Effect of Laurus nobilis on bacteria and human transforming growth factor-ß1.

OBJECTIVE: In this study, we aimed to determine the phenolic compounds, the antibacterial activity of extract from Laurus nobilis leaves, and its possible effect on transforming growth factor-ß1 expression level in peripheral blood mononuclear cells. METHODS: The phenolic components of Laurus nobilis were identified by the high-performance liquid chromatography method. The antibacterial activity of this extract was determined by disk diffusion and broth microdilution methods. The transforming growth factor-ß1 expression was analyzed using the RT-qPCR method. RESULTS: Epicatechin was found in the highest amount and o-coumaric acid in the lowest amount. The half-maximal inhibitory concentration (IC50) was determined to be 55.17 µg/mL. The zones of inhibition and minimum inhibitory concentration for Staphylococcus aureus, Enterococcus faecalis, and Klebsiella pneumoniae were 15, 14, and 8 mm and 125, 250, and 1000 µg/mL, respectively. The change in transforming growth factor-ß1 expression levels was found to be statistically significant compared with the control groups (p<0.0001). CONCLUSION: Laurus nobilis extract was found to be effective against bacteria and altered the expression level of transforming growth factor-ß1 in peripheral blood mononuclear cells.

Leaf Extracts of Moringa oleifera Cultivated in Baghdad: Characterization and Antimicrobial Potential against Endodontic Pathogens.

The use of medicinal plant preparations to clean and disinfect root canal infection is gaining popularity. The aim of this study was to evaluate the bioactive composition of leaf extracts of Moringa oleifera plants cultivated in Iraq (specifically Baghdad) and their antimicrobial activity against selected root canal pathogens for potential application in endodontic treatment. Materials and Methods. Moringa leaf extracts were prepared either through cold maceration or warm digestion techniques to perform an ethanolic or aqueous extraction, respectively. Phytochemical detection was performed before thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) to measure flavonoids and phenolic compounds within both extracts. Then, their antimicrobial activities were investigated against Streptococcus mutans, Enterococcus faecalis, and Candida albicans through minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), and agar well diffusion assay in comparison to NaOCl and Ca(OH)2. Results. Phytochemical screening showed several active ingredients but with higher expression of flavonoids and phenolic compounds. Also, different types of these compounds were detected through TLC and quantified by HPLC. MIC values for ethanolic extract against Streptococcus mutans, Enterococcus faecalis, and Candida albicans were 60, 65, and 55, respectively, while for aqueous extract, MIC values were 70, 80, and 50, respectively. Aqueous extract showed a higher inhibition zone than ethanolic extract for both Streptococcus mutans and Enterococcus faecalis with a statistically significant difference (p ≤ 0.001) for all tested materials except with NaOCl and Ca(OH)2 in Streptococcus mutans and Enterococcus faecalis, respectively. The ethanolic extract showed a higher inhibition zone against Candida albicans, with a statistically significant difference (p ≤ 0.001) for all tested materials. Conclusion. Ethanolic and aqueous extracts of Moringa oleifera leaves cultivated in Baghdad contain considerable quantities of phytochemicals, especially flavonoid and phenolic compounds, and demonstrated antimicrobial activities against selected endodontic pathogens. Therefore, Moringa leaf extracts could be suggested as an alternative antimicrobial material in endodontic treatment.

IS6 family insertion sequences promote optrA dissemination between plasmids varying in transfer abilities.

Plasmids are the primary vectors for intercellular transfer of the oxazolidinone and phenicol cross-resistance gene optrA, while insertion sequences (ISs) are mobile genetic elements that can mobilize plasmid-borne optrA intracellularly. However, little is known about how the IS-mediated intracellular mobility facilitates the dissemination of the optrA gene between plasmid categories that vary in transfer abilities, including non-mobilizable, mobilizable, and conjugative plasmids. Here, we performed a holistic genomic study of 52 optrA-carrying plasmids obtained from searches guided by the Comprehensive Antibiotic Resistance Database. Among the 132 ISs identified within 10 kbp from the optrA gene in the plasmids, IS6 family genes were the most prevalent (86/132). Homologous gene arrays containing IS6 family genes were shared between different plasmids, especially between mobilizable and conjugative plasmids. All these indicated the central role of IS6 family genes in disseminating plasmid-borne optrA. Thirty-three of the 52 plasmids were harbored by Enterococcus faecalis found mainly in humans and animals. By Nanopore sequencing and inverse PCR, the potential of the enterococcal optrA to be transmitted from a mobilizable plasmid to a conjugative plasmid mediated by IS6 family genes was further confirmed in Enterococcus faecalis strains recovered from the effluents of anaerobic digestion systems for treating chicken manure. Our findings highlight the increased intercellular transfer abilities and dissemination risk of plasmid-borne optrA gene caused by IS-mediated intracellular mobility, and underscore the importance of routinely monitoring the dynamic genetic contexts of clinically important antibiotic resistance genes to effectively control this critical public health threat. KEY POINTS: • IS6 was prevalent in optrA-plasmids varying in intercellular transfer abilities. • Enterococcal optrA-plasmids were widespread among human, animal, and the environment. • IS6 elevated the dissemination risk of enterococcal optrA-plasmids.

