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Hospital surfaces are known to contribute to the spread of healthcare-associated antimicrobial pathogens. Environmental sampling can help locate reservoirs and determine intervention strategies, although sampling and detection can be labor intensive. Composite approaches may help reduce time and costs associated with sampling and detection. We investigated optimum surface areas for sampling antimicrobial-resistant organisms (AROs) with a single side of cellulose sponge, created theoretical composites (TC) by adding recovery results from multiple optimum areas, then compared the TC to the standard Centers for Disease Control and Prevention sampling method (one sponge using all sides, whole tool; (WT)). Five AROs were evaluated: carbapenemase-producing KPC+ Klebsiella pneumoniae (KPC), Acinetobacter baumannii (AB), methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecalis (VRE) and Clostridioides difficile spores (CD). Steel coupons comprising four surface areas (323; 645; 1,290 and 2,258 cm2) were inoculated, dried, and sampled with one sampling pass using the larger side (face) or the smaller side (edge) of a pre-moistened cellulose sponge tool. Based on the optimum areas determined for each organism, composite areas were 1,290 cm2 for MRSA and VRE, 1,936 cm2 for AB, 2,580 cm2 for CD spores and 3,870 cm2 for KPC. Total colony forming units (CFU) recovered using a composite approach was greater or comparable than using multiple WT samplings (over the same area as the composite) for MRSA, VRE and AB (130%; 144% and 95%) yet less than if using multiple WT samplings for KP and CD (47% and 66%). We propose a conservative composite sampling strategy if the target organism is unknown; 323 cm2 sampling area for each of the four sides of the sponge, (1290 cm2 total). The conservative composite sampling strategy improved the recovery of KP (from 47% to 85% of multiple WT samplings), while MRSA, VRE, AB and CD (131%; 144%; 97% and 66%) remained within 5% to that of the optimum area TC. [ABSTRACT FROM AUTHOR]

Background: Identifying therapeutic inhibitors of crucial enzymes involved in the peptidoglycan biosynthesis pathway is pivotal for developing new treatments against multidrug-resistant Enterococcus faecalis V583. MurM, an essential enzyme in this pathway, plays a significant role in the bacterium's cell wall synthesis, making it an attractive druggable target for novel antimicrobial strategies. This study explored the potential of natural compounds as inhibitors of MurM, aiming to discover promising drug candidates that could serve as the foundation for future therapeutic development. Methods: The three-dimensional structure of MurM was predicted, optimized, and its binding pocket was analyzed by comparing it with related structures. Over 4,70,000 natural compounds from the COCONUT database were subjected to virtual high-throughput screening (vHTS). The top lead candidates were selected based on their Lipinski's profile, ADME profile, toxicity profile, estimated binding free energy (ΔG) and estimated inhibition constant (Ki). Interaction pattern analysis was used to evaluate the non-covalent interactions between the inhibitors and key residues in MurM's binding pocket. Molecular dynamics simulations were performed over 300 ns to assess the structural stability and impact of these inhibitors on MurM's enzyme. Results: Three lead compounds—CNP0056520, CNP0126952, and CNP0248480—were identified and prioritized with estimated ΔG ranging from − 9.35 to -7.9 kcal/mol. Molecular dynamics simulations revealed minimal impact on MurM's overall structure and dynamics, with the candidate inhibitors forming stable protein-ligand complexes. These interactions were supported by several non-covalent interactions between the candidate inhibitors and key residues within MurM's binding pocket. Conclusion: These findings suggest that the identified natural product candidates could serve as promising inhibitors of MurM, potentially leading to novel therapeutics targeting cell wall biosynthesis in multidrug-resistant E. faecalis. [ABSTRACT FROM AUTHOR]

Bacterial cellulose (BC) is a highly versatile biopolymer renowned for its exceptional mechanical strength, water retention, and biocompatibility. These properties make it a valuable material for various industrial and biomedical applications. In this study, Enterococcus faecalis synthesized extracellular BC, utilizing Phoenix dactylifera and Musa acuminata fruit extracts as sustainable carbon sources. LC-MS analysis identified glucose as the primary carbohydrate in these extracts, providing a suitable substrate for BC production. Scanning Electron Microscopy (SEM) revealed a network of BC nanofibers on Congo red agar plates. ATR-FTIR spectroscopy confirmed the presence of characteristic cellulose functional groups, further supporting BC synthesis. X-ray diffraction (XRD) analysis indicated a high crystallinity index of 71%, consistent with the cellulose I structure, as evidenced by peaks at 16.22°, 21.46°, 22.52°, and 34.70°. Whole-genome sequencing of E. faecalis identified vital genes involved in BC biosynthesis, including bcsA, bcsB, diguanylate cyclase (DGC), and 6-phosphofructokinase (pfkA). Antibiotic susceptibility tests revealed resistance to cefotaxime, ceftazidime, and ceftriaxone, while susceptibility to imipenem was observed. Quantitative assessment demonstrated that higher concentrations of fruit extracts (5.0–20 mg/mL) significantly enhanced BC production. Cytotoxicity testing via the MTT assay confirmed excellent biocompatibility with NIH/3T3 fibroblast cells, showing high cell viability (97–105%). Unlike commonly studied Gram-negative bacteria like Acetobacter xylinum for BC production, this research focuses on Gram-positive Enterococcus faecalis and utilizes Phoenix dactylifera and Musa acuminata fruit extracts as carbon sources. This approach offers a sustainable and promising avenue for BC production. [ABSTRACT FROM AUTHOR]

Disease resistance (defined as the host capacity to limit systemic infection intensity) and disease tolerance (defined as the host capacity to limit infection-induced damage) are 2 complementary defense strategies that help the hosts maximize their survival and fitness when infected with pathogens and parasites. In addition to the underlying physiological mechanisms, the existing theory postulates that these 2 strategies differ in terms of the conditions under which each strategy evolves in the host populations, their evolutionary dynamics, and the ecological and epidemiological consequences of their evolution. Here, we explored if one or both of these strategies evolve when host populations are subjected to selection for increased postinfection survival. We experimentally evolved Drosophila melanogaster populations, selecting for the flies that survived an infection with the entomopathogen Enterococcus faecalis. We found that the host populations evolved increased disease resistance in response to selection for increased survival. This was despite the physiological costs associated with increased resistance, the expression of which varied with the phase of infection. We did not find evidence of any change in disease tolerance in the evolved host populations. [ABSTRACT FROM AUTHOR]

The persistence of bacterial infections and antibiotic resistance in secondary root canal infections necessitate effective antimicrobial approaches utilizing natural compounds. This study aimed to evaluate the extraction efficiency, phytochemical composition, and antibacterial activity of the ethanol extract of Aceh patchouli leaves (Pogostemon cablin Benth.) against Enterococcus faecalis. The maceration method was employed for extraction and phytochemical analysis was conducted using gas chromatography-mass spectrometry (GC-MS). Antibacterial activity was assessed using the Kirby-Bauer method, and the minimum inhibitory concentration (MIC) was determined using UV-vis spectrophotometry. The results indicated a total extract yield of 17.25% and significant antibacterial activity against E. faecalis. The active compounds in the extract were primarily sesquiterpenes and fatty acids, with patchouli alcohol as the most prominent. The MIC value was 12.5%, suggesting the potential of Aceh patchouli extract as an alternative or adjunctive treatment for root canal infections. This study has the potential to make Aceh's indigenous Pogostemon cablin Benth. plant as an alternative or adjunctive treatment for root canal infections leading to safe and effective treatments that minimize the risk of antibiotic resistance. [ABSTRACT FROM AUTHOR]

Two monoterpenoid lactones, loliolide (1) and epi-loliolide (2), were isolated from the crude dichloromethane extract of a microalga, Thalassiosira sp.). The structures of loliolide (1) and epi-loliolide (2) were elucidated by 1D and 2D NMR analysis, as well as a comparison of their 1H or/and 13C NMR data with those reported in the literature. In the case of loliolide (1), the absolute configurations of its stereogenic carbons were confirmed by X-ray analysis, whereas those of epi-loliolide (2) were determined by NOESY correlations. Loliolide (1) and epi-loliolide (2) were tested for their growth inhibitory activity against two Gram-positive (Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212) and two Gram-negative (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853) bacteria, as well as one clinical isolate (E. coli SA/2, an extended-spectrum β-lactamase producer-ESBL) and two environmental isolates, S. aureus 74/24, a methicillin-resistant (MRSA), and E. faecalis B3/101, a vancomycin-resistant (VRE) isolates. The results showed that none of the tested compounds exhibited antibacterial activity at the highest concentrations tested (325 μM), and both revealed low antioxidant activity, with ORAC values of 2.786 ± 0.070 and 2.520 ± 0.319 µmol TE/100 mg for loliolide (1) and epi-loliolide (2), respectively. [ABSTRACT FROM AUTHOR]

