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Sample records for aeruginosa enterococcus faecalis

  1. Chlorine disinfection of Pseudomonas aeruginosa, total coliforms, Escherichia coli and Enterococcus faecalis: revisiting reclaimed water regulations.

    PubMed

    Coronel-Olivares, Claudia; Reyes-Gómez, Lidia María; Hernández-Muñoz, Aurelio; Martínez-Falcón, Ana Paola; Vázquez-Rodríguez, Gabriela A; Iturbe, Ulises

    2011-01-01

    Pathogenic organisms can be transmitted orally through drinking water or through skin and mucosae by both direct and indirect contact, and their presence in water thus has a negative impact on public health. In wastewater treatment plants (WWTP), water is disinfected to inactivate pathogens. The quantification of several microbial indicators in aquatic systems is required to estimate the biological quality of such systems. So far, coliform bacteria have been used as traditional indicators world-wide. This study has assessed the resistance of total coliforms, Escherichia coli, Pseudomonas aeruginosa and Enterococcus faecalis to three dosages of sodium hypochlorite (NaClO) at two exposure times. The bacteria were isolated from secondary effluents of a WWTP located in Hidalgo, Mexico. The results show that the number of colony-forming units of all studied bacterial types decreased when both the NaClO concentration and exposure times increased. However, they were not eliminated. The inclusion of the species Pseudomonas aeruginosa in regulations for treated wastewater quality as a new indicator is highly recommended due to its importance as an opportunistic pathogen. The detection of this species along with the traditional organisms could be particulary significant for reclaimed water to be used with direct human contact.

  2. Quantitative detection of Staphylococcus aureus, Enterococcus faecalis and Pseudomonas aeruginosa in human oral epithelial cells from subjects with periodontitis and periodontal health.

    PubMed

    Colombo, Andrea V; Barbosa, Graziela M; Higashi, Daniela; di Micheli, Giorgio; Rodrigues, Paulo H; Simionato, Maria Regina L

    2013-10-01

    Epithelial cells in oral cavities can be considered reservoirs for a variety of bacterial species. A polymicrobial intracellular flora associated with periodontal disease has been demonstrated in buccal cells. Important aetiological agents of systemic and nosocomial infections have been detected in the microbiota of subgingival biofilm, especially in individuals with periodontal disease. However, non-oral pathogens internalized in oral epithelial cells and their relationship with periodontal status are poorly understood. The purpose of this study was to detect opportunistic species within buccal and gingival crevice epithelial cells collected from subjects with periodontitis or individuals with good periodontal health, and to associate their prevalence with periodontal clinical status. Quantitative detection of total bacteria and Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis in oral epithelial cells was determined by quantitative real-time PCR using universal and species-specific primer sets. Intracellular bacteria were visualized by confocal microscopy and fluorescence in situ hybridization. Overall, 33% of cell samples from patients with periodontitis contained at least one opportunistic species, compared with 15% of samples from healthy individuals. E. faecalis was the most prevalent species found in oral epithelial cells (detected in 20.6% of patients with periodontitis, P = 0.03 versus healthy individuals) and was detected only in cells from patients with periodontitis. Quantitative real-time PCR showed that high levels of P. aeruginosa and S. aureus were present in both the periodontitis and healthy groups. However, the proportion of these species was significantly higher in epithelial cells of subjects with periodontitis compared with healthy individuals (P = 0.016 for P. aeruginosa and P = 0.047 for S. aureus). Although E. faecalis and P. aeruginosa were detected in 57% and 50% of patients, respectively, with probing depth and

  3. Quantitative detection of Staphylococcus aureus, Enterococcus faecalis and Pseudomonas aeruginosa in human oral epithelial cells from subjects with periodontitis and periodontal health.

    PubMed

    Colombo, Andrea V; Barbosa, Graziela M; Higashi, Daniela; di Micheli, Giorgio; Rodrigues, Paulo H; Simionato, Maria Regina L

    2013-10-01

    Epithelial cells in oral cavities can be considered reservoirs for a variety of bacterial species. A polymicrobial intracellular flora associated with periodontal disease has been demonstrated in buccal cells. Important aetiological agents of systemic and nosocomial infections have been detected in the microbiota of subgingival biofilm, especially in individuals with periodontal disease. However, non-oral pathogens internalized in oral epithelial cells and their relationship with periodontal status are poorly understood. The purpose of this study was to detect opportunistic species within buccal and gingival crevice epithelial cells collected from subjects with periodontitis or individuals with good periodontal health, and to associate their prevalence with periodontal clinical status. Quantitative detection of total bacteria and Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis in oral epithelial cells was determined by quantitative real-time PCR using universal and species-specific primer sets. Intracellular bacteria were visualized by confocal microscopy and fluorescence in situ hybridization. Overall, 33% of cell samples from patients with periodontitis contained at least one opportunistic species, compared with 15% of samples from healthy individuals. E. faecalis was the most prevalent species found in oral epithelial cells (detected in 20.6% of patients with periodontitis, P = 0.03 versus healthy individuals) and was detected only in cells from patients with periodontitis. Quantitative real-time PCR showed that high levels of P. aeruginosa and S. aureus were present in both the periodontitis and healthy groups. However, the proportion of these species was significantly higher in epithelial cells of subjects with periodontitis compared with healthy individuals (P = 0.016 for P. aeruginosa and P = 0.047 for S. aureus). Although E. faecalis and P. aeruginosa were detected in 57% and 50% of patients, respectively, with probing depth and

  4. Genetic Diversity among Enterococcus faecalis

    PubMed Central

    McBride, Shonna M.; Fischetti, Vincent A.; LeBlanc, Donald J.; Moellering, Robert C.; Gilmore, Michael S.

    2007-01-01

    Enterococcus faecalis, a ubiquitous member of mammalian gastrointestinal flora, is a leading cause of nosocomial infections and a growing public health concern. The enterococci responsible for these infections are often resistant to multiple antibiotics and have become notorious for their ability to acquire and disseminate antibiotic resistances. In the current study, we examined genetic relationships among 106 strains of E. faecalis isolated over the past 100 years, including strains identified for their diversity and used historically for serotyping, strains that have been adapted for laboratory use, and isolates from previously described E. faecalis infection outbreaks. This collection also includes isolates first characterized as having novel plasmids, virulence traits, antibiotic resistances, and pathogenicity island (PAI) components. We evaluated variation in factors contributing to pathogenicity, including toxin production, antibiotic resistance, polymorphism in the capsule (cps) operon, pathogenicity island (PAI) gene content, and other accessory factors. This information was correlated with multi-locus sequence typing (MLST) data, which was used to define genetic lineages. Our findings show that virulence and antibiotic resistance traits can be found within many diverse lineages of E. faecalis. However, lineages have emerged that have caused infection outbreaks globally, in which several new antibiotic resistances have entered the species, and in which virulence traits have converged. Comparing genomic hybridization profiles, using a microarray, of strains identified by MLST as spanning the diversity of the species, allowed us to identify the core E. faecalis genome as consisting of an estimated 2057 unique genes. PMID:17611618

  5. Inactivation of Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli present in treated urban wastewater by coagulation-flocculation and photo-Fenton processes.

    PubMed

    Rodríguez-Chueca, J; Morales, M; Mosteo, R; Ormad, M P; Ovelleiro, J L

    2013-05-01

    The purpose of the current study is to evaluate the inactivation of three different kinds of bacteria usually present in municipal wastewater treatment effluents (Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli) using a coagulation-flocculation-decantation (CFD) process combined with photo-Fenton treatment at pH 5. Different concentrations of Fe(3+)-H2O2 (0.4/25, 5/25 and 15/25 mg L(-1)), and H2O2 (25 mg L(-1)) were evaluated for 210 minutes under artificial solar irradiation in a solar chamber ATLAS SUNTEST CPS+. The results were compared applying the CFD process before or after the disinfection treatment. The results of the bacteria inactivation show that the highest rate was observed using CFD-photo-Fenton treatment with 15 mg L(-1) of Fe(3+) and 25 mg L(-1) of H2O2, obtaining the total inactivation of Pseudomonas sp., a 5.64-log inactivation of Enterococcus sp. and a 4.61-log inactivation of E. coli. In addition, turbidity and suspended solids decreased more than 90% with the combined treatments. The treated wastewater samples could be reused in urban, agricultural, industrial, recreational and environmental uses according to current Spanish legislation (RD 1620/2007). PMID:23411627

  6. Inactivation of Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli present in treated urban wastewater by coagulation-flocculation and photo-Fenton processes.

    PubMed

    Rodríguez-Chueca, J; Morales, M; Mosteo, R; Ormad, M P; Ovelleiro, J L

    2013-05-01

    The purpose of the current study is to evaluate the inactivation of three different kinds of bacteria usually present in municipal wastewater treatment effluents (Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli) using a coagulation-flocculation-decantation (CFD) process combined with photo-Fenton treatment at pH 5. Different concentrations of Fe(3+)-H2O2 (0.4/25, 5/25 and 15/25 mg L(-1)), and H2O2 (25 mg L(-1)) were evaluated for 210 minutes under artificial solar irradiation in a solar chamber ATLAS SUNTEST CPS+. The results were compared applying the CFD process before or after the disinfection treatment. The results of the bacteria inactivation show that the highest rate was observed using CFD-photo-Fenton treatment with 15 mg L(-1) of Fe(3+) and 25 mg L(-1) of H2O2, obtaining the total inactivation of Pseudomonas sp., a 5.64-log inactivation of Enterococcus sp. and a 4.61-log inactivation of E. coli. In addition, turbidity and suspended solids decreased more than 90% with the combined treatments. The treated wastewater samples could be reused in urban, agricultural, industrial, recreational and environmental uses according to current Spanish legislation (RD 1620/2007).

  7. Targeting Enterococcus faecalis biofilms with phage therapy.

    PubMed

    Khalifa, Leron; Brosh, Yair; Gelman, Daniel; Coppenhagen-Glazer, Shunit; Beyth, Shaul; Poradosu-Cohen, Ronit; Que, Yok-Ai; Beyth, Nurit; Hazan, Ronen

    2015-04-01

    Phage therapy has been proven to be more effective, in some cases, than conventional antibiotics, especially regarding multidrug-resistant biofilm infections. The objective here was to isolate an anti-Enterococcus faecalis bacteriophage and to evaluate its efficacy against planktonic and biofilm cultures. E. faecalis is an important pathogen found in many infections, including endocarditis and persistent infections associated with root canal treatment failure. The difficulty in E. faecalis treatment has been attributed to the lack of anti-infective strategies to eradicate its biofilm and to the frequent emergence of multidrug-resistant strains. To this end, an anti-E. faecalis and E. faecium phage, termed EFDG1, was isolated from sewage effluents. The phage was visualized by electron microscopy. EFDG1 coding sequences and phylogeny were determined by whole genome sequencing (GenBank accession number KP339049), revealing it belongs to the Spounavirinae subfamily of the Myoviridae phages, which includes promising candidates for therapy against Gram-positive pathogens. This analysis also showed that the EFDG1 genome does not contain apparent harmful genes. EFDG1 antibacterial efficacy was evaluated in vitro against planktonic and biofilm cultures, showing effective lytic activity against various E. faecalis and E. faecium isolates, regardless of their antibiotic resistance profile. In addition, EFDG1 efficiently prevented ex vivo E. faecalis root canal infection. These findings suggest that phage therapy using EFDG1 might be efficacious to prevent E. faecalis infection after root canal treatment.

  8. Characterization of Enterococcus faecalis and Enterococcus faecium from wild flowers.

    PubMed

    Sánchez Valenzuela, Antonio; Benomar, Nabil; Abriouel, Hikmate; Pérez Pulido, Rubén; Martínez Cañamero, Magdalena; Gálvez, Antonio

    2012-05-01

    Wild flowers in the South of Spain were screened for Enterococcus faecalis and Enterococcus faecium. Enterococci were frequently associated with prickypear and fieldpoppy flowers. Forty-six isolates, from 8 different flower species, were identified as E. faecalis (28 isolates) or E. faecium (18 isolates) and clustered in well-defined groups by ERIC-PCR fingerprinting. A high incidence of antibiotic resistance was detected among the E. faecalis isolates, especially to quinupristin/dalfopristin (75%), rifampicin (68%) and ciprofloxacin (57%), and to a lesser extent to levofloxacin (35.7%), erythromycin (28.5%), tetracycline (3.5%), chloramphenicol (3.5%) and streptomycin (3.5%). Similar results were observed for E. faecium isolates, except for a higher incidence of resistance to tetracycline (17%) and lower to erythromycin (11%) or quinupristin/dalfopristin (22%). Vancomycin or teicoplanin resistances were not detected. Most isolates (especially E. faecalis) were proteolytic and carried the gelatinase gene gelE. Genes encoding other potential virulence factors (ace, efaA (fs), ccf and cpd) were frequently detected. Cytolysin genes were mainly detected in a few haemolytic E. faecium isolates, three of which also carried the collagen adhesin acm gene. Hyaluronidase gene (hyl ( Efm )) was detected in two isolates. Many isolates produced bacteriocins and carried genes for enterocins A, B, and L50 mainly. The similarities found between enterococci from wild flowers and those from animal and food sources raise new questions about the puzzling lifestyle of these commensals and opportunistic pathogens.

  9. Transcriptional response of Enterococcus faecalis to sunlight.

    PubMed

    Sassoubre, Lauren M; Ramsey, Matthew M; Gilmore, Michael S; Boehm, Alexandria B

    2014-01-01

    Microarrays were used to investigate the transcriptional response of Enterococcus faecalis to photostress. E. faecalis are Gram-positive bacteria used as indicators of water quality and have been shown to vary diurnally in response to sunlight. E. faecalis in filtered seawater microcosms were exposed to artificial sunlight for 12h and then placed in the dark for 12h. Transcript abundance was measured at 0, 2, 6, 12, and 24h in the sunlit microcosm and a dark control using microarrays. Culturable E. faecalis concentrations decreased 6-7 orders of magnitude within the first 6h of light exposure. After 12h in the dark, no evidence of dark-repair was observed. Expression data collected after 12h of sunlight exposure revealed a difference in transcript abundance in the light relative to dark microcosms for 35 unique ORFs, 33 ORFs showed increased transcript abundance and 2 ORFs showed reduced transcript abundance. A majority (51%) of the ORFs with increased transcript abundance in the sunlit relative to dark microcosms encoded hypothetical proteins; others were associated with protein synthesis, oxidative stress and DNA repair. Results suggest that E. faecalis exposed to sunlight actively transcribe RNA in response to photostress.

  10. Pheromone-inducible conjugation in Enterococcus faecalis

    PubMed Central

    Kozlowicz, Briana K.; Dworkin, Martin; Dunny, Gary M.

    2009-01-01

    Pheromone-inducible transfer of the plasmid pCF10 in Enterococcus faecalis is regulated using a complicated network of proteins and RNAs. The plasmid itself has been assembled from parts garnered from a variety of sources, and many aspects of the system resemble a biological kluge. Recently several new functions of various pCF10 gene products that participate in regulation of plasmid transfer have been identified. The results indicate that selective pressures controlling the evolution of the plasmid have produced a highly complex regulatory network with multiple biological functions that may serve well as a model for the evolution of biological complexity. PMID:16503196

  11. Antimicrobial Susceptibility Patterns of Enterococcus faecalis and Enterococcus faecium Isolated from Poultry Flocks in Germany.

    PubMed

    Maasjost, J; Mühldorfer, K; Cortez de Jäckel S; Hafez, H M

    2015-03-01

    Between 2010 and 2011, 145 Enterococcus isolates (Enterococcus faecalis, n = 127; Enterococcus faecium, n = 18) were collected during routine bacteriologic diagnostics from broilers, layers, and fattening turkeys in Germany showing various clinical signs. The susceptibility to 24 antimicrobial agents was investigated by broth microdilution test to determine minimum inhibitory concentrations (MICs). All E. faecalis isolates (n = 127) were susceptible to the beta-lactam antibiotics ampicillin, amoxicillin-clavulanic acid, and penicillin. Corresponding MIC with 50% inhibition (MIC50) and MIC with 90% inhibition (MIC90) values of these antimicrobial agents were at the lower end of the test range (≤ 4 μg/ml). In addition, no vancomycin-resistant enterococci (VRE) were found. High resistance rates were identified in both Enterococcus species for lincomycin (72%-99%) and tetracycline (67%-82%). Half or more than half of Enterococcus isolates were resistant to gentamicin (54%-72%) and the macrolide antibiotics erythromycin (44%-61%) and tylosin-tartate (44%-56%). Enterococcus faecalis isolated from fattening turkeys showed the highest prevalence of antimicrobial resistance compared to other poultry production systems. Eighty-nine out of 145 Enterococcus isolates were resistant to three or more antimicrobial classes. Again, turkeys stood out with 42 (8 1%) multiresistant isolates. The most-frequent resistance patterns of E. faecalis were gentamicin, lincomycin, and tetracycline in all poultry production systems. PMID:26292548

  12. Antimicrobial Susceptibility Patterns of Enterococcus faecalis and Enterococcus faecium Isolated from Poultry Flocks in Germany.

    PubMed

    Maasjost, J; Mühldorfer, K; Cortez de Jäckel S; Hafez, H M

    2015-03-01

    Between 2010 and 2011, 145 Enterococcus isolates (Enterococcus faecalis, n = 127; Enterococcus faecium, n = 18) were collected during routine bacteriologic diagnostics from broilers, layers, and fattening turkeys in Germany showing various clinical signs. The susceptibility to 24 antimicrobial agents was investigated by broth microdilution test to determine minimum inhibitory concentrations (MICs). All E. faecalis isolates (n = 127) were susceptible to the beta-lactam antibiotics ampicillin, amoxicillin-clavulanic acid, and penicillin. Corresponding MIC with 50% inhibition (MIC50) and MIC with 90% inhibition (MIC90) values of these antimicrobial agents were at the lower end of the test range (≤ 4 μg/ml). In addition, no vancomycin-resistant enterococci (VRE) were found. High resistance rates were identified in both Enterococcus species for lincomycin (72%-99%) and tetracycline (67%-82%). Half or more than half of Enterococcus isolates were resistant to gentamicin (54%-72%) and the macrolide antibiotics erythromycin (44%-61%) and tylosin-tartate (44%-56%). Enterococcus faecalis isolated from fattening turkeys showed the highest prevalence of antimicrobial resistance compared to other poultry production systems. Eighty-nine out of 145 Enterococcus isolates were resistant to three or more antimicrobial classes. Again, turkeys stood out with 42 (8 1%) multiresistant isolates. The most-frequent resistance patterns of E. faecalis were gentamicin, lincomycin, and tetracycline in all poultry production systems.

  13. An antimicrobial peptidoglycan hydrolase for treating Enterococcus faecalis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enterococcus faecalis is an intestinal bacteria species that can become an opportunistic pathogen in humans and farm animals with antibiotic resistant strains becoming increasingly common. In farm animals, strong antimicrobials, such as Vancomycin, should not be used due to the risk of propagation ...

  14. Enterococcus faecalis promotes osteoclastogenesis and semaphorin 4D expression.

    PubMed

    Wang, Shuai; Deng, Zuhui; Seneviratne, Chaminda J; Cheung, Gary S P; Jin, Lijian; Zhao, Baohong; Zhang, Chengfei

    2015-10-01

    Enterococcus faecalis is considered a major bacterial pathogen implicated in endodontic infections and contributes considerably to periapical periodontitis. This study aimed to investigate the potential mechanisms by which E. faecalis accounts for the bone destruction in periapical periodontitis in vitro. Osteoclast precursor RAW264.7 cells were treated with E. faecalis ATCC 29212 and a wild strain of E. faecalis derived clinically from an infected root canal. The results showed that, to some extent, E. faecalis induced the RAW264.7 cells to form tartrate-resistant acid phosphatase (TRAP)-positive multinucleated osteoclast-like cells. This pathogen markedly stimulated RAW264.7 cells to express semaphorin 4D (Sema4D), which inhibits bone formation. Once RAW264.7 cells were primed by low-dose receptor activator of nuclear factor-kappa B ligand (RANKL), E. faecalis could significantly increase the production of TRAP-positive multinucleated cells and up-regulate the expression of osteoclast-specific markers, including NFATc1, TRAP and cathepsin K. Both p38 and ERK1/2 MAPK signaling pathways were activated by E. faecalis in RANKL-primed RAW264.7 cells, and meanwhile the expression of Sema4D was highly increased. In conclusion, E. faecalis may greatly contribute to the bone resorption in periapical periodontitis by promoting RANKL-dependent osteoclastogenesis and expression of Sema4D through activation of p38 and ERK1/2 MAPK signaling pathways.

  15. Transcriptome analysis of Enterococcus faecalis in response to alkaline stress.

    PubMed

    Ran, Shujun; Liu, Bin; Jiang, Wei; Sun, Zhe; Liang, Jingping

    2015-01-01

    Enterococcus faecalis is the most commonly isolated species from endodontic failure root canals; its persistence in treated root canals has been attributed to its ability to resist high pH stress. The goal of this study was to characterize the E. faecalis transcriptome and to identify candidate genes for response and resistance to alkaline stress using Illumina HiSeq 2000 sequencing. We found that E. faecalis could survive and form biofilms in a pH 10 environment and that alkaline stress had a great impact on the transcription of many genes in the E. faecalis genome. The transcriptome sequencing results revealed that 613 genes were differentially expressed (DEGs) for E. faecalis grown in pH 10 medium; 211 genes were found to be differentially up-regulated and 402 genes differentially down-regulated. Many of the down-regulated genes found are involved in cell energy production and metabolism and carbohydrate and amino acid metabolism, and the up-regulated genes are mostly related to nucleotide transport and metabolism. The results presented here reveal that cultivation of E. faecalis in alkaline stress has a profound impact on its transcriptome. The observed regulation of genes and pathways revealed that E. faecalis reduced its carbohydrate and amino acid metabolism and increased nucleotide synthesis to adapt and grow in alkaline stress. A number of the regulated genes may be useful candidates for the development of new therapeutic approaches for the treatment of E. faecalis infections.

  16. Transcriptome analysis of Enterococcus faecalis in response to alkaline stress

    PubMed Central

    Ran, Shujun; Liu, Bin; Jiang, Wei; Sun, Zhe; Liang, Jingping

    2015-01-01

    Enterococcus faecalis is the most commonly isolated species from endodontic failure root canals; its persistence in treated root canals has been attributed to its ability to resist high pH stress. The goal of this study was to characterize the E. faecalis transcriptome and to identify candidate genes for response and resistance to alkaline stress using Illumina HiSeq 2000 sequencing. We found that E. faecalis could survive and form biofilms in a pH 10 environment and that alkaline stress had a great impact on the transcription of many genes in the E. faecalis genome. The transcriptome sequencing results revealed that 613 genes were differentially expressed (DEGs) for E. faecalis grown in pH 10 medium; 211 genes were found to be differentially up-regulated and 402 genes differentially down-regulated. Many of the down-regulated genes found are involved in cell energy production and metabolism and carbohydrate and amino acid metabolism, and the up-regulated genes are mostly related to nucleotide transport and metabolism. The results presented here reveal that cultivation of E. faecalis in alkaline stress has a profound impact on its transcriptome. The observed regulation of genes and pathways revealed that E. faecalis reduced its carbohydrate and amino acid metabolism and increased nucleotide synthesis to adapt and grow in alkaline stress. A number of the regulated genes may be useful candidates for the development of new therapeutic approaches for the treatment of E. faecalis infections. PMID:26300863

  17. Abundance of Enterococcus species, Enterococcus faecalis and Enterococcus faecium, essential indicators of fecal pollution, in river water.

    PubMed

    Suzuki, Yoshihiro; Kanda, Naoki; Furukawa, Takashi

    2012-01-01

    Enterococci such as Enterococcus faecalis and E. faecium are considered as the most suitable indicators of fecal pollution in an aquatic environment, and new methods for Enterococcus determination have been developed, namely, membrane filtration (MF) using membrane-Enterococcus indoxyl-β-D-glucoside agar (mEI) and defined substrate technology (DST) using Enterolert®. This study used PCR analysis to identify E. faecalis and E. faecium among Enterococcus strains in river water isolated using both mEI plates and Enterolert® trays. There was a significantly high correlation between MF and DST in terms of enterococcal counts for river water samples. The combined percentages of E. faecalis and E. faecium with respect to the total number of all strains obtained using mEI plates and Enterolert® trays were approximately 30 % and >30 %, respectively. Other than E. faecalis and E. faecium, a large number of Enterococcus species were unspecified in the actual urban river samples. A comparison of the predominance of E. faecalis and E. faecium found that the abundance of a species depended on the sampling river and date. E. faecium is a non-predominant species in intestinal and fecal Enterococci, and it was one of the main Enterococcus species detected in surface water.

  18. Comparison of risk factors and outcome in patients with Enterococcus faecalis vs Enterococcus faecium bacteraemia.

    PubMed

    Suppola, J P; Kuikka, A; Vaara, M; Valtonen, V V

    1998-01-01

    The purpose of our study was to determine retrospectively the risk factors for the acquisition of Enterococcus faecalis vs E. faecium bacteraemia, as well as the clinical outcomes of these patients. 62 patients with Enterococcus faecalis bacteraemia were compared to 31 patients with E. faecium bacteraemia. Haematologic malignancies, neutropenia, high-risk source and previous use of aminoglycosides, carbapenems, cephalosporins and clindamycin were significantly associated with E. faecium bacteraemia. Instead, urinary catheterization was found to be related to Enterococcus faecalis bacteraemia. The mortality rates within 7 d and 30 d were 13% and 27%, respectively, in patients with E. faecalis bacteraemia and 6% and 29%, respectively, in patients with E. faecium bacteraemia. There was no difference in mortality between E. faecalis and E. faecium bacteraemia, nor was there a difference in seriousness of disease at the time of bacteraemia. In the subgroups of patients with monomicrobial or clinically significant E. faecalis vs E. faecium bacteraemia, the mortality rates were similar to the results of all subjects. Our results do not support the theory that E. faecium would be a more virulent organism than E. faecalis.