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 effect of Pentaclethra Macroloba plant extract against Enterococcus Faecalis.

This study evaluated the antimicrobial efficacy of a new intracanal drug based on Pentaclethra macroloba extract, a plant of Amazonian origin, against Enterococcus faecalis using macrodilution test and intratubular evaluation with Confocal Laser Scanning Microscopy (CLSM). The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the pure extract of Pentaclethra macroloba andits association with calcium hydroxide and ultracall were determined. Then, thirty-three dentin cylinders were prepared and inoculated with E. faecalis, to evaluate the antimicrobial effect of the medications on the dentinal tubules with CLSM. The data was analyzed using the Kruskal-Wallis and Dunn tests. The extract in association with calcium hydroxide showed a lower CBM, and in the intratubular test all tested medications were effective against E. faecalis (P >0.05). The new intracanal drug based on P. macroloba extract has an antimicrobial effect against E. faecalis and further studies are needed for its clinical use.

Antimicrobial efficacy of different herbal extracts against root canal pathogens - An in vitro study.

Background: The antimicrobial activity exhibited by the plant extracts against various pathogens has previously been demonstrated to a limited extent. However, the antibacterial effects of most of them have not yet been clarified in endodontics. Aim: This study aimed to determine the antimicrobial activity of some medicinal plants' methanolic extracts against some root canal microorganisms. Methods and Materials: In this in vitro study, after gathering the five plants (pomegranate peel, clove, Zhumeria majdae, Eucalyptus galbie and green tea), their methanolic extracts were obtained by the maceration method, and the antimicrobial activity was determined using micro- and macro-dilution methods. The microorganisms tested in this study were Enterococcus (E.) faecalis, Porphyromonas (P.) gingivalis and Fusobacterium (F.) nucleatum. Non-ready-to-use calcium hydroxide (CH), ready-to-use Calcipex II CH and methanol were used as control materials. Statistical Analysis: The nonparametric Kruskal-Wallis and Friedman tests were used to analyse and evaluate the data and variables. Results: In all concentrations and time intervals studied, eucalyptus extract showed the highest antimicrobial activity against E. faecalis. All the extracts showed growth-inhibitory effects against P. gingivalis and F. nucleatum. There were no significant differences between the anti-enterococcus effects of eucalyptus extracts and non-ready-to-use CH (P > 0.05). Conclusion: There was a greater efficacy of Eucalyptus galbie extract than other extracts.

First Report of a Wastewater Treatment-Adapted Enterococcus faecalis ST21 Harboring vanA Gene in Brazil.

Enterococcus faecalis has emerged as an important opportunistic pathogen due to its increasing resistance to antimicrobials, mainly to vancomycin, which leads substantial cases of therapeutic failures. Wastewater treatment plants (WWTP), in turn, are considered hotpots in the spread of antimicrobial resistance according to One Health perspective. In this study, we present the first report of a vancomycin-resistant E. faecalis strain recovered from treated effluent in Brazil. For this purpose, the whole-genome sequencing (WGS) was carried out aiming to elucidate its molecular mechanisms of antimicrobial resistance and its phylogenetic relationships amongst strains from other sources and countries. According to Multilocus Sequence Typing (MLST) analysis this strain belongs to ST21. The WGS pointed the presence of vanA operon, multiple antibiotic resistance and virulence genes, and a significant pathogenic potential for humans. The phylogenomic analysis of E. faecalis 6805 was performed with ST21 representatives from the PubMLST database, including the E. faecalis IE81 strain from clinical sample in Brazil, which had its genome sequenced in this study. Our results demonstrated a strain showing resistance to vancomycin in treated effluent. To the best of our knowledge, this is an unprecedented report of vanA-carrying E. faecalis ST21. Furthermore, it is the first description of a vanA-harboring strain of this species from environmental sample in Brazil. Our data highlight the role of WWTP in the spread of AMR, since these environments are favorable for the selection of multidrug-resistant pathogens and the treated effluents, carrying antibiotic resistance genes, are directed to receiving water bodies.