Background/Objectives: To assess the cytocompatibility, antibacterial and anti-biofilm efficacy of grape seed extract (GSE) and quercetin hydrogels versus calcium hydroxide (CH) as intracanal medications (ICMs) against an endodontic ex vivo biofilm model. Methods: Single-rooted teeth (n = 50) were prepared and sterilized before being infected with E. faecalis to develop a mature biofilm. They were divided into five equal groups according to the ICM used: G1: medicated with CH paste, G2: medicated with GSE hydrogel, G3: medicated with quercetin hydrogel, G4: positive control group that was infected and not medicated, and G5: negative control group that was neither infected nor medicated. After 1 week, the ICM was removed, and the root canals were cultured to assess the antibacterial efficacy by counting the colony-forming units and the anti-biofilm efficacy by the crystal violet assay. Dead/live bacterial viability was assessed by CFLSM examination, while the cytocompatibility was assessed using the MTT assay. Results: CH had the best antibacterial efficacy, followed by GSE and quercetin hydrogels (p < 0.001). Regarding the anti-biofilm efficacy, GSE was superior, followed by quercetin and CH (p < 0.001). CFLSM examination showed CH and GSE hydrogel to be highly effective in comparison to the positive control (p < 0.0001), with no statistical difference between them (p > 0.05). CH showed significantly higher cell viability percentages using a 500 μg/mL, while quercetin and GSE started to show cell viability > 70% at concentrations of 125 μg/mL and 62.5 μg/mL. Conclusions: CH fulfilled the ideal requirements of ICM as being both antibacterial and non-cytotoxic compared to the other materials tested. [ABSTRACT FROM AUTHOR]

Purpose: Few data have been published on the efficacy of linezolid in enterococcal urinary tract infection (e-UTI). The aims of this study were to describe the characteristics of patients with enterococci UTI treated with linezolid, and to evaluate the efficacy and the tolerance of linezolid treatment. Methods: An observational multicentre retrospective study was conducted in 5 hospitals in France. Patients were included if they met the following criteria: ≥18 years, clinical and microbiological criteria for enterococcal UTI and linezolid treatment > 48 h. Primary outcome was clinical failure. Results: Eighty-one patients were included between January 2015 and December 2021. The median age was 73.0 [64; 83] years and 47 (58%) were men. The median Charlson comorbidity index was 3.00 [2; 6]. E. faecium was reported in 65 (80%) cases and E. faecalis in 26 cases (32%). Polymicrobial infections occurred in 41 (51%) cases. No enterococci was resistant to vancomycin. Before linezolid prescription an empiric antimicrobial treatment was started in 48 (59%) cases and was effective against enterococci in 19/48 (39.5%) patients for a median of 3.5 days [2.0; 4.0]. The median duration of linezolid antibiotic treatment was 13 days [10; 14]. Three adverse events were reported, none were serious but one led to discontinuation of treatment. Treatment failure was reported in 2 cases (2.5%). Conclusion: This study provides evidence for efficacy and safety of linezolid in enterococcal UTI. [ABSTRACT FROM AUTHOR]

Background Vancomycin-resistant enterococci (VRE) represent a public health threat due to the few available treatments. Such alarm has triggered worldwide initiatives to develop effective antimicrobial compounds and novel delivery and therapeutic strategies. vanA operon is responsible for most cases of acquired vancomycin resistance in enterococci. Objectives Development of a transcription factor decoy (TFD) system as a vanA gene transcription-inhibitor. Methods Vancomycin MIC was determined in the presence of TFD-lipoplexes. Additionally, the effect of TFD-lipoplexes on the expression level of the vanA gene and the growth pattern of E. faecalis was evaluated. The haemolytic activity of the developed TFD-lipoplexes and their cytotoxicity were examined. TFD-lipoplexes efficiency in treating vancomycin-resistant E. faecalis (VREF) infection was tested in vivo using a systemic mice infection model. Results A reduction in vancomycin MIC against VRE from 256 mg/L (resistant) to 16 mg/L (intermediate susceptible), in the presence of TFD-lipoplexes, was recorded. The developed TFD-lipoplexes lacked any effect on E. faecalis growth and significantly reduced the transcription level of the vanA gene by about 3-fold. In an initial evaluation of the safety of TFD-lipoplexes, they were found not to be overtly haemolytic to human blood or cytotoxic to human skin fibroblast cells. The co-administration of TFD-lipoplexes and vancomycin efficiently eradicated VREF infection in vivo. Conclusions The developed TFD-lipoplexes successfully restored vancomycin activity against VREF. They offer a safe effective unconventional therapy against this stubborn organism and present a revolution in gene therapy that can be applied to other resistance-encoding genes in various organisms. [ABSTRACT FROM AUTHOR]

Different Streptococcal species including Streptococcus mutans, Streptococcus sobrinus and Enterococcus faecalis are commonly isolated in root canal infections including refractory, recurrent, and persistent cases. Calcium hydroxide (Ca (OH)2) has been widely used in endodontics as an intracanal medicament. However, using new antimicrobial herbal alternatives offers promising potentials which can be additionally enhanced by using nanoparticles (NPs). In this study, we evaluate the antimicrobial efficacy and antibiofilm effect of Neem oil including its NPs preparations and we compare the effect of conventional Ca (OH)2 to Ca (OH)2 NPs using standard disc diffusion method and quantitative microtitre dish biofilm formation assay against common pathogens isolated from root canal samples. Molecular docking was used to test the binding of 10 Streptococcal macromolecules to 5 candidate neem active constituents. Neem NPs 0.125 mg/ml showed better antibacterial effect than both Neem 15 mg/ml and Neem 0.15 mg/ml. Ca (OH)2 NPs 0.125 mg/ml also showed better antibacterial effect than each of Ca (OH)2 10 mg/ml and Ca (OH)2 0.1 mg/ml. Best biofilm mass inhibition was achieved by Neem oil 0.15 mg/ml at 74.55% (IQ: 67.36–87.65) and Neem NPs 0.0125 mg/ml at 59.33% (IQ: 51-–75.27). For Ca (OH)2, the best biofilm mass inhibition was observed with Ca (OH)2 NPs 0.125 mg/ml at 54.7% (IQ: 42.37– 77.25). Both neem oil and neem NPs show promising antibacterial and antibiofilm potential against Mutans Streptococci group at low concentrations and hence are good candidates for use as endodontic medications. In silico analysis shows that both Sitosterol and Gedunin appear to be important active constituents of neem and possible drug candidates. Additionally, Ca (OH)2 NPs showed significantly higher antimicrobial effect against Mutans streptococci group than conventional Ca (OH)2 preparations. [ABSTRACT FROM AUTHOR]

The purpose of this study was to establish a porcine model of urinary tract infection (UTI) with gram‐positive uropathogens. Ten female domestic pigs were experimentally inoculated with human UTI isolates of Enterococcus faecalis (n = 3), Staphylococcus saprophyticus (n = 3), or Staphylococcus aureus (n = 4) and followed with regular urine samples. Bladders and kidneys were aseptically removed at termination (5–7 days post infection) and assessed by gross pathology and bacterial enumeration. Enterococcus faecalis (n = 3 of 3) and S. aureus (n = 2 of 4) successfully colonized the pig bladders. Inoculation with S. saprophyticus never resulted in detectable bacteriuria. All infected pigs had cleared the infection spontaneously before termination. Surprisingly, three (of four) pigs inoculated with S. aureus led to spontaneous infection with opportunistic pathogens. Also, one pig colonized with E. faecalis resulted in spontaneous infection with E. coli. In conlusion, the pig supports experimental UTI with E. faecalis for up to 24 h but not prolonged infection. S. aureus and S. saprophyticus fails to cause UTI in pigs and other animals should be considered for studying these pathogens. [ABSTRACT FROM AUTHOR]

This study investigates the antimicrobial properties of leukocyte‐ and platelet‐rich fibrin (L‐PRF) against Enterococcus faecalis (ATCC 29212), Pseudomonas aeruginosa (ATCC 27853), and Candida albicans (ATCC 90028). Infections can hinder wound healing posing challenges. L‐PRF's potential for regeneration and antimicrobial action has been studied. Considering the increasing concern about antibiotic resistance, assessing the antimicrobial properties of L‐PRF provides valuable insights into its potential as a therapeutic agent in postoperative infections. Twenty volunteers were enrolled in the study, following ethical guidelines, and obtaining informed consent. Blood samples were collected and L‐PRF was prepared. Microbial suspensions were prepared, and susceptibility testing was performed using the Kirby‐Bauer agar diffusion method. The study revealed significant heterogeneity in the susceptibility to L‐PRF. All L‐PRF membrane samples exhibited antimicrobial activity against P. aeruginosa, with inhibition zones of 13 mm ± 3.85 SD. Enterococcus faecalis displayed inhibition diameter of 7.25 mm ± 5.15 SD. Candida albicans susceptibility to L‐PRF varied among samples, with both inhibitory and non‐inhibitory results. Results showed varying degrees of antimicrobial activity, particularly against P. aeruginosa, and highlight the complexity of the L‐PRF‐microorganism interaction. Further investigations are needed to elucidate the clinical implications and optimize the use of L‐PRF. [ABSTRACT FROM AUTHOR]