  19. Probiotic potential of Enterococcus faecalis strains isolated from meconium

    PubMed Central

    Al Atya, Ahmed K.; Drider-Hadiouche, Karima; Ravallec, Rozenn; Silvain, Amadine; Vachee, Anne; Drider, Djamel

    2015-01-01

    107 bacterial isolates with Gram positive staining and negative catalase activity, presumably assumed as lactic acid bacteria, were isolated from samples of meconium of 6 donors at Roubaix hospital, in the north of France. All these bacterial isolates were identified by MALDI-TOF mass spectrometry as Enterococcus faecalis. However, only six isolates among which E. faecalis 14, E. faecalis 28, E. faecalis 90, E. faecalis 97, and E. faecalis 101 (obtained from donor 3), and E. faecalis 93 (obtained from donor 5) were active against some Gram-negative bacteria and Gram-positive bacteria , through production of lactic acid, and bacteriocin like inhibitory substances. The identification of these isolates was confirmed by 16rDNA sequencing and their genetic relatedness was established by REP-PCR and pulsed field gel electrophoresis methods. Importantly, the aforementioned antagonistic isolates were sensitive to various classes of antibiotics tested, exhibited high scores of coaggregation and hydrophobicity, and were not hemolytic. Taken together, these properties render these strains as potential candidates for probiotic applications. PMID:25883590

  20. Identification of Enterococcus faecalis antigens specifically expressed in vivo

    PubMed Central

    Shet, Uttom K.; Park, Sang-Won; Lim, Hyun-Pil; Yun, Kwi-Dug; Kang, Seong Soo; Kim, Se Eun

    2015-01-01

    Objectives Molecular mechanism of the pathogenicity of Enterococcus faecalis (E. faecalis), a suspected endodontic pathogen, has not yet been adequately elucidated due to limited information on its virulence factors. Here we report the identification of in vivo expressed antigens of E. faecalis by using a novel immunoscreening technique called change-mediated antigen technology (CMAT) and an experimental animal model of endodontic infection. Materials and Methods Among 4,500 E. coli recombinant clones screened, 19 positive clones reacted reproducibly with hyperimmune sera obtained from rabbits immunized with E. faecalis cells isolated from an experimental endodontic infection. DNA sequences from 16 of these in vivo-induced (IVI) genes were determined. Results Identified protein antigens of E. faecalis included enzymes involved in housekeeping functions, copper resistance protein, putative outer membrane proteins, and proteins of unknown function. Conclusions In vivo expressed antigens of E. faecalis could be identified by using a novel immune-screening technique CMAT and an experimental animal model of endodontic infection. Detailed analysis of these IVI genes will lead to a better understanding of the molecular mechanisms involved in the endodontic infection of E. faecalis. PMID:26587417

  1. Postneurosurgical Central Nervous System Infection Due to Enterococcus faecalis Successfully Treated With Intraventricular Vancomycin

    PubMed Central

    Patel, Trisha; Lewis, Mark E.; Niesley, Michelle L.; Chowdhury, Mashiul

    2016-01-01

    Abstract Infections from Enterococcus faecalis and Enterococcus faecium are uncommon in the post-neurosurgical intervention setting., [1, 2, 3, 4] Intraventricular antibiotics are recommended when standard intravenous therapy fails. [5] Here we present a case of post-neurosurgical ventriculitis, meningitis, and cerebritis in an oncology patient caused by refractory Enterococcus faecalis successfully treated with intraventricular vancomycin. PMID:27226704

  2. Mature biofilms of Enterococcus faecalis and Enterococcus faecium are highly resistant to antibiotics.

    PubMed

    Holmberg, Anna; Rasmussen, Magnus

    2016-01-01

    Enterococcus faecalis and Enterococcus faecium are important nosocomial pathogens that form biofilms on implanted materials. We compare the antibiotic sensitivity of bacteria in new (established during 24 hours) and mature (established during 120 hours) enterococcal biofilms. Mature biofilms contained more bacteria and were much more tolerant to antibiotics, including rifampicin-containing combinations, as judged by determination of minimal biofilm eradication concentrations and by time-kill experiments of bacteria in biofilms formed on beads of bone cement.

  3. Enterococcus faecium and Enterococcus faecalis in blood of newborns with suspected nosocomial infection.

    PubMed

    Furtado, Isabela; Xavier, Paula Cristhina Niz; Tavares, Luciana Venhofen Martinelli; Alves, Fabiana; Martins, Sarah Fonseca; Martins, Almir de Sousa; Palhares, Durval Batista

    2014-01-01

    Enterococci are Gram-positive cocci saprophyte of the human gastrointestinal tract, diners who act as opportunistic pathogens. They can cause infections in patients hospitalized for a long time or who have received multiple antibiotic therapy. Enterococcus faecalis and Enterococcus faecium are the most common species in human infections. To evaluate the possibility of rapid detection of these species and their occurrence in the blood of newborns with suspected nosocomial infection, blood samples were collected from 50 newborns with late infections, admitted to the Neonatal Care Unit of the University Hospital Federal de Mato Grosso do Sul (UFMS-HU), from September 2010 to January 2011. The samples were subjected to conventional PCR and real time PCR (qPCR) to search for Enterococcus faecium and Enterococcus faecalis, respectively. The PCR results were compared with respective blood cultures from 40 patients. No blood cultures were positive for Enterococci, however, eight blood samples were identified as genomic DNA of Enterococcus faecium by qPCR and 22 blood samples were detected as genomic DNA of Enterococcus faecalis by conventional PCR. These findings are important because of the clinical severity of the evaluated patients who were found positive by conventional PCR and not through routine microbiological methods.

  4. Endocarditis and biofilm-associated pili of Enterococcus faecalis

    PubMed Central

    Nallapareddy, Sreedhar R.; Singh, Kavindra V.; Sillanpää, Jouko; Garsin, Danielle A.; Höök, Magnus; Erlandsen, Stanley L.; Murray, Barbara E.

    2006-01-01

    Increasing multidrug resistance in Enterococcus faecalis, a nosocomial opportunist and common cause of bacterial endocarditis, emphasizes the need for alternative therapeutic approaches such as immunotherapy or immunoprophylaxis. In an earlier study, we demonstrated the presence of antibodies in E. faecalis endocarditis patient sera to recombinant forms of 9 E. faecalis cell wall–anchored proteins; of these, we have now characterized an in vivo–expressed locus of 3 genes and an associated sortase gene (encoding sortase C; SrtC). Here, using mutation analyses and complementation, we demonstrated that both the ebp (encoding endocarditis and biofilm-associated pili) operon and srtC are important for biofilm production of E. faecalis strain OG1RF. In addition, immunogold electron microscopy using antisera against EbpA–EbpC proteins as well as patient serum demonstrated that E. faecalis produces pleomorphic surface pili. Assembly of pili and their cell wall attachment appeared to occur via a mechanism of cross-linking of the Ebp proteins by the designated SrtC. Importantly, a nonpiliated, allelic replacement mutant was significantly attenuated in an endocarditis model. These biologically important surface pili, which are antigenic in humans during endocarditis and encoded by a ubiquitous E. faecalis operon, may be a useful immunotarget for studies aimed at prevention and/or treatment of this pathogen. PMID:17016560

  5. Neuroinfections due to Enterococcus faecalis in children.

    PubMed

    Benca, J; Ondrusova, A; Huttova, M; Rudinsky, B; Kisac, P; Bauer, F

    2007-06-01

    Enterococcal meningitis is a rare complication of neurosurgical procedure or high technology treatment of children and occurs mainly imunocompromised neonates with very low birth weight, severe prematurity and complicates sometime ventriculoperitoneal shunt insertion or perinatal trauma. E. faecalis caused 10 nosocomial meningitis and all strains were susceptible to vancomycin and chloramphenicol, and in our database 90% also to gentamicin and ampicillin. Mortality in our group of 10 children was 20% what is insignificantly higher than overall mortality in the whole cohort of meningitis within last 15 years in our database (15.1%). Early empiric therapy should include also ampicillin or vancomycin, if enterococcal etiology is suspected.

  6. [High level of aminoglycoside resistance among Enterococcus faecalis and Enterococcus faecium strains].

    PubMed

    Kozuszko, Sylwia; Białucha, Agata; Bogiel, Tomasz; Gospodarek, Eugenia

    2011-01-01

    Enterococcus sp. strains are believed as important reason of serious nosocomial infections currently. These infections are cured by using combination of beta-lactams and aminoglycosides for their treatment. Enterococcus sp. resistant to high-level doses of aminoglycosides, beta-lactams and vancomycin are responsible for therapeutic failure. The aim of our study was to evaluate the incidence of isolation and susceptibility to antibiotics of HLAR Enterococcus sp. strains isolated between 2007 and 2010 from the patients of University Hospital No. 1 of dr A. Jurasz Collegium Medicum of L. Rydygier in Bydgoszcz Nicolaus Copernicus University in Toruń. Amongst 6137 Enterococcus sp. strains 1124 (18,3%) presented HLAR phenotype; 53,1% of them was identified as E. faecalis and 46,9% as E. faecium. The highest percentage of all examined strains was isolated from the patients of different surgery clinics, Intensive Care Units, and Pediatrics, Hematology and Oncology Clinic. HLAR and HLSR phenotypes were noted in E. faecalis, for 45,7% and 27,5% strains, in E. faecium - 29,8% and 9,5%, respectively. HLGR phenotype was presented twice more often in E. faecium than E. faecalis. Highest percentages of E. faecium resistant to glycopeptides and rifampicin were observed when compared with E. faecalis. The highest percentages of strains intermediate, resistant to vancomycin and resistant to glycopeptides were noted for E. faecium strains with phenotypes HLAR, HLGR and HLSR.

  7. vanA in Enterococcus faecium, Enterococcus faecalis, and Enterococcus casseliflavus detected in French cattle.

    PubMed

    Haenni, Marisa; Saras, Estelle; Châtre, Pierre; Meunier, Danièle; Martin, Sylvie; Lepage, Gérard; Ménard, Marie-Françoise; Lebreton, Patricia; Rambaud, Thomas; Madec, Jean-Yves

    2009-11-01

    The goal of this study was to assess the presence of enterococci species presenting van-mediated glycopeptide resistance in French cattle. Fecal samples were collected from healthy and sick animals, and enterococci were screened for vancomycin resistance. Vancomycin resistance was principally encountered in Enterococcus gallinarum and Enterococcus casseliflavus strains. However, glycopeptide resistance was detected in three different species of enterococci (E. faecalis, E. faecium, and E. casseliflavus). Molecular characterization of the genetic support proved that they all presented the prototypic VanA element. Interestingly, the E. casseliflavus strain displayed a remarkable VanB phenotype/vanA-vanC genotype. Transferability, associated resistances, and factors of vanA cotransfer were sought. This study proved that acquired vanA genes can still be detected in food-producing animals more than a decade after the avoparcin ban. Indeed, calves, which are recurrently exposed to antibiotics in France, may allow the re-emergence of glycopeptide resistance through coselection factors, and this might potentially be concerning for human health. PMID:19694552

  8. Presence of virulence factors in Enterococcus faecalis and Enterococcus faecium susceptible and resistant to vancomycin.

    PubMed

    Comerlato, Carolina Baldisserotto; Resende, Mariah Costa Carvalho de; Caierão, Juliana; d'Azevedo, Pedro Alves

    2013-08-01

    Despite the increasing importance of Enterococcus as opportunistic pathogens, their virulence factors are still poorly understood. This study determines the frequency of virulence factors in clinical and commensal Enterococcus isolates from inpatients in Porto Alegre, Brazil. Fifty Enterococcus isolates were analysed and the presence of the gelE, asa1 and esp genes was determined. Gelatinase activity and biofilm formation were also tested. The clonal relationships among the isolates were evaluated using pulsed-field gel electrophoresis. The asa1, gelE and esp genes were identified in 38%, 60% and 76% of all isolates, respectively. The first two genes were more prevalent in Enterococcus faecalis than in Enterococcus faecium, as was biofilm formation, which was associated with gelE and asa1 genes, but not with the esp gene. The presence of gelE and the activity of gelatinase were not fully concordant. No relationship was observed among any virulence factors and specific subclones of E. faecalis or E. faecium resistant to vancomycin. In conclusion, E. faecalis and E. faecium isolates showed significantly different patterns of virulence determinants. Neither the source of isolation nor the clonal relationship or vancomycin resistance influenced their distribution.

  9. Wide distribution of virulence genes among Enterococcus faecium and Enterococcus faecalis clinical isolates.

    PubMed

    Soheili, Sara; Ghafourian, Sobhan; Sekawi, Zamberi; Neela, Vasanthakumari; Sadeghifard, Nourkhoda; Ramli, Ramliza; Hamat, Rukman Awang

    2014-01-01

    Enterococcus, a Gram-positive facultative anaerobic cocci belonging to the lactic acid bacteria of the phylum Firmicutes, is known to be able to resist a wide range of hostile conditions such as different pH levels, high concentration of NaCl (6.5%), and the extended temperatures between 5(°)C and 65(°)C. Despite being the third most common nosocomial pathogen, our understanding on its virulence factors is still poorly understood. The current study was aimed to determine the prevalence of different virulence genes in Enterococcus faecalis and Enterococcus faecium. For this purpose, 79 clinical isolates of Malaysian enterococci were evaluated for the presence of virulence genes. pilB, fms8, efaAfm, and sgrA genes are prevalent in all clinical isolates. In conclusion, the pathogenicity of E. faecalis and E. faecium could be associated with different virulence factors and these genes are widely distributed among the enterococcal species.

  10. Enterococcus faecalis as multidrug resistance strains in clinical isolates in Imam Reza Hospital in Kermanshah, Iran.

    PubMed

    Mohammadi, F; Ghafourian, S; Mohebi, R; Taherikalani, M; Pakzad, I; Valadbeigi, H; Hatami, V; Sadeghifard, N

    2015-01-01

    The current study aimed to investigate the prevalence of vancomycin-resistant Enterococcus in E. faecalis and E. faecium and antimicrobial susceptibility patterns, then dominant genes responsible for vancomycin resistance were determined. For this propose, 180 clinical isolates of Enterococcus were subjected for identification and antibiotic susceptibility assay. Then, the gene responsible vancomycin resistant strains were determined. The results demonstrated the E. faecalis as a dominant Enterococcus. Resistance to erythromycin was dominant and multidrug resistance strains observed in E. faecalis. vanA was responsible for vancomycin resistance. In conclusion, a high rate of resistance to antibiotics in Enterococcus is clearly problematic, and a novel strategy is needed to decrease resistance in Enterococcus.

  11. First Japanese case of infectious endocarditis due to Enterococcus faecalis small-colony variants.

    PubMed

    Ogihara, Shinji; Saito, Ryoichi; Sawabe, Etsuko; Hagihara, Michio; Tohda, Shuji

    2016-10-01

    A male patient was admitted to our hospital due to infectious endocarditis. He had been treated with levofloxacin for 6 weeks, sulbactam/cefoperazone for 4 weeks, and benzylpenicillin for 2 days prior to valve replacement surgery. Gram-positive cocci, with morphology consistent with γ-Streptococcus, were detected in blood cultures obtained at admission, as well as in vegetation obtained from the aortic valve. However, the strain could not be identified using biochemical methods. Sequencing of the 16S rRNA gene indicated that the culture was a small-colony variant of Enterococcus faecalis. This is the first case in Japan of infectious endocarditis due to E. faecalis small-colony variants. Small-colony variants are subpopulations of bacteria with slow growth, reduced sugar fermentation, and unstable phenotype. As a result, these strains tend to be misidentified. Further, small-colony variants are associated with recurrent and persistent infections such as prosthetic joint infection and infectious endocarditis. These strains are found in various bacterial species, including Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa, but rarely in Enterococcus species. The case highlights the need to be vigilant of E. faecalis small-colony variants, especially in patients who received long-term courses of antibiotics.

  12. First Japanese case of infectious endocarditis due to Enterococcus faecalis small-colony variants.

    PubMed

    Ogihara, Shinji; Saito, Ryoichi; Sawabe, Etsuko; Hagihara, Michio; Tohda, Shuji

    2016-10-01

    A male patient was admitted to our hospital due to infectious endocarditis. He had been treated with levofloxacin for 6 weeks, sulbactam/cefoperazone for 4 weeks, and benzylpenicillin for 2 days prior to valve replacement surgery. Gram-positive cocci, with morphology consistent with γ-Streptococcus, were detected in blood cultures obtained at admission, as well as in vegetation obtained from the aortic valve. However, the strain could not be identified using biochemical methods. Sequencing of the 16S rRNA gene indicated that the culture was a small-colony variant of Enterococcus faecalis. This is the first case in Japan of infectious endocarditis due to E. faecalis small-colony variants. Small-colony variants are subpopulations of bacteria with slow growth, reduced sugar fermentation, and unstable phenotype. As a result, these strains tend to be misidentified. Further, small-colony variants are associated with recurrent and persistent infections such as prosthetic joint infection and infectious endocarditis. These strains are found in various bacterial species, including Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa, but rarely in Enterococcus species. The case highlights the need to be vigilant of E. faecalis small-colony variants, especially in patients who received long-term courses of antibiotics. PMID:27094238

  13. Ecology of Enterococcus faecalis and niche adapted or non-niche-adapted Enterococcus faecium in continuous-flow anaerobic cultures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Objectives: To study the survivability of niche adapted Enterococcus faecium I.3rif (I.3rif) vs. non-niche adapted Enterococcus faecium (GRE47) in cultures that contain Enterococcus faecalis I.2. Methods: An anaerobic continuous-flow culture of chicken microflora (CCF) that models the chicken gastr...

  14. Antibiotic Resistance in Enterococcus faecalis Isolated from Hospitalized Patients

    PubMed Central

    Balaei Gajan, Esrafil; Shirmohammadi, Adileh; Aghazadeh, Mohammad; Alizadeh, Mohammad; Sighari Deljavan, Alireza; Ahmadpour, Farzin

    2013-01-01

    Background and aims Enterococci are Gram-positive cocci that often occur in pairs (diplococci) or short chains. Be-side developing high level of antibiotic resistance, these bacteria can cause wide range of disease in human, thus to help provide an effective treatment for infections caused by this genus, this study was conceived to provide information on Enterococcus faecalis Antibiotic resistance to widely used antibiotics in hospitalized patients. Materials and methods Disk diffusion agar and Broth dilution methods were used to perform Antibiogram test on isolated Enterococcus faecalis. Culture medium used for Disk diffusion agar test was Muller Hinton agar, and for Broth dilution methods, Muller Hinton broth culture medium was utilized. In disk diffusion agar method, different commercial antibiotics disks produced by Pharmaceutical companies were used. Microsoft Excel software was used to perform statistical analysis. Results Based on antibiograms of 105 cases, a high resistance to Synercid, Nalidixic acid, Oxacillin and Teofilin was de-tected whereas the lowest resistance observed in Nitrofurantoin, Vancomycin, Linezolid and Teicoplanin antibiotics. Conclusion According to the results, Teicoplanin, Vancomycin, Linezolid and Nitrofurantoin are recommended against E. faecalis species. PMID:23875089

  15. Molecular characterization of Rifr mutations in Enterococcus faecalis and Enterococcus faecium.

    PubMed

    Du, Xiaoxing; Hua, Xiaoting; Qu, Tingting; Jiang, Yan; Zhou, Zhihui; Yu, Yunsong

    2014-08-01

    Mutation rate is an important factor affecting the appearance and spread of acquired antibiotic resistance. The frequencies and types of enterococci mutations were determined in this study. The MICs of rifampicin in enterococci and their rifampicin-resistant mutants were determined by the Clinical and Laboratory Standards Institute (CLSI) agar dilution method. The Enterococcus faecalis isolates A15 and 18165 showed no significant differences in mutation frequencies or mutation rates. In Enterococcus faecium, the mutation frequency and mutation rate were both 6·4-fold lower than in E. faecalis. The spectrum of mutations characterized in E. faecium B42 differed significantly from that of E. faecalis. The types and rate of mutations indicated that E. faecalis had a higher potential to develop linezolid resistance. Rifampicin resistance was associated with mutations in the rpoB gene. Rifampicin MICs for the E. faecalis mutant were 2048 mg/l, but rifampicin MICs for E. faecium mutants ranged from 64 to 1024 mg/l.

  16. Phage therapy against Enterococcus faecalis in dental root canals.

    PubMed

    Khalifa, Leron; Shlezinger, Mor; Beyth, Shaul; Houri-Haddad, Yael; Coppenhagen-Glazer, Shunit; Beyth, Nurit; Hazan, Ronen

    2016-01-01

    Antibiotic resistance is an ever-growing problem faced by all major sectors of health care, including dentistry. Recurrent infections related to multidrug-resistant bacteria such as methicillin-resistant Staphylococcus aureus, carbapenem-resistant Enterobacteriaceae, and vancomycin-resistant enterococci (VRE) in hospitals are untreatable and question the effectiveness of notable drugs. Two major reasons for these recurrent infections are acquired antibiotic resistance genes and biofilm formation. None of the traditionally known effective techniques have been able to efficiently resolve these issues. Hence, development of a highly effective antibacterial practice has become inevitable. One example of a hard-to-eradicate pathogen in dentistry is Enterococcus faecalis, which is one of the most common threats observed in recurrent root canal treatment failures, of which the most problematic to treat are its biofilm-forming VRE strains. An effective response against such infections could be the use of bacteriophages (phages). Phage therapy was found to be highly effective against biofilm and multidrug-resistant bacteria and has other advantages like ease of isolation and possibilities for genetic manipulations. The potential of phage therapy in dentistry, in particular against E. faecalis biofilms in root canals, is almost unexplored. Here we review the efforts to develop phage therapy against biofilms. We also focus on the phages isolated against E. faecalis and discuss the possibility of using phages against E. faecalis biofilm in root canals. PMID:27640530

  17. Biohybrid polymer-antimicrobial peptide medium against Enterococcus faecalis.

    PubMed

    Eckhard, Lea H; Sol, Asaf; Abtew, Ester; Shai, Yechiel; Domb, Abraham J; Bachrach, Gilad; Beyth, Nurit

    2014-01-01

    Antimicrobial peptides (AMPs) are conserved evolutionary components of the innate immune system that are being tested as alternatives to antibiotics. Slow release of AMPs using biodegradable polymers can be advantageous in maintaining high peptide levels for topical treatment, especially in the oral environment in which dosage retention is challenged by drug dilution with saliva flow and by drug inactivation by salivary enzymatic activity. Enterococcus faecalis is a multidrug resistant nosocomial pathogen and a persistent pathogen in root canal infections. In this study, four ultra-short lipopeptides (C16-KGGK, C16-KLLK, C16-KAAK and C16-KKK) and an amphipathic α-helical antimicrobial peptide (Amp-1D) were tested against E. faecalis. The antibacterial effect was determined against planktonic bacteria and bacteria grown in biofilm. Of the five tested AMPs, C16-KGGK was the most effective. Next C16-KGGK was formulated with one of two polymers poly (lactic acid co castor oil) (DLLA) or ricinoleic acid-based poly (ester-anhydride) P(SA-RA). Peptide-synthetic polymer conjugates, also referred to as biohybrid mediums were tested for antibacterial activity against E. faecalis grown in suspension and in biofilms. The new formulations exhibited strong and improved anti-E. faecalis activity.

  18. Biohybrid Polymer-Antimicrobial Peptide Medium against Enterococcus faecalis

    PubMed Central

    Eckhard, Lea H.; Sol, Asaf; Abtew, Ester; Shai, Yechiel; Domb, Abraham J.

    2014-01-01

    Antimicrobial peptides (AMPs) are conserved evolutionary components of the innate immune system that are being tested as alternatives to antibiotics. Slow release of AMPs using biodegradable polymers can be advantageous in maintaining high peptide levels for topical treatment, especially in the oral environment in which dosage retention is challenged by drug dilution with saliva flow and by drug inactivation by salivary enzymatic activity. Enterococcus faecalis is a multidrug resistant nosocomial pathogen and a persistent pathogen in root canal infections. In this study, four ultra-short lipopeptides (C16-KGGK, C16-KLLK, C16-KAAK and C16-KKK) and an amphipathic α-helical antimicrobial peptide (Amp-1D) were tested against E. faecalis. The antibacterial effect was determined against planktonic bacteria and bacteria grown in biofilm. Of the five tested AMPs, C16-KGGK was the most effective. Next C16-KGGK was formulated with one of two polymers poly (lactic acid co castor oil) (DLLA) or ricinoleic acid-based poly (ester-anhydride) P(SA-RA). Peptide-synthetic polymer conjugates, also referred to as biohybrid mediums were tested for antibacterial activity against E. faecalis grown in suspension and in biofilms. The new formulations exhibited strong and improved anti- E. faecalis activity. PMID:25279943

  19. Phage therapy against Enterococcus faecalis in dental root canals

    PubMed Central

    Khalifa, Leron; Shlezinger, Mor; Beyth, Shaul; Houri-Haddad, Yael; Coppenhagen-Glazer, Shunit; Beyth, Nurit; Hazan, Ronen

    2016-01-01

    Antibiotic resistance is an ever-growing problem faced by all major sectors of health care, including dentistry. Recurrent infections related to multidrug-resistant bacteria such as methicillin-resistant Staphylococcus aureus, carbapenem-resistant Enterobacteriaceae, and vancomycin-resistant enterococci (VRE) in hospitals are untreatable and question the effectiveness of notable drugs. Two major reasons for these recurrent infections are acquired antibiotic resistance genes and biofilm formation. None of the traditionally known effective techniques have been able to efficiently resolve these issues. Hence, development of a highly effective antibacterial practice has become inevitable. One example of a hard-to-eradicate pathogen in dentistry is Enterococcus faecalis, which is one of the most common threats observed in recurrent root canal treatment failures, of which the most problematic to treat are its biofilm-forming VRE strains. An effective response against such infections could be the use of bacteriophages (phages). Phage therapy was found to be highly effective against biofilm and multidrug-resistant bacteria and has other advantages like ease of isolation and possibilities for genetic manipulations. The potential of phage therapy in dentistry, in particular against E. faecalis biofilms in root canals, is almost unexplored. Here we review the efforts to develop phage therapy against biofilms. We also focus on the phages isolated against E. faecalis and discuss the possibility of using phages against E. faecalis biofilm in root canals. PMID:27640530

  20. Candida albicans and Enterococcus faecalis in the gut

    PubMed Central

    Garsin, Danielle A; Lorenz, Michael C

    2013-01-01

    The fungus Candida albicans and the gram-positive bacterium Enterococcus faecalis are both normal residents of the human gut microbiome and cause opportunistic disseminated infections in immunocompromised individuals. Using a nematode infection model, we recently showed that co-infection resulted in less pathology and less mortality than infection with either species alone and this was partly explained by an interkingdom signaling event in which a bacterial-derived product inhibits hyphal morphogenesis of C. albicans. In this addendum we discuss these findings in the contest of other described bacterial-fungal interactions and recent data suggesting a potentially synergistic relationship between these two species in the mouse gut as well. We suggest that E. faecalis and C. albicans promote a mutually beneficial association with the host, in effect choosing a commensal lifestyle over a pathogenic one. PMID:23941906

  1. Antibiotic resistance and virulence traits in clinical and environmental Enterococcus faecalis and Enterococcus faecium isolates.