Low-dose perinatal supplementation with Enterococcus faecalis increases concentrations of short-chain fatty acids in the offspring but does not protect against allergic asthma.

Childhood allergic asthma is associated with a dysbiotic gut microbiome in early life, and maternal perinatal treatment with probiotics is a potential way alter the infant microbiome, which may improve asthma outcomes. This study used a mouse model to examine the effect of maternal supplementation with the probiotic Enterococcus faecalis on faecal short-chain fatty acid (SCFA) concentrations and asthma risk in the offspring. Pregnant/lactating mice were treated daily, from gestation day 6 to postnatal day 21, with an oral suspension of 106, 107 or 108 colony-forming units of a live preparation of the probiotic E. faecalis (Symbioflor®1). At weaning, offspring were subjected to an ovalbumin-induced experimental asthma protocol. Faeces were collected from the mothers and offspring at several different time points to determine SCFA concentrations. It was found that maternal supplementation with E. faecalis did not alter litter size, sex ratio or offspring weight, and was associated with an increase in SCFAs in offspring faeces at weaning and after allergy induction. However, allergic offspring from E. faecalis supplemented mothers showed no difference in asthma severity when compared with allergic offspring from control mothers. In conclusion, although maternal perinatal supplementation with low-dose E. faecalis was associated with increased faecal SCFAs in the offspring, it did not protect against offspring asthma. This is may be because SCFA concentrations were not increased to an immunoprotective level. We recommend that future studies concentrate on probiotic supplementation in high-risk cases, for instance, to repair gut dysbiosis resulting from antibiotic use in pregnant mothers or their infants.

Growth of Enterococcus faecalis ∆plsX strains is restored by increased saturated fatty acid synthesis.

The Enterococcus faecalis acyl-acyl carrier protein (ACP) phosphate acyltransferase PlsX plays an important role in phospholipid synthesis and exogenous fatty acid incorporation. Loss of plsX almost completely blocks growth by decreasing de novo phospholipid synthesis, which leads to abnormally long-chain acyl chains in the cell membrane phospholipids. The ∆plsX strain failed to grow without supplementation with an appropriate exogenous fatty acid. Introduction of a ∆fabT mutation into the ∆plsX strain to increase fatty acid synthesis allowed very weak growth. The ∆plsX strain accumulated suppressor mutants. One of these encoded a truncated ß-ketoacyl-ACP synthase II (FabO) which restored normal growth and restored de novo phospholipid acyl chain synthesis by increasing saturated acyl-ACP synthesis. Saturated acyl-ACPs are cleaved by a thioesterase to provide free fatty acids for conversion to acyl-phosphates by the FakAB system. The acyl-phosphates are incorporated into position sn1 of the phospholipids by PlsY. We report the tesE gene encodes a thioesterase that can provide free fatty acids. However, we were unable to delete the chromosomal tesE gene to confirm that it is the responsible enzyme. TesE readily cleaves unsaturated acyl-ACPs, whereas saturated acyl-ACPs are cleaved much more slowly. Overexpression of an E. faecalis enoyl-ACP reductase either FabK or FabI which results in high levels of saturated fatty acid synthesis also restored the growth of the ∆plsX strain. The ∆plsX strain grew faster in the presence of palmitic acid than in the presence of oleic acid with improvement in phospholipid acyl chain synthesis. Positional analysis of the acyl chain distribution in the phospholipids showed that saturated acyl chains dominate the sn1-position indicating a preference for saturated fatty acids at this position. High-level production of saturated acyl-ACPs is required to offset the marked preference of the TesE thioesterase for unsaturated acyl-ACPs and allow the initiation of phospholipid synthesis.

Nano-antibiotic based on silver nanoparticles functionalized to the vancomycin-cysteamine complex for treating Staphylococcus aureus and Enterococcus faecalis.