Enterococci are Gram-positive coccus bacteria that are normally present in the gastrointestinal tract and ordinarily function commensally with humans. Very few studies have investigated the characteristics of enterococcal infections. We aimed to characterize patients with urinary tract infections (UTIs) due to Enterococci and their outcomes. This was a retrospective cohort study between June 2012–November 2022. Patients who had clinically and microbiologically confirmed Enterococcal UTI based on a urine culture positive for E. faecalis or E. faecium with a count of ≥105 CFU/mL and having urinary tract symptoms were included. A total of 396 patients were eligible and included. The patients had a median age of 61 years and were mostly females (56.8 %). The most common characteristics were hospitalization in a non-ICU ward, having a urinary catheter, and recent use of antibiotics within the last 3 months (66.4 %, 59.3 %, and 51.8 %, respectively). Infection with E. faecalis was more common than E. faecium (77.3 % vs. 22.7 %). However, the latter exhibited higher rates of antibiotic resistance (P < 0.001 to several antibiotics) and was associated with significantly higher median C-reactive protein level (26.7 vs. 13 mg/dL; P = 0.025), mortality (23 % vs. 10.1 %; P = 0.002), and median length of stay (25 vs. 11.5 days; P < 0.001). We found that most patients with enterococcal UTIs had a history of having a urinary catheter and recent antibiotic use and were mostly females and hospitalized in non-ICU wards. E. faecium -infected patients experienced more severe episodes and poorer outcomes compared to patients infected with E. faecalis ; thus, would need more aggressive therapy. [ABSTRACT FROM AUTHOR]

Oral diseases are often associated with bacterial and fungal pathogens such as Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis, and Candida albicans. This research explored a novel approach to addressing these pathogens by synthesizing zinc oxide nanoparticles (ZnO NPs) coated with rutin (RT), a plant‐derived compound. The synthesized ZnO‐RT NPs were comprehensively characterized using UV‐Vis spectrophotometer, SEM, and EDAX techniques to confirm their structural composition. The antioxidant potential was assessed through free radical scavenging assays. Additionally, the antimicrobial activity of ZnO‐RT NPs was evaluated using a zone of inhibition assay against oral pathogens. Molecular docking studies with the Autodock tool were performed to elucidate the interactions between RT and the receptors of oral pathogens. The findings demonstrated that ZnO‐RT NPs exhibited robust free radical scavenging activity. Furthermore, they showed significant antimicrobial activity with a minimal inhibitory concentration of 40 μg/mL against oral pathogens. ZnO‐RT NPs also displayed dose‐dependent anticancer effects on human oral cancer cells at concentrations of 10, 20, 40, and 80 μg/mL. Mechanistic insights into the anticancer activity on KB cells revealed the upregulation of apoptotic genes. This study underscores the promising potential of ZnO‐RT NPs for dental applications due to their strong antioxidant, anticancer, and antimicrobial properties. These nanoparticles offer a hopeful prospect for addressing oral pathogen challenges and enhancing overall oral health. Summary: Zinc oxide‐tutin nanoparticle (ZnO‐RT NPs) exhibited robust free radical scavenging activity.It showed significant antimicrobial activity against oral pathogens.ZnO‐RT NPs also displayed dose‐dependent anticancer effects on human oral cancer cells.Mechanism action of ZnO‐RT NPs revealed the upregulation of apoptotic genes. [ABSTRACT FROM AUTHOR]

BACKGROUND: In pursuit of innovative wastewater treatment solutions, this study investigates the use of thermally treated palygorskite (TP) as an adsorbent to remove Escherichia coli and Enterococcus faecalis from synthetic wastewater. The goal is to explore a natural alternative to chlorine‐based disinfectants by utilizing TP's antimicrobial properties. Columns were packed with two granulometries (G1: 0.25–0.6 mm; G2: 1.40–2.36 mm) of TP and arranged in three different configurations (CA1, CA2 and CA3) to assess their bacterial removal efficiency, kinetic behavior and potential for reuse after dry heat sterilization. RESULTS: The CA3 column configuration, with its multilayer arrangement of TP, achieved the highest bacterial removal efficiency, reaching 99.1% for E. coli and 98.1% for E. faecalis. Kinetic experiments revealed that most bacterial adsorption occurred within the first 3 min, with E. coli requiring up to 10 min to reach maximum removal. TP's antibacterial effectiveness remained above 90% after two reuses. Additionally, dry heat sterilization allowed for repeated use of TP, showing stable removal efficiencies for E. faecalis and a slight decline for E. coli with each successive reuse. CONCLUSIONS: TP demonstrates significant potential as an adsorbent for wastewater disinfection, particularly in the CA3 multilayer configuration. Its rapid adsorption kinetics and resilience to heat sterilization underscore its reusability, making it a viable natural alternative to chemical disinfectants. Further research should focus on scaling this method to real wastewater treatment applications to validate its functionality in real‐world conditions. © 2024 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

Despite the intensive research on gut microbiome-associated diseases over the past 20 years, pharmacological methods for effectively eliminating pathobionts remain unsatisfactory. This study investigated the therapeutic potential of bacteriophages against Enterococcus faecalis, in which bacterial tyrosine decarboxylase (TDC) converts orally administered levodopa (L-DOPA) to dopamine, in an MPTP mouse model of Parkinson's disease (PD). E. faecalis bacteriophages PBEF62, PBEF66, and PBEF67 (4 × 1010 PFU total/200 µl/day), and E. faecalis cells (2 × 109 CFU/200 µl/day) were orally administered at 2-h intervals before every MPTP (i.p.) and/or L-DOPA (p.o.) treatments for 13 days. The relative abundances of E. faecalis cells and bacteriophages in the feces peaked at 4 and 12 h after administration and gradually decreased by 12 and 48 h, respectively. While the administration of E. faecalis cells eliminated the beneficial effect of L-DOPA on MPTP-induced behavioral deficits, as assessed by cylinder and rotarod tests, the co-administration of bacteriophages with bacterial cells restored this effect. The modulating effects of L-DOPA, E. faecalis, and bacteriophages on PD behavior were closely associated with choline acetyltransferase expression levels in the striatum but not with tyrosine hydroxylase in the substantia nigra of each group. Recurrence and extinction of PD behaviors following treatment with E. faecalis and/or bacteriophages were also coincident with the dopamine levels in the blood and brain tissues of PD mice. The effectiveness of L-DOPA was restored after the three types of E. faecalis bacteriophages selectively eliminated E. faecalis cells, along with the TDC gene copies and transcripts responsible for converting L-DOPA to dopamine in the gastrointestinal tract. In conclusion, a combination of bacteriophages PBEF62, PBEF66, and PBEF67 targeting E. faecalis demonstrates potential as a valuable supplement to L-DOPA therapy for PD. [ABSTRACT FROM AUTHOR]

Background: Chronic wound infections caused by Enterococcus faecalis pose formidable challenges in clinical management, exacerbated by the emergence of vancomycin-resistant strains. Phage therapy offers a targeted approach but encounters delivery hurdles. Due to their biocompatibility and controlled release properties, hydrogels hold promise as carriers. Objective: This study aimed to fabricate phage-containing hydrogels using sodium alginate (SA), carboxymethyl cellulose (CMC), and hyaluronic acid (HA) to treat E. faecalis-infected wounds. We assessed the efficacy of these hydrogels both in vitro and in vivo. Methods: The hydrogel was prepared using SA-CMC-HA polymers. Phage SAM-E.f 12 was incorporated into the SA-CMC-HA hydrogel. The hydrogel's swelling index was measured after 24 h, and degradation was assessed over seven days. Surface morphology and composition were analyzed using Scanning Electron Microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). Antibacterial activity was tested via optical density (OD) and disk diffusion assays. Phage release and stability were evaluated over a month. In vivo efficacy was tested in mice through wound healing and bacterial count assays, with histopathological analysis. Results: Hydrogels exhibited a swelling index of 0.43, a water absorption rate of %30, and 23% degradation over seven days. FTIR confirmed successful polymer incorporation. In vitro studies demonstrated that phage-containing hydrogels significantly inhibited bacterial growth, with an OD of 0.3 compared to 1.1 for the controls. Hydrogels remained stable for four weeks. In vivo, phage-containing hydrogels reduced bacterial load and enhanced wound healing, as shown by improved epithelialization and tissue restoration. Conclusion: Phage-containing hydrogels effectively treat wounds infected with E. faecalis-infected wounds, promoting wound healing through controlled phage release. These hydrogels can improve clinical outcomes in the treatment of infected wounds. [ABSTRACT FROM AUTHOR]

Background: Urinary tract infections (UTIs) represent one of the most prevalent bacterial infections, with Enterococcus species now recognized as the second leading cause of these infections. This study focused on symptomatic UTI cases to investigate the risk factors associated with Enterococcus faecalis clinical isolates in patients from Tehran, Iran. Methods: Urine samples were collected from patients presenting with symptomatic UTIs. The identification of E. faecalis isolates was performed using standard microbiological techniques, with confirmation via polymerase chain reaction (PCR). Antibiotic susceptibility testing was conducted using the Kirby--Bauer disc diffusion method. The presence of virulence genes was determined through PCR, and biofilm formation was assessed using the microtiter plate method. Additionally, multi-locus sequence typing (MLST) was utilized to genotype linezolid-resistant isolates. Results: Out of 300 UTI cases, E. faecalis was identified as the causative agent in 160 instances. Notably, a high proportion of these isolates exhibited resistance to tetracycline (83.8%) and minocycline (82.5%). Linezolid resistance was observed in 1.3% (n = 2) of the isolates. Conversely, the highest susceptibility rates were observed for vancomycin, penicillin G, ampicillin, and nitrofurantoin, each demonstrating a 98.8% susceptibility rate. Biofilm formation was detected in 25% of the E. faecalis isolates. A significant majority (93.8%) of the isolates harbored the efbA and ace genes, with varying frequencies of esp (72.5%), asa1 (61.2%), cylA (52.5%), and gelE (88.8%) genes. MLST analysis demonstrated that both linezolid-resistant isolates, characterized by strong biofilm formation and the presence of virulence genes, were assigned to the ST150 lineage, which has not been previously documented in clinical settings. Conclusion: The emergence of the ST150 clonal lineage, underscores its clinical significance, particularly in relation to linezolid resistance in E. faecalis. This study adds to the growing body of evidence linking specific clonal lineages with antibiotic resistance, highlighting the critical need for ongoing surveillance and molecular characterization of resistant pathogens. [ABSTRACT FROM AUTHOR]