    PubMed

    Rathnayake, I U; Hargreaves, M; Huygens, F

    2012-07-01

    This study compared virulence and antibiotic resistance traits in clinical and environmental Enterococcus faecalis and Enterococcus faecium isolates. E. faecalis isolates harboured a broader spectrum of virulence determinants compared to E. faecium isolates. The virulence traits Cyl-A, Cyl-B, Cyl-M, gel-E, esp and acm were tested and environmental isolates predominantly harboured gel-E (80% of E. faecalis and 31.9% of E. faecium) whereas esp was more prevalent in clinical isolates (67.8% of E. faecalis and 70.4% of E. faecium). E. faecalis and E. faecium isolated from water had different antibiotic resistance patterns compared to those isolated from clinical samples. Linezolid resistance was not observed in any isolates tested and vancomycin resistance was observed only in clinical isolates. Resistance to other antibiotics (tetracycline, gentamicin, ciprofloxacin and ampicillin) was detected in both clinical and water isolates. Clinical isolates were more resistant to all the antibiotics tested compared to water isolates. Multi-drug resistance was more prevalent in clinical isolates (71.2% of E. faecalis and 70.3% of E. faecium) compared to water isolates (only 5.7% E. faecium). tet L and tet M genes were predominantly identified in tetracycline-resistant isolates. All water and clinical isolates resistant to ciprofloxacin and ampicillin contained mutations in the gyrA, parC and pbp5 genes. A significant correlation was found between the presence of virulence determinants and antibiotic resistance in all the isolates tested in this study (p<0.05). The presence of antibiotic resistant enterococci, together with associated virulence traits, in surface recreational water could be a public health risk.

  2. Antimicrobial effect of alexidine and chlorhexidine against Enterococcus faecalis infection

    PubMed Central

    Kim, Hyun-Shik; Woo Chang, Seok; Baek, Seung-Ho; Han, Seung Hyun; Lee, Yoon; Zhu, Qiang; Kum, Kee-Yeon

    2013-01-01

    A previous study demonstrated that alexidine has greater affinity for the major virulence factors of bacteria than chlorhexidine. The aim of this study was to compare the antimicrobial activity of 1% alexidine with that of 2% chlorhexidine using Enterococcus faecalis-infected dentin blocks. Sixty bovine dentin blocks were prepared and randomly divided into six groups of 10 each. E. faecalis was inoculated on 60 dentin blocks using the Luppens apparatus for 24 h and then the dentin blocks were soaked in 2% chlorhexidine or 1% alexidine solutions for 5 and 10 min, respectively. Sterile saline was used as a control. The antimicrobial efficacy was assessed by counting the number of bacteria adhering to the dentin surface and observing the degradation of bacterial shape or membrane rupture under a scanning electron microscope. Significantly fewer bacteria were observed in the 2% chlorhexidine- or 1% alexidine-soaked groups than in the control group (P<0.05). However, there was no significant difference in the number of bacteria adhering to the dentinal surface between the two experimental groups or between the two soaking time groups (P>0.05). Ruptured or antiseptic-attached bacteria were more frequently observed in the 10-min-soaked chlorhexidine and alexidine groups than in the 5-min-soaked chlorhexidine and alexidine groups. In conclusion, 10-min soaking with 1% alexidine or 2% chlorhexidine can be effective against E. faecalis infection. PMID:23492900

  3. In vitro effectiveness of Brazilian brown propolis against Enterococcus faecalis.

    PubMed

    Pimenta, Hévelin Couto; Violante, Ivana Maria Povoa; Musis, Carlo Ralph de; Borges, Álvaro Henrique; Aranha, Andreza Maria Fábio

    2015-01-01

    The aim of this study was to evaluate the in vitro antimicrobial activity of Brazilian brown propolis as an intracanal medication against Enterococcus faecalis. Thirty dentin discs prepared from intact freshly extracted bovine maxillary central incisors were infected with E. faecalis for 21 days. The specimens were distributed into six groups according to the medicament used as follows: G1- calcium hydroxide paste; G2- Carbowax 400 (control group); G3- 20% brown propolis paste; G4- 40% brown propolis paste; G5- 20% brown propolis paste + calcium hydroxide paste; and G6- 40% brown propolis paste + calcium hydroxide paste. The experimental pastes were placed into the canal lumen and left for 14 days. After each period, irrigation was performed with sterile saline to remove the medicament, and the canals were dried with sterile paper points. The dentin chips were removed from the canals with sequential sterile round burs at low speed and were immediately collected in separate test tubes containing BHI broth. The tubes were incubated at 37°C, and microbial growth was analyzed by spectrophotometry after 15 days. All the experimental medications significantly reduced the number of viable bacteria. The G4 and G5 pastes were more effective than the G1 paste, with 35.8%, 41%, and 21.3% antibacterial activity, respectively. Brazilian brown propolis shows antibacterial capacity against E. faecalis.

  4. Citrate Metabolism by Enterococcus faecalis FAIR-E 229

    PubMed Central

    Sarantinopoulos, Panagiotis; Kalantzopoulos, George; Tsakalidou, Effie

    2001-01-01

    Citrate metabolism by Enterococcus faecalis FAIR-E 229 was studied in various growth media containing citrate either in the presence of glucose or lactose or as the sole carbon source. In skim milk (130 mM lactose, 8 mM citrate), cometabolism of citrate and lactose was observed from the first stages of the growth phase. Lactose was stoichiometrically converted into lactate, while citrate was converted into acetate, formate, and ethanol. When de Man-Rogosa-Sharpe (MRS) broth containing lactose (28 mM) instead of glucose was used, E. faecalis FAIR-E 229 catabolized only the carbohydrate. Lactate was the major end product, and small amounts of ethanol were also detected. Increasing concentrations of citrate (10, 40, 70, and 100 mM) added to MRS broth enhanced both the maximum growth rate of E. faecalis FAIR-E 229 and glucose catabolism, although citrate itself was not catabolized. Glucose was converted stoichiometrically into lactate, while small amounts of ethanol were produced as well. Finally, when increasing initial concentrations of citrate (10, 40, 70, and 100 mM) were used as the sole carbon sources in MRS broth without glucose, the main end products were acetate and formate. Small amounts of lactate, ethanol, and acetoin were also detected. This work strongly supports the suggestion that enterococcal strains have the metabolic potential to metabolize citrate and therefore to actively contribute to the flavor development of fermented dairy products. PMID:11722896

  5. Antimicrobial resistance and virulence of Enterococcus faecalis isolated from retail food

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Although enterococci are considered opportunistic nosocomial pathogens, their contribution to food-borne illnesses via dissemination through retail food remains undefined. In this study, prevalence and association of antimicrobial resistance and virulence factors of 80 Enterococcus faecalis isolate...

  6. Draft Genome Sequence of an Enterococcus faecalis Strain Isolated from a Neonatal Blood Sepsis Patient.

    PubMed

    Kropp, K A; Lucid, A; Carroll, J; Belgrudov, V; Walsh, P; Kelly, B; Smith, C; Dickinson, P; O'Driscoll, A; Templeton, K; Ghazal, P; Sleator, R D

    2014-01-01

    Herein, we report the draft genome sequence of Enterococcus faecalis ED-NGS-1009, cultivated from a blood sample taken from a neonatal sepsis patient at the Royal Infirmary in Edinburgh, Scotland, United Kingdom. PMID:25212626

  7. Complete genome sequencing and comparative analysis of the linezolid-resistant Enterococcus faecalis strain DENG1.

    PubMed

    Yu, Zhijian; Chen, Zhong; Cheng, Hang; Zheng, Jinxin; Li, Duoyun; Deng, Xiangbin; Pan, Weiguang; Yang, Weizhi; Deng, Qiwen

    2014-07-01

    Genome level analysis of bacterial strains provides information on genetic composition and resistance mechanisms to clinically relevant antibiotics. To date, whole genome characterization of linezolid-resistant Enterococcus faecalis isolated in the clinic is lacking. In this study, we report the entire genome sequence, genomic characteristics and virulence factors of a pathogenic E. faecalis strain, DENG1. Our results showed considerable differences in genomic characteristics and virulence factors compared with other E. faecalis strains (V583 and OG1RF). The genome of this LZD-resistant E. faecalis strain can be used as a reference to study the mechanism of LZD resistance and the phylogenetic relationship of E. faecalis strains worldwide.

  8. Purification and characterization of enterocin 4, a bacteriocin produced by Enterococcus faecalis INIA 4.

    PubMed Central

    Joosten, H M; Nunez, M; Devreese, B; Van Beeumen, J; Marugg, J D

    1996-01-01

    A simple two-step procedure was developed to obtain pure enterocin 4, a bacteriocin produced by Enterococcus faecalis INIA 4. Chemical and genetic characterization revealed that the primary structure of enterocin 4 is identical to that of peptide antibiotic AS-48 from Enterococcus faecalis S-48. In contrast to the reported inhibitory spectrum of AS-48, enterocin 4 displayed no activity against gram-negative bacteria. PMID:8900014

  9. The effect of berberine hydrochloride on Enterococcus faecalis biofilm formation and dispersion in vitro.

    PubMed

    Chen, Lihua; Bu, Qianqian; Xu, Huan; Liu, Yuan; She, Pengfei; Tan, Ruichen; Wu, Yong

    2016-01-01

    Enterococcus faecalis (E. faecalis) is one of the major causes of biofilm infections. Berberine hydrochloride (BBH) has diverse pharmacological effects; however, the effects and mechanisms of BBH on E. faecalis biofilm formation and dispersion have not been reported. In this study, 99 clinical isolates from the urine samples of patients with urinary tract infections (UTIs) were collected and identified. Ten strains of E. faecalis with biofilm formation ability were studied. BBH inhibited E. faecalis biofilm formation and promoted the biofilm dispersion of E. faecalis. In addition, sortase A and esp expression levels were elevated during early E. faecalis biofilm development, whereas BBH significantly reduced their expression levels. The results of this study indicated that BBH effectively prevents biofilm formation and promotes biofilm dispersion in E. faecalis, most likely by inhibiting the expressions of sortase A and esp. PMID:27242142

  10. REAL-TIME PCR METHOD TO DETECT ENTEROCOCCUS FAECALIS IN WATER

    EPA Science Inventory

    A 16S rDNA real-time PCR method was developed to detect Enterococcus faecalis in water samples. The dynamic range for cell detection spanned five logs and the detection limit was determined to be 6 cfu/reaction. The assay was capable of detecting E. faecalis cells added to biof...

  11. Mechanisms for Photoinactivation of Enterococcus faecalis in Seawater

    PubMed Central

    Sassoubre, Lauren M.; Nelson, Kara L.

    2012-01-01

    Field studies in fresh and marine waters consistently show diel fluctuations in concentrations of enterococci, indicators of water quality. We investigated sunlight inactivation of Enterococcus faecalis to gain insight into photoinactivation mechanisms and cellular responses to photostress. E. faecalis bacteria were exposed to natural sunlight in clear, filtered seawater under both oxic and anoxic conditions to test the relative importance of oxygen-mediated and non-oxygen-mediated photoinactivation mechanisms. Multiple methods were used to assess changes in bacterial concentration, including cultivation, quantitative PCR (qPCR), propidium monoazide (PMA)-qPCR, LIVE/DEAD staining using propidium iodide (PI), and cellular activity, including ATP concentrations and expression of the superoxide dismutase-encoding gene, sodA. Photoinactivation, based on numbers of cultivable cells, was faster in oxic than in anoxic microcosms exposed to sunlight, suggesting that oxygen-mediated photoinactivation dominated. There was little change in qPCR signal over the course of the experiment, demonstrating that the nucleic acid targets were not damaged to a significant extent. The PMA-qPCR signal was also fairly stable, consistent with the observation that the fraction of PI-permeable cells was constant. Thus, damage to the membrane was minimal. Microbial ATP concentrations decreased in all microcosms, particularly the sunlit oxic microcosms. The increase in relative expression of the sodA gene in the sunlit oxic microcosms suggests that cells were actively responding to oxidative stress. Dark repair was not observed. This research furthers our understanding of photoinactivation mechanisms and the conditions under which diel fluctuations in enterococci can be expected in natural and engineered systems. PMID:22941072

  12. Rapid Kill—Novel Endodontic Sealer and Enterococcus faecalis

    PubMed Central

    Zaltsman, Nathan; Houri-Haddad, Yael; Abramovitz, Itzhak; Davidi, Michael Perez; Weiss, Ervin I.

    2013-01-01

    With growing concern over bacterial resistance, the identification of new antimicrobial means is paramount. In the oral cavity microorganisms are essential to the development of periradicular diseases and are the major causative factors associated with endodontic treatment failure. As quaternary ammonium compounds have the ability to kill a wide array of bacteria through electrostatic interactions with multiple anionic targets on the bacterial surface, it is likely that they can overcome bacterial resistance. Melding these ideas, we investigated the potency of a novel endodontic sealer in limiting Enterococcus faecalis growth. We used a polyethyleneimine scaffold to synthesize nano-sized particles, optimized for incorporation into an epoxy-based endodontic sealer. The novel endodontic sealer was tested for its antimicrobial efficacy and evaluated for biocompatibility and physical eligibility. Our results show that the novel sealer foundation affixes the nanoparticles, achieving surface bactericidal properties, but at the same time impeding nanoparticle penetration into eukaryotic cells and thereby mitigating a possible toxic effect. Moreover, adequate physical properties are maintained. The nanosized quaternary amine particles interact within minutes with bacteria, triggering cell death across wide pH values. Throughout this study we demonstrate a new antibacterial perspective for endodontic sealers; a novel antibacterial, effective and safe antimicrobial means. PMID:24223159

  13. Covalent immobilization of Enterococcus faecalis Esawy dextransucrase and dextran synthesis.

    PubMed

    Hashem, Amal M; Gamal, Amira A; Hassan, Mohamed E; Hassanein, Naziha M; Esawy, Mona A

    2016-01-01

    Enterococcus faecalis Esawy dextransucrase was immobilized in Fe(3+)-cross-linked alginate/carboxymethyl cellulose (AC) beads. The gel beads were modified with polyethylenimine (PEI) followed by glutaraldehyde (GA) to form Fe(3+) (ACPG) beads. Fe(3+) (ACPG) was characterized using FTIR and DSC techniques. GA activated beads showed new two peaks. The first was at 1,717 cm(-1) which refers to (CO) group of a free aldehyde end of glutaraldehyde, and another peak was at 1,660 cm(-1) referring to (CN) group. The immobilization process improved the optimum temperature from 35 to 45°C. The immobilized enzyme showed its optimum activity in wide pH range (4.5-5.4) compared to pH 5.4 in case of free form. Also, the immobilization process improved the thermal and pH enzyme stability to great extent. Reusability test proved that the enzyme activity retained 60% after 15 batch reactions. Immobilized enzyme was applied successfully in the synthesis of oligosaccharides and different molecular weights of dextran.

  14. Mathematical models for Enterococcus faecalis recovery after microwave water disinfection.

    PubMed

    Benjamin, Earl; Reznik, Aron; Benjamin, Ellis; Pramanik, Saroj K; Sowers, Louise; Williams, Arthur L

    2009-12-01

    Microwave water disinfection is a rapid purification technique which can give billions of people access to clean drinking water. However, better understanding of bacterial recovery after microwave heating over time is necessary to determine parameters such as delayed bacterial growth rates and maximum bacterial yields. Mathematical models for Enterococcus faecalis recovery after microwave treatment in optimum growth conditions were developed for times up to 5 minutes using an optical absorbance method. Microwave times below 3 minutes (2,450 MHz, 130W) showed that bacterial recovery maintained a time-dependent sigmoidal form which included a maximum value. At microwave times greater than three minutes, bacterial recovery, with a time-dependent exponential form, significantly decreased and did not reach the maximum value within the interval of observance (0-8 hours). No bacterial growth was found after 6 minutes of microwave treatment. The prepared mathematical models were produced by transforming the given variables to the logistic or exponential functions. We found that time-dependent maximum growth rates and lag times could be approximated with second order polynomial functions. The determined models can be used as a template to illustrate bacterial survival during water purification using microwave irradiation, in both commercial and industrial processes.

  15. Biocide and antibiotic resistance of Enterococcus faecalis and Enterococcus faecium isolated from the swine meat chain.

    PubMed

    Rizzotti, Lucia; Rossi, Franca; Torriani, Sandra

    2016-12-01

    In this study nine strains of Enterococcus faecalis and 12 strains of Enterococcus faecium, isolated from different sample types in the swine meat chain and previously characterized for the presence of antibiotic resistance genes, were examined for phenotypic tolerance to seven biocides (chlorexidine, benzalkonium chloride, triclosan, sodium hypochlorite, 2-propanol, formaldehyde and hydrogen peroxide) and resistance to nine antibiotics (ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, erythromycin, clindamycin, tetracycline and chloramphenicol). Moreover, the presence of efflux system encoding genes qacA/B, qacC, qacE, qacEΔ1, emeA, and stress response genes, sigV and gsp65, involved in the tolerance to biocides, was analysed. Most strains were not tolerant to the biocides, but showed minimum inhibitory concentrations (MICs) higher than the recommended cut-off values for all the antibiotics tested, except for vancomycin and chloramphenicol. Only weak correlations, if any, were found between biocide and antibiotic resistance data. One E. faecalis strain was tolerant to triclosan and one E. faecium strain, with higher tolerance to chlorexidine than the other strains tested, was found to carry a qacA/B gene. Our results indicated that phenotypic resistance to antibiotics is very frequent in enterococcal isolates from the swine meat chain, but phenotypic tolerance to biocides is not common. On the other hand, the gene qacA/B was found for the first time in the species E. faecium, an indication of the necessity to adopt measures suitable to control the spread of biocide resistance determinants among enterococci. PMID:27554158

  16. Conjugal transfer of plasmid-borne bacteriocin production in Enterococcus faecalis 226 NWC.

    PubMed

    Salzano, G; Villani, F; Pepe, O; Sorrentino, E; Moschetti, G; Coppola, S

    1992-11-15

    Enterococcus faecalis 226 NWC, isolated from natural whey cultures utilized as starter in water-buffalo Mozzarella cheese manufacture, produces a bacteriocin, designated Enterocin 226 NWC, which is inhibitory to Listeria monocytogenes. Plasmid analysis of E. faecalis 226 NWC showed a single 5.2-kb plasmid, pEF226. In conjugation experiments, pEF226 was transferred into a plasmid-free strain of E. faecalis JH2-2. The transfer required direct cell-to-cell contact and was not inhibited by DNase. The identity of conjugation was confirmed by digestion with SmaI restriction endonuclease and subsequent pulsed-field gel electrophoresis (PFGE) of the genomic DNA of E. faecalis 226, E. faecalis JH2-2 and of the isolates after the mating. The data indicate that the ability of E. faecalis 226 NWC to produce the bacteriocin is linked to the 5.2-kb conjugative plasmid pEF226.

  17. Dissemination of Enterococcus faecalis and Enterococcus faecium in a ricotta processing plant and evaluation of pathogenic and antibiotic resistance profiles.

    PubMed

    Fernandes, Meg da Silva; Fujimoto, Graciela; de Souza, Leandro Pio; Kabuki, Dirce Yorika; da Silva, Márcio José; Kuaye, Arnaldo Yoshiteru

    2015-04-01

    In this work, the sources of contamination by Enterococcus spp. in a ricotta processing line were evaluated. The isolated strains were tested for virulence genes (gelE, cylA,B, M, esp, agg, ace, efaA, vanB), expression of virulence factors (hemolysin and gelatinase), and the resistance to 10 different antibiotics. Enterococcus faecium and Enterococcus faecalis were subjected to discriminatory identification by intergenic spacer region (ITS)-polymerase chain reaction and sequencing of the ITS region. The results showed that Enterococcus spp. was detected in the raw materials, environment samples and the final product. None of the 107 Enterococcus isolates were completely free from all virulence genes considered. A fraction of 21.5% of the isolates containing all of the genes of the cylA, B, M operon also expressed β-hemolysis. Most of the isolates showed the gelE gene, but only 9.3% were able to hydrolyze gelatin. In addition, 23.5% of the observed Enterococcus isolates had the vanB gene but were susceptible to vancomycin in vitro. The dissemination of antibiotic-resistant enterococci was revealed in this study: 19.3% of the E. faecium samples and 78.0% of the E. faecalis samples were resistant to at least one of the antibiotics tested. Sequencing of region discriminated 5 and 7 distinct groups among E. faecalis and E. faecium, respectively. Although some similarity was observed among some of the isolates, all E. faecalis and E. faecium isolates had genetic differences both in the ITS region and in the virulence profile, which makes them different from each other.

  18. Draft Genome Sequence of Enterococcus faecalis Strain F165 Isolated from a Urinary Tract Infection

    PubMed Central

    Pieta, Luiza; Prichula, Janira; Sambrano, Gustavo E.; Soares, Renata; Bello, Aline Dall; Frazzon, Jeverson; d’Azevedo, Pedro A.

    2016-01-01

    We report here a draft genome sequence of Enterococcus faecalis strain F165 isolated from a urine specimen in South Brazil. The genome size was 3,049,734 bp, with a G+C content of 37.38%, and genes related to antimicrobial resistance and adherence were found in the strain. These findings are consistent with pathogenesis of E. faecalis species. PMID:27795252

  19. Riboflavin-shuttled extracellular electron transfer from Enterococcus faecalis to electrodes in microbial fuel cells.

    PubMed

    Zhang, Enren; Cai, Yamin; Luo, Yue; Piao, Zhe

    2014-11-01

    Great attention has been focused on Gram-negative bacteria in the application of microbial fuel cells. In this study, the Gram-positive bacterium Enterococcus faecalis was employed in microbial fuel cells. Bacterial biofilms formed by E. faecalis ZER6 were investigated with respect to electricity production through the riboflavin-shuttled extracellular electron transfer. Trace riboflavin was shown to be essential for transferring electrons derived from the oxidation of glucose outside the peptidoglycan layer in the cell wall of E. faecalis biofilms formed on the surface of electrodes, in the absence of other potential electron mediators (e.g., yeast extract).

  20. Global transcriptional analysis of the stringent response in Enterococcus faecalis

    PubMed Central

    Gaca, Anthony O.; Abranches, Jacqueline; Kajfasz, Jessica K.

    2012-01-01

    In Enterococcus faecalis, production of guanosine tetraphosphate/guanosine pentaphosphate [(p)ppGpp], the effector molecule of the stringent response, is controlled by the bifunctional synthetase/hydrolase RelA and the monofunctional synthetase RelQ. Previously, the (p)ppGpp profiles of strains lacking relA, relQ or both genes indicated that RelA is the primary enzyme responsible for (p)ppGpp synthesis under stress conditions, while the contributions of RelQ to the stringent response and cell homeostasis remained elusive. Here, survival within the mouse-derived macrophage cell line J774A.1 and killing of Galleria mellonella supported initial evidence that virulence was attenuated in the (p)ppGpp0 ΔrelAΔrelQ strain but not in the ΔrelA or ΔrelQ strains. We performed, for the first time to our knowledge, global transcriptome analysis in a documented (p)ppGpp0 Gram-positive bacterium and provided the first insights into the role of a Gram-positive monofunctional (p)ppGpp synthetase in transcriptional regulation. Transcription profiling after mupirocin treatment confirmed that RelA is the major enzyme responsible for the (p)ppGpp-mediated transcriptional repression of genes associated with macromolecular biosynthesis, but also revealed that RelQ is required for full and timely stringent response induction. The delayed transcriptional response of ΔrelQ could not be correlated with reduced or slower production of (p)ppGpp, in part because RelA-dependent (p)ppGpp accumulation occurred very rapidly. Comparisons of the transcriptional responses of ΔrelA or ΔrelAΔrelQ strains with the parent strain under starvation conditions revealed upregulation of operons involved in energy metabolism in the (p)ppGpp0 strain. Thus, while ΔrelA and ΔrelAΔrelQ cannot use (p)ppGpp to sense and respond to stresses, fitness of ΔrelAΔrelQ may be further impaired due to an unbalanced metabolism. PMID:22653948

  1. Crystal structure of enterococcus faecalis sly A-like transcriptional factor.

    SciTech Connect

    Wu, R.; Zhang, R.; Zagnitko, O.; Dementieva, I.; Maltsev, N.; Watson, J. D.; Laskowski, R.; Gornicki, P.; Joachimiak, A.; Univ. of Chicago; European Bioinformatics Inst.

    2003-05-30

    The crystal structure of a SlyA transcriptional regulator at 1.6 {angstrom} resolution is presented, and structural relationships between members of the MarR/SlyA family are discussed. The SlyA family, which includes SlyA, Rap, Hor, and RovA proteins, is widely distributed in bacterial and archaeal genomes. Current evidence suggests that SlyA-like factors act as repressors, activators, and modulators of gene transcription. These proteins have been shown to up-regulate the expression of molecular chaperones, acid-resistance proteins, and cytolysin, and down-regulate several biosynthetic enzymes. The structure of SlyA from Enterococcus faecalis, determined as a part of an ongoing structural genomics initiative (www.mcsg.anl.gov), revealed the same winged helix DNA-binding motif that was recently found in the MarR repressor from Escherichia coli and the MexR repressor from Pseudomonas aeruginosa, a sequence homologue of MarR. Phylogenetic analysis of the MarR/SlyA family suggests that Sly is placed between the SlyA and MarR subfamilies and shows significant sequence similarity to members of both subfamilies.

  2. Fluorescent in situ hybridization with specific DNA probes offers adequate detection of Enterococcus faecalis and Enterococcus faecium in clinical samples.

    PubMed

    Waar, Karola; Degener, John E; van Luyn, Marja J; Harmsen, Hermie J M

    2005-10-01

    Enterococcus faecalis and Enterococcus faecium are among the leading causes of hospital-acquired infections. Reliable and quick identification of E. faecalis and E. faecium is important for accurate treatment and understanding their role in the pathogenesis of infections. Fluorescent in situ hybridization (FISH) of whole bacterial cells with oligonucleotides targeted at the 16S rRNA molecule leads to a reduced time to identification. In clinical practice, FISH therefore can be used in situations in which quick identification is necessary for optimal treatment of the patient. Furthermore, the abundance, spatial distribution and bacterial cell morphology can be observed in situ. This report describes the design of two fluorescent-labelled oligonucleotides that, respectively, detect the 16S rRNA of E. faecalis and the 16S rRNA of E. faecium, Enterococcus hirae, Enterococcus mundtii, Enterococcus villorum and Enterococcus saccharolyticus. Different protocols for the application of these oligonucleotides with FISH in different clinical samples such as faeces or blood cultures are given. Enterococci in a biofilm attached to a biomaterial were also visualized. Embedding of the biomaterial preserved the morphology and therefore the architecture of the biofilm could be observed. The usefulness of other studies describing FISH for detection of enterococci is generally hampered by the fact that they have only focused on one material and one protocol to detect the enterococci. However, the results of this study show that the probes can be used both in the routine laboratory to detect and determine the enterococcal species in different clinical samples and in a research setting to enumerate and detect the enterococci in their physical environment.