BACKGROUND: Bacterial resistance is defined as a microorganism's capacity to develop mechanisms for resisting a determined antimicrobial. Gram-positive bacteria, such as Staphylococcus aureus (S. aureus) and Enterococcus faecalis (E. faecalis), are internationally recognized among the isolates with this resistance profile. In this context, the demand for new medicines has risen, and silver nanoparticles (AgNPs) have been highlighted, especially for their anti-bacterial effects. To develop a nano-antibiotic for treating these Gram-positive strains, we herein report synthesizing and characterizing a nano-antibiotic based on AgNPs functionalized with the complex vancomycin-cysteamine. METHODS: AgNPs were produced using the bottom-up methodology and functionalized with vancomycin modified by the carbodiimide chemistry, forming Ag@vancomycin. Susceptibility tests were performed using S. aureus and E. faecalis strains to assess the bacteriostatic and bactericidal potential of the developed nano-antibiotic. RESULTS: Fourier transform infrared spectroscopy measurements showed the efficacy of vancomycin chemical modification, and the characteristic bands of AgNPs functionalization with the antibiotic. The increase in the nano-antibiotic average hydrodynamic diameter observed by dynamic light scattering proved the presence of vancomycin at the surface of AgNPs. The data from the minimum inhibitory concentration and minimal bactericidal concentration assays tested on standard and clinical planktonic strains of S. aureus and E. faecalis presented excellent performance. CONCLUSION: The results indicate the promising development of a new nano-antibiotic in which the functionalization potentiates the bacteriostatic action of AgNPs and vancomycin with greater efficacy against Gram-positive strains.

Ex Vivo Evaluation of the Efficacy of Photodynamic Therapy in Eliminating Enterococcus faecalis from Dentinal Tubules by Confocal Laser Scanning Microscopy.

INTRODUCTION: The aim of the present study was to investigate ex vivo by confocal laser scanning microscopy the antibacterial effect of photodynamic therapy (PDT) on dentinal tubules in the apical 5 mm of human mandibular premolars contaminated with Enterococcus faecalis. METHODS: Thirty-four teeth were standardized to 20 mm and foraminal anatomic diameters using a #20 K-file (Dentsply Maillefer). Samples were contaminated for 21 days and divided into the following 3 experimental groups (n = 10): the PDT group (instrumented canals and PDT), the passive ultrasonic irrigation (PUI) group (instrumented canals and PUI), and the PUI-PDT group (instrumented canals, PUI, and PDT), along with a control group (n = 4) (noninstrumented canals). The canals in the experimental groups were instrumented with ProTaper Next (Dentsply Maillefer) up to X3 and rinsed with EDTA and sodium hypochlorite. The photosensitizer used was 0.01% methylene blue with a preirradiation time of 5 minutes and a diode laser with 4 J energy and a 660-nm wavelength. Cross sections were made 5 mm from the apex of all samples, which were analyzed using confocal laser scanning microscopy. The results were analyzed using the Shapiro-Wilk and Kruskal-Wallis (Dunn) tests. RESULTS: There was a lower percentage of live bacteria in the PUI-PDT group, with a statistical difference compared with the control and PDT groups (P < .05). There was no statistical difference in the percentage of live bacteria between PUI-PDT and PUI (P > .05). CONCLUSIONS: It was concluded that the PUI-PDT association was most effective in disinfecting root canals compared with the control group and PDT.

In Vitro Antibacterial and Phytochemical Screening of Hypericum perforatum Extract as Potential Antimicrobial Agents against Multi-Drug-Resistant (MDR) Strains of Clinical Origin.