Clostridioides difficile is a bacterial pathogen that has been implicated in severe gastrointestinal infections. C. difficile has intrinsic green autofluorescence and the level of this autofluorescence is known to be increased by growth time and oxygen. Currently, it is unclear if dietary compounds or metabolites from the gut microbiota are able to enhance C. difficile autofluorescence. Here, we aimed to determine potential factors that affect C. difficile autofluorescence. After screening a large repertoire of compounds, we identified several substances, like L-lysine and pantothenate, that led to an increased C. difficile autofluorescence. We also found that several members of the gut microbiota, such as Enterococcus faecalis, Klebsiella aerogenes and K. pneumoniae, can increase C. difficile autofluorescence through their secreted compounds. We further focused on the effect of K. pneumoniae on C. difficile autofluorescence and found that multiple enteric strains of K. pneumoniae could enhance C. difficile's autofluorescence. We used this enhanced autofluorescence to identify C. difficile in K. pneumoniae cocultures by flow cytometry. Our findings shed light on the relationship between C. difficile and other members of the gut microbiota, as well as different factors that can affect C. difficile autofluorescence. [ABSTRACT FROM AUTHOR]

Aim: To compare shock wave-enhanced emission photoacoustic streaming (SWEEPS) with sonic- and ultrasonically activated irrigation systems in removing Enterococcus faecalis biofilm from the root canal system. Methodology: Fifty human single-canalled mandibular premolars were included in the study. After access cavity preparation, the root canals were prepared to a standardized size and taper. Then, the entire root surface was covered with two layers of resin, and the root apices were sealed before sterilization. All root canals were inoculated with E. faecalis biofilm, and the samples were incubated aerobically for 2 weeks at 37 °C. Biofilm formation was confirmed by scanning electron microscopy. All samples were randomly divided into five groups (n = 10 each) based on their irrigation activation method as A (no treatment or negative control), B (no irrigation or positive control), C (sonically activated irrigation (SAI)), D (ultrasonically activated irrigation (UAI)), and E (needle irrigation activated by an Er: YAG laser device using a SWEEPS quartz tip (SWEEPS)). Then, dentine chips were retrieved, vortexed, and diluted for colony-forming unit counts. Data were analysed using analysis of variance and post-hoc Tukey tests (α = 5%). Results: All methods could significantly reduce E. faecalis biofilm compared with control so that the UAI, SWEEPS, and SAI groups indicated a 23.54%, 14.89%, and 7.81% biofilm reduction, respectively. UAI demonstrated a significantly more effective reduction of E. faecalis biofilm than SAI (p = 0.004). Conclusions: All irrigation activation methods significantly reduced E. faecalis biofilm, with ultrasonic use being the most effective. [ABSTRACT FROM AUTHOR]

Burn wounds are associated with infections, drug resistance, allergic reactions, odour, bleeding, excess exudates, and scars, requiring prolonged hospital stay. It is crucial to develop wound dressings that can effectively combat allergic reactions and drug resistance, inhibit infections, and absorb excess exudates to accelerate wound healing. To overcome the above-mentioned problems associated with burn wounds, SA/PVA/PLGA/Capparis sepiaria and SA/PVA/Capparis sepiaria nanofibers incorporated with Capparis sepiaria plant extract were prepared using an electrospinning technique. Fourier-transform infrared spectroscopy confirmed the successful incorporation of the extract into the nanofibers without any interaction between the extract and the polymers. The nanofibers displayed porous morphology and a rough surface suitable for cellular adhesion and proliferation. SA/PVA/PLGA/Capparis sepiaria and SA/PVA/Capparis sepiaria nanofibers demonstrated significant antibacterial effects against wound infection-associated bacterial strains: Pseudomonas aeruginosa, Enterococcus faecalis, Mycobaterium smegmatis, Escherichia coli, Enterobacter cloacae, Proteus vulgaris, and Staphylococcus aureus. Cytocompatibility studies using HaCaT cells revealed the non-toxicity of the nanofibers. SA/PVA/PLGA/Capparis sepiaria and SA/PVA/Capparis sepiaria nanofibers exhibited hemostatic properties, resulting from the synergistic effect of the plant extract and polymers. The in vitro scratch wound healing assay showed that the SA/PVA/Capparis sepiaria nanofiber wound-healing capability is more than the plant extract and a commercially available wound dressing. The wound-healing potential of SA/PVA/Capparis sepiaria nanofiber is attributed to the synergistic effect of the phytochemicals present in the extract, their porosity, and the ECM-mimicking structure of the nanofibers. The findings suggest that the electrospun nanofibers loaded with Capparis sepiaria extract are promising wound dressings that should be explored for burn wounds. [ABSTRACT FROM AUTHOR]

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One new 2,5-DKP derivative O-dihydroxycyclopenol (1) and seven known congeners 2–8 were isolated from the marine fungus Penicillium sp. ZJUT-34 cultured on rice medium. The planar structure of 1 was established by extensive spectroscopic analysis, including 1D, 2D NMR and HR-ESI-MS, while the relative configuration of 1 was determined by quantum chemical calculation. In the QS inhibitory assay, 1 significantly inhibited the production of violacein in Chromobacterium violaceum ATCC12472 (20.65%) at a concentration of 6.25 μg/mL without affecting the growth of the strain, as compared with norharmane (22.14%), a quorum sensing inhibitor (QSI) identified in our previous study. It represented the first report on the QS inhibitory activity of the seven-membered 2,5-DKPs. In addition, compounds 1–8 were subjected to antibacterial assay against six pathogenic bacteria Compound 8 exhibited comparable antibacterial activity against Enterococcus faecalis FA2-2 (MIC = 96 μg/mL) with the positive control gentamicin (MIC = 80 μg/mL). [ABSTRACT FROM AUTHOR]

Novel drug like chloroquinoline‐benzylamine hybrids were synthesized and subjected to comprehensive antibacterial evaluation in the present investigation. The pharmacological evaluation comprises determination of minimum inhibitory concentration (MIC), disk diffusion assay, hemolysis and combination assays against both Gram‐negative (Pseudomonas aeruginosa) and Gram‐positive (Bacillus subtilis, Enterococcus faecalis, and Staphylococcus aureus) bacterial strains. Among all, 7‐chloroquinoline‐benzylamine hybrid bearing p‐bromophenyl substituent (SA11) demonstrated significant antibacterial efficacy (MIC=128μg/mL) against the tested panel of Gram‐positive strains with no sign of toxicity towards human red blood cells (hRBCs). Furthermore, another hybrid with o‐hydroxyphenyl substitution (SA12) exhibited notable activity against the tested isolates with MIC values ranging from 64 to 256μg/mL. Molecular docking studies suggested compound SA11 binds within the active site of the biofilm causing protein (PDB: 7C7U) further supported by the molecular dynamics (MD) simulation studies at 100 ns. Compound SA11 exhibited significant efficacy in inhibiting S. aureus growth in the disk diffusion assay. Moreover, it displayed a synergistic effect when combined with ciprofloxacin (CIP), implying its potential utility in addressing antibiotic‐resistant bacterial strains through combination therapy. The pkCSM‐Pharmacokinetics assessment indicated favourable ADME profiles for SA11, supporting its potential as a viable drug candidate. [ABSTRACT FROM AUTHOR]

Vancomycin-resistant enterococci (VRE) are a public health concern as they lead to therapeutic impasses and play a pivotal role in the dissemination of vancomycin resistance genes. As recent evidence suggests that wildlife can play a role in the dissemination of bacterial resistomes, this study explored the potential role of Algerian wild birds as a reservoir of VRE. A total of 222 cloacal and fecal samples were collected from various wild bird species and screened for VRE using a selective medium. Of the 47 isolated strains, 22 were identified as Enterococcus casseliflavus with the van C2/C3 gene, 24 as Enterococcus gallinarum (19 carrying van C1 and five carrying van C2/C3), and one strain as Enterococcus faecalis with the van C1 gene. Twenty-four (24) strains were multidrug-resistant with 61.7% resistant to rifampicin, while no resistance to teicoplanin, linezolid, and gentamicin was found. Additionally, 53.20% of the strains exhibited at least one virulence factor. To our knowledge, this study represents the first documentation of the van C1 gene in E. faecalis isolated from wild birds. Furthermore, this gene was found to be carried by a conjugative plasmid, highlighting its ability to spread among bacterial populations and lead to the emergence of novel resistance phenotypes. [ABSTRACT FROM AUTHOR]