  3. Phenotypic and molecular antibiotic resistance profile of Enterococcus faecalis and Enterococcus faecium isolated from different traditional fermented foods.

    PubMed

    Sánchez Valenzuela, Antonio; Lavilla Lerma, Leyre; Benomar, Nabil; Gálvez, Antonio; Pérez Pulido, Rubén; Abriouel, Hikmate

    2013-02-01

    A collection of 55 enterococci (41 Enterococcus faecium and 14 E. faecalis strains) isolated from various traditional fermented foodstuffs of both animal and vegetable origins, and water was evaluated for resistance against 15 antibiotics. Lower incidence of resistance was observed with gentamicin, ampicillin, penicillin and teicoplanin. However, a high incidence of antibiotic resistance was detected for rifampicin (12 out of 14 of isolates), ciprofloxacin (9/14), and quinupristin/dalfopristin (8/14) in E. faecalis strains. Enterococcus faecium isolates were resistant to rifampicin (25/41), ciprofloxacin (23/41), erythromycin (18/41), levofloxacin (16/41), and nitrofurantoin (15/41). One Enterococcus faecalis and two E. faecium strains were resistant to vancomycin (MIC>16 μg/mL). Among 55 isolates, 27 (19 E. faecium and eight E. faecalis) were resistant to at least three antibiotics. High level of multidrug resistance to clinically important antibiotics was detected in E. faecalis strains (57% of E. faecalis versus 46% of E. faecium), which showed resistance to six to seven antibiotics, especially those isolated from foods of animal origin. So, it is necessary to re-evaluate the use of therapeutic antibiotics in stock farms at both regional and international levels due to the high number of multiple resistant (MR) bacteria. Fifty-six MR E. faecalis and E. faecium strains selected from this and previous studies (Valenzuela et al., 2008, 2010) were screened by polymerase chain reaction for antibiotic resistance genes, revealing the presence of tet(L), tet(M), ermB, cat, efrA, efrB, mphA, or msrA/B genes. The ABC Multidrug Efflux Pump EfrAB was detected in 96% of E. faecalis strains and also in 13% of E. faecium strains; this is the first report describing EfrAB in this enterococcal species. The efflux pump-associated msrA/B gene was detected in 66.66% of E. faecium strains, but not in E. faecalis strains.

  4. Complete Genome Sequence of a New Enterococcus faecalis Bacteriophage, vB_EfaS_IME197

    PubMed Central

    Cheng, Shi; Xing, Shaozhen; Zhang, Xianglilan; Pei, Guangqian; An, Xiaoping; Mi, Zhiqiang; Huang, Yong

    2016-01-01

    We report here the whole-genome sequence of a new Enterococcus faecalis phage, vB_EfaS_IME197, which has a linear double-stranded DNA genome of 41,307 bp with 34% G+C content. We describe the main features of the genome of vB_EfaS_IME197. PMID:27634987

  5. Complete Genome Sequence of Enterococcus faecalis Strain W11 Isolated from an Algal Food Product

    PubMed Central

    Takizawa, Noboru

    2016-01-01

    Here, we report the complete genome sequence of Enterococcus faecalis strain W11 isolated from an algal food product in Japan. This study should facilitate the identification of a novel mechanism of glycerol metabolic control in lactic acid bacteria. PMID:27688337

  6. Effects of ionophores on Enterococcus faecalis and E. faecium growth in pure and mixed ruminal culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enterococcus faecalis and faecium are Gram-positive human pathogens that can live in the gastrointestinal tract of food animals. Vancomycin-resistant enterococci (VRE) are an increasing threat to humans as a nosocomial infection, as well as a reservoir of antibiotic resistance genes. Ionophores ar...

  7. An indigenous gut bacterium, Enterococcus faecalis (Lactobacillales: Enterococcaceae), increases seed consumption by Harpalus pensylvanicus (Coleoptera: Carabidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Harpalus pensylvanicus is a beneficial beetle contributing to insect control and seed predation in North American cropland. The bacterial endosymbiont Enterococcus faecalis is found in the intestinal tract of H. pensylvanicus and is thought to contribute to the digestion of the insect's seed diet. W...

  8. Complete Genome Sequence of a New Enterococcus faecalis Bacteriophage, vB_EfaS_IME197.

    PubMed

    Cheng, Shi; Xing, Shaozhen; Zhang, Xianglilan; Pei, Guangqian; An, Xiaoping; Mi, Zhiqiang; Huang, Yong; Tong, Yigang

    2016-01-01

    We report here the whole-genome sequence of a new Enterococcus faecalis phage, vB_EfaS_IME197, which has a linear double-stranded DNA genome of 41,307 bp with 34% G+C content. We describe the main features of the genome of vB_EfaS_IME197. PMID:27634987

  9. Complete Genome Sequence of Enterococcus faecalis Strain W11 Isolated from an Algal Food Product.

    PubMed

    Doi, Yuki; Takizawa, Noboru

    2016-01-01

    Here, we report the complete genome sequence of Enterococcus faecalis strain W11 isolated from an algal food product in Japan. This study should facilitate the identification of a novel mechanism of glycerol metabolic control in lactic acid bacteria. PMID:27688337

  10. Effects Of Myrcia Ovata Cambess. Essential Oil On Planktonic Growth Of Gastrointestinal Microorganisms and Biofilm Formation Of Enterococcus Faecalis

    PubMed Central

    Cândido, Cinthya S.; Portella, Cadmo Silton A.; Laranjeira, Bruno J.; da Silva, Sérgio S.; Arriaga, Angela M.C.; Santiago, Gilvandete M. P.; Gomes, Geovany A.; Almeida, Paulo César; Carvalho, Cibele B. M.

    2010-01-01

    The essential oil from the leaves of Myrcia ovata Cambess., commonly used in Brazil for the treatment of gastric illnesses, was screened for antimicrobial activity and action in the formation of microbial biofilms by Enterococcus faecalis. The oil was obtained by hydrodistillation using a clevenger-type system. Its chemical composition was analyzed using GC and GC-MS. Both MIC and MBC of the essential oil were determined by broth microdilution techniques and agar dilution method. The essential oil showed antimicrobial activity against E. faecalis, Escherichia coli, Pseudomonas aeruginosa, Salmonella choleraesuis, Staphylococcus aureus, Streptococcus pneumoniae and Candida parapsilosis. The results showed that the essential oil of M. ovata Cambess. was effective against the formation of biofilm by E. faecalis when compared with the control. Four volatile compounds, representing 92.1 % of the oil, were identified and geranial was the major component (50.4 %). At the best of our knowledge, this is the first report of the chemical composition and antimicrobial activity of the essential oil from leaves of M. ovata. PMID:24031537

  11. Genetic variability of vancomycin-resistant Enterococcus faecium and Enterococcus faecalis isolates from humans, chickens, and pigs in Malaysia.

    PubMed

    Getachew, Yitbarek; Hassan, Latiffah; Zakaria, Zunita; Abdul Aziz, Saleha

    2013-08-01

    Vancomycin-resistant enterococci (VRE) have been reported to be present in humans, chickens, and pigs in Malaysia. In the present study, representative samples of VRE isolated from these populations were examined for similarities and differences by using the multilocus sequence typing (MLST) method. Housekeeping genes of Enterococcus faecium (n = 14) and Enterococcus faecalis (n = 11) isolates were sequenced and analyzed using the MLST databases eBURST and goeBURST. We found five sequence types (STs) of E. faecium and six STs of E. faecalis existing in Malaysia. Enterococcus faecium isolates belonging to ST203, ST17, ST55, ST79, and ST29 were identified, and E. faecium ST203 was the most common among humans. The MLST profiles of E. faecium from humans in this study were similar to the globally reported nosocomial-related strain lineage belonging to clonal complex 17 (CC17). Isolates from chickens and pigs have few similarities to those from humans, except for one isolate from a chicken, which was identified as ST203. E. faecalis isolates were more diverse and were identified as ST4, ST6, ST87, ST108, ST274, and ST244, which were grouped as specific to the three hosts. E. faecalis, belonging to the high-risk CC2 and CC87, were detected among isolates from humans. In conclusion, even though one isolate from a chicken was found clonal to that of humans, the MLST analysis of E. faecium and E. faecalis supports the findings of others who suggest VRE to be predominantly host specific and that clinically important strains are found mainly among humans. The infrequent detection of a human VRE clone in a chicken may in fact suggest a reverse transmission of VRE from humans to animals.

  12. Prevalence of Enterococcus faecalis in saliva and filled root canals of teeth associated with apical periodontitis

    PubMed Central

    Wang, Qian-Qian; Zhang, Cheng-Fei; Chu, Chun-Hung; Zhu, Xiao-Fei

    2012-01-01

    To investigate the prevalence of Enterococcus faecalis in saliva and filled root canals of patients requiring endodontic retreatment for apical periodontitis. Patients with apical periodontitis who were referred for endodontic retreatment were examined. The type and quality of the restoration, symptoms, quality of obturation were recorded. During retreatment, an oral rinse sample and root canal sample were cultured using brain-heart infusion agar and bile esculinazide agar to select for E. faecalis. The 16S rRNA technique was used to identify E. faecalis. A total of 32 women and 22 men (mean age: 38 years; s.d.: 11 years) and 58 teeth were studied. The prevalence of E. faecalis was 19% in the saliva and 38% in the root canals. The odds that root canals harbored E. faecalis were increased if the saliva habored this bacterium (odds ratio=9.7; 95% confidence interval=1.8–51.6; P<0.05). Teeth with unsatisfactory root obturation had more cultivable bacterial species in root canals than teeth with satisfactory root obturation (P<0.05). E. faecalis is more common in root canals of teeth with apical periodontitis than in saliva. The prevalence of E. faecalis in root canals is associated with the presence of E. faecalis in saliva. PMID:22422085

  13. Efficacy of Atmospheric Pressure Plasma as an Antibacterial Agent Against Enterococcus Faecalis in Vitro

    NASA Astrophysics Data System (ADS)

    Cao, Yingguang; Yang, Ping; Lu, Xinpei; Xiong, Zilan; Ye, Tao; Xiong, Qing; Sun, Ziyong

    2011-02-01

    Enterococcus faecalis (E. faecalis) is a microorganism that can survive extreme challenges in obturated root canals. The aim of this study was to evaluate the efficacy of a non-thermal atmospheric pressure plasma plume against E. faecalis in vitro. A non-thermal atmospheric pressure plasma jet device which could generate a cold plasma plume carrying a peak current of 300 mA was used. The antibacterial efficacy of this device against E. faecalis and its biofilm under different conditions was detected. The antibacterial efficacy of the plasma against E. faecalis and Staphylococcus aureus (S. aureus) was also evaluated. After plasma treatment, the average diameter of inhibition zone on S. aureus and E. faecalis was 2.62±0.26 cm and 1.06±0.30 cm, respectively (P < 0.05). The diameter was increased with prolongation of the treatment duration. The diameters of inhibition zone of the sealed Petri dishes were larger than those of the uncovered Petri dishes. There was significant difference in colony-forming units between plasma group and control group on E. faecalis biofilm (P < 0.01). The transmission electron microscopy revealed that the ultrastructural changes cytoderm of E. faecalis were observed after treatment for 2 min. It is concluded that the non-thermal atmospheric pressure plasma could serve as an effective adjunct to standard endodontic microbial treatment.

  14. Biological changes of Enterococcus faecalis in the viable but nonculturable state.

    PubMed

    E, J; Jiang, Y T; Yan, P F; Liang, J P

    2015-11-23

    Enterococcus faecalis may enter a viable but nonculturable (VBNC) state under adverse conditions. E. faecalis, the major bacterial species present in failed root canal treatments, is thought to survive after endodontic treatment by entering a VBNC state. In this study, we characterized the VBNC state of E. faecalis. We designed 3 different protocols to successfully induce the VBNC state. Approximately one-third of bacteria entered a VBNC state after 15-30 days, and all remained viable for at least 2 months. The morphology, glycometabolism, and adhesion capabilities of VBNC cells differed from those of E. faecalis during the exponential growth phase. Specifically, VBNC E. faecalis cells could not decompose lactose, D-mannitol, or D-sorbitol, although they were able to metabolize sucrose. Transmission electron microscopy showed that the morphology of the VBNC E. faecalis cells changed significantly; the cytoplasmic matrix was unevenly condensed and the overall morphology of the cells became irregular, but the cell membranes remained intact. Although the adhesion ability of the bacteria decreased, VBNC E. faecalis could still adhere to collagen fiber type I and tooth dentine. The persistence of this adhesion ability may be important in the virulence of VBNC E. faecalis.

  15. Nonclinical and clinical Enterococcus faecium strains, but not Enterococcus faecalis strains, have distinct structural and functional genomic features.

    PubMed

    Kim, Eun Bae; Marco, Maria L

    2014-01-01

    Certain strains of Enterococcus faecium and Enterococcus faecalis contribute beneficially to animal health and food production, while others are associated with nosocomial infections. To determine whether there are structural and functional genomic features that are distinct between nonclinical (NC) and clinical (CL) strains of those species, we analyzed the genomes of 31 E. faecium and 38 E. faecalis strains. Hierarchical clustering of 7,017 orthologs found in the E. faecium pangenome revealed that NC strains clustered into two clades and are distinct from CL strains. NC E. faecium genomes are significantly smaller than CL genomes, and this difference was partly explained by significantly fewer mobile genetic elements (ME), virulence factors (VF), and antibiotic resistance (AR) genes. E. faecium ortholog comparisons identified 68 and 153 genes that are enriched for NC and CL strains, respectively. Proximity analysis showed that CL-enriched loci, and not NC-enriched loci, are more frequently colocalized on the genome with ME. In CL genomes, AR genes are also colocalized with ME, and VF are more frequently associated with CL-enriched loci. Genes in 23 functional groups are also differentially enriched between NC and CL E. faecium genomes. In contrast, differences were not observed between NC and CL E. faecalis genomes despite their having larger genomes than E. faecium. Our findings show that unlike E. faecalis, NC and CL E. faecium strains are equipped with distinct structural and functional genomic features indicative of adaptation to different environments.

  16. Detection of vanC1 gene transcription in vancomycin-susceptible Enterococcus faecalis.

    PubMed

    Moura, Tiane Martin de; Cassenego, Ana Paula Vaz; Campos, Fabrício Souza; Ribeiro, Andrea Machado Leal; Franco, Ana Cláudia; d'Azevedo, Pedro Alves; Frazzon, Jeverson; Frazzon, Ana Paula Guedes

    2013-06-01

    Here we report the presence and expression levels of the vanC1 and vanC(2/3) genes in vancomycin-susceptible strains of Enterococcus faecalis. The vanC1 and vanC(2/3) genes were located in the plasmid DNA and on the chromosome, respectively. Specific mRNA of the vanC1 gene was detected in one of these strains. Additionally, analysis of the vanC gene sequences showed that these genes are related to the vanC genes of Enterococcus gallinarum and Enterococcus casseliflavus. The presence of vanC genes is useful for the identification of E. gallinarum and E. casseliflavus. Moreover, this is the first report of vanC mRNA in E. faecalis.

  17. Enterococcus Faecalis Biofilm. Formation and Development in Vitro Observed by Scanning Electron Microscopy.

    PubMed

    Bulacio, María de Los Á; Galván, Lucas R; Gaudioso, Cristina; Cangemi, Rosa; Erimbaue, Marta I

    2015-12-01

    Biofilm produced by Enterococcus faecalis isolated from root canals was detected by growing it on microplates and using 10% crystal violet stain, elution with alcohol and three procedures: no fixation, heat fixation and 10% formaldehyde fixation. The biofilm was evaluated using a Versamax Microplate Reader (USA). Twenty sterile root portions were incubated in TS broth with E. faecalis (108) for 48 hours, 4, 7, 14 and 30 days, after which they were processed and observed by scanning electron microscopy (SEM). Significantly more biofilm was found on the microplates for formaldehyde fixation than for heat fixation or no fixation (ANOVA p<0.0001). SEM showed E. faecalis growth at all times and biofilm development as from 14 days' incubation. Fixation with 10% formaldehyde was the most appropriate technique for detecting E. faecalis biofilm development on microplates. SEM confirmed biofilm formation after 14 days incubation.

  18. The surface rhamnopolysaccharide epa of Enterococcus faecalis is a key determinant of intestinal colonization.

    PubMed

    Rigottier-Gois, Lionel; Madec, Clément; Navickas, Albertas; Matos, Renata C; Akary-Lepage, Elodie; Mistou, Michel-Yves; Serror, Pascale

    2015-01-01

    Enterococcus faecalis is a commensal bacterium of the human intestine and a major opportunistic pathogen in immunocompromised and elderly patients. The pathogenesis of E. faecalis infection relies in part on its capacity to colonize the gut. Following disruption of intestinal homeostasis, E. faecalis can overgrow, cross the intestinal barrier, and enter the lymph and bloodstream. To identify and characterize E. faecalis genes that are key to intestinal colonization, our strategy consisted in screening mutants for the following phenotypes related to intestinal lifestyle: antibiotic resistance, overgrowth, and competition against microbiota. From the identified colonization genes, epaX encodes a glycosyltransferase located in a variable region of the enterococcal polysaccharide antigen (epa) locus. We demonstrated that EpaX acts on sugar composition, promoting resistance to bile salts and cell wall integrity. Given that EpaX is enriched in hospital-adapted isolates, this study points to the importance of the epa variability as a key determinant for enterococcal intestinal colonization.

  19. Increased Enterococcus faecalis infection is associated with clinically active Crohn disease.

    PubMed

    Zhou, Youlian; Chen, Huiting; He, Hanchang; Du, Yanlei; Hu, Jiaqi; Li, Yingfei; Li, Yuyuan; Zhou, Yongjian; Wang, Hong; Chen, Ye; Nie, Yuqiang

    2016-09-01

    This study was performed to investigate the relationship between the abundance of pathogenic gut microbes in Chinese patients with inflammatory bowel disease (IBD) and disease severity.We collected clinical data and fecal samples from 47 therapy-naive Chinese patients with ulcerative colitis (UC), 67 patients with Crohn disease (CD), and 48 healthy volunteers. Bacteria levels of Fusobacterium species (spp), enterotoxigenic Bacteroides fragilis (B fragilis), enteropathogenic Escherichia coli (E coli), and Enterococcus faecalis (E faecalis) were assessed by quantitative real-time PCR (qRT-PCR). Spearman correlation coefficients were calculated to test associations between bacterial content and clinical parameters.Compared to healthy controls, the levels of both Fusobacterium spp and E faecalis were significantly increased in the feces of patients with IBD (P < 0.01). B fragilis levels were higher (P < 0.05) and E faecalis levels lower (P < 0.05) in patients with CD compared to those with UC. Increased E faecalis colonization in CD associated positively with disease activity (P = 0.015), Crohn disease activity index (CDAI; R = 0.3118, P = 0.0108), and fecal calprotectin (P = 0.016).E faecalis and Fusobacterium spp are significantly enriched in patients with IBD, and increased E faecalis infection is associated with clinically active CD. PMID:27684872

  20. Synergistic Antibacterial Effect of the Combination of ε-Polylysine and Nisin against Enterococcus faecalis.

    PubMed

    Liu, Fang; Liu, Mei; Du, Lihui; Wang, Daoying; Geng, Zhiming; Zhang, Muhan; Sun, Chong; Xu, Xiaoxi; Zhu, Yongzhi; Xu, Weimin

    2015-12-01

    This study evaluated the antibacterial effect of the combination of ε-polylysine (ε-PL) and nisin against Enterococcus faecalis strains. The combination of ε-PL and nisin showed synergistic antibacterial activity against three Enterococcus strains. Scanning electron microscopy and a membrane permeability assay revealed that the combined treatment with ε-PL and nisin synergistically damaged the cell morphology of E. faecalis strain R612Z1 cells. Both ε-PL and nisin can dissipate the transmembrane electric potential of E. faecalis R612Z1 cells, but these peptides did not affect the transmembrane pH gradient. The combination of ε-PL and nisin can produce a high reactive oxygen species level in E. faecalis R612Z1 cells. The results indicated that the uptake of ε-PL into cells was promoted through nisin and that the combination of ε-PL and nisin could produce a high reactive oxygen species level in E. faecalis R612Z1 cells, leading to cell growth inhibition.

  1. Genetic relationships among Enterococcus faecalis isolates from different sources as revealed by multilocus sequence typing.

    PubMed

    Chen, X; Song, Y Q; Xu, H Y; Menghe, B L G; Zhang, H P; Sun, Z H

    2015-08-01

    Enterococcus faecalis is part of the natural gut flora of humans and other mammals; some isolates are also used in food production. So, it is important to evaluate the genetic diversity and phylogenetic relationships among E. faecalis isolates from different sources. Multilocus sequence typing protocol was used to compare 39 E. faecalis isolates from Chinese traditional food products (including dairy products, acidic gruel) and 4 published E. faecalis isolates from other sources including human-derived isolates employing 5 housekeeping genes (groEL, clpX, recA, rpoB, and pepC). A total of 23 unique sequence types were identified, which were grouped into 5 clonal complexes and 10 singletons. The value of standardized index of association of the alleles (IA(S)=0.1465) and network structure indicated a high frequency of intraspecies recombination across these isolates. Enterococcus faecalis lineages also exhibited clearly source-clustered distributions. The isolates from dairy source were clustered together. However, the relationship between isolates from acidic gruel and one isolate from a human source was close. The MLST scheme presented in this study provides a sharable and continuously growing sequence database enabling global comparison of strains from different sources, and will further advance our understanding of the microbial ecology of this important species.

  2. Identification of Polyketide Inhibitors Targeting 3-Dehydroquinate Dehydratase in the Shikimate Pathway of Enterococcus faecalis

    PubMed Central

    Hernandez-Valladares, Maria; Go, Maybelle Kho; Tung, Alvin; Aguda, Adeleke H.; Robinson, Robert C.; Yew, Wen Shan

    2014-01-01

    Due to the emergence of resistance toward current antibiotics, there is a pressing need to develop the next generation of antibiotics as therapeutics against infectious and opportunistic diseases of microbial origins. The shikimate pathway is exclusive to microbes, plants and fungi, and hence is an attractive and logical target for development of antimicrobial therapeutics. The Gram-positive commensal microbe, Enterococcus faecalis, is a major human pathogen associated with nosocomial infections and resistance to vancomycin, the “drug of last resort”. Here, we report the identification of several polyketide-based inhibitors against the E. faecalis shikimate pathway enzyme, 3-dehydroquinate dehydratase (DHQase). In particular, marein, a flavonoid polyketide, both inhibited DHQase and retarded the growth of Enterococcus faecalis. The purification, crystallization and structural resolution of recombinant DHQase from E. faecalis (at 2.2 Å resolution) are also reported. This study provides a route in the development of polyketide-based antimicrobial inhibitors targeting the shikimate pathway of the human pathogen E. faecalis. PMID:25072253

  3. High Frequency of Vancomycin-Resistant Enterococcus Faecalis in an Iranian Referral Children Medical Hospital

    PubMed Central

    POURAKBARI, Babak; AGHDAM, Mojtaba Kamali; MAHMOUDI, Shima; ASHTIANI, Mohammad Taghi Haghi; SABOUNI, Farah; MOVAHEDI, Zahra; ALYARI, Amir Esmael; SADEGHI, Reihane Hosseinpour; MAMISHI, Setareh

    2012-01-01

    ABSTRACT Background: Enterococci have emerged in recent years as important nosocomial pathogens. Although most enterococcal human infections are caused by Enterococcus faecalis, studies on vancomycin resistance are usually limited to Enterococcus faecium isolates and a little is known about E. faecalis. Therefore we undertook this study to obtain information about the prevalence of vancomycin -resistant E. faecalis (VREF) and genes responsible for resistance. Material and methods: Ninety-one E. faecalis isolates of different patients admitted at Children's Medical Center from August 2009 to June 2010 were included in this cross-sectional study. Antimicrobial testing was performed by Kirby-Bauer disk diffusion method according to Clinical Laboratories Standards Institute (CLSI). Results: Among all isolates, 15 (16%) were identified as VR E. faecalis. PCR analysis revealed that all VREF isolates were positive for the vanA gene. Conclusion: The present study reports the highest range of VREF in Iran. The increased frequency of VREF, as seen with rapid rise in the number of VanA isolates should be considered in infection control practices. PMID:23400108

  4. Genetic relationships among Enterococcus faecalis isolates from different sources as revealed by multilocus sequence typing.