Background: The perennial plant Hypericum perforatum is widely distributed around the world. It has been used for many years in conventional medicine to treat a variety of illnesses, including stress, mild to moderate depression, and minor injuries. This study examined the antimicrobial activity of the H. perforatum total extract and its fractions (n-hexane, ethyl acetate, chloroform, and aqueous) against multi-drug-resistant (MDR) isolates that were gathered from clinical samples, including methicillin-resistant Staphylococcus aureus (MRSA), Enterococcus faecalis, Escherichia coli, and Klebsiella pneumonia. Materials and Methods: Aerial parts of H. perforatum were collected and extracted using various solvents and were tested versus different isolated bacterial species. The inhibition zone of tested extracts was detected using an agar diffusion assay, and MICs were measured. Phytochemical analysis of promising H. perforatum extract was done using LC-ESI-MS/MS. Ultrastructure examination for the most altered bacteria used transmission electron microscopy. Antioxidant assays were done using DPPH and ABTS scavenging capacity methods. Cytotoxicity was reported versus Vero cells. Results: Different extracts of H. perforatum showed promising antibacterial activity against the pathogens. While the subfractions of the total extract were observed to show lesser inhibition zones and higher MIC values than the total extract of H. perforatum against MDR strains, the total extract of H. perforatum demonstrated the most potent antimicrobial action with an inhibition zone range of 17.9-27.9 mm. MDR-K. pneumoniae was discovered to be the most susceptible strain, which is consistent with the antibacterial inhibitory action of H. perforatum whole extract. Additionally, after treatment at the minimum inhibitory concentration (MIC 3.9 µg/ml), the transmission electron microscope showed alterations in the ultrastructure of the K. pneumoniae cells. Methanol extract from H. perforatum has a CC50 value of 976.75 µg/ml. Conclusion: Future inhibitors that target MDR strains may be revealed by these findings. Additionally, the extracts that were put to the test demonstrated strong antioxidant effects as shown by DPPH or ABTS radical-scavenging assays.

Identification of Multiple Iron Uptake Mechanisms in Enterococcus faecalis and Their Relationship to Virulence.

Among the unfavorable conditions bacteria encounter within the host is restricted access to essential trace metals such as iron. To overcome iron deficiency, bacteria deploy multiple strategies to scavenge iron from host tissues, with abundant examples of iron acquisition systems being implicated in bacterial pathogenesis. Yet the mechanisms utilized by the major nosocomial pathogen Enterococcus faecalis to maintain intracellular iron balance are poorly understood. In this study, we conducted a systematic investigation to identify and characterize the iron acquisition mechanisms of E. faecalis and to determine their contribution to virulence. Bioinformatic analysis and literature surveys revealed that E. faecalis possesses three conserved iron uptake systems. Through transcriptomics, we discovered two novel ABC-type transporters that mediate iron uptake. While inactivation of a single transporter had minimal impact on the ability of E. faecalis to maintain iron homeostasis, inactivation of all five systems (Δ5Fe strain) disrupted intracellular iron homeostasis and considerably impaired cell growth under iron deficiency. Virulence of the Δ5Fe strain was generally impaired in different animal models but showed niche-specific variations in mouse models, leading us to suspect that heme can serve as an iron source to E. faecalis during mammalian infections. Indeed, heme supplementation restored growth of Δ5Fe under iron depletion and virulence in an invertebrate infection model. This study revealed that the collective contribution of five iron transporters promotes E. faecalis virulence and that the ability to acquire and utilize heme as an iron source is critical to the systemic dissemination of E. faecalis.

Antibacterial activity and physicochemical properties of a sealer containing copaiba oil.

This study aimed to evaluate the antimicrobial activity and physicochemical properties of a novel dual-cure endodontic sealer containing copaiba oil. The copaiba oil was obtained and characterized by gas chromatography (GC), and the minimum inhibitory concentration (MIC) was performed. The experimental sealers were formulated with copaiba oil concentrations of 0, 0.5, 1, and 2%, and the RealSeal™ (Sybron endo, Orange, USA) and AH Plus (Dentsply De Trey Gmbh, Konstanz, Germany) were used as the commercial references. The antimicrobial activity of the sealers was evaluated by the direct contact test for 1h and 24h. To evaluate the physicochemical properties of the sealers, the degree of conversion, setting time, film thickness, dimensional stability, and radiopacity tests were performed. The data were statistically analyzed by one-way ANOVA and Tukey's test (α = 0.05). Concerning the results, the sealers containing copaiba oil showed antimicrobial activity without harming the physicochemical properties.

Depleting Cationic Lipids Involved in Antimicrobial Resistance Drives Adaptive Lipid Remodeling in Enterococcus faecalis.

