mm of PTFE, IRM or Coltosol F placed on the bottom of the pulp chamber and into radicular dentinal tubules in extracted maxillary third molars. The results demonstrated that E.faecalis grows on and penetrates through PTFE significantly more than it does with IRM and Coltosol F, revealing its comparably reduced overall antimicrobial sealing ability when placed as the base part of temporary restorations.

Attenuation of Enterococcus faecalis biofilm formation by Rhodethrin: A combinatorial study with an antibiotic

The nosocomial pathogen Enterococcus faecalis critically implicated in the hospital environment. Its major virulence attributes biofilm formation and antibiotic resistance. The novel therapeutics are required to inhibit E. faecalis biofilm formation and virulence. Thus combinatorial and drug repurposing has been promising approaches to tackling biofilm-associated infections. Here, we have used a bacterium that produced indole terpenoid Rhodethrin (Rdn) with a combination of known antibiotic chloramphenicol (Chpl) against E. faecalis (ATCC 19433). The fractional inhibitory concentration index (FICI) values showed between 0.25 and 0.33 synergistic activities. The exopolysaccharides (EPSs) production significant decrease with Rdn (34.6 ± 4.6%), Chpl (31.0 ± 5.2%), and combination (Rdn-Chpl) (76.0 ± 4.5%) (p > 0.05). However, the biofilm interruption can attenuate of total biofilm was shown with Rdn (39.7 ± 5.1%), Chpl (32.6 ± 4.7%), and Rdn-Chpl (69.0 ± 5.3%), (p > 0.05). The microscopic observations reveal that the gradually unstructured biofilm architecture in E. faecalis. Furthermore, in silico, studies on biofilm-associated proteins (GelE, LuxS), virulence regulating (SprE), and cell division (FtsZ) have resulted in high and reasonable binding affinity, respectively. Thus, our results suggested that the synergism of Rdn-Chpl has the potential to function as a combinatorial antibiotic accelerates in treating vancomycin-resistant Enterococcus faecalis infections.

The inhibition of leaf extract Moringaoleifera on the formation biofilm bacteria Enterococcus faecalis

Background: Enterococcus faecalisis the most common bacteria that cause failure to root canal treatment, which can become very resistant under biofilms. Moringaoleifera has antibacterial properties and may affect the multidrug-resistant bacteria. Purpose: This study aimed to observe the inhibition of Moringaoleifera leaf extract on the biofilm formation of bacteria Enterococcus faecalis. Method: This study was true experimental laboratory research with post-test only control group design and tested using biofilm method, divided into six groups, each group consisted of eight samples. The control groups were: K- (CMC 0,1%), K+ (ChKM), and four treatment groups were: P1 (Moringaoleifera 20%), P2 (Moringaoleifera 40%), P3 (Moringaoleifera 60%), P4 (Moringaoleifera 80%). Antibacterial inhibition was determined by the value of Optical Density in the ELISA Reader. Data analysis using Kruskal-wallis followed by Mann-Whitney test. Results: There were significant differences (p &lt;0.05) seen from the percentage value of biofilm inhibition, on the K - (0 %) group compared with K+ (47,69%), P1(7,68%), P2 (21,13%), P3 (42,33%) and P4 (55,78%), as well on K + group (ChKM) compared with P4 group (Moringaoleifera 80%). Conclusion: Moringaoleiferaleaf extract has inhibition effect for the formation of bacteria Enterococcus faecalis biofilm and the effect is 80% greater than ChKM.