Oak bark, which is commonly used in the wood industry, has by-products often repurposed as fuel. Its extracts are rich in compounds with anticancer, antibacterial, antifungal, and anti-inflammatory properties. This study synthesized platinum nanoparticles (PtNPs) using aqueous extracts from Quercus dalechampii (QD), Q. frainetto (QF), and Q. petraea (QP). Key factors during nanoparticle formation included reaction time, metal ion concentration, pH, extract-to-metal ion ratio, and temperature. The PtNPs were characterized by dynamic light scattering, Fourier transform infrared spectroscopy, and transmission electron microscopy. The average diameters were 58.5 ± 7.6 nm for QD-PtNPs, 41.6 ± 5.4 nm for QF-PtNPs, and 41 ± 5.3 nm for QP-PtNPs. Antioxidant and antimicrobial activities were also analyzed. The QP-PtNPs had the highest DPPH (2,2-Diphenyl- 1-picrylhydrazyl), FRAP (Ferric Reducing Antioxidant Power), and CUPRAC (Cupric Reducing Antioxidant Capacity) free radical scavenging activities, while QD-PtNPs excelled in ABTS (2,2'-azinobis-(3- ethylbenzothiazoline-6-sulfonic acid)) scavenging. All PtNPs showed strong antimicrobial properties, particularly against Enterococcus faecalis, Escherichia coli, Candida krusei, and Candida auris. These findings suggest that Quercus-mediated PtNPs have significant potential for developing treatments against bacterial and fungal infections, with promising applications in medicine. [ABSTRACT FROM AUTHOR]

Cinnamon is a plant with significant medicinal value that is used extensively as a spice, flavoring, and fragrance ingredient. Phytochemical characterization via high-performance liquid chromatography, antimicrobial activity against various microorganisms, anti-diabetic properties by α-glucosidase and α-amylase assessments, antioxidant activity via 2,2-diphenyl-1-picrylhydrazyl-hydrate, and total antioxidant capacities of cinnamon bark were evaluated. Moreover, antitumor potential against human breast cancer (MCF-7) and human fetal lung fibroblast (WI-38) cells of the bark extract were detected. A promising antimicrobial action towards Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Salmonella typhi, Candida albicans, and Mucor circinelloides where the inhibition areas were 25, 23, 18, 19, 29, and 16 mm, respectively, was recorded. Cinnamon bark has IC50 of 5.01 and 2.58 µg/mL compared to standard acrobose with their IC50 values 4.32 and 1.99 µg/mL, respectively for α-amylase and α-glucosidase inhibitions. Cinnamon bark extract has IC50 of 77.39 ± 0.84 and 162.67 ± 0.28 µg/mL toward cell lines of MCF-7 and WI-38, respectively. The protective effect of cinnamon in accelerating the apoptosis of MCF-7 cells has been verified by flow cytometric evaluation employing Annexin-V and cell cycle kits, as well as the increasing levels of malondialdehyde, hydrogen peroxide, and nitric oxide with decreasing glutathione and catalase levels. [ABSTRACT FROM AUTHOR]

In this study, ethanol extracts of some wild mushroom species, in the phylum Basidiomycota, (Armillaria mellea (Vahl) P. Kumm., Infundibulicybe geotropa (Bull.) Harmaja, Leucopaxillus gentianeus (Quél.) Kotl. and Trametes versicolor (L.) Lloyd) were tested for their antioxidant, antimicrobial, antibiofilm, and cytotoxic activities. Mushroom samples showed low antimicrobial activity on Enterococcus faecalis and Staphylococcus aureus, while biofilm inhibitory activity on test microorganisms ranged from 24.6% to 80.5%. At the end of the antioxidant activity studies, A. mellea was the mushroom sample having the highest DPPH radical scavenging capacity (0.105±0.001 mg/mL) whereas T. versicolor was the one having the highest iron (III) ion reducing power (40.709±0.003 µg TE/100g). The highest polyphenol content was observed in T. versicolor (29.916±0.002 mg GAE/100g) samples, and the lowest in A. mellea (9.5±0.006 mg GAE/100g) samples. The cytotoxic effects of the samples were tested on MCF-7, MDA-MB-231 (breast cancer) and L929 (mouse fibroblast) cell lines using the MTT method. As a result, it was observed that A. mellea and T. versicolor samples were more effective on the MCF-7 cell line, and A. mellea on MDA-MB-231 cell line. [ABSTRACT FROM AUTHOR]

Objective: The crucial aim of irrigation solutions used in root canal treatments is the elimination of biofilm which is considered an important virulence factor of Enterococcus faecalis. Essential oils have been investigated to understand their efficacy for biofilm elimination in recent years. This study aimed to evaluate the antimicrobial and antibiofilm effects of Thymus vulgaris essential oils (TEO) and Origanum onites essential oils (OEO) on E. faecalis. Methods: The antimicrobial effectiveness of TEO and OEO against E. faecalis (ATCC 29212) was investigated by broth microdilution method, and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. The effect of TEO and OEO on preventing biofilm formation was evaluated by measuring biofilm biomass using crystal-violet method, and its effect on biofilm viability was evaluated by determining the number of living cells in the biofilm as colony-forming units. Biofilm viability was analyzed with one-sample t-test with statistical significance accepted as p<0.05. Results: The MIC values of TEO and OEO were determined as 0.078 µL/mL, and MBC values were determined as 0.156 µL/mL for OEO and 0.078 µL/mL for TEO. The percentage inhibition of biofilm formation at MIC value for OEO and TEO was determined as 53.9% and 55.6%, respectively. Both essential oils caused a significant reduction in the number of viable cells within the biofilm. Conclusion: It is concluded that TEO and OEO show high antimicrobial and antibiofilm activity against E. faecalis biofilm. Therefore, these essential oils can be considered an alternative irrigation solution for eliminating resistant root canal infections. [ABSTRACT FROM AUTHOR]

In this in vitro study, we aimed to evaluate and compare the antibacterial efficacy of a novel erbium-doped yttrium aluminum garnet laser modality, shock wave enhanced emission of photoacoustic streaming (SWEEPS), ultrasonically activated irrigation (UAI), and single needle irrigation (SNI) against old bacterial biofilm. A two-week-old Enterococcus faecalis biofilm was cultivated on transversal dentinal discs made from the middle third of the roots of single-rooted, single-canal premolars. Biofilm growth was confirmed using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The dentine samples were randomly distributed into three experimental groups and one control group based on the irrigation protocol used: Group 1, SWEEPS; Group 2, UAI; and Group 3, SNI. The root canals were irrigated with a 3% sodium hypochlorite solution. Antibacterial efficacy was evaluated quantitatively through bacterial culture and qualitatively through CLSM and SEM. Both SWEEPS and UAI demonstrated a statistically significant reduction in Enterococcus faecalis colony-forming units (CFUs) (p < 0.001), while SNI did not show a statistically significant reduction (p = 0.553). No significant difference was observed between the efficacy of SWEEPS and UAI (p > 0.05). The SWEEPS and UAI techniques were equally effective in eliminating mature E. faecalis biofilm. [ABSTRACT FROM AUTHOR]

Background/Objectives: The World Health Organization has selected enterococci as one of the priority multidrug-resistant microorganisms for the development of new antibacterial drugs. Bacteriophages are promising antibacterial agents, but the biology of bacteriophages requires deeper understanding. Methods: The vB_EfS_SE phage which is capable of infecting four species of the genus Enterococci was isolated from sewage plant. The complete genome of the vB_EfS_SE phage was sequenced using illumina technology. The endolysin gene was cloned into pBAD18 expression vector. Two chimeric endolysins were engineered using the vB_EfS_SE carbohydrate-binding domain (CBD) and replacing its enzymatically active domain (EAD). Results: The bacteriophage exhibits promising lytic properties and persists at temperatures of 40 °C and below, and under pH conditions ranging from 5 to 11. The genome sequence is 57,904 bp in length. The vB_EfS_SE endolysin PlySE and chimeric endolysins PlyIME-SE and PlySheep-SE were found to have the same range of specificity, but different thermostability properties and a different pH range for enzyme activity. Conclusions: Taking together the results obtained in this work and other published studies, we can highly appreciate the potential of Saphexavirus phages and their endolysins as novel antibacterial compounds. [ABSTRACT FROM AUTHOR]

Background and Objective: Eliminating intracanal Enterococcus faecalis (E. faecalis) is challenging because of its ability to penetrate deep dentinal tubules and its high resistance to many chemicals. This study evaluated the effectiveness of conventional needle irrigation and three different irrigant activation methods in reducing E. faecalis. Methods: The root canals of extracted teeth were shaped, contaminated with E. faecalis, and incubated for three weeks. They were randomly allocated to four experimental groups of 15 teeth each according to the final irrigation Method: group 1, conventional needle irrigation; group 2, passive ultrasonic (PU) irrigation; group 3, XP-endo Finisher (XPF); and group 4, laser-activated (LA) irrigation. Bacterial samples were taken and cultured before and after these final irrigation procedures. The colony-forming units were counted, and the bacterial reduction percentages of each group were calculated. The Kruskal-Wallis and Dunnet tests were used for statistical analysis. Results: All irrigant activation methods were significantly more effective than conventional needle irrigation. Although the LA group generated more negative samples than PU, there was no statistically significant difference between the LA and PU groups. LA was significantly more effective than the XPF, whereas PU and XPF were statistically similar. Conclusions: Within the limitations of this study, the final irrigation with LA and PU showed the best reductive effect on E. faecalis colonies. Considering that the LA group had more negative samples, it may be chosen as an alternative to enhance root canal disinfection, especially in difficult cases. [ABSTRACT FROM AUTHOR]