    PubMed

    Chen, X; Song, Y Q; Xu, H Y; Menghe, B L G; Zhang, H P; Sun, Z H

    2015-08-01

    Enterococcus faecalis is part of the natural gut flora of humans and other mammals; some isolates are also used in food production. So, it is important to evaluate the genetic diversity and phylogenetic relationships among E. faecalis isolates from different sources. Multilocus sequence typing protocol was used to compare 39 E. faecalis isolates from Chinese traditional food products (including dairy products, acidic gruel) and 4 published E. faecalis isolates from other sources including human-derived isolates employing 5 housekeeping genes (groEL, clpX, recA, rpoB, and pepC). A total of 23 unique sequence types were identified, which were grouped into 5 clonal complexes and 10 singletons. The value of standardized index of association of the alleles (IA(S)=0.1465) and network structure indicated a high frequency of intraspecies recombination across these isolates. Enterococcus faecalis lineages also exhibited clearly source-clustered distributions. The isolates from dairy source were clustered together. However, the relationship between isolates from acidic gruel and one isolate from a human source was close. The MLST scheme presented in this study provides a sharable and continuously growing sequence database enabling global comparison of strains from different sources, and will further advance our understanding of the microbial ecology of this important species. PMID:26074239

  5. Synergistic Antibacterial Effect of the Combination of ε-Polylysine and Nisin against Enterococcus faecalis.

    PubMed

    Liu, Fang; Liu, Mei; Du, Lihui; Wang, Daoying; Geng, Zhiming; Zhang, Muhan; Sun, Chong; Xu, Xiaoxi; Zhu, Yongzhi; Xu, Weimin

    2015-12-01

    This study evaluated the antibacterial effect of the combination of ε-polylysine (ε-PL) and nisin against Enterococcus faecalis strains. The combination of ε-PL and nisin showed synergistic antibacterial activity against three Enterococcus strains. Scanning electron microscopy and a membrane permeability assay revealed that the combined treatment with ε-PL and nisin synergistically damaged the cell morphology of E. faecalis strain R612Z1 cells. Both ε-PL and nisin can dissipate the transmembrane electric potential of E. faecalis R612Z1 cells, but these peptides did not affect the transmembrane pH gradient. The combination of ε-PL and nisin can produce a high reactive oxygen species level in E. faecalis R612Z1 cells. The results indicated that the uptake of ε-PL into cells was promoted through nisin and that the combination of ε-PL and nisin could produce a high reactive oxygen species level in E. faecalis R612Z1 cells, leading to cell growth inhibition. PMID:26613915

  6. Role of house flies in the ecology of Enterococcus faecalis from wastewater treatment facilities.

    PubMed

    Doud, C W; Scott, H M; Zurek, L

    2014-02-01

    Enterococci are important nosocomial pathogens, with Enterococcus faecalis most commonly responsible for human infections. In this study, we used several measures to test the hypothesis that house flies, Musca domestica (L.), acquire and disseminate antibiotic-resistant and potentially virulent E. faecalis from wastewater treatment facilities (WWTF) to the surrounding urban environment. House flies and sludge from four WWTF (1-4) as well as house flies from three urban sites close to WWTF-1 were collected and cultured for enterococci. Enterococci were identified, quantified, screened for antibiotic resistance and virulence traits, and assessed for clonality. Of the 11 antibiotics tested, E. faecalis was most commonly resistant to tetracycline, doxycycline, streptomycin, gentamicin, and erythromycin, and these traits were intra-species horizontally transferrable by in vitro conjugation. Profiles of E. faecalis (prevalence, antibiotic resistance, and virulence traits) from each of WWTF sludge and associated house flies were similar, indicating that flies successfully acquired these bacteria from this substrate. The greatest number of E. faecalis with antibiotic resistance and virulence factors (i.e., gelatinase, cytolysin, enterococcus surface protein, and aggregation substance) originated from WWTF-1 that processed meat waste from a nearby commercial meat-processing plant, suggesting an agricultural rather than human clinical source of these isolates. E. faecalis from house flies collected from three sites 0.7-1.5 km away from WWTF-1 were also similar in their antibiotic resistance profiles; however, antibiotic resistance was significantly less frequent. Clonal diversity assessment using pulsed-field gel electrophoresis revealed the same clones of E. faecalis from sludge and house flies from WWTF-1 but not from the three urban sites close to WWTF-1. This study demonstrates that house flies acquire antibiotic-resistant enterococci from WWTF and potentially

  7. Role of house flies in the ecology of Enterococcus faecalis from wastewater treatment facilities.

    PubMed

    Doud, C W; Scott, H M; Zurek, L

    2014-02-01

    Enterococci are important nosocomial pathogens, with Enterococcus faecalis most commonly responsible for human infections. In this study, we used several measures to test the hypothesis that house flies, Musca domestica (L.), acquire and disseminate antibiotic-resistant and potentially virulent E. faecalis from wastewater treatment facilities (WWTF) to the surrounding urban environment. House flies and sludge from four WWTF (1-4) as well as house flies from three urban sites close to WWTF-1 were collected and cultured for enterococci. Enterococci were identified, quantified, screened for antibiotic resistance and virulence traits, and assessed for clonality. Of the 11 antibiotics tested, E. faecalis was most commonly resistant to tetracycline, doxycycline, streptomycin, gentamicin, and erythromycin, and these traits were intra-species horizontally transferrable by in vitro conjugation. Profiles of E. faecalis (prevalence, antibiotic resistance, and virulence traits) from each of WWTF sludge and associated house flies were similar, indicating that flies successfully acquired these bacteria from this substrate. The greatest number of E. faecalis with antibiotic resistance and virulence factors (i.e., gelatinase, cytolysin, enterococcus surface protein, and aggregation substance) originated from WWTF-1 that processed meat waste from a nearby commercial meat-processing plant, suggesting an agricultural rather than human clinical source of these isolates. E. faecalis from house flies collected from three sites 0.7-1.5 km away from WWTF-1 were also similar in their antibiotic resistance profiles; however, antibiotic resistance was significantly less frequent. Clonal diversity assessment using pulsed-field gel electrophoresis revealed the same clones of E. faecalis from sludge and house flies from WWTF-1 but not from the three urban sites close to WWTF-1. This study demonstrates that house flies acquire antibiotic-resistant enterococci from WWTF and potentially

  8. Dichotomous Metabolism of Enterococcus faecalis Induced by Hematin Starvation Modulates Colonic Gene Expression

    PubMed Central

    Allen, Toby D.; Moore, Danny R.; Wang, Xingmin; Casu, Viviana; May, Randal; Lerner, Megan R.; Houchen, Courtney; Brackett, Daniel J.; Huycke, Mark M.

    2009-01-01

    Summary Enterococcus faecalis is an intestinal commensal that cannot synthesize porphyrins and only expresses a functional respiratory chain when provided exogenous hematin. In the absence of hematin, E. faecalis reverts to fermentative metabolism and produces extracellular superoxide that can damage epithelial cell DNA. The acute response of the colonic mucosa to hematin-starved E. faecalis was identified by gene array. E. faecalis was inoculated into murine colons using a surgical ligation model that preserved tissue architecture and homeostasis. The mucosa was exposed to hematin-starved E. faecalis and compared to a control consisting of the same strain grown with hematin. At 1 hour post-inoculation six mucosal genes were differentially regulated and this increased to 42 genes at 6 hours. At 6 hours a highly significant biological interaction network was identified with functions that included NF-κB signaling, apoptosis, and cell cycle regulation. Colon biopsies showed no histological abnormalities by hematoxylin and eosin staining. Immunohistochemical staining, however, detected NF-κB activation in tissue macrophages using antibodies to the nuclear localization sequence for p65 and the F4/80 marker for murine macrophages. Similarly, hematin-starved E. faecalis strongly activated NF-κB in murine macrophages in vitro. Furthermore, primary and transformed colonic epithelial cells activated the G2/M checkpoint in vitro following exposure to hematin-starved E. faecalis. Modulation of this cell cycle checkpoint was due to extracellular superoxide produced as a result of the respiratory block in hematin-starved E. faecalis. These results demonstrate that the uniquely dichotomous metabolism of E. faecalis can significantly modulate gene expression in the colonic mucosa for pathways associated with inflammation, apoptosis, and cell cycle regulation. PMID:18809545

  9. Enterococcus faecalis Gelatinase Mediates Intestinal Permeability via Protease-Activated Receptor 2

    PubMed Central

    Maharshak, Nitsan; Huh, Eun Young; Paiboonrungruang, Chorlada; Shanahan, Michael; Thurlow, Lance; Herzog, Jeremy; Djukic, Zorka; Orlando, Roy; Pawlinski, Rafal; Ellermann, Melissa; Borst, Luke; Patel, Siten; Dotan, Iris; Sartor, Ryan B.

    2015-01-01

    Microbial protease-mediated disruption of the intestinal epithelium is a potential mechanism whereby a dysbiotic enteric microbiota can lead to disease. This mechanism was investigated using the colitogenic, protease-secreting enteric microbe Enterococcus faecalis. Caco-2 and T-84 epithelial cell monolayers and the mouse colonic epithelium were exposed to concentrated conditioned media (CCM) from E. faecalis V583 and E. faecalis lacking the gelatinase gene (gelE). The flux of fluorescein isothiocyanate (FITC)-labeled dextran across monolayers or the mouse epithelium following exposure to CCM from parental or mutant E. faecalis strains indicated paracellular permeability. A protease-activated receptor 2 (PAR2) antagonist and PAR2-deficient (PAR2−/−) mice were used to investigate the role of this receptor in E. faecalis-induced permeability. Gelatinase (GelE) purified from E. faecalis V583 was used to confirm the ability of this protease to induce epithelial cell permeability and activate PAR2. The protease-mediated permeability of colonic epithelia from wild-type (WT) and PAR2−/− mice by fecal supernatants from ulcerative colitis patients was assessed. Secreted E. faecalis proteins induced permeability in epithelial cell monolayers, which was reduced in the absence of gelE or by blocking PAR2 activity. Secreted E. faecalis proteins induced permeability in the colonic epithelia of WT mice that was absent in tissues from PAR2−/− mice. Purified GelE confirmed the ability of this protease to induce epithelial cell permeability via PAR2 activation. Fecal supernatants from ulcerative colitis patients induced permeability in the colonic epithelia of WT mice that was reduced in tissues from PAR2−/− mice. Our investigations demonstrate that GelE from E. faecalis can regulate enteric epithelial permeability via PAR2. PMID:25916983

  10. Enterococcus faecalis Gelatinase Mediates Intestinal Permeability via Protease-Activated Receptor 2.

    PubMed

    Maharshak, Nitsan; Huh, Eun Young; Paiboonrungruang, Chorlada; Shanahan, Michael; Thurlow, Lance; Herzog, Jeremy; Djukic, Zorka; Orlando, Roy; Pawlinski, Rafal; Ellermann, Melissa; Borst, Luke; Patel, Siten; Dotan, Iris; Sartor, Ryan B; Carroll, Ian M

    2015-07-01

    Microbial protease-mediated disruption of the intestinal epithelium is a potential mechanism whereby a dysbiotic enteric microbiota can lead to disease. This mechanism was investigated using the colitogenic, protease-secreting enteric microbe Enterococcus faecalis. Caco-2 and T-84 epithelial cell monolayers and the mouse colonic epithelium were exposed to concentrated conditioned media (CCM) from E. faecalis V583 and E. faecalis lacking the gelatinase gene (gelE). The flux of fluorescein isothiocyanate (FITC)-labeled dextran across monolayers or the mouse epithelium following exposure to CCM from parental or mutant E. faecalis strains indicated paracellular permeability. A protease-activated receptor 2 (PAR2) antagonist and PAR2-deficient (PAR2(-/-)) mice were used to investigate the role of this receptor in E. faecalis-induced permeability. Gelatinase (GelE) purified from E. faecalis V583 was used to confirm the ability of this protease to induce epithelial cell permeability and activate PAR2. The protease-mediated permeability of colonic epithelia from wild-type (WT) and PAR2(-/-) mice by fecal supernatants from ulcerative colitis patients was assessed. Secreted E. faecalis proteins induced permeability in epithelial cell monolayers, which was reduced in the absence of gelE or by blocking PAR2 activity. Secreted E. faecalis proteins induced permeability in the colonic epithelia of WT mice that was absent in tissues from PAR2(-/-) mice. Purified GelE confirmed the ability of this protease to induce epithelial cell permeability via PAR2 activation. Fecal supernatants from ulcerative colitis patients induced permeability in the colonic epithelia of WT mice that was reduced in tissues from PAR2(-/-) mice. Our investigations demonstrate that GelE from E. faecalis can regulate enteric epithelial permeability via PAR2. PMID:25916983

  11. Different extracts of Zingiber officinale decrease Enterococcus faecalis infection in Galleria mellonella.

    PubMed

    Maekawa, Lilian Eiko; Rossoni, Rodnei Dennis; Barbosa, Júnia Oliveira; Jorge, Antonio Olavo Cardoso; Junqueira, Juliana Campos; Valera, Marcia Carneiro

    2015-01-01

    Dried, fresh and glycolic extracts of Zingiber officinale were obtained to evaluate the action against G. mellonella survival assay against Enterococcus faecalis infection. Eighty larvae were divided into: 1) E. faecalis suspension (control); 2) E. faecalis + fresh extract of Z. officinale (FEO); 3) E. faecalis + dried extract of Z. officinale (DEO); 4) E. faecalis + glycolic extract of Z. officinale (GEO); 5) Phosphate buffered saline (PBS). For control group, a 5 μL inoculum of standardized suspension (107 cells/mL) of E. faecalis (ATCC 29212) was injected into the last left proleg of each larva. For the treatment groups, after E. faecalis inoculation, the extracts were also injected, but into the last right proleg. The larvae were stored at 37 °C and the number of dead larvae was recorded daily for 168 h (7 days) to analyze the survival curve. The larvae were considered dead when they did not show any movement after touching. E. faecalis infection led to the death of 85% of the larvae after 168 h. Notwithstanding, in treatment groups with association of extracts, there was an increase in the survival rates of 50% (GEO), 61% (FEO) and 66% (DEO) of the larvae. In all treatment groups, the larvae exhibited a survival increase with statistically significant difference in relation to control group (p=0.0029). There were no statistically significant differences among treatment groups with different extracts (p=0.3859). It may be concluded that the tested extracts showed antimicrobial activity against E. faecalis infection by increasing the survival of Galleria mellonella larvae.

  12. Enterococcus faecalis 6-Phosphogluconolactonase Is Required for Both Commensal and Pathogenic Interactions with Manduca sexta

    PubMed Central

    Holt, Jonathan F.; Frank, Kristi L.; Du, Jing; Guan, Changhui; Handelsman, Jo

    2014-01-01

    Enterococcus faecalis is a commensal and pathogen of humans and insects. In Manduca sexta, E. faecalis is an infrequent member of the commensal gut community, but its translocation to the hemocoel results in a commensal-to-pathogen switch. To investigate E. faecalis factors required for commensalism, we identified E. faecalis genes that are upregulated in the gut of M. sexta using recombinase-based in vivo expression technology (RIVET). The RIVET screen produced 113 clones, from which we identified 50 genes that are more highly expressed in the insect gut than in culture. The most frequently recovered gene was locus OG1RF_11582, which encodes a 6-phosphogluconolactonase that we designated pglA. A pglA deletion mutant was impaired in both pathogenesis and gut persistence in M. sexta and produced enhanced biofilms compared with the wild type in an in vitro polystyrene plate assay. Mutation of four other genes identified by RIVET did not affect persistence in caterpillar guts but led to impaired pathogenesis. This is the first identification of genetic determinants for E. faecalis commensal and pathogenic interactions with M. sexta. Bacterial factors identified in this model system may provide insight into colonization or persistence in other host-associated microbial communities and represent potential targets for interventions to prevent E. faecalis infections. PMID:25385794

  13. Detection of Enterococcus faecalis in Necrotic Teeth Root Canals by Culture and Polymerase Chain Reaction Methods

    PubMed Central

    Cogulu, Dilsah; Uzel, Atac; Oncag, Ozant; Aksoy, Semiha C.; Eronat, Cemal

    2007-01-01

    Objectives The aim of this study was to investigate the presence of Enterococcus faecalis in endodontic infections in both deciduous and permanent teeth by culture and polymerase chain reaction (PCR) methods. Methods A total of 145 children aged 5–13 years old were involved in this study. The presence of E. faecalis in necrotic deciduous and permanent teeth root canals was studied using culture and polymerase chain reaction methods. Results Among 145 molar teeth, 57% (n=83) presented necrotic asymptomatic pulp tissues and were included in this study. Culture and PCR methods detected the test species in 18 and 22 of 83 teeth involved, respectively. E. faecalis was cultured from 8 (18%) of 45 necrotic deciduous teeth and from 10 (26%) of 38 necrotic permanent teeth. PCR detection identified the target species in 10 (22%) and 12 (32%) of necrotic deciduous and permanent teeth respectively. Statistically significant difference in the presence of E. faecalis in deciduous and permanent teeth was found by culture and PCR methods (P=0.03 and 0.02, respectively). The difference in the presence of E. faecalis between two different methods was not statistically significant (P>.05). Conclusions The results of the present study confirm that both culture and PCR methods are sensitive to detect E. faecalis in root canals. PMID:19212470

  14. Efflux Pump Inhibitor Potentiates Antimicrobial Photodynamic Inactivation of Enterococcus faecalis Biofilm

    PubMed Central

    Kishen, Anil; Upadya, Megha; Tegos, George P.; Hamblin, Michael R.

    2010-01-01

    Microbial biofilm architecture contains numerous protective features including extracellular polymeric material that render biofilms impermeable to conventional antimicrobial agents. This study evaluated the efficacy of antimicrobial photodynamic inactivation (aPDI) of Enterococcus faecalis biofilms. The ability of a cationic, phenothiazinium photosensitizer, methylene blue (MB) and an anionic, xanthene photosensitizer, rose bengal (RB) to inactivate biofilms of E. faecalis (OGIRF and FA 2-2) and disrupt the biofilm structure was evaluated. Bacterial cells were tested as planktonic suspensions, intact biofilms and biofilm-derived suspensions obtained by the mechanical disruption of biofilms. The role of a specific microbial efflux pump inhibitor (EPI), verapamil hydrochloride in the MB-mediated aPDI of E. faecalis biofilms was also investigated. The results showed that E. faecalis biofilms exhibited significantly higher resistance to aPDI when compared to E. faecalis in suspension (P < 0.001). aPDI with cationic MB produced superior inactivation of E. faecalis strains in a biofilm along with significant destruction of biofilm structure when compared to anionic RB (P < 0.05). The ability to inactivate biofilm bacteria was further enhanced when the EPI was used with M B (P < 0.001). These experiments demonstrated the advantage of a cationic phenothiazinium photosensitizer combined with an EPI to inactivate biofilm bacteria and disrupt biofilm structure. PMID:20860692

  15. Draft Genome Sequences of the Probiotic Enterococcus faecalis Symbioflor 1 Clones DSM16430 and DSM16434

    PubMed Central

    Fritzenwanker, Moritz; Chakraborty, Anindita; Hain, Torsten; Zimmermann, Kurt

    2016-01-01

    The probiotic Symbioflor 1 is a historical concoction of 10 isolates of Enterococcus faecalis. Pulsed-field gel electrophoresis revealed two groups: one comprising eight identical clones (DSM16430, DSM16432, DSM16433, DSM16435 to DSM16439) and a further two isolates (DSM16431, DSM16434) with marginally different profiles. Here, we report a comparative analysis of the draft genome sequences of representative isolates. PMID:27688319

  16. A rare case of Enterococcus faecalis-induced orbital cellulitis and myositis

    PubMed Central

    Kohli, Piyush; Ichhpujani, Parul; Bansal, Rakesh Kumar; Kumar, Suresh

    2016-01-01

    Orbital cellulitis is an infection of soft tissue behind the orbital septum. Common pathogens isolated include Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus pneumoniae. It is a straightforward diagnosis and usually responds to empirical treatment without any sequela. We report a case of orbital cellulitis caused by Enterococcus faecalis, which was complicated by myositis of levator palpebrae superioris. To the best of our knowledge, only one case report exists dating way back to 1986. PMID:27688288

  17. Synergy characterization for Enterococcus faecalis strains displaying moderately high-level gentamicin and streptomycin resistance.

    PubMed Central

    Bantar, C E; Micucci, M; Fernandez Canigia, L; Smayevsky, J; Bianchini, H M

    1993-01-01

    Synergy of 14 Enterococcus faecalis strains displaying moderately high-level aminoglycoside resistance (MICs, 500 and 256 to 1,000 micrograms/ml for gentamicin and streptomycin, respectively) was characterized by time-kill studies. All strains proved resistant to penicillin plus the respective aminoglycoside. Strains with moderately high-level aminoglycoside resistance should be considered to exhibit high-level resistance in severe infections. PMID:8349776

  18. Synergy characterization for Enterococcus faecalis strains displaying moderately high-level gentamicin and streptomycin resistance.

    PubMed

    Bantar, C E; Micucci, M; Fernandez Canigia, L; Smayevsky, J; Bianchini, H M

    1993-07-01

    Synergy of 14 Enterococcus faecalis strains displaying moderately high-level aminoglycoside resistance (MICs, 500 and 256 to 1,000 micrograms/ml for gentamicin and streptomycin, respectively) was characterized by time-kill studies. All strains proved resistant to penicillin plus the respective aminoglycoside. Strains with moderately high-level aminoglycoside resistance should be considered to exhibit high-level resistance in severe infections.

  19. A rare case of Enterococcus faecalis-induced orbital cellulitis and myositis.

    PubMed

    Kohli, Piyush; Ichhpujani, Parul; Bansal, Rakesh Kumar; Kumar, Suresh

    2016-08-01

    Orbital cellulitis is an infection of soft tissue behind the orbital septum. Common pathogens isolated include Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus pneumoniae. It is a straightforward diagnosis and usually responds to empirical treatment without any sequela. We report a case of orbital cellulitis caused by Enterococcus faecalis, which was complicated by myositis of levator palpebrae superioris. To the best of our knowledge, only one case report exists dating way back to 1986. PMID:27688288

  20. Deactivation of Enterococcus Faecalis Bacteria by an Atmospheric Cold Plasma Brush

    NASA Astrophysics Data System (ADS)

    Chen, Wei; Huang, Jun; Du, Ning; Liu, Xiao-Di; Lv, Guo-Hua; Wang, Xing-Quan; Zhang, Guo-Ping; Guo, Li-Hong; Yang, Si-Ze

    2012-07-01

    An atmospheric cold plasma brush suitable for large area and low-temperature plasma-based sterilization is designed and used to treat enterococcus faecalis bacteria. The results show that the efficiency of the inactivation process by helium plasma is dependent on applied power and exposure time. After plasma treatments, the cell structure and morphology changes can be observed by scanning electron microscopy. Optical emission measurements indicate that reactive species such as O and OH play a significant role in the sterilization process.

  1. Complete genome sequence of Enterococcus faecalis LD33, a bacteriocin-producing strain.

    PubMed

    Yuehua, Jiao; Lanwei, Zhang; Fei, Liu; Huaxi, Yi; Xue, Han

    2016-06-10

    Enterococcus faecalis LD33 strain was originally isolated from traditional naturally fermented cream in Inner Mongolia of China. Its complete genome sequence was carried out using the Illumina Hiseq and the PacBio RSII platform. The genome only has a circular chromosome and a GC content of 37.58%. Other core information shown in the genome sequencing results further insight on this bacterium's genetic elements for bacteriocin production and the genes related to respiratory chain.

  2. Draft Genome Sequences of the Probiotic Enterococcus faecalis Symbioflor 1 Clones DSM16430 and DSM16434.

    PubMed

    Fritzenwanker, Moritz; Chakraborty, Anindita; Hain, Torsten; Zimmermann, Kurt; Domann, Eugen

    2016-01-01

    The probiotic Symbioflor 1 is a historical concoction of 10 isolates of Enterococcus faecalis Pulsed-field gel electrophoresis revealed two groups: one comprising eight identical clones (DSM16430, DSM16432, DSM16433, DSM16435 to DSM16439) and a further two isolates (DSM16431, DSM16434) with marginally different profiles. Here, we report a comparative analysis of the draft genome sequences of representative isolates. PMID:27688319

  3. Effect of NaCl Treatments on Tyramine Biosynthesis of Enterococcus faecalis.

    PubMed

    Liu, Fang; Wang, Xinxin; Du, Lihui; Wang, Daoying; Zhu, Yongzhi; Geng, Zhiming; Xu, Xiaoxi; Xu, Weimin

    2015-05-01

    The effect of NaCl stress (0 to 8%, wt/vol) on the growth and tyramine production in two Enterococcus faecalis strains was examined during culture time. The growth of E. faecalis was inhibited by the increase in NaCl concentration, but tyramine production was unaffected. Tyramine accumulated rapidly during the logarithmic phase of the strains, and the final tyramine levels were approximately 800 μg/ml. Relative gene expression of four genes in the tyrosine decarboxylase locus, namely, tyrRS, tyrDC, tyrP, and nhaC, was evaluated at different incubation times. The results showed that NaCl stress could upregulate the expression of tyrDC and tyrP to improve the tyramine production of a single E. faecalis strain under certain conditions, and TyrS could act as a negative regulator on the genetic regulation of the tyramine cluster.

  4. Bactericidal Effects of Diode Laser Irradiation on Enterococcus faecalis Using Periapical Lesion Defect Model

    PubMed Central

    Nagayoshi, Masato; Nishihara, Tatsuji; Nakashima, Keisuke; Iwaki, Shigetsugu; Chen, Ker-Kong; Terashita, Masamichi; Kitamura, Chiaki

    2011-01-01

    Objective. Photodynamic therapy has been expanded for use in endodontic treatment. The aim of this study was to investigate the antimicrobial effects of diode laser irradiation on endodontic pathogens in periapical lesions using an in vitro apical lesion model. Study Design. Enterococcus faecalis in 0.5% semisolid agar with a photosensitizer was injected into apical lesion area of in vitro apical lesion model. The direct effects of irradiation with a diode laser as well as heat produced by irradiation on the viability of microorganisms in the lesions were analyzed. Results. The viability of E. faecalis was significantly reduced by the combination of a photosensitizer and laser irradiation. The temperature caused by irradiation rose, however, there were no cytotoxic effects of heat on the viability of E. faecalis. Conclusion. Our results suggest that utilization of a diode laser in combination with a photosensitizer may be useful for clinical treatment of periapical lesions. PMID:21991489

  5. Ag-loaded mesoporous bioactive glasses against Enterococcus faecalis biofilm in root canal of human teeth.

    PubMed

    Fan, Wei; Wu, Daming; Ma, Tengjiao; Fan, Bing

    2015-01-01

    Ag-loaded mesoporous bioactive glass (Ag-MBG) powders were synthesized and characterized. The ions release of Ag-MBGs in Tris-HCl and the pH stability of simulated body fluids after immersing Ag-MBGs were tested. Root canals were inoculated with Enterococcus faecalis for 4 weeks, and the antibacterial activity of MBGs, Ag-MBGs and calcium hydroxide against E. faecalis biofilm were evaluated. Results showed that Ag-MBGs possessed highly ordered mesoporous structure with silver nanoparticles deposited in the mesopores, which enabled a sustained Ag ions released. The biofilms treated with Ag-MBGs showed a significant structural disruption compared with MBGs. These results indicated that Ag-MBGs possess a potent antibacterial effect against E.faecalis biofilm in root canal, and the antibacterial activity was induced by the release of Ag ions from Ag-MBGs.

  6. The antimicrobial effect of MTAD, sodium hypochlorite, doxycycline, and citric acid on Enterococcus faecalis.

    PubMed

    Krause, Trisha A; Liewehr, Frederick R; Hahn, Chin-Lo

    2007-01-01

    This study compared the antimicrobial effect of MTAD, two of its components, doxycycline and citric acid, and sodium hypochlorite (NaOCl) in two in vitro models on Enterococcus faecalis. In the bovine tooth model, the lumens of 30 bovine dentin discs were infected with E. faecalis for 2 weeks before treating with either one of the experimental irrigants or saline. Bacteria in the shavings were collected with two sizes of burs and enumerated after overnight culturing. Zones of inhibition were recorded in the agar diffusion model for each irrigant. In the tooth model, NaOCl and doxycycline were more effective than control in killing E. faecalis at the shallow bur depth, but at the deeper bur depth only NaOCl was superior. In the agar diffusion model, NaOCl produced less inhibition than MTAD or doxycycline.