The bacterial cell membrane is an interface for cell envelope synthesis, protein secretion, virulence factor assembly, and a target for host cationic antimicrobial peptides (CAMPs). To resist CAMP killing, several Gram-positive pathogens encode the multiple peptide resistance factor (MprF) enzyme that covalently attaches cationic amino acids to anionic phospholipids in the cell membrane. While E. faecalis encodes two mprF paralogs, MprF2 plays a dominant role in conferring resistance to killing by the CAMP human ß-defensin 2 (hBD-2) in E. faecalis strain OG1RF. The goal of the current study is to understand the broader lipidomic and functional roles of E. faecalis mprF. We analyzed the lipid profiles of parental wild-type and mprF mutant strains and show that while ΔmprF2 and ΔmprF1 ΔmprF2 mutants completely lacked cationic lysyl-phosphatidylglycerol (L-PG), the ΔmprF1 mutant synthesized ~70% of L-PG compared to the parent. Unexpectedly, we also observed a significant reduction of PG in ΔmprF2 and ΔmprF1 ΔmprF2. In the mprF mutants, particularly ΔmprF1 ΔmprF2, the decrease in L-PG and phosphatidylglycerol (PG) is compensated by an increase in a phosphorus-containing lipid, glycerophospho-diglucosyl-diacylglycerol (GPDGDAG), and D-ala-GPDGDAG. These changes were accompanied by a downregulation of de novo fatty acid biosynthesis and an accumulation of long-chain acyl-acyl carrier proteins (long-chain acyl-ACPs), suggesting that the suppression of fatty acid biosynthesis was mediated by the transcriptional repressor FabT. Growth in chemically defined media lacking fatty acids revealed severe growth defects in the ΔmprF1 ΔmprF2 mutant strain, but not the single mutants, which was partially rescued through supplementation with palmitic and stearic acids. Changes in lipid homeostasis correlated with lower membrane fluidity, impaired protein secretion, and increased biofilm formation in both ΔmprF2 and ΔmprF1 ΔmprF2, compared to the wild type and ΔmprF1. Collectively, our findings reveal a previously unappreciated role for mprF in global lipid regulation and cellular physiology, which could facilitate the development of novel therapeutics targeting MprF. IMPORTANCE The cell membrane plays a pivotal role in protecting bacteria against external threats, such as antibiotics. Cationic phospholipids such as lysyl-phosphatidyglycerol (L-PG) resist the action of cationic antimicrobial peptides through electrostatic repulsion. Here we demonstrate that L-PG depletion has several unexpected consequences in Enterococcus faecalis, including a reduction of phosphatidylglycerol (PG), enrichment of a phosphorus-containing lipid, reduced fatty acid synthesis accompanied by an accumulation of long-chain acyl-acyl carrier proteins (long chain acyl-ACPs), lower membrane fluidity, and impaired secretion. These changes are not deleterious to the organism as long as exogenous fatty acids are available for uptake from the culture medium. Our findings suggest an adaptive mechanism involving compensatory changes across the entire lipidome upon removal of a single phospholipid modification. Such adaptations must be considered when devising antimicrobial strategies that target membrane lipids.

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

OBJECTIVE: 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. MATERIALS AND METHODS: E. faecalis at planktonic and biofilm stages was treated with the photosensitizer HP, followed by no irradiation or irradiation with a power of 2.5 W (ʎ = 980 nm). The cell viability, anti-biofilm, anti-metabolic potential, and temperature changes were evaluated. RESULTS: The combination of HP and 980 nm 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. CONCLUSIONS: It could be concluded that the HP could improve anti-biofilm efficacy as a photosensitizer in a-PDT.

Antimicrobial effect of grape seed extract as a potential intracanal medicament combined with Nd:YAG laser.

This study evaluated the antimicrobial efficacy of grape seed extract medicament combined with Nd:YAG laser, against Enterococcus faecalis, Staphylococcus aureus and Candida albicans biofilms. Root canals infected with 4-week-old biofilms were divided into five groups (n = 11): calcium hydroxide, 6.5% GSE, Nd:YAG laser (1064 nm, 1.5 w, 15 Hz and 100 mj) and 6.5% GSE followed by Nd:YAG laser and normal saline (control). Dentin chips were collected using Gates-Glidden and cultured to obtain colony-forming units. Statistical analysis was performed using Kruskal-Wallis and Mann-Whitney U test. GSE showed higher antibacterial activity against all species investigated compared to Ca(OH)2 . However, the lowest microbial count was obtained using a combination of GSE and Nd:YAG laser (p < 0.001). No significant difference in the susceptibility of tested pathogens to each of the protocols was observed (p > 0.05). Application of Nd:YAG laser following GSE medicament is efficient against endodontic biofilms; also, GSE can be considered as an alternative to Ca(OH)2 dressing.