Background: Probiotics, which are live microorganisms that, when given in sufficient quantities, promote the host's health, have drawn a lot of interest for their ability to enhance gut health. Enterococcus faecalis, a member of the human gut microbiota, has shown promise as a probiotic candidate due to its functional attributes. However, safety concerns associated with certain strains warrant comprehensive evaluation before therapeutic application. Materials and Methods: In this study, E. faecalis EF-2001, originally isolated from fecal samples of a healthy human infant, was subjected to a multi-faceted assessment for its safety and probiotic potential. In silico analysis, CAZyme, biosynthetic, and stress-responsive proteins were identified. Results: The genome lacked biogenic amine genes but contained some essential amino acid and vitamin synthetic genes, and carbohydrate-related enzymes essential for probiotic properties. The negligible difference of 0.03% between the 1st and 25th generations indicates that the genetic information of the E. faecalis EF-2001 genome remained stable. The live E. faecalis EF-2001 (E. faecalis EF-2001L) demonstrated low or no virulence potential, minimal D-Lactate production, and susceptibility to most antibiotics except some aminoglycosides. No bile salt deconjugation or biogenic amine production was observed in an in vitro assay. Hemolytic activity assessment showed a β-hemolytic pattern, indicating no red blood cell lysis. Furthermore, the EF-2001L did not produce gelatinase and tolerated simulated gastric and intestinal fluids in an in vitro study. Similarly, heat-killed E. faecalis EF-2001 (E. faecalis EF-2001HK) exhibits tolerance in both acid and base conditions in vitro. Further, no cytotoxicity of postbiotic EF-2001HK was observed in human colorectal adenocarcinoma HT-29 cells. Conclusions: These potential properties suggest that probiotic and postbiotic E. faecalis EF-2001 could be considered safe and retain metabolic activity suitable for human consumption. [ABSTRACT FROM AUTHOR]

With antimicrobial resistance (AMR) rapidly evolving in pathogens, quick and accurate identification of genetic determinants of phenotypic resistance is essential for improving surveillance, stewardship, and clinical mitigation. Machine learning (ML) models show promise for AMR prediction in diagnostics but require a deep understanding of internal processes to use effectively. Our study utilised AMR gene, pangenomic, and predicted plasmid features from 647 Enterococcus faecium and Enterococcus faecalis genomes across the One Health continuum, along with corresponding resistance phenotypes, to develop interpretive ML classifiers. Vancomycin resistance could be predicted with 99% accuracy with AMR gene features, 98% with pangenome features, and 96% with plasmid clusters. Top pangenome features overlapped with the resistance genes of the vanA operon, which are often laterally transmitted via plasmids. Doxycycline resistance prediction achieved approximately 92% accuracy with pangenome features, with the top feature being elements of Tn916 conjugative transposon, a tet(M) carrier. Erythromycin resistance prediction models achieved about 90% accuracy, but top features were negatively correlated with resistance due to the confounding effect of population structure. This work demonstrates the importance of reviewing ML models' features to discern biological relevance even when achieving high-performance metrics. Our workflow offers the potential to propose hypotheses for experimental testing, enhancing the understanding of AMR mechanisms, which are crucial for combating the AMR crisis. [ABSTRACT FROM AUTHOR]

The rise of vancomycin-resistant enterococci (VRE) due to antibiotic overuse poses a significant threat to long-term care patients and those with impaired immune systems. Therefore, it is imperative to seek alternatives to overcome multidrug resistance. This study aimed to evaluate totarol, a natural compound derived from Podocarpus totara, for its antibacterial activity against vancomycin-resistant Enterococcus faecalis (VREF). Totarol exhibited potent antibacterial activity at a very low concentration of 0.25 µg/mL and demonstrated antibiofilm effects through biofilm inhibitory concentration and biofilm eradication concentration assays. Confocal laser scanning microscopy confirmed that totarol inhibited not only biofilm mass but also bacterial cell viability. The combinatorial use of sublethal concentrations of totarol and vancomycin showed antibacterial activity, as observed in the time-kill assay. Quantitative polymerase chain reaction assays revealed a concentration-dependent downregulation of key virulence genes (vanA, ace, asa, efaA, and esp) in VREF when exposed to totarol. In summary, totarol emerges as a promising adjuvant with vancomycin for inhibiting VREF, addressing vancomycin resistance and biofilm formation—critical challenges associated with VRE infection. Since this was an in vitro study, the role of totarol in the clinical implications of VREF treatment remains to be demonstrated. [ABSTRACT FROM AUTHOR]

This study presents the empirical findings of an in-depth genomic analysis of Enterococcus faecalis and Enterococcus lactis isolates from South Africa. It offers valuable insights into their genetic characteristics and their significant implications for public health. The study uncovers nuanced variations in the gene content of these isolates, despite their similar GC contents, providing a comprehensive view of the evolutionary diversity within the species. Genomic islands are identified, particularly in E. faecalis, emphasizing its propensity for horizontal gene transfer and genetic diversity, especially in terms of antibiotic resistance genes. Pangenome analysis reveals the existence of a core genome, accounting for a modest proportion of the total genes, with 2157 core genes, 1164 shell genes, and 4638 cloud genes out of 7959 genes in 52 South African E. faecalis genomes (2 from this study, 49 south Africa genomes downloaded from NCBI, and E. faecalis reference genome). Detecting large-scale genomic rearrangements, including chromosomal inversions, underscores the dynamic nature of bacterial genomes and their role in generating genetic diversity. The study uncovers an array of antibiotic resistance genes, with trimethoprim, tetracycline, glycopeptide, and multidrug resistance genes prevalent, raising concerns about the effectiveness of antibiotic treatment. Virulence gene profiling unveils a diverse repertoire of factors contributing to pathogenicity, encompassing adhesion, biofilm formation, stress resistance, and tissue damage. These empirical findings provide indispensable insights into these bacteria's genomic dynamics, antibiotic resistance mechanisms, and virulence potential, underlining the pressing need to address antibiotic resistance and implement robust control measures. [ABSTRACT FROM AUTHOR]

Purposes: Enterococcal BSI is associated with significant morbidity and mortality, with fatality rates of approximately 20–30%. There are microbiological and clinical differences between E. faecalis and E. faecium infections. The aim of this study was to investigate differences in predisposing factors for E. faecalis and E. faecium BSI and to explore prognostic factors. Methods: This study was a post-hoc analysis of PROBAC, a Spanish prospective, multicenter, cohort in 2016–2017. Patients with E. faecalis or E. faecium BSI were eligible. Independent predictors for BSI development in polymicrobial and monomicrobial BSI and in-hospital mortality in the monomicrobial group were identified by logistic regression. Results: A total of 431 patients were included. Independent factors associated with E. faecium BSI were previous use of penicillins (aOR 1.99 (95% CI 1.20–3.32)) or carbapenems (2.35 (1.12–4.93)), hospital-acquired BSI (2.58 (1.61–4.12)), and biliary tract source (3.36 (1.84–6.13)), while congestive heart failure (0.51 (0.27–0.97)), cerebrovascular disease (0.45 (0.21–0.98)), and urinary tract source (0.49 (0.26–0.92)) were associated with E. faecalis BSI. Independent prognostic factors for in-hospital mortality in E. faecalis BSI were Charlson Comorbidity Index (1.27 (1.08–1.51)), SOFA score (1.47 (1.24–1.73)), age (1.06 (1.02–1.10)), and urinary/biliary source (0.29 (0.09–0.90)). For E. faecium BSI, only SOFA score (1.34 (1.14–1.58) was associated with in-hospital mortality. Conclusions: The factors associated with E. faecium and E. faecalis BSI are different. These variables may be helpful in the suspicion of one or other species for empiric therapeutic decisions and provide valuable information on prognosis. [ABSTRACT FROM AUTHOR]

Considering the correlation between survival microenvironment of E. faecalis and acidic pH value, this study aimed to investigate the potential of utilizing pH-responsive DMAEM monomers and their copolymers with resin-based root canal sealers to inhibit E. faecalis. Broth microdilution assay, crystal violet staining and qPCR were performed to evaluate antibacterial effects of DMAEM monomers against E. faecalis at different pH. Methacrylate-resin based root canal sealers were prepared and copolymerized with DMAEM. The flow, solubility, water sorption, apical sealing ability and cytotoxicity of sealers were investigated to optimize formulation. The anti -E. faecalis effects of DMAEM copolymers with sealers were evaluated by direct contact test, colony-forming unit counting and live/dead staining. DMAEM monomers inhibited the growth, biofilm formation and virulence factors expression of E. faecalis in a concentration- and pH-dependent manner. Incorporation of 1.25 % and 2.5 % DMAEM into experimental sealers would not affect the flowability, solubility and periapical sealing ability (P > 0.05), but increased the water sorption of sealers (P < 0.01). Cells viability was higher than 90 % in both 1.25 % and 2.5 % DMAEM groups at pH 7.0. DMAEM copolymers with sealers reduced E. faecalis counts, inhibited biofilm formation and decreased live cells within the biofilm in response to pH values. DMAEM monomers and their copolymers with resin-based sealers possessed antibacterial and antibiofilm effects on E. faecalis in response to pH values. DMAEM is promising to inhibit intraradicular E. faecalis in response to its acidic survival environment and maintain low cytotoxicity under neutral conditions, ensuring their biosafety in case of inadvertent entry into periapical tissues. [Display omitted] • DMAEM monomers inhibit E. faecalis in response to acidic pH values. • DMAEM incorporated resin-based root canal sealers achieve good physical properties. • DMAEM incorporated sealers maintain good biocompatibilities at neutral pH. • DMAEM incorporated sealers inhibit E. faecalis in response to acidic pH values. [ABSTRACT FROM AUTHOR]