  7. In vitro antibacterial effect of different irrigating solutions on Enterococcus faecalis.

    PubMed

    Bulacio, María de los Angeles; Cangemi, Rosa; Cecilia, Marta; Raiden, Guillermo

    2006-01-01

    Was evaluated the minimum inhibitory concentration (MIC) and the antibacterial effect (AE) of 2.5% NaOCl, 0.2% chlorhexidine gluconate (CHX) and 17% EDTA on Enterococcus faecalis. The antibacterial capacity was assessed by difusion in agar. The AE was evaluated on contaminated root dentin, employing apical and middle portions of human roots, sterilized and contaminated with Enterococcus faecalis, immersed in the irrigation solutions and incubated at 37 degrees C. Viable cells were counted at 0, 4, 8 and 24 hours. MIC: NaOCl and CHX: 0.2%, EDTA below 5%. Diffusion in agar: NaOCl 2.5% = 21 mm. CHX 0.2% = 14 mm. EDTA 17% = 20 mm. Effect on root dentin: NaOCl 2.5%: Enterococcus faecalis was totally inhibited for 24 hours in the apical area, and for 8 hours in the middle area. CHX 0.2% elicited a reduction of more than 5 log CFU and EDTA 17% induced a reduction of more than 3 log CFU at all the time points examined in the apical and middle areas.

  8. Prevalence and characterization of antibiotic resistant Enterococcus faecalis in French cheeses.

    PubMed

    Jamet, Emmanuel; Akary, Elodie; Poisson, Marie-Ange; Chamba, Jean-François; Bertrand, Xavier; Serror, Pascale

    2012-09-01

    Prevalence of enterococci and antibiotic resistance profiles of Enterococcus faecalis was analyzed in 126 French cheeses from retail stores. Forty-four percent of pasteurized or thermised-milk cheeses, and up to 92% of raw-milk cheeses contained detectable enterococci. A total of 337 antibiotic resistant enterococci were isolated in 29% and 60% of pasteurized-milk and raw-milk cheeses, respectively. E. faecalis was the predominant antibiotic resistant species recovered (81%), followed by Enterococcus faecium (13%), and Enterococcus durans (6%). The most prevalent antibiotic resistances were tetracycline (Tet) and minocycline (Min), followed by erythromycin (Ery), kanamycin (Kan) and chloramphenicol (Cm). The most common multiple antibiotic resistance phenotype was Cm Ery Kan Min Tet. The occurrence of antibiotic genes, as searched by PCR, was 100 % for aph3'IIIa, 96 % for ermB, 90 % for tetM and 80 % for catA in isolates resistant to Kan, Ery, Tet or Cm, respectively. MLST analysis of 30 multidrug resistant E. faecalis revealed that ST19, CC21, CC25 and CC55 isolates were the most common in cheeses. In conclusion, as in many other European countries, French cheeses do contain enterococci with multiple antibiotics resistances. However, low occurrence of high-level gentamicin resistant or sulfamethoxazole/trimethoprim-resistant enterococci and absence of vancomycin- or ampicillin- resistant enterococci indicate that cheeses cannot be considered as a major reservoir for nosocomial multi-drug resistant enterococci.

  9. EFFICACY OF SODIUM HYPOCHLORITE AND CHLORHEXIDINE AGAINST Enterococcus faecalis – A SYSTEMATIC REVIEW

    PubMed Central

    Estrela, Carlos; Silva, Julio Almeida; de Alencar, Ana Helena Gonçalves; Leles, Claudio Rodrigues; Decurcio, Daniel Almeida

    2008-01-01

    The efficacy of the sodium hypochlorite (NaOCl) and chlorhexidine (CHX) on Enterococcus faecalis was evaluated by systematic review and meta-analysis. The search strategies included search in electronic biomedical journal databases (MEDLINE, EMBASE, CENTRAL) and handsearching records, using different matches of keywords for NaOCl, CHX and Enterococcus faecalis. From 41 in vivo studies, 5 studies met the inclusion criteria. In a sample containing 159 teeth, E. faecalis was detected initially in 16 (10%) teeth by polymerase chain reaction (PCR) and 42 (26.4%) teeth by microbial culture techniques. After root canal disinfection, this species was observed in 11 (6.9%) teeth by PCR and 12 (7.5%) teeth by culture. Risk differences of included studies were combined as generic inverse variance data type (Review Manager Version 5.0 – Cochrane Collaboration, http://www.cc-ims.net, accessed 15 May 2008), taking into account the separate tracking of positive and negative cultures/PCR. The level of statistical significance was set at p<0.05. In conclusion, NaOCl or CHX showed low ability to eliminate E. faecalis when evaluated by either PCR or culture techniques. PMID:19082392

  10. Investigation of mechanism and molecular epidemiology of linezolid-resistant Enterococcus faecalis in China.

    PubMed

    Wang, Lipeng; He, Yunyan; Xia, Yun; Wang, Huijuan; Liang, Shumei

    2014-08-01

    Enterococcus is a major cause of important nosocomial infections. Linezolid, the first member of an entirely new class of antibiotics (oxazolidinones), is effective against serious infections caused by Enterococcus. However, resistance to linezolid has been discovered throughout the world rapidly. From 2011 to 2013, nine linezolid-resistant E. faecalis isolates were collected and the possible mechanisms of linezolid resistance, including mutations in domain V of 23S rRNA genes and in ribosomal proteins L3 and L4, and the multiresistance gene cfr, were investigated. Furthermore, an epidemiological survey of the nine linezolid-resistant E. faecalis isolates was performed by pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and DiversiLab. The three methods were compared to evaluate their merits and demerits, respectively. We failed to find the resistance mechanisms that have been revealed in recent years by PCR and sequencing analysis in the linezolid-resistant E. faecalis. Epidemiological investigation suggested that a small-scale outbreak of linezolid-resistant E. faecalis emerged in neurosurgery ICU from March to May of 2013. DiversiLab was a reliable typing tool and a suitable alternative to PFGE because it was as discriminatory as PFGE and better than MLST.

  11. Antibacterial Effect of All-in-one Self-etch Adhesives on Enterococcus faecalis

    PubMed Central

    Ebrahimi Chaharom, Mohammad Esmaeel; Ajami, Amir Ahmad; Abed Kahnamouei, Mehdi; Jafari Navimipour, Elmira; Tehranchi, Pardis; Zand, Vahid; Sadeghi, Mohammad Reza; Sohrabi, Aydin

    2014-01-01

    Background and aims. The aim of this study was to evaluate the antibacterial activity of one-step self-etch adhesives on Enterococcus faecalis on days 1, 7 and 14 with the use of modified direct contact test. Materials and methods. The modified directcontact test was used to evaluate the antibacterial effect of Adper Easy One, Bond Force, Clearfil S3 Bond, Futurabond M, G-Bond, iBond and OptiBond All-in-one adhesives on Enterococcus faecalis after aging the samples in phosphate-buffered saline for one, seven and fourteen days. Data were analyzed using one-way ANOVA and post hoc Tukey tests. Aging effect of each adhesive was evaluated by paired-sample test. In this study, P<0.05 was considered significant. Results. All the tested adhesives exhibited antibacterial activity after one day and had significant differences with the positive control group (P<0.05). After one week, OptiBond All-in-one, iBond and Futurabond M exhibited significant differences in bacterial growth from other groups (P<0.05). There were no significant differences between the groups in two weeks (P>0.05). Conclusion. iBond exhibited the highest antibacterial effect on E. faecalis after one week. Futurabond and OptiBond All-in-one exhibited antibacterial effects against E. faecalis for one week. PMID:25587384

  12. Antimicrobial activity of essential oils and chloroform alone and combinated with cetrimide against Enterococcus faecalis biofilm

    PubMed Central

    Ferrer Luque, Carmen Maria; González-Rodríguez, Maria Paloma; Arias-Moliz, Maria Teresa; Baca, Pilar

    2013-01-01

    Abstract The Enterococcus faecalis bacteria have been identified as the most commonly recovered species from teeth with persistent endodontic infections. The antimicrobial activity of essential oils and chloroform (CHL), alone and in association with various concentrations of cetrimide (CTR), against biofilm of Enterococcus faecalis was investigated. Solutions of CHL, eucalyptus oil (EO) and orange oil (OO) associated with CTR at 0.3%, 0.2%, 0.1%, and 0.05% were used to determine antimicrobial activity by exposing treated bovine dentine blocks to E. faecalis. Biofilms grown in the dentine blocks for 7 days were exposed to solutions for 2 and 5 min. Biofilm reduction between OO and EO at 2 min did not show any significant differences; however, OO had a higher kill percentage of biofilms than did the eucalyptus oil at 5 min (p < 0.01). Combinations with CTR at all concentrations achieved a 100% kill rate at 2 and 5 min. The association of CTR with solvent agents achieved the maximum antimicrobial activity against E. faecalis biofilms in dentine. PMID:24265917

  13. Bio-preservation of ground beef meat by Enterococcus faecalis CECT7121

    PubMed Central

    Sparo, M.D.; Confalonieri, A.; Urbizu, L.; Ceci, M.; Bruni, S.F. Sánchez

    2013-01-01

    Meat and particularly ground beef is frequently associated with Food Poisoning episodes and breeches in Food Safety. The main goal of this research was to evaluate the bactericide effect of the probiotic Enterococcus faecalis CECT7121, against different pathogens as: Escherichia coli O157:H7, Staphylococcus aureus, Clostridium perfringens and Listeria monocytogenes, inoculated in ground beef meat. Three studies were performed to evaluate the inhibition of E. faecalis CECT7121 on ground beef meat samples inoculated with pathogens: Study I: Samples (100 g meat) were inoculated with pathogens (103 CFU/g)) and E. faecalis CECT7121 (104 CFU/g) simultaneously. Study II: Samples were inoculated with E. faecalis CECT7121 24 h before the pathogens. Study III: E. faecalis CECT7121were inoculated 24 h after pathogens. The viable counts were performed at 0, 24, 48 and 72 h post-inoculation. The simultaneous inoculation of E. faecalis CECT7121 with E. coli O157:H7 strains resulted in the absence of viable counts of bacteria at 72 h post-treatment. However, when the probiotic was added 24 h before and 24 h after the pathogen E. coli O157:H7, viable cells were not detected at 24 h and 48 h post-treatment, respectively. Consistently, neither S. aureus nor Cl. perfringens viable bacteria were detected at 48 h in whole assays when inoculated with E. faecalis CECT7121. The same trend than described before was obtained after applying the 3 models assayed for L. monocytogenes. The current assays demonstrated the bactericide activity of E. faecalis CECT7121 strain on bacterial pathogens in ground beef meat. PMID:24159282

  14. Comparative Analysis of the Orphan CRISPR2 Locus in 242 Enterococcus faecalis Strains

    PubMed Central

    Hullahalli, Karthik; Rodrigues, Marinelle; Schmidt, Brendan D.; Li, Xiang; Bhardwaj, Pooja; Palmer, Kelli L.

    2015-01-01

    Clustered, Regularly Interspaced Short Palindromic Repeats and their associated Cas proteins (CRISPR-Cas) provide prokaryotes with a mechanism for defense against mobile genetic elements (MGEs). A CRISPR locus is a molecular memory of MGE encounters. It contains an array of short sequences, called spacers, that generally have sequence identity to MGEs. Three different CRISPR loci have been identified among strains of the opportunistic pathogen Enterococcus faecalis. CRISPR1 and CRISPR3 are associated with the cas genes necessary for blocking MGEs, but these loci are present in only a subset of E. faecalis strains. The orphan CRISPR2 lacks cas genes and is ubiquitous in E. faecalis, although its spacer content varies from strain to strain. Because CRISPR2 is a variable locus occurring in all E. faecalis, comparative analysis of CRISPR2 sequences may provide information about the clonality of E. faecalis strains. We examined CRISPR2 sequences from 228 E. faecalis genomes in relationship to subspecies phylogenetic lineages (sequence types; STs) determined by multilocus sequence typing (MLST), and to a genome phylogeny generated for a representative 71 genomes. We found that specific CRISPR2 sequences are associated with specific STs and with specific branches on the genome tree. To explore possible applications of CRISPR2 analysis, we evaluated 14 E. faecalis bloodstream isolates using CRISPR2 analysis and MLST. CRISPR2 analysis identified two groups of clonal strains among the 14 isolates, an assessment that was confirmed by MLST. CRISPR2 analysis was also used to accurately predict the ST of a subset of isolates. We conclude that CRISPR2 analysis, while not a replacement for MLST, is an inexpensive method to assess clonality among E. faecalis isolates, and can be used in conjunction with MLST to identify recombination events occurring between STs. PMID:26398194

  15. A Rex Family Transcriptional Repressor Influences H2O2 Accumulation by Enterococcus faecalis

    PubMed Central

    Vesić, Dušanka

    2013-01-01

    Rex factors are bacterial transcription factors thought to respond to the cellular NAD+/NADH ratio in order to modulate gene expression by differentially binding DNA. To date, Rex factors have been implicated in regulating genes of central metabolism, oxidative stress response, and biofilm formation. The genome of Enterococcus faecalis, a low-GC Gram-positive opportunistic pathogen, encodes EF2638, a putative Rex factor. To study the role of E. faecalis Rex, we purified EF2638 and evaluated its DNA binding activity in vitro. EF2638 was able to bind putative promoter segments of several E. faecalis genes in an NADH-responsive manner, indicating that it represents an authentic Rex factor. Transcriptome analysis of a ΔEF2638 mutant revealed that genes likely to be involved in anaerobic metabolism were upregulated during aerobic growth, and the mutant exhibited an altered NAD+/NADH ratio. The ΔEF2638 mutant also exhibited a growth defect when grown with aeration on several carbon sources, suggesting an impaired ability to cope with oxidative stress. Inclusion of catalase in the medium alleviated the growth defect. H2O2 measurements revealed that the mutant accumulates significantly more H2O2 than wild-type E. faecalis. In summary, EF2638 represents an authentic Rex factor in E. faecalis that influences the production or detoxification of H2O2 in addition to its more familiar role as a regulator of anaerobic gene expression. PMID:23417491

  16. Proteolytic activity of Enterococcus faecalis VB63F for reduction of allergenicity of bovine milk proteins.

    PubMed

    Biscola, V; Tulini, F L; Choiset, Y; Rabesona, H; Ivanova, I; Chobert, J-M; Todorov, S D; Haertlé, T; Franco, B D G M

    2016-07-01

    With the aim of screening proteolytic strains of lactic acid bacteria to evaluate their potential for the reduction of allergenicity of the major bovine milk proteins, we isolated a new proteolytic strain of Enterococcus faecalis (Ent. faecalis VB63F) from raw bovine milk. The proteases produced by this strain had strong activity against caseins (αS1-, αS2-, and β-casein), in both skim milk and sodium caseinate. However, only partial hydrolysis of whey proteins was observed. Proteolysis of Na-caseinate and whey proteins, observed after sodium dodecyl sulfate-PAGE, was confirmed by analysis of peptide profiles by reversed-phase HPLC. Inhibition of proteolysis with EDTA indicated that the proteases produced by Ent. faecalis VB63F belonged to the group of metalloproteases. The optimal conditions for their activity were 42°C and pH 6.5. The majority of assessed virulence genes were absent in Ent. faecalis VB63F. The obtained results suggest that Ent. faecalis VB63F could be efficient in reducing the immunoreactivity of bovine milk proteins. PMID:27179865

  17. Vitality of Enterococcus faecalis inside dentinal tubules after five root canal disinfection methods

    PubMed Central

    Vatkar, Niranjan Ashok; Hegde, Vivek; Sathe, Sucheta

    2016-01-01

    Aim: To compare the vitality of Enterococcus faecalis within dentinal tubules after subjected to five root canal disinfection methods. Materials and Methods: Dentin blocks (n = 60) were colonized with E. faecalis. After 4 weeks of incubation, the dentin blocks were divided into one control and five test groups (n = 10 each). The root canals of test groups were subjected to one of the disinfection methods, namely, normal saline (NS), sodium hypochlorite (NaOCl), chlorhexidine digluconate (CHX), neodymium-doped yttrium aluminum garnet (Nd: YAG) laser, and diode laser. The effect of disinfection methods was assessed by LIVE/DEAD BacLight stain under the confocal laser scanning microscopy to determine the “zone of dead bacteria” (ZDB). Mean values were calculated for ZDB and the difference between groups was established. Results: Penetration of E. faecalis was seen to a depth of >1000 μm. Viable bacteria were detected with NS irrigation. NaOCl and CHX showed partial ZDB. When the root canals were disinfected with Nd: YAG and diode lasers, no viable bacteria were found. Conclusion: E. faecalis has the ability to colonize inside dentinal tubules to a depth of >1000 μm. In contrast to conventional irrigants, both Nd: YAG and diode lasers were effective in eliminating the vitality of E. faecalis. NS, NaOCl, and CHX showed viable bacteria remaining in dentinal tubules.

  18. Identification of malic and soluble oxaloacetate decarboxylase enzymes in Enterococcus faecalis.

    PubMed

    Espariz, Martín; Repizo, Guillermo; Blancato, Víctor; Mortera, Pablo; Alarcón, Sergio; Magni, Christian

    2011-06-01

    Two paralogous genes, maeE and citM, that encode putative malic enzyme family members were identified in the Enterococcus faecalis genome. MaeE (41 kDa) and CitM (42 kDa) share a high degree of homology between them (47% identities and 68% conservative substitutions). However, the genetic context of each gene suggested that maeE is associated with malate utilization whereas citM is linked to the citrate fermentation pathway. In the present work, we focus on the biochemical characterization and physiological contribution of these enzymes in E. faecalis. With this aim, the recombinant versions of the two proteins were expressed in Escherichia coli, affinity purified and finally their kinetic parameters were determined. This approach allowed us to establish that MaeE is a malate oxidative decarboxylating enzyme and CitM is a soluble oxaloacetate decarboxylase. Moreover, our genetic studies in E. faecalis showed that the citrate fermentation phenotype is not affected by citM deletion. On the other hand, maeE gene disruption resulted in a malate fermentation deficient strain indicating that MaeE is responsible for malate metabolism in E. faecalis. Lastly, it was demonstrated that malate fermentation in E. faecalis is associated with cytoplasmic and extracellular alkalinization which clearly contributes to pH homeostasis in neutral or mild acidic conditions. PMID:21518252

  19. Vitality of Enterococcus faecalis inside dentinal tubules after five root canal disinfection methods

    PubMed Central

    Vatkar, Niranjan Ashok; Hegde, Vivek; Sathe, Sucheta

    2016-01-01

    Aim: To compare the vitality of Enterococcus faecalis within dentinal tubules after subjected to five root canal disinfection methods. Materials and Methods: Dentin blocks (n = 60) were colonized with E. faecalis. After 4 weeks of incubation, the dentin blocks were divided into one control and five test groups (n = 10 each). The root canals of test groups were subjected to one of the disinfection methods, namely, normal saline (NS), sodium hypochlorite (NaOCl), chlorhexidine digluconate (CHX), neodymium-doped yttrium aluminum garnet (Nd: YAG) laser, and diode laser. The effect of disinfection methods was assessed by LIVE/DEAD BacLight stain under the confocal laser scanning microscopy to determine the “zone of dead bacteria” (ZDB). Mean values were calculated for ZDB and the difference between groups was established. Results: Penetration of E. faecalis was seen to a depth of >1000 μm. Viable bacteria were detected with NS irrigation. NaOCl and CHX showed partial ZDB. When the root canals were disinfected with Nd: YAG and diode lasers, no viable bacteria were found. Conclusion: E. faecalis has the ability to colonize inside dentinal tubules to a depth of >1000 μm. In contrast to conventional irrigants, both Nd: YAG and diode lasers were effective in eliminating the vitality of E. faecalis. NS, NaOCl, and CHX showed viable bacteria remaining in dentinal tubules. PMID:27656064

  20. Monoclonal antibodies recognizing the Enterococcus faecalis collagen-binding MSCRAMM Ace: conditional expression and binding analysis.

    PubMed

    Hall, Andrea E; Gorovits, Elena L; Syribeys, Peter J; Domanski, Paul J; Ames, Brenda R; Chang, Cathy Y; Vernachio, John H; Patti, Joseph M; Hutchins, Jeff T

    2007-01-01

    Enterococci are opportunistic pathogens known to cause numerous clinical infections and complications in humans. Adhesin-mediated binding to extracellular matrix (ECM) proteins of the host is thought to be a crucial step in the pathogenesis of these bacterial infections. Adhesin of collagen from Enterococcus faecalis (Ace) is a cell-wall anchored protein of E. faecalis that has been shown to be important for bacterial binding to the ECM. In this report, we characterize the conditions for Ace expression and demonstrate Ace binding to mammalian epithelial and endothelial cells as well as to collagens found in the ECM. To further characterize Ace expression and function, we report the generation of a panel of monoclonal antibodies (mAbs) directed against this important E. faecalis virulence factor. Through the use of multiple in vitro assays, surface plasmon resonance and flow cytometry, we have characterized this panel of mAbs which may prove to be not only beneficial in studies that address the precise biological role of adhesion of E. faecalis, but may also serve as beneficial therapeutic agents against E. faecalis infections. PMID:17521860

  1. Transfer of tetracycline resistance genes with aggregation substance in food-borne Enterococcus faecalis.

    PubMed

    Choi, Jong-Mi; Woo, Gun-Jo

    2015-04-01

    Enterococcus faecalis has the ability to conjugate with the aid of aggregation substance (AS) and inducible sex pheromones to exchange genetic elements in food matrix. To evaluate the food safety condition and the transferable factor, 250 tetracycline-resistant food-borne E. faecalis were collected in Korea. Among the isolates, a majority of tetracycline-resistant isolates (49.6 %) harbored both the tet(M) and tet(L) genes together, followed by tet(M) (19.6 %), and tet(L) (6.8 %) alone. Also, we found the combination of tet(L)/tet(M)/tet(O) or tet(M)/tet(O). We identified two tet(S) genes including the isolate carrying tet(M) + tet(S) genes. Additionally, most E. faecalis were positive for cpd and ccf (both 96.8 %) followed by cob (57.2 %). Through mating experiments, we confirmed E. faecalis possessing the Int-Tn gene and/or any AS gene successfully transferred tet genes to JH2-2 E. faecalis, whereas neither E. faecalis carrying AS genes nor the Int-Tn gene showed the conjugation. Pulsed-field gel electrophoresis results supported a distinct pattern, implying transfer of genetic information. Our study revealed a high occurrence of tetracycline resistance genes in E. faecalis from various foods. The widespread dissemination of tetracycline resistance genes would be promoted to transfer tetracycline resistance genes by pheromone-mediated conjugation systems.

  2. Collagen degradation and MMP9 activation by Enterococcus faecalis contributes to intestinal anastomotic leak

    PubMed Central

    Shogan, B. D.; Belogortseva, N.; Luong, P. M.; Zaborin, A.; Lax, S.; Bethel, Cindy; Ward, M.; Muldoon, J. P.; Singer, M.; An, G.; Umanskiy, K.; Konda, V.; Shakhsheer, B.; Luo, J.; Klabbers, R.; Hancock, L. E.; Gilbert, J.; Zaborina, O.; Alverdy, J. C.

    2016-01-01

    Even under the most expert care, a properly constructed intestinal anastomosis can fail to heal resulting in leakage of its contents, peritonitis and sepsis. The cause of anastomotic leak remains unknown and its incidence has not changed in decades. Here, we demonstrate that the commensal bacterium Enterococcus faecalis contributes to the pathogenesis of anastomotic leak through its capacity to degrade collagen and to activate tissue matrix metalloprotease-9 (MMP9) in host intestinal tissues. We demonstrate in rats that leaking anastomotic tissues were colonized by E. faecalis strains that showed an increased collagen-degrading activity and also an increased ability to activate host MMP9, both of which contributed to anastomotic leakage. We demonstrate that the E. faecalis genes gelE and sprE were required for E. faecalis-mediated MMP9 activation. Either elimination of E. faecalis strains through direct topical antibiotics applied to rat intestinal tissues or pharmacological suppression of intestinal MMP9 activation prevented anastomotic leak in rats. In contrast, the standard recommended intravenous antibiotics used in patients undergoing colorectal surgery did not eliminate E. faecalis at anastomotic tissues nor did they prevent leak in our rat model. Finally, we show in humans undergoing colon surgery and treated with the standard recommended intravenous antibiotics, that their anastomotic tissues still contained E. faecalis and other bacterial strains with collagen-degrading/MMP9 activity. We suggest that intestinal microbes with the capacity to produce collagenases and to activate host metalloproteinase MMP9 may break down collagen in the gut tissue contributing to anastomotic leak. PMID:25947163

  3. Characterization of Enterococcus faecalis phage IME-EF1 and its endolysin.

    PubMed

    Zhang, Wenhui; Mi, Zhiqiang; Yin, Xiuyun; Fan, Hang; An, Xiaoping; Zhang, Zhiyi; Chen, Jiankui; Tong, Yigang

    2013-01-01

    Enterococcus faecalis is increasingly becoming an important nosocomial infection opportunistic pathogen. E. faecalis can easily obtain drug resistance, making it difficult to be controlled in clinical settings. Using bacteriophage as an alternative treatment to drug-resistant bacteria has been revitalized recently, especially for fighting drug-resistant bacteria. In this research, an E. faecalis bacteriophage named IME-EF1 was isolated from hospital sewage. Whole genomic sequence analysis demonstrated that the isolated IME-EF1 belong to the Siphoviridae family, and has a linear double-stranded DNA genome consisting of 57,081 nucleotides. The IME-EF1 genome has a 40.04% G+C content and contains 98 putative coding sequences. In addition, IME-EF1 has an isometric head with a width of 35 nm to 60 nm and length of 75 nm to 90 nm, as well as morphology resembling a tadpole. IME-EF1 can adsorb to its host cells within 9 min, with an absorbance rate more than 99% and a latent period time of 25 min. The endolysin of IME-EF1 contains a CHAP domain in its N-terminal and has a wider bactericidal spectrum than its parental bacteriophage, including 2 strains of vancomycin-resistant E. faecalis. When administrated intraperitoneally, one dose of IME-EF1 or its endolysin can reduce bacterial count in the blood and protected the mice from a lethal challenge of E. faecalis, with a survival rate of 60% or 80%, respectively. Although bacteriophage could rescue mice from bacterial challenge, to the best of our knowledge, this study further supports the potential function of bacteriophage in dealing with E. faecalis infection in vivo. The results also indicated that the newly isolated bacteriophage IME-EF1 enriched the arsenal library of lytic E. faecalis bacteriophages and presented another choice for phage therapy in the future.