Efficacy of disinfection procedures performed prior to regenerative endodontic therapy: An integrative review.

The aim of this integrative review was to identify whether the disinfection procedures performed prior to regenerative endodontic treatment were effective on biofilm removal from the root canals. The research was based on PubMed, Latin American and Caribbean Health Sciences Literature (Lilacs) and Scientific Electronic Library Online (SciELO) databases. Four articles were selected; one of the studies was in vivo and the others ex vivo. Different disinfection procedures were studied, characterised mainly by the use of intracanal medication, highlighting the double antibiotic paste, triple antibiotic paste and calcium hydroxide paste. Disinfection ability was evaluated against Enterococcus faecalis and multispecies biofilms by using the fluorescence technique and colony forming unit counting, for 7 to 21 days. Double antibiotic paste and triple antibiotic paste demonstrated excellent antibiofilm activity, unlike CH paste that showed limited disinfection, even when associated with different antimicrobial agents. Triple antibiotic paste was the most effective medication against biofilm.

Comparative metabolite profiling of four polyphenol rich Morus leaves extracts in relation to their antibiofilm activity against Enterococcus faecalis.

Enterococci are a common cause of urinary tract infections. The severity of enterococcal infections is associated with their ability to form biofilms. Morus leaves are known as a natural antibacterial, however, their antibiofilm activity against Enterococcus remains unveiled. This study aimed to evaluate the ability of four polyphenol-rich Morus leaves extracts (Morus nigra, M. rubra, M. macroura, and M. alba) to inhibit biofilm formed by enterococcal clinical isolates in relation to their metabolic profiling. Results revealed that 48% of the isolates formed strong biofilm, 28% formed moderate biofilm, 20% formed weak biofilm, and only 4% did not form a biofilm. The strong biofilm-forming isolates were E. faecalis, and hence were chosen for this study. The antibiofilm activity of the four polyphenol-rich Morus leaves extracts revealed that the M. nigra extract exhibited the highest percentage of biofilm inhibition followed by M. rubra then M. macroura and the least inhibition was detected in M. alba, and these results were in accordance with the phenolic and flavonoid contents of each extract. UPLC-ESI-MS/MS identified 61 polyphenolic compounds in the four extracts. Further, multivariate analysis confirmed clear segregation of M. nigra from the other species suggesting disparity in its metabolome, with accumulation of flavonoids, anthocyanidins, phenolic acids and coumarin derivatives. Quercetin and kaempferol glycosides were found to be positively and significantly correlated to the antibiofilm activity. In conclusion, M. nigra ethanolic extracts showed the highest phenolic content and antibiofilm activity and they could be developed as a complementary treatment for the development of antimicrobial agents.

Enterococcus faecalis thrives in dual-species biofilm models under iron-rich conditions.

Escherichia coli (E. coli) and Enterococcus faecalis (E. faecalis) are pathogenic strains that often coexist in intestinal flora of humans and are prone to cause biofilm-associated infections, such as gastrointestinal tract and urinary tract infections. Earlier studies have demonstrated that E. faecalis biofilm can metabolize ferrous ions in iron-rich environments and promote biofilm growth under in-vivo conditions. However, the influence of iron transporters on dual-species biofilm growth and the nature of molecular-level interactions between iron transporter proteins and Fe2+ remains unknown. Therefore, in this work, co-culture studies were performed and the study indicates that Fe2+ at concentrations of 50-150 µM promotes the colonization of E. coli, and Fe2+ concentrations of 50-200 µM promote the growth of E. faecalis and dual-species colonies. Atomic absorption spectroscopy results reveal that Fe2+ ion augmentation in bacterial cells was increased to 4 folds in the single-species model and 11 folds in the dual-species model under iron-supplemented conditions. Furthermore, Fe2+ augmentation increased the antibiotic resistance of E. faecalis in both single- and dual-species bacterial cultures. In addition, in-silico docking were performed to determine a three-dimensional (3D) structure of ferrous iron-transporter proteins FeoB of E. faecalis and its affinity to extracellular Fe2+. Our model suggests that the FeoB facilitates the Fe2+ uptake in E. faecalis cells in the absence of iron chelator, 2,2-bipyridyl.