Objectives: The mannose phosphotransferase system (Man-PTS) plays crucial roles in the adaptive metabolic activity of Enterococcus faecalis (E. faecalis) in adverse environments. The aim of this study was to evaluate the role of Man-PTS in the alkaline resistance of E. faecalis against calcium hydroxide (CH) and the effect of metformin (Met) on the alkaline resistance of E. faecalis to CH. Materials and methods: The regulatory role of Man-PTS EII in the alkaline resistance of E. faecalis was firstly investigated using a wild-type highly alkaline-resistant E. faecalis XS 003, standard ATCC 29212 and Man-PTS EIID gene deficient (△mptD) and overexpressing (+mptD) strains of E. faecalis. RNA sequencing of Met-treated E. faecalis was performed to further validate the effect of Met on Man-PTS. The effect of Met on CH resistance of E. faecalis was verified by evaluating the survival, membrane potential and permeability, intracellular pH and ATP, and the expression of Man-PTS EII and membrane transporter-related genes of E. faecalis. The effect of Met on the ability of CH to remove E. faecalis biofilm on the dentin surface was also tested. The in vivo therapeutic effect of Met plus CH (CHM) was further investigated in a rat apical periodontitis model induced by E. faecalis XS 003. Results: Man-PTS EII significantly promoted the survival ability of E. faecalis in CH and enhanced its resistance to CH. The inhibition of Man-PTS EII by Met resulted in reduced alkaline resistance of E. faecalis in the presence of CH, while also enhancing the antimicrobial properties of CH against E. faecalis biofilm on dentin. Additionally, Met plus CH showed the synergistically promoted intra-canal E. faecalis infection control and healing of periapical lesion in rats. Conclusions: Met could significantly reduce the alkaline resistance of E. faecalis against CH through the modulation of Man-PTS EII, and improved the antibacterial effect of CH against E. faecalis infection both in vitro and in vivo. Clinical relevance: Met could significantly enhance the ability of CH to control E. faecalis infection through reducing the alkaline resistance of E. faecalis. [ABSTRACT FROM AUTHOR]

In the present work, an effective, ecofriendly and novel photocatalytic composite of Ni-ZnO/g-C3N4 (NZG) has been successfully prepared via a hydrothermal method. The crystalline structure and phase purity of the produced g-C3N4, Ni-ZnO, and Ni-ZnO/g-C3N4 nanocomposite were ascertained using XRD analysis. Both the g-C3N4, and Ni-ZnO retained peaks showed minor changes that suggested component interaction. Determine the functional groups and validate the composite's development using FTIR analysis of the combined g-C3N4, and Ni-ZnO characteristics, along with the addition of new Zn-O and C-N stretching bands. The optical properties and bandgap energy were observed for g-C3N4, Ni-ZnO, and Ni-ZnO/g-C3N4 nanocomposite were 360, 380, and 470 nm−1 with 2.74, 2.94 and 2.86 eV. The homogeneous distribution of Ni-ZnO nanoparticles on g-C3N4, sheets, with strong contact at the interface and consistent elemental composition, was shown by using TEM analysis to investigate the morphological and elemental composition. The photocatalytic degradation efficiency was investigated against RB5 dyes. The Ni-ZnO/g-C3N4 nanocomposite showed excellent than pure Ni-ZnO and g-C3N4 catalyst reached 89.7% degradation for RB5 after 120 min under UV light. Due to this enhanced stability, in addition to the improved electron hole separations and synergistic photocatalytic mechanism between Ni-ZnO and g-C3N4 is an excellent photocatalytic activity against waste water management. The positive control chloramphenicol was showed the inhibition zone was against Sterptococcus aureus and Enterococcus faecalis for 16 nm and 18 nm, for negative control there was no zone of inhibition, and the Ni-ZnO/g-C3N4 NCs showed the maximum zone of inhibition was observed in Enterococcus faecalis for 21 nm at and 100 µg/mL respectively. [ABSTRACT FROM AUTHOR]

The aim of this study was to test the hypothesis that a combination of D-amino acids (DAAs) and trans -cinnamaldehyde (TC) demonstrates superior antibiofilm activity to calcium hydroxide (CH) and untreated controls. In this 3-part in vitro study, the concentration of DAAs (D-methionine, D-leucine, D-tyrosine, and D-tryptophan) that would significantly decrease Enterococcus faecalis and Actinomyces naeslundii biofilm biomass was first determined. Then, the effect of TC + selected DAAs on polymicrobial biofilms was characterized by quantifying the biomass and biofilm viability. Finally, the antibiofilm effects of TC + DAA was compared with CH and untreated controls by (i) determining bacterial viability and (ii) quantifying biofilm matrix composition using selective fluorescence-binding analysis. Statistical analysis was performed using one-way ANOVA and appropriate multiple comparisons test, with P <.05 considered as statistically significant. TC (0.06%) + D-tyrosine (1 mM) + D-tryptophan (25 mM) significantly reduced the biomass and biofilm viability compared to the control (P <.05). While no significant difference was observed between TC + DAA and CH in the cultivable bacterial counts (P >.05), confocal microscopy demonstrated a significantly greater percentage of dead bacteria in TC + DAA-treated biofilms compared to CH and the control (P <.05). TC + DAA significantly decreased the biovolume and all the examined components of the biofilm matrix quantity compared to the control, while CH significantly reduced only the exopolysaccharide quantity (P <.05). The combination of TC + D-tyrosine + D-tryptophan demonstrated superior antibiofilm activity (biofilm bacterial killing and reduction of matrix quantity) to CH and has potential to be developed as an intracanal medicament. [ABSTRACT FROM AUTHOR]

The demand for safe and effective endodontic medicaments to control Enterococcus faecalis biofilms, a contributor to apical periodontitis, is increasing. Recently, lipoteichoic acid (LTA) of family Lactobacillaceae has been shown to have anti-biofilm effects against various oral pathogens. Preliminary experiments showed that LTA purified from Lacticaseibacillus rhamnosus GG (Lgg.LTA) was the most effective against E. faecalis biofilms among LTAs from three Lactobacillaceae including L. rhamnosus GG, Lacticaseibacillus casei, and Lactobacillus acidophilus. Therefore, in this study, we investigated the potential of Lgg.LTA as an intracanal medicament in human root canals infected with E. faecalis. Twenty eight dentinal cylinders were prepared from extracted human teeth, where two-week-old E. faecalis biofilms were formed followed by intracanal treatment with sterile distilled water (SDW), N-2 methyl pyrrolidone (NMP), calcium hydroxide (CH), or Lgg.LTA. Bacteria and biofilms that formed in the root canals were analyzed by scanning electron microscopy and confocal laser scanning microscopy. The remaining E. faecalis cells in the root canals after intracanal medicament treatment were enumerated by culturing and counting. When applied to intracanal biofilms, Lgg.LTA effectively inhibited E. faecalis biofilm formation as much as CH, while SDW and NMP had little effect. Furthermore, Lgg.LTA reduced both live and dead bacteria within the dentinal tubules, indicating the possibility of minimal re-infection in the root canals. Collectively, intracanal application of Lgg.LTA effectively inhibited E. faecalis biofilm formation, implying that Lgg.LTA can be used as a novel endodontic medicament. [ABSTRACT FROM AUTHOR]

Enterococcus faecalis is considered a probiotic, commensal lactic acid bacterium in human breast milk (HBM), but there are circulating antibiotic resistant and virulence determinants that could pose a risk in some strains. The study aimed to conduct genomic analysis of E. faecalis isolates recovered from HBM and animal milk and to evaluate their probiotic and pathogenic features through comparative genomics with isolates from clinical specimens (e.g., urine, wound, and blood). Genomic analysis of 61 isolates was performed, including E. faecalis isolates recovered from HBM in Saudi Arabia. Genome sequencing was conducted using the MiSeq system. The fewest antibiotic resistance genes (lsaA, tetM, ermB) were identified in isolates from HBM and animal milk compared to clinical isolates. Several known and unknown mutations in the gyrA and parC genes were observed in clinical isolates. However, 11 virulence genes were commonly found in more than 95% of isolates, and 13 virulence genes were consistently present in the HBM isolates. Phylogenetically, the HBM isolates from China clustered with the probiotic reference strain Symbioflor 1, but all isolates from HBM and animal milk clustered separately from the clinical reference strain V583. Subsystem functions 188 of 263 were common in all analyzed genome assemblies. Regardless of the source of isolation, genes associated with carbohydrate metabolism, fatty acid, and vitamin biosynthesis were commonly found in E. faecalis isolates. In conclusion, comparative genomic analysis can help distinguish the probiotic or pathogenic potential of E. faecalis based on genomic features. [ABSTRACT FROM AUTHOR]