  4. Transmission and genetic diversity of Enterococcus faecalis among layer chickens during hatch

    PubMed Central

    2011-01-01

    Background Studies on transmission of Enterococcus faecalis among chickens during hatch have not been carried out so far. Information about vertical transmission and subsequent spreading and colonization of the cloacal mucosa through cloacal 'drinking' during hatch are important to understand the epidemiology of E. faecalis infections. In the present investigation vertical transmission and subsequent spreading and colonization of the cloacal mucosa of chickens by E. faecalis through cloacal 'drinking' were examined. Methods Two different batches of layer chickens originating from 45 weeks old Brown and White Lohmann parents, respectively from the same farm were sampled in the hatcher. Isolates were confirmed to be E. faecalis by polymerase chain reaction (PCR) and further by multilocus sequence typing (MLST) to state their population structure and comparison made to sequence types previously obtained from chicken. Results A total of 480 chickens were swabbed from the cloacae just after hatch and after 24 hours. A total of 101 isolates were confirmed as E. faecalis by a species specific PCR. The prevalence of E. faecalis increased from 14% at 0 h to 97% after 24 h for the Brown Lohmann chickens and from 0.5% to 23% for the White Lohmann flock. The 84 isolates analysed by MLST were distributed on 14 sequence types (ST). Three ST (401, 82 and 249) accounted for 64% of all isolates analysed by MLST after 24 h. ST 82 has previously been reported from amyloid arthropathy and other lesions in poultry. Conclusions The present findings demonstrated a high potential of a few contaminated eggs or embryos to rapidly facilitate the spread of E. faecalis to almost all chickens during hatch. PMID:22017822

  5. Surotomycin demonstrates low in vitro frequency of resistance and rapid bactericidal activity in Clostridium difficile, Enterococcus faecalis, and Enterococcus faecium.

    PubMed

    Mascio, Carmela T M; Chesnel, Laurent; Thorne, Grace; Silverman, Jared A

    2014-07-01

    Surotomycin (CB-183,315) is an orally administered, minimally absorbed, selective bactericidal cyclic lipopeptide in phase 3 development for the treatment of Clostridium difficile-associated diarrhea. The aim of this study was to evaluate the emergence of resistance in C. difficile (ATCC 700057 and three recent clinical isolates from the restriction endonuclease analysis groups BI, BK, and K), vancomycin-susceptible (VS) Enterococcus faecalis (ATCC 49452), vancomycin-resistant (VR) E. faecalis (ATCC 700802), VS Enterococcus faecium (ATCC 6569), and VR E. faecium (ATCC 51559) under anaerobic conditions. The rate of spontaneous resistance was below the limit of detection (<10(-8) to <10(-9)) for surotomycin at 16 and 32× the MIC for all isolates tested. Under selective pressure by serial passage, C. difficile grew in a maximum of 4 μg/ml surotomycin (final MICs of 2 to 8 μg/ml [4- to 16-fold higher than those of the naive control]) at day 15, with the exception of the C. difficile BK strain, which grew in 16 to 32 μg/ml (final MICs of 8 to 32 μg/ml [16- to 64-fold higher than those of the naive control]). Enterococci remained relatively unchanged over 15 days, growing in a maximum of 8 μg/ml surotomycin (final MICs of 2 to 16 μg/ml [8- to 64-fold higher than those of the naive control]). Of the isolates tested, no cross-resistance to vancomycin, rifampin, ampicillin, metronidazole, or moxifloxacin was observed. Surotomycin at 20× MIC demonstrated equally rapid bactericidal activity (≥ 3-log-unit reduction in CFU/ml in ≤ 8 h) against naive and reduced-susceptibility isolates of C. difficile, VS Enterococcus (VSE), and VR Enterococcus (VRE), except for C. difficile BK (2.6-log-unit reductions for both). These results suggest that emergence of resistance to surotomycin against C. difficile, E. faecalis, and E. faecium is likely to be rare.

  6. Multilevel selection of bcrABDR-mediated bacitracin resistance in Enterococcus faecalis from chicken farms

    PubMed Central

    Chen, Mu-Ya; Lira, Felipe; Liang, Hua-Qing; Wu, Rui-Ting; Duan, Jia-Hong; Liao, Xiao-Ping; Martínez, José L.; Liu, Ya-Hong; Sun, Jian

    2016-01-01

    In this study we isolated 109 Enterococcus faecalis from chicken faecal samples in 6 provinces of China to investigate the prevalence and transmission mechanism of the bacitracin resistance locus bcrABDR in E. faecalis. Thirty-seven bcrABDR-positive E. faecalis were detected with 26 different PFGE clusters. The MLST of 14 positive strains belonged to ST16 and we also detected three new sequence types. S1-PFGE analysis indicated that the locus was located on plasmids presenting different sizes, with the most prevalent size being ~50 kb (13/37). Sequence analysis revealed that 17 out of the 37 strains harbored a 5400-bp central region, in which locus bcrABDR was bracketed by two ISEnfa1 of the same orientation. Two types of bcrABDR alleles, differing in around 10% of their sequence were found. In silico analysis showed that bcrABDR is present in a variety of bacteria including the chicken commensal Enterococcus cecorum. Our results indicate that the use of bacitracin at farms might trigger the emergence and spread of the bacitracin resistance determinant bcrABDR among human bacterial pathogens. The finding of bcrABDR in the chicken commensal E. cecorum indicates that farm animals microbiota can be an important reservoir of resistance genes with relevance for human health. PMID:27731342

  7. First detection of the antiseptic resistance gene qacA/B in Enterococcus faecalis.

    PubMed

    Bischoff, Meike; Bauer, Johann; Preikschat, Petra; Schwaiger, Karin; Mölle, Gabriele; Hölzel, Christina

    2012-02-01

    Resistance to disinfectants is well investigated in staphylococci and pseudomonads but nearly unexplored in bacteria of the genus Enterococcus, despite their rising significance as nosocomial pathogens. In this study, Enterococcus faecalis (n=585) from blood (n=42) and stool (n=109) of hospitalized humans, from faeces of farm animals (n=226), and from food (milk and dairy products, n=96; meat and meat products, n=112) were screened for the presence of qac-genes (qacA, qacB, qacC, smr [qacC+qacD], qacEΔ1, qacG, qacH, qacJ) via PCR. The isolates' susceptibility to a quaternary ammonium compound (didecyldimethylammoniumchloride, DDAC) and antibiotics was assessed by microdilution. Four E. faecalis strains were positive for qac-genes: qacA/B was found in one isolate from cattle and one isolate from human blood; smr (qacC+qacD) was detected in one isolate from human stool and in one isolate from cheese ("Camembert"). The sequences of the qacA/B-amplicons differed in two basepairs. DDAC had an elevated minimum inhibitory concentration (MIC) of 2.45-3.5 mg/L in one qacA/B-positive strain from human blood, whereas the other qac-gene carriers had wild-type MIC-values for DDAC (1.05 mg/L). This is the first detection of qacA/B in the genus Enterococcus.

  8. Enterococcus species diversity and molecular characterization of biomarker genes in Enterococcus faecalis in Port Blair Bay, Andaman and Nicobar Islands, India.

    PubMed

    Meena, Balakrishnan; Anburajan, Lawrance; Sathish, Thadikamala; Raghavan, Rangamaran Vijaya; Jha, Dilip Kumar; Venkateshwaran, Pitchiah; Das, Apurba Kumar; Dheenan, Palaiya Sukumaran; Vinithkumar, Nambali Valsalan; Dharani, Gopal; Kirubagaran, Ramalingam

    2015-05-15

    This study was performed to evaluate the abundance and diversity of Enterococcus sp. and the distribution of biomarker genes in Enterococcus faecalis in Port Blair Bay, Andaman and Nicobar Islands. The Enterococcus sp. densities at the seven sampling stations were highly influenced by tidal fluctuations and season. The distributions and diversities of species varied in the inner and outer regions of Port Blair Bay. Among the 1816 total isolates, the occurrence of fecal Enterococcus was high (1.78×10(4) CFU/100 mL) in Phoenix Bay. Moreover, 67.76% of the isolates were identified as Enterococcus, and the most frequently identified species were E. hirae, E. avium and E. faecalis. Assessments of antibiotic resistance and biomarker genes revealed the maximum occurrence in the Aberdeen Bay isolates. The most prevalent biomarker genes observed in the E. faecalis isolates were gelE and asa1, whereas cyl was not found among the isolates. In silico sequence analysis of biomarker genes of E. faecalis also revealed that they are evolutionarily well conserved with those of earlier reports. Further, multivariate analysis distinguished the JB, PB and OS stations from the other stations according to distinctive microbial densities and compositions. In addition, the Shannon-Wiener diversity indices and box-whisker plots further facilitated and supported the multivariate results.

  9. Antimicrobial Activity of Nanoparticle Calcium Hydroxide against Enterococcus Faecalis: An In Vitro Study

    PubMed Central

    Dianat, Omid; Saedi, Sara; Kazem, Majid; Alam, Mostafa

    2015-01-01

    Introduction: Enterococcus faecalis (E. faecalis) has the ability to invade the dentinal tubules and resist high pH levels. As a result, calcium hydroxide (CH) is not much effective on this bacterium. In theory, nanoparticle calcium hydroxide (NCH) has smaller size and high surface area that enables it to penetrate into the deeper layers of dentin and be more effective on E. faecalis. This in vitro study was designed to compare the antimicrobial activity of NCH and CH against E. faecalis. Methods and Materials: The antimicrobial activity of NCH against E. faecalis was evaluated by two independent tests: the minimum inhibitory concentration (MIC) of intracanal medicament and agar diffusion test (ADT). The efficiency of the medicament in dentinal tubules was evaluated on 23 human tooth blocks that were inoculated with E. faecalis. The tooth blocks were assigned to one control group (saline irrigation) and two experimental groups receiving CH and NCH as intracanal medication. The optical density in each group was assessed with spectrophotometer after collecting samples from dentin depths of 0, 200 and 400 µm. Data were analyzed by SPSS software ANOVA, Kruskal-Wallis and Dunnett’s test. Results: The MIC for NCH was 1/4 of the MIC for CH. NCH with distilled water (DW) produced the greatest inhibition zone in agar diffusion test. NCH had greater antimicrobial activity in dentin samples from depths of 200 and 400 µm compared to CH. Conclusion: The antimicrobial activity of NCH was superior to CH in culture medium. In dentinal tubules the efficacy of NCH was again better than CH on the 200- and 400-µm samples. PMID:25598808

  10. Erythromycin- and copper-resistant Enterococcus hirae from marine sediment and co-transfer of erm(B) and tcrB to human Enterococcus faecalis.

    PubMed

    Pasquaroli, Sonia; Di Cesare, Andrea; Vignaroli, Carla; Conti, Giulia; Citterio, Barbara; Biavasco, Francesca

    2014-09-01

    An erythromycin-, copper- and cadmium-resistant isolate of Enterococcus hirae from marine sediment was shown to harbor the plasmid pRE25 and to co-transfer erm(B) and tcrB to Enterococcus faecalis JH2-2. These data highlight the scope for antibiotic resistance selection by the marine environment through heavy metals and its possible involvement in antibiotic-resistant enterococcal infections.

  11. Evaluation of the presence of Enterococcus Faecalis in root cementum: A confocal laser scanning microscope analysis

    PubMed Central

    Halkai, Rahul; Hegde, Mithra N; Halkai, Kiran

    2014-01-01

    Aim: The aim of this study is to address the cause of persistent infection of root cementum by Enterococcus faecalis. Materials and Methods: A sample of 60 human single-rooted teeth were divided into three groups. Group I (control group) had no access opening and one-third of the apical root cementum was sealed using varnish. Group II had no preparation of teeth samples. In group III, apical root cementum was exposed to organic acid and roughened using diamond point to mimic apical resorption. After access opening in groups II and III, all teeth samples were sterilized using gamma irradiation (25 kGy). E. faecalis broth was placed in the root canal and apical one-third of the tooth was immersed in the broth for 8 weeks with alternate day refreshment followed by biomechanical preparation, obturation and coronal seal. Apical one-third of all teeth samples were again immersed in the broth for 8 weeks with alternate day refreshment to mimic secondary infection. The samples were observed under a confocal microscope after splitting the teeth into two halves. Results: E. faecalis penetrated 160 μm deep into the root cementum in group III samples and only showed adhesion in group II samples. Conclusion: Penetration and survival of E. faecalis deep inside the cementum in extreme conditions could be the reason for persistent infection. PMID:24778505

  12. Link between Culture Zeta Potential Homogeneity and Ebp in Enterococcus faecalis

    PubMed Central

    Tariq, Muhammad; Bruijs, Chissa; Krom, Bastiaan P.

    2012-01-01

    Enterococcus faecalis, a commensal of the gastrointestinal tract and an opportunistic pathogen, has the ability to adhere to surfaces and form biofilms. It has been shown earlier that only 10 to 20% of an E. faecalis OG1RF culture expresses endocarditis- and biofilm-associated pili (Ebp), which are involved in biofilm formation. Another study revealed that E. faecalis clinical isolates, as well as OG1RF, are heterogeneous with respect to their apparent zeta potential, which was also correlated with increased ability to form biofilm. The aim of this study was to demonstrate that the heterogeneity in the presence of Ebp is correlated to that in apparent zeta potential. Heterogeneous cultures of OG1RF showed two distinct subpopulations with the most (−38 mV) and least (−26 mV) negative zeta potential. Deletion of EbpR, the activator of the ebp operon, or the structural genes ebpABC resulted in homogeneous culture with the most negative zeta potential. Conversely, overexpression of EbpR or the structural genes ebpABC resulted in homogeneous culture with the least negative zeta potential. The results show that ebp operon expression in E. faecalis, as measured by using Pebp-gfp promoter fusion, is the cause of heterogeneity in zeta potential and that pilus production causes the cells to behave as the least negative particle in an electric field. PMID:22267669

  13. Sodium chloride and potassium sorbate: a synergistic combination against Enterococcus faecalis biofilms: an in vitro study.

    PubMed

    van der Waal, Suzette V; Jiang, Lei-Meng; de Soet, Johannes J; van der Sluis, Lucas W M; Wesselink, Paul R; Crielaard, Wim

    2012-10-01

    Incomplete disinfection of the root canal system is a major cause of post-treatment disease. This study aimed to investigate the disinfecting property of organic acid salts and sodium chloride (NaCl), in a double-hurdle strategy, on Enterococcus faecalis biofilms. First of all, the high-throughput resazurin metabolism assay (RMA) was used to test a range of organic acid salts. Then, to gain more insight into the efficacy of sorbate salt solutions, 48-h E. faecalis biofilms were evaluated in colony-forming unit (CFU) assays. Chlorhexidine (CHX) and calcium hydroxide [Ca(OH)(2) ] were tested in parallel as controls. Sorbate salt produced the largest and most significant reduction of fluorescence intensity in the RMA assay. Neither NaCl nor potassium sorbate (KS) alone induced a clinically relevant reduction of CFU counts after 1 h. Surprisingly, the combination of the two in a single solution had a synergistic effect on the inactivation of E. faecalis. Potassium sorbate amplified the efficacy of NaCl. Of the salts tested, NaCl with KS eradicated E. faecalis biofilms within 1 h. This study showed that the double-hurdle strategy indeed leads to synergistic efficacy and is a possible next step in the complete disinfection of endodontic infections. PMID:22985004

  14. Genetic analysis of faropenem-resistant Enterococcus faecalis in urinary isolates.

    PubMed

    Hiraga, Noriyuki; Muratani, Tetsuro; Naito, Seiji; Matsumoto, Tetsuro

    2008-04-01

    We isolated faropenem-resistant Enterococcus faecalis in urine specimens and studied the mechanisms of resistance to faropenem in these isolates. Three mechanisms of penicillin resistance have been reported in E. faecalis; (1) beta-lactamase production, (2) overproduction of penicillin-binding protein (PBP) 4 or PBP5, and (3) decreasing affinities of penicillins for PBP4 by the occurrence of point mutations of the penicillin-binding domain. None of the E. faecalis isolates examined produced beta-lactamase or overproduced any PBPs, but the affinities of faropenem for PBP4 were decreased in faropenem-insensitive and -resistant strains. We found single amino acid substitutions at positions 475, 520 or 605 in PBP4 in the insensitive strains and two amino acid substitutions at positions 520 and 605 in PBP4 in the resistant strains by sequencing the entire pbp4 gene from each isolate. We conclude that development of resistance to faropenem in E. faecalis is due to decreasing affinities for PBP4 that are the result of the occurrence of one or two point mutations. PMID:18503200

  15. Antimicrobial Activity of Calcium Hydroxide and Betamethasone on Enterococcus faecalis; An in vitro Assessment

    PubMed Central

    Tabrizizadeh, Mahdi; Rasti, Mojtaba; Ayatollahi, Fatemeh; Mossadegh, Mohammad Hossein; Zandi, Hengameh; Dehghan, Farzad; Mousavi, Zohreh

    2015-01-01

    Introduction: Calcium hydroxide (CH) is one of the most common intracanal medications. Corticosteroids (CS) are used in endodontics because of their anti-inflammatory activity. This study aimed to evaluate the antimicrobial effect of CH+betamethasone and CH+saline against Enterococcus faecalis (E. faecalis) using agar diffusion test and measuring the microbial zone of inhibition (ZOI). Methods and Materials: Four plates containing Mueller-Hinton broth and E. faecalis culture media, were prepared. In each plate, 5 holes (5×3 mm) were created and a creamy mixture of CH+betamethasone was inserted into the holes (10 holes for each material). Two holes with ampicillin disks and two empty holes were used as negative and positive controls, respectively. Plates were incubated for 24 h and then the diameter of microbial ZOI was measured. The pH of each mixture was measured by pH meter. Data were analyzed using the Mann-Whitney U test. Results: The mean diameter of ZOI for CH+betamethasone and CH+saline was 3.4 and 3 mm, respectively. The difference was not significant (P=0.143). The pH was 12.5 for CH+saline and 12.3 CH+betamethasone, respectively. Conclusion: The mixture of CH+betamethasone had good antimicrobial effects against E. faecalis. Further studies are needed to confirm the value of this mixture in clinical settings. PMID:26213541

  16. Transcriptomic and Functional Analysis of NaCl-Induced Stress in Enterococcus faecalis

    PubMed Central

    Solheim, Margrete; La Rosa, Sabina Leanti; Mathisen, Thomas; Snipen, Lars G.; Nes, Ingolf F.; Brede, Dag Anders

    2014-01-01

    The robust physiology of Enterococcus faecalis facilitates tolerance to various stresses. We here report the transcriptional response of E. faecalis V583 to growth in the presence of 6.5% NaCl. Among the early responses observed was an immediate down-regulation of mscL, accompanied by an up-regulation of genes predicted to be involved in uptake of extracellular potassium and glycine betaine. The high NaCl concentration also induced expression of chaperons and cell envelope related traits, such as the enterococcal polysaccharide antigen (epa) locus. Functional genetic analysis revealed reduced salt stress resistance in both epaB and epaE mutants. The reduced salt resistance phenotype associated with the epaB mutant was restored by complementation, hence demonstrating a role of Epa in the physiological robustness of E. faecalis. Furthermore, we demonstrate that Epa confers increased resistance towards multiple cell envelope stress-inducing factors. Accordingly, these findings delineate a potential link between the robust nature of E. faecalis and its ability to perform as a human pathogen, and provide a new perspective on the mechanisms by which Epa contributes to virulence. Notably, the high NaCl concentration also resulted in strict repression of the gelE-sprE operon and impaired gelatinase activity. We demonstrate that NaCl antagonize the GBAP-pheromone dependent induction in a concentration dependent manner. PMID:24755907

  17. Transcriptomic and functional analysis of NaCl-induced stress in Enterococcus faecalis.

    PubMed

    Solheim, Margrete; La Rosa, Sabina Leanti; Mathisen, Thomas; Snipen, Lars G; Nes, Ingolf F; Brede, Dag Anders

    2014-01-01

    The robust physiology of Enterococcus faecalis facilitates tolerance to various stresses. We here report the transcriptional response of E. faecalis V583 to growth in the presence of 6.5% NaCl. Among the early responses observed was an immediate down-regulation of mscL, accompanied by an up-regulation of genes predicted to be involved in uptake of extracellular potassium and glycine betaine. The high NaCl concentration also induced expression of chaperons and cell envelope related traits, such as the enterococcal polysaccharide antigen (epa) locus. Functional genetic analysis revealed reduced salt stress resistance in both epaB and epaE mutants. The reduced salt resistance phenotype associated with the epaB mutant was restored by complementation, hence demonstrating a role of Epa in the physiological robustness of E. faecalis. Furthermore, we demonstrate that Epa confers increased resistance towards multiple cell envelope stress-inducing factors. Accordingly, these findings delineate a potential link between the robust nature of E. faecalis and its ability to perform as a human pathogen, and provide a new perspective on the mechanisms by which Epa contributes to virulence. Notably, the high NaCl concentration also resulted in strict repression of the gelE-sprE operon and impaired gelatinase activity. We demonstrate that NaCl antagonize the GBAP-pheromone dependent induction in a concentration dependent manner.

  18. Sodium chloride and potassium sorbate: a synergistic combination against Enterococcus faecalis biofilms: an in vitro study.

    PubMed

    van der Waal, Suzette V; Jiang, Lei-Meng; de Soet, Johannes J; van der Sluis, Lucas W M; Wesselink, Paul R; Crielaard, Wim

    2012-10-01

    Incomplete disinfection of the root canal system is a major cause of post-treatment disease. This study aimed to investigate the disinfecting property of organic acid salts and sodium chloride (NaCl), in a double-hurdle strategy, on Enterococcus faecalis biofilms. First of all, the high-throughput resazurin metabolism assay (RMA) was used to test a range of organic acid salts. Then, to gain more insight into the efficacy of sorbate salt solutions, 48-h E. faecalis biofilms were evaluated in colony-forming unit (CFU) assays. Chlorhexidine (CHX) and calcium hydroxide [Ca(OH)(2) ] were tested in parallel as controls. Sorbate salt produced the largest and most significant reduction of fluorescence intensity in the RMA assay. Neither NaCl nor potassium sorbate (KS) alone induced a clinically relevant reduction of CFU counts after 1 h. Surprisingly, the combination of the two in a single solution had a synergistic effect on the inactivation of E. faecalis. Potassium sorbate amplified the efficacy of NaCl. Of the salts tested, NaCl with KS eradicated E. faecalis biofilms within 1 h. This study showed that the double-hurdle strategy indeed leads to synergistic efficacy and is a possible next step in the complete disinfection of endodontic infections.

  19. Biochemical and Genetic Characterization of the Enterococcus faecalis Oxaloacetate Decarboxylase Complex

    PubMed Central

    Repizo, Guillermo D.; Blancato, Víctor S.; Mortera, Pablo; Lolkema, Juke S.

    2013-01-01

    Enterococcus faecalis encodes a biotin-dependent oxaloacetate decarboxylase (OAD), which is constituted by four subunits: E. faecalis carboxyltransferase subunit OadA (termed Ef-A), membrane pump Ef-B, biotin acceptor protein Ef-D, and the novel subunit Ef-H. Our results show that in E. faecalis, subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex (termed Ef-AHD) which is also associated with the membrane. In order to characterize the role of the novel Ef-H subunit, coexpression of oad genes was performed in Escherichia coli, showing that this subunit is vital for Ef-A and Ef-D interaction. Diminished growth of the oadA and oadD single deletion mutants in citrate-supplemented medium indicated that the activity of the complex is essential for citrate utilization. Remarkably, the oadB-deficient strain was still capable of growing to wild-type levels but with a delay during the citrate-consuming phase, suggesting that the soluble Ef-AHD complex is functional in E. faecalis. These results suggest that the Ef-AHD complex is active in its soluble form, and that it is capable of interacting in a dynamic way with the membrane-bound Ef-B subunit to achieve its maximal alkalinization capacity during citrate fermentation. PMID:23435880

  20. Biochemical and genetic characterization of the Enterococcus faecalis oxaloacetate decarboxylase complex.

    PubMed

    Repizo, Guillermo D; Blancato, Víctor S; Mortera, Pablo; Lolkema, Juke S; Magni, Christian

    2013-05-01

    Enterococcus faecalis encodes a biotin-dependent oxaloacetate decarboxylase (OAD), which is constituted by four subunits: E. faecalis carboxyltransferase subunit OadA (termed Ef-A), membrane pump Ef-B, biotin acceptor protein Ef-D, and the novel subunit Ef-H. Our results show that in E. faecalis, subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex (termed Ef-AHD) which is also associated with the membrane. In order to characterize the role of the novel Ef-H subunit, coexpression of oad genes was performed in Escherichia coli, showing that this subunit is vital for Ef-A and Ef-D interaction. Diminished growth of the oadA and oadD single deletion mutants in citrate-supplemented medium indicated that the activity of the complex is essential for citrate utilization. Remarkably, the oadB-deficient strain was still capable of growing to wild-type levels but with a delay during the citrate-consuming phase, suggesting that the soluble Ef-AHD complex is functional in E. faecalis. These results suggest that the Ef-AHD complex is active in its soluble form, and that it is capable of interacting in a dynamic way with the membrane-bound Ef-B subunit to achieve its maximal alkalinization capacity during citrate fermentation.

  1. Effect of the quorum-sensing luxS gene on biofilm formation by Enterococcus faecalis.

    PubMed

    He, Zhiyan; Liang, Jingping; Zhou, Wei; Xie, Qian; Tang, Zisheng; Ma, Rui; Huang, Zhengwei

    2016-06-01

    Enterococcus faecalis is the species of bacterium most frequently isolated from the root canals of teeth that exhibit chronic apical periodontitis refractory to endodontic treatment. In this study, we evaluated the effect of the S-ribosylhomocysteine lyase (luxS) quorum-sensing gene on E. faecalis biofilm formation by constructing a knockout mutant. The biofilms formed by both E. faecalis and its luxS mutant strain were evaluated using the MTT method. Important parameters that influence biofilm formation, including cell-surface hydrophobicity and the nutrient content of the growth medium, were also studied. Biofilm structures were observed using confocal laser scanning microscopy (CLSM), and expression of biofilm-related genes was investigated using RT-PCR. The results showed that the luxS gene can affect biofilm formation, whereas it does not affect the bacterial growth rate. Deletion of the luxS gene also increased cell-surface hydrophobicity. Biofilm formation was accelerated by the addition of increasing concentrations of glucose. The CLSM images revealed that the luxS mutant strain tends to aggregate into distinct clusters and relatively dense structures, whereas the wild-type strain appears confluent and more evenly distributed. All genes examined were up-regulated in the biofilms formed by the luxS mutant strain. The quorum-sensing luxS gene can affect E. faecalis biofilm formation.