Background: The management of endodontic infections is a complex challenge, mainly due to the involvement of diverse microorganisms and their by-products. This study aimed to evaluate the efficacy of N-acetylcysteine (NAC), calcium hydroxide (Ca(OH)2), and their combined application as intracanal medications in combating Enterococcus faecalis, Escherichia coli, and lipopolysaccharides (LPS) from E. coli. Methods: A total of 60 single-rooted human teeth were carefully selected and divided into six groups. These tooth canals were deliberately exposed to E. faecalis (ATCC 29212) and E. coli (ATCC 25922) to induce biofilm formation. Subsequently, the specimens were treated with NAC, Ca(OH)2, or a combination of both substances. Three samples of the root canals were collected at three moments: the first sample (S1) was to confirm the initial contamination, the second sample (S2) was immediately post-instrumentation, and the third sample (S3) was collected after the use of the intracanal medication. The antimicrobial efficacy of these intracanal medications was assessed by enumerating colony-forming units per milliliter (CFU/mL). In addition to this, the kinetic chromogenic Limulus Amebocyte Lysate (LAL) assay by Lonza was used to quantify LPS from E. coli. Data tested for normality; then, Kruskal-Wallis and Friedman tests were used, and Dunn's for multiple comparisons. Results: The findings of this study showed significant reductions in the microbial load of E. faecalis and E. coli by S3. Notably, there were no statistically significant differences among the treatment groups concerning these microorganisms. However, it was observed that only the combination of NAC and Ca(OH)2 led to a noteworthy decrease in the quantity of E. coli's LPS after 7-days, demonstrating a statistically significant difference from the other treatment groups. NAC + Ca(OH)2 combination, applied for a duration of 7-days, proved to be more suitable in reducing the presence of E. faecalis, E. coli, and LPS from E. coli within the context of endodontic infections. [ABSTRACT FROM AUTHOR]

Background: Treating tooth root canal systems with Er:YAG laser together with irrigants has been shown to be effective in reducing biofilms formed by Enterococcus faecalis. This study investigated whether laser-activated irrigation (LAI) with side-firing Endo tip (LiteTouch™; Light Instruments, Yokneam, Israel) affects dentin mineral composition when used with common endodontic irrigants. Methods: Root canals of extracted human teeth were treated with Er:YAG laser using a side-firing Endo tip combined with 17% ethylenediaminetetraacetic acid (EDTA) and/or 2.5% NaOCl in continuous or intermittent mode for 60 s. Dentin mineral composition (Ca, P, O) in coronal, middle, and apical regions of root canals was examined by energy dispersive X-ray spectroscopy. Results: The use of LAI with continuous EDTA resulted in the largest reduction in Ca and P levels. A final NaOCl rinse mitigated the EDTA-mediated mineral loss in all root canal regions and increased the O content. Likewise, the reduced Ca/O and Ca/P ratios caused by continuous EDTA irrigation were reversed when combined with a final NaOCl rinse. Conclusions: LAI with Er:YAG Endo tip using continuous EDTA irrigation followed by NaOCl caused minimal dentin mineral loss and can therefore be considered to be a safe treatment module for cleaning root canals. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to evaluate the photothermal effect of a 970 nm diode laser on Enterococcus faecalis biofilms. Methods: 72 extracted human single-rooted teeth were prepared, sterilized, and inoculated with Enterococcus faecalis to establish a two-week-old biofilm. The specimens were divided into six groups (n = 12): Group 1 (G1)—negative control (PBS—no laser), Group 2 (G2)—positive control (1% NaOCl rinse—no laser), Group 3 (G3)—a 970 nm laser in 1.5 W pulse mode, Group 4 (G4)—a 970 nm laser in 2 W pulse mode, Group 5 (G5)—a 970 nm laser in 1.5 W continuous mode, Group 6 (G6)—a 970 nm laser in 2 W continuous mode. Bacterial viability was evaluated using the LIVE/DEAD BacLight kit and analyzed by flow cytometry (FCM). Temperature changes on the root surface during irradiation were analyzed using a K-type thermocouple. Data were statistically analyzed using one-way ANOVA and Tukey's multiple comparison test (α = 0.05). Results: Bacterial viability was significantly reduced after laser irradiation in continuous mode using 1.5 W (21% of live bacteria) and 2 W (14% of live bacteria). When the pulsed mode was applied, the reduction in bacterial viability was less, with a mean survival of 53% (1.5 PF, whereas 29% of bacteria survived after 2 W irradiation). Conclusions: The 970 nm diode laser at 2 W continuous mode effectively reduced the viability of E. faecalis biofilms in root canals without causing unacceptable temperature rises at the root surface. [ABSTRACT FROM AUTHOR]

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Pelargonium plants are very popular and well-known for their essential oils (EOs), which are used for medicinal purposes and in food. This study focused on the EO of Pelargonium odoratissimum. First, its composition and antioxidant and antimicrobial activity were evaluated, and finally, its efficacy as a natural preservative in ground beef was tested. The main EO constituents were citronellol (40.0%), nerol (15.3%), and citronellyl formate (12.6%). The antibacterial activity of POEO showed that Enterococcus faecalis ATCC 29212 was the most susceptible strain compared to the other eight strains tested. The antioxidant activity, as measured by the DPPH assay, showed a dose-dependent effect with an IC50 comparable to the standard used, gallic acid. Aerobic plate count, psychotropic bacteria, and Enterobacteriaceae, including Salmonella, were reduced by the addition of Pelargonium odoratissimum essential oils. The oxidative stability was significantly improved compared to the untreated sample. Additionally, the results for metmyoglobin demonstrated a notable preservative effect on sensory properties, including appearance, odor, color, and overall acceptability. The ability to discriminate between all samples and correlate protein and lipid oxidation processes, microbiological characteristics, and sensory measurements was made possible by principal component analysis and heat maps. This research shows the potential benefits of using POEO in the preservation of ground beef by effectively extending shelf life and improving product safety. [ABSTRACT FROM AUTHOR]

The main objective of this study is to isolate and identify lactic acid bacteria (LAB) from various Egyptian dairy products, examine their antibacterial and hemolysis potential, and ensure their safety when used as starter cultures in different dairy industries. Egyptian dairy products are often made without the use of commercial starter cultures, using raw milk and artisanal methods. The most popular traditional dairy products are Laban Rayeb and Zabady, as well as the cheese varieties of Ras, Domiati, and Karish. The microbial communities used for fermentation and the diversity of lactic acid bacteria are the most important factors that can affect the quality of these products. In order to investigate the diversity of cocci lactic acid bacteria in Egyptian dairy products, 70 samples of raw or fermented milk and cheeses were collected from traditional cheese-making factories, local markets, and farmhouses located in the Delta area of Egypt. Following this, the LAB were isolated from the samples. One hundred fifty-seven isolates of Gram-positive, catalase-negative, and cocci bacterial species were identified via rep-PCR, and some isolates were confirmed using pheS and 16S rRNA gene sequencing, as follows: Streptococcus infantarius subsp. infantarius (three isolates), Enterococcus hirae (three isolates), Enterococcus faecium (ninety-six isolates), Enterococcus faecalis (forty isolates), Enterococcus durans (six isolates), Lactococcus garvieae (one isolate), Pediococcus acidilactici (seven isolates), and Lactococcus lactis subsp. Lactis (one isolate). These findings validate that five strains have strong antibacterial activity against Escherichia coli, Salmonella typhimurium, and Listeria monocytogenes, and one hundred thirty-four strains were safe for hemolysis. The five strains were selected as protective cultures, including Pediococcus acidilactici, Lactococcus lactis subsp. lactis, E. faecalis, and E. faecium. [ABSTRACT FROM AUTHOR]

Vancomycin-resistant enterococci (VRE) are considered one of the main nosocomial pathogens due to their increasing antibiotic resistance and ability to cause life-threatening infections in humans. This study included VRE isolates obtained from various specimens including urine, blood, faeces, wounds, sputum, and oral cavity wash. Of the 37 strains, 30 (81.1%) and 7 (18.9%) were identified by MALDI TOF as Enterococcus faecium and Enterococcus faecalis, respectively. The clinical vancomycin-resistant enterococci exhibited multi-drug resistance (MDR). Apart from vancomycin, the enterococci exhibited resistance to penicillins (89.1 to 100%), fluoroquinolones (100%), rifampicin (86.5%), tetracycline (27%), aminoglycosides (56.8 to 86.5%), quinupristin–dalfopristin (35.1%), and chloramphenicol (10.8%). Moreover, resistance to linezolid and tigecycline emerged among the tested vancomycin-resistant enterococci. The analysis of aminoglycoside modifying enzyme (AME) genes showed the presence of bifunctional aac(6′)-Ie-aph(2″)-Ia genes contributed to high-level aminoglycoside resistance (HLAR) in the E. faecalis and E. faecium isolates. The other AME gene, i.e., aph(3′)-IIIa, was also found in the VRE isolates. All strains carried the vanA gene. Enterococci from colonised gastrointestinal tracts (1/2.7%) and from infection (6/16.2%) showed cytotoxic activity against the human epithelial cell line HEp-2. [ABSTRACT FROM AUTHOR]