  2. A Cluster of Genes Involved in Polysaccharide Biosynthesis from Enterococcus faecalis OG1RF

    PubMed Central

    Xu, Yi; Murray, Barbara E.; Weinstock, George M.

    1998-01-01

    Our previous work identified a cosmid clone containing a 43-kb insert from Enterococcus faecalis OG1RF that produced a nonprotein antigen in Escherichia coli. In the present work, we studied this clone in detail. Periodate treatment of lysates of the clone confirmed that the antigen was carbohydrate in nature. Analysis of DNA sequences and transposon insertion mutants suggested that the insert contained a multicistronic gene cluster. Database comparison showed that the cluster contained genes similar to genes involved in the biosynthesis of dTDP-rhamnose, glycosyltransferases, and ABC transporters involved in the export of sugar polymers from both gram-positive and gram-negative organisms. Insertions in several genes within the cluster abolished the immunoreactivity of the clone. This is the first report on a gene cluster of E. faecalis involved in the biosynthesis of an antigenic polysaccharide. PMID:9712783

  3. [In vitro activity of ampicillin-ceftriaxone against Enterococcus faecalis isolates recovered from invasive infections].

    PubMed

    Burguer Moreira, Noelia; Nastro, Marcela; Vay, Carlos; Famiglietti, Ángela; Rodríguez, Carlos Hernán

    2016-01-01

    In vitro activity of the combination of ampicillin- ceftriaxone against 30 Enterococcus faecalis isolates recovered from invasive infections in patients admitted to Hospital de Clínicas José de San Martin in the city of Buenos Aires was assessed. Ampicillin- ceftriaxone synergies were determined by microdilution in Müeller-Hinton (MH) broth with and without subinhibitory concentrations of ceftriaxone. Synergy was detected in 22/30 isolates. A decrease in both minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) was observed in 14/30 isolates, whereas in 6/30 isolates the decrease was observed in the MIC value and only in the MBC value in the 2 remaining isolates. The bactericidal activity of the combination showed to be higher at low concentrations of ampicillin (< 1 μg/ml). We detected in vitro synergy using the ampicillin-ceftriaxone combination and thus, its efficacy was confirmed in the treatment of severe infections by E. faecalis.

  4. Cloning, purification, crystallization and preliminary crystallographic analysis of SecA from Enterococcus faecalis

    SciTech Connect

    Meining, Winfried; Scheuring, Johannes; Fischer, Markus; Weinkauf, Sevil

    2006-06-01

    SecA ATPase from E. faecalis has been cloned, overexpressed, purified and crystallized. Crystals belong to space group C2 and diffract to 2.4 Å resolution. The gene coding for SecA from Enterococcus faecalis was cloned and overexpressed in Escherichia coli. In this protein, the lysine at position 6 was replaced by an asparagine in order to reduce sensitivity towards proteases. The modified protein was purified and crystallized. Crystals diffracting to 2.4 Å resolution were obtained using the vapour-diffusion technique. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 203.4, b = 49.8, c = 100.8 Å, α = γ = 90.0, β = 119.1°. A selenomethionine derivative was prepared and is currently being tested in crystallization trials.

  5. Enterococcus faecalis cytolysin without effect on the intestinal growth of susceptible enterococci in mice.

    PubMed

    Huycke, M M; Joyce, W A; Gilmore, M S

    1995-07-01

    A murine model was developed to determine whether the Enterococcus faecalis cytolysin, through its bacteriolytic action on gram-positive bacteria, could promote intestinal overgrowth of cytolytic strains. Sets of E. faecalis strains with varying cytolytic production and susceptibility to cytolytic activity were mixed 1:1 and allowed to compete in vitro in broth or in vivo after orogastric administration in mice pretreated with antibiotics. In general, cytolytic strains outgrew, by as much as 2000-fold, competing cytolysin-susceptible or -hypersusceptible strains in vitro. In contrast, no growth advantage was observed in vivo, despite similar transient colonization of the murine intestinal tract by both cytolytic and cytolysin-susceptible strains. These data suggest that cytolysin plays little role in promoting intestinal overgrowth of enterococci through bacteriolytic activity. PMID:7797930

  6. Effect of peptide AS-48 on Enterococcus faecalis subsp. liquefaciens S-47.

    PubMed Central

    Galvez, A; Valdivia, E; Martinez, M; Maqueda, M

    1989-01-01

    The enterococcal peptide AS-48 exerts a concentration-dependent bactericidal effect on Enterococcus faecalis subsp. liquefaciens S-47; cell rescue by cardiolipin and trypsin can be effected only in the first few minutes after antibiotic addition. Gramicidin-exposed cells are protected from killing by AS-48. Long-term and pulse incorporation of radiolabeled substrates into trichloroacetic acid-precipitable material, O2 consumption, and the ability to maintain intracellular potassium levels are impaired shortly after addition of AS-48. PMID:2473710

  7. Enterocin 226NWC, a bacteriocin produced by Enterococcus faecalis 226, active against Listeria monocytogenes.

    PubMed

    Villani, F; Salzano, G; Sorrentino, E; Pepe, O; Marino, P; Coppola, S

    1993-04-01

    Enterococcus faecalis 226, isolated from natural whey cultures utilized as starters in the manufacture of mozzarella cheese from water-buffalo milk, produces a bacteriocin designated enterocin 226NWC. The bacteriocin was isolated from culture supernatant fluids of the producer strain and was active against strains of the same species and Listeria monocytogenes, but not against useful lactic acid bacteria. Enterocin 226NWC is a protein with an apparent molecular weight of about 5800; it is relatively heat-stable and has a bactericidal mode of action. Listeria monocytogenes, growing in the presence of the enterocin 226NWC producer strain in broth and in reconstituted skim milk, was inhibited.

  8. Characterization of antimicrobial resistance and virulence genotypes of Enterococcus faecalis recovered from a pork processing plant.

    PubMed

    Aslam, Mueen; Diarra, Moussa S; Masson, Luke

    2012-08-01

    The objective of this study was to assess the antimicrobial resistance and virulence genotypes of Enterococcus faecalis isolated from samples obtained from a commercial pork processing plant. A total of 200 samples were randomly obtained from carcasses after bleeding (BC; 50 samples) and pasteurization (PC; 100 samples) and from retail pork products (RP; 50 samples). One isolate from each E. faecalis -positive sample was analyzed for antimicrobial susceptibility and characterized using a enterococcal microarray for analysis of resistance and virulence genes. E. faecalis was isolated from 79.5% of BC samples, 2% of PC samples, and 72.7% of RP samples. Resistance to the clinically important drugs ciprofloxacin (one isolate each from BC and RP samples) and daptomycin (one isolate each from PC and RP samples) was found. Multiresistance (to five or more antimicrobials) was more common in E. faecalis isolates from BC (77.4% of isolates) samples than those from PC (25%) and RP (37.6%) samples. Resistance to kanamycin (43.5%) and streptomycin (69.2%) was noted mostly in E. faecalis from BC samples. The most common resistance genes (>5% prevalence) found in E. faecalis were those for aminoglycosides (aac(6), aphA3, and aadE), macrolides-lincosamide (ermB, ermA, sat(4), and linB), and tetracyclines (tetL, tetM, and tetO ). The virulence genes expressing adhesion (ace, efaAfs, and agrBfs), gelatinase (gelE), and pheromone (cAM, ccF10, cob, and cpd1) factors were found in the majority of isolates. Significant associations were found between resistance and virulence genes, suggesting their possible relationship. These data suggest that carcasses entering the final product processing area are mostly free of E. faecalis but are recontaminated with antimicrobial-resistant strains during processing. The source of these contaminants remains to be identified; however, these results underscore the importance of E. faecalis as a reservoir of resistance and virulence genes.

  9. The Viable but Nonculturable State and Starvation Are Different Stress Responses of Enterococcus faecalis, as Determined by Proteome Analysis

    PubMed Central

    Heim, Sabina; Del Mar Lleo, Maria; Bonato, Barbara; Guzman, Carlos A.; Canepari, Pietro

    2002-01-01

    The protein expression patterns of exponentially growing, starved, and viable but nonculturable (VBNC) Enterococcus faecalis cells were analyzed to establish whether differences exist between the VBNC state and other stress responses. The results indicate that the protein profile of VBNC cells differs from that of either starved or exponentially growing bacteria. This demonstrates that the VBNC state is a distinct physiological phase within the life cycle of E. faecalis, which is activated in response to multiple environmental stresses. PMID:12426365

  10. Sub-lethal stress effects on virulence gene expression in Enterococcus faecalis.

    PubMed

    Lenz, Christian A; Hew Ferstl, Carrie M; Vogel, Rudi F

    2010-05-01

    Enterococci are ubiquitous lactic acid bacteria commonly associated with the human digestive tract as commensal organisms. Additionally, these organisms have a long history of use in foods improving flavor as well as providing protective mechanisms as either a probiotic or antimicrobial additive. However, Enterococcus faecalis accounts for up to 10% of all nosocomial infections of the bloodstream, wounds, urinary tract and heart. Knowledge about the regulation of virulence factors is limited and the involvement of environmental signals contributing to E. faecalis pathogenicity is poorly documented. In this study, two clinical E. faecalis isolates, TMW 2.63 and OG1RF, as well as one food isolate, TMW 2.629, were subjected to six sub-lethal food- and host-related stresses including 6.8% NaCl, 200 ppm nitrite, 51 degrees C, 80 MPa, pH 4.1 and 0.08% bile salts (cholic acid:chenodeoxycholic acid 1:1), respectively, reducing their growth rate to 10%. Relative gene expression of 15 stress and virulence-associated genes including dnaK, groEL, ctsR, clpPBCEX, gls24, efaAfs, ace, fsrB, gelE, sprE and cylB, was quantified by using real time PCR and Lightcycler((R)) technology (reference conditions: BHI broth, 37 degrees C, pH = 7.4). Apart from strain-dependent differences, sub-lethal environmental stress was capable of provoking significant alterations in the expression of virulence-associated genes in E. faecalis from clinical as well as food origins of isolation. These results help to avoid preconditioning enterococci in food production processes and to understand the complex mechanisms in E. faecalis' switch to pathogenicity. PMID:20227595

  11. Enterococcus faecalis overcomes foreign body-mediated inflammation to establish urinary tract infections.

    PubMed

    Guiton, Pascale S; Hannan, Thomas J; Ford, Bradley; Caparon, Michael G; Hultgren, Scott J

    2013-01-01

    Urinary catheterization elicits major histological and immunological changes that render the bladder susceptible to microbial invasion, colonization, and dissemination. However, it is not understood how catheters induce these changes, how these changes act to promote infection, or whether they may have any protective benefit. In the present study, we examined how catheter-associated inflammation impacts infection by Enterococcus faecalis, a leading cause of catheter-associated urinary tract infection (CAUTI), a source of significant societal and clinical challenges. Using a recently optimized murine model of foreign body-associated UTI, we found that the implanted catheter itself was the primary inducer of inflammation. In the absence of the silicone tubing implant, E. faecalis induced only minimal inflammation and was rapidly cleared from the bladder. The catheter-induced inflammation was only minimally altered by subsequent enterococcal infection and was not suppressed by inhibitors of the neurogenic pathway and only partially by dexamethasone. Despite the robust inflammatory response induced by urinary implantation, E. faecalis produced biofilm and high bladder titers in these animals. Induction of inflammation in the absence of an implanted catheter failed to promote infection, suggesting that the presence of the catheter itself is essential for E. faecalis persistence in the bladder. Immunosuppression prior to urinary catheterization enhanced E. faecalis colonization, suggesting that implant-mediated inflammation contributes to the control of enterococcal infection. Thus, this study underscores the need for novel strategies against CAUTIs that seek to reduce the deleterious effects of implant-mediated inflammation on bladder homeostasis while maintaining an active immune response that effectively limits bacterial invaders.

  12. Antibacterial Efficacy of Different Concentrations of Sodium Hypochlorite Gel and Solution on Enterococcus faecalis Biofilm

    PubMed Central

    Zand, Vahid; Lotfi, Mehrdad; Soroush, Mohammad Hosein; Abdollahi, Amir Ardalan; Sadeghi, Mehdi; Mojadadi, Ali

    2016-01-01

    Introduction: This in vitro study compared the antibacterial efficacy of 2.5% sodium hypochlorite gel and 2.5% and 5.25% sodium hypochlorite solutions on Enterococcus faecalis (E. faecalis) biofilm. Methods and Materials: The root canals of 60 extracted human single-rooted teeth were contaminated with E. faecalis and incubated for 6 weeks. The samples were randomly assigned to three experimental groups and one control group (n=15). The study protocol in the experimental groups consisted of injection of 5 mL of each irrigant into the root canals. Samples were collected from the root canal walls and 1:10 serial dilutions were prepared and added to Muller Hinton Agar (MHA) plates and incubated at 37°C for 48 h. A classic colony counting technique was used for determining vital E. faecalis bacterial counts in MHA plates. The Kruskal-Wallis test was used for statistical analysis of the data. The level of significance was set at 0.05. Results: The antibacterial effect of the irrigants in all three experimental groups was significantly greater than the control group (P<0.05), with no significant difference between 2.5% and 5.25% NaOCl solutions (P>0.05). The effect of 2.5% and 5.25% NaOCl solutions were significantly superior to 2.5% NaOCl gel (P<0.05). Conclusion: Under the limitations of this study, 2.5% NaOCl gel was effective in reducing E. faecalis counts; however this effect was less than that of NaOCl solutions. PMID:27790262

  13. Effect of Nanosilver Gel, Chlorhexidine Gluconate, and Camphorated Phenol on Enterococcus faecalis Biofilm.

    PubMed

    Bo, Dong; Kayombo, Cecilia Marcellino

    2014-01-01

    Aim. To assess the effectiveness of nanosilver gel (NSG) in comparison to chlorhexidine gluconate (CHX) and camphorated phenol (CP) against Enterococcus faecalis (E.f) biofilm. Methods and Materials. Two tests were done, methyl thiazolyl tetrazolium (MTT) assay and confocal laser scanning microscopy (CLSM) analysis, to determine the effectiveness of NSG, CHX, and CP on E.f biofilm. Polystyrene microtiter 96- and 6-well plates were used for MTT and CLSM, respectively. Nanosilver gel was in three concentrations (0.05%, 0.1%, and 0.2%), chlorhexidine gluconate used was 2%, and camphorated phenol and normal saline were as control. Analysis was done using one-way ANOVA; the post hoc test was run for multiple comparisons. The level of statistical significance was set at P < 0.05. Results. One-way ANOVA showed significant differences among groups (0.05% NSG and CP, 0.1% NSG and CP, 0.2% NSG and CP, 0.1% NSG and 2% CHX, 0.2% and NSG and 2% CHX) (P < 0.001) and also showed significant difference between groups (P < 0.001), f-ratio 87.823. A post hoc Tukey's test revealed no significant difference between chlorhexidine gluconate and 0.05% nanosilver gel (P > 0.05). Conclusions. 0.1% and 0.2% nanosilver gel is more effective on Enterococcus faecalis biofilm as compared to chlorhexidine gluconate and camphorated phenol.

  14. High genetic diversity of Enterococcus faecium and Enterococcus faecalis clinical isolates by pulsed-field gel electrophoresis and multilocus sequence typing from a hospital in Malaysia.

    PubMed

    Weng, Poh Leng; Ramli, Ramliza; Shamsudin, Mariana Nor; Cheah, Yoke-Kqueen; Hamat, Rukman Awang

    2013-01-01

    Little is known on the genetic relatedness and potential dissemination of particular enterococcal clones in Malaysia. We studied the antibiotic susceptibility profiles of Enterococcus faecium and Enterococcus faecalis and subjected them to pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). E. faecium and E. faecalis displayed 27 and 30 pulsotypes, respectively, and 10 representative E. faecium and E. faecalis isolates (five each) yielded few different sequence types (STs): ST17 (2 isolates), ST78, ST203, and ST601 for E. faecium, and ST6, ST16, ST28, ST179, and ST399 for E. faecalis. Resistance to tazobactam-piperacillin and ampicillin amongst E. faecium isolates was highly observed as compared to E. faecalis isolates. All of the isolates were sensitive to vancomycin and teicoplanin. The presence of epidemic and nosocomial strains of selected E. faecium STs: 17, 78, and 203 and E. faecalis ST6 as well as high rates of resistance to multiple antibiotics amongst E. faecium isolates is of a particular concern.

  15. High Genetic Diversity of Enterococcus faecium and Enterococcus faecalis Clinical Isolates by Pulsed-Field Gel Electrophoresis and Multilocus Sequence Typing from a Hospital in Malaysia

    PubMed Central

    Weng, Poh Leng; Ramli, Ramliza; Shamsudin, Mariana Nor; Cheah, Yoke-Kqueen; Hamat, Rukman Awang

    2013-01-01

    Little is known on the genetic relatedness and potential dissemination of particular enterococcal clones in Malaysia. We studied the antibiotic susceptibility profiles of Enterococcus faecium and Enterococcus faecalis and subjected them to pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). E. faecium and E. faecalis displayed 27 and 30 pulsotypes, respectively, and 10 representative E. faecium and E. faecalis isolates (five each) yielded few different sequence types (STs): ST17 (2 isolates), ST78, ST203, and ST601 for E. faecium, and ST6, ST16, ST28, ST179, and ST399 for E. faecalis. Resistance to tazobactam-piperacillin and ampicillin amongst E. faecium isolates was highly observed as compared to E. faecalis isolates. All of the isolates were sensitive to vancomycin and teicoplanin. The presence of epidemic and nosocomial strains of selected E. faecium STs: 17, 78, and 203 and E. faecalis ST6 as well as high rates of resistance to multiple antibiotics amongst E. faecium isolates is of a particular concern. PMID:23819125

  16. Transcriptome profiling of TDC cluster deletion mutant of Enterococcus faecalis V583.

    PubMed

    Perez, Marta; Ladero, Victor; Del Rio, Beatriz; Redruello, Begoña; de Jong, Anne; Kuipers, Oscar P; Kok, Jan; Martin, M Cruz; Fernandez, Maria; Alvarez, Miguel A

    2016-09-01

    The species Enterococcus faecalis is able to catabolise the amino acid tyrosine into the biogenic amine tyramine by the tyrosine decarboxilase (TDC) pathway Ladero et al. (2012) [1]. The TDC cluster comprises four genes: tyrS, an aminoacyl-tRNA synthetase-like gene; tdcA, which encodes the tyrosine decarboxylase; tyrP, a tyrosine/tyramine exchanger gene and nhaC-2, which encodes an Na(+)/H(+) antiporter and whose role in the tyramine biosynthesis remains unknown [2]. In E. faecalis V583 the last three genes are co-transcribed as a single polycistronic mRNA forming the catabolic operon, while tyrS is transcribed independently of the catabolic genes as a monocistronic mRNA [2]. The catabolic operon is transcriptionally induced by tyrosine and acidic pH. On the opposite, the tyrS expression is repressed by tyrosine concentrations [2]. In this work we report the transcriptional profiling of the TDC cluster deletion mutant (E. faecalis V583 ΔTDC) [2] compared to the wild-type strain, both grown in M17 medium supplemented with tyrosine. The transcriptional profile data of TDC cluster-regulated genes were deposited in the Gene Expression Omnibus (GEO) database under accession no. GSE77864. PMID:27408815

  17. Capsular polysaccharide production in Enterococcus faecalis and contribution of CpsF to capsule serospecificity.

    PubMed

    Thurlow, Lance R; Thomas, Vinai Chittezham; Hancock, Lynn E

    2009-10-01

    Many bacterial species produce capsular polysaccharides that contribute to pathogenesis through evasion of the host innate immune system. The gram-positive pathogen Enterococcus faecalis was previously reported to produce one of four capsule serotypes (A, B, C, or D). Previous studies describing the four capsule serotypes of E. faecalis were based on immunodetection methods; however, the underlying genetics of capsule production did not fully support these findings. Previously, it was shown that capsule production for serotype C (Maekawa type 2) was dependent on the presence of nine open reading frames (cpsC to cpsK). Using a novel genetic system, we demonstrated that seven of the nine genes in the cps operon are essential for capsule production, indicating that serotypes A and B do not make a capsular polysaccharide. In support of this observation, we showed that serotype C and D capsule polysaccharides mask lipoteichoic acid from detection by agglutinating antibodies. Furthermore, we determined that the genetic basis for the difference in antigenicity between serotypes C and D is the presence of cpsF in serotype C strains. High-pH anion-exchange chromatography with pulsed amperometric detection analysis of serotype C and D capsules indicated that cpsF is responsible for glucosylation of serotype C capsular polysaccharide in E. faecalis.

  18. Persistence of Enterococcus faecalis in Aquatic Environments via Surface Interactions with Copepods

    PubMed Central

    Signoretto, Caterina; Burlacchini, Gloria; Pruzzo, Carla; Canepari, Pietro

    2005-01-01

    Several human pathogens and fecal-pollution indicators may persist as viable organisms in natural environments, owing to their ability to activate different types of survival strategies. These strategies include adhesion on both abiotic and biotic surfaces and the entrance to the so-called viable but nonculturable (VBNC) state. In an 18-month survey for the detection of enterococci in both lake water and seawater, C. Signoretto et al. (Appl. Environ. Microbiol. 70:6892-6896, 2004) have shown that Enterococcus faecalis was detected mostly bound to plankton and in the VBNC state. In the present study, we show that in vitro adhesion of E. faecalis to copepods accelerated the entry of cells into the VBNC state relative to that of planktonic bacteria. VBNC E. faecalis cells maintained their adhesive properties to copepods and chitin (the main component of the copepod carapace), though to a reduced extent in comparison with growing cells. Sugar competition experiments showed interference with adhesion to both copepods and chitin by GlcNAc and only to copepods by d-mannose. Four enterococcal cell wall proteins present in both growing and VBNC cells and lipoteichoic acid were shown to be capable of binding chitin. The results indicate that copepods may represent an additional environmental reservoir of enterococci, thus suggesting the advisability of redesigning the protocols currently used for microbial detection during the evaluation of the microbiological quality of environmental samples. PMID:15870369

  19. Competitive polymerase chain reaction for quantification of nonculturable Enterococcus faecalis cells in lake water.

    PubMed

    del Mar Lleó M; Tafi; Signoretto; Dal Cero C; Canepari

    1999-12-01

    Among the survival strategies developed by bacteria when faced with adverse environmental conditions, the viable but nonculturable (VNC) state has been described. In this state, bacteria are unable to form colonies but are still alive and capable of metabolic activity. The VNC state has been described in numerous Gram-negative species, but recently also in Enterococcus faecalis, a Gram-positive species which can be found in the environment. In this study we describe a competitive PCR (cPCR) protocol to detect and quantify a specific sequence of DNA from culturable and nonculturable E. faecalis cells present in water samples. The protocol was found to be specific and capable of detecting amounts of DNA up to 0.1 pg corresponding to approximately 2 cells ml(-1). Moreover, it allows an internal standard to be used to quantify the amount of specific DNA present in samples from different environments. The application of this cPCR method to water samples from Lake Garda enabled us to demonstrate the presence of nonculturable forms of E. faecalis in lake water and to quantify their DNA and the corresponding concentration of nonculturable cells.

  20. Identification of the Enterococcus faecalis Tyrosine Decarboxylase Operon Involved in Tyramine Production

    PubMed Central

    Connil, Nathalie; Le Breton, Yoann; Dousset, Xavier; Auffray, Yanick; Rincé, Alain; Prévost, Hervé

    2002-01-01

    Screening of a library of Enterococcus faecalis insertional mutants allowed isolation of a mutant affected in tyramine production. The growth of this mutant was similar to that of the wild-type E. faecalis JH2-2 strain in Maijala broth, whereas high-performance liquid chromatography analyses showed that tyramine production, which reached 1,000 μg ml−1 for the wild-type strain, was completely abolished. Genetic analysis of the insertion locus revealed a gene encoding a decarboxylase with similarity to eukaryotic tyrosine decarboxylases. Sequence analysis revealed a pyridoxal phosphate binding site, indicating that this enzyme belongs to the family of amino acid decarboxylases using this cofactor. Reverse transcription-PCR analyses demonstrated that the gene (tdc) encoding the putative tyrosine decarboxylase of E. faecalis JH2-2 is cotranscribed with the downstream gene encoding a putative tyrosine-tyramine antiporter and with the upstream tyrosyl-tRNA synthetase gene. This study is the first description of a tyrosine decarboxylase gene in prokaryotes. PMID:12089039

  1. Inhibition of initial adhesion of uropathogenic Enterococcus faecalis by biosurfactants from Lactobacillus isolates.

    PubMed Central

    Velraeds, M M; van der Mei, H C; Reid, G; Busscher, H J

    1996-01-01

    In this study, 15 Lactobacillus isolates were found to produce biosurfactants in the mid-exponential and stationary growth phases. The stationary-phase biosurfactants from lactobacillus casei subsp. rhamnosus 36 and ATCC 7469, Lactobacillus fermentum B54, and Lactobacillus acidophilus RC14 were investigated further to determine their capacity to inhibit the initial adhesion of uropathogenic Enterococcus faecalis 1131 to glass in a parallel-plate flow chamber. The initial deposition rate of E. faecalis to glass with an adsorbed biosurfactant layer from L. acidophilus RC14 or L. fermentum B54 was significantly decreased by approximately 70%, while the number of adhering enterococci after 4 h of adhesion was reduced by an average of 77%. The surface activity of the biosurfactants and their activity inhibiting the initial adhesion of E. faecalis 1131 were retained after dialysis (molecular weight cutoff, 6,000 to 8,000) and freeze-drying. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy revealed that the freeze-dried biosurfactants from L. acidophilus RC14 and L. fermentum B54 were richest in protein, while those from L. casei subsp. rhamnosus 36 and ATCC 7469 had relatively high polysaccharide and phosphate contents. PMID:8787394

  2. Enterococcus faecalis zinc-responsive proteins mediate bacterial defence against zinc overload, lysozyme and oxidative stress.

    PubMed

    Abrantes, Marta C; Kok, Jan; Silva Lopes, Maria de Fátima

    2014-12-01

    Two Enterococcus faecalis genes encoding the P-type ATPase EF1400 and the putative SapB protein EF0759 were previously shown to be strongly upregulated in the presence of high concentrations of zinc. In the present work, we showed that a Zn(2+)-responsive DNA-binding motif (zim) is present in the promoter regions of these genes. Both proteins were further studied with respect to their involvement in zinc homeostasis and invasion of the host. EF0759 contributed to intramacrophage survival by an as-yet unknown mechanism(s). EF1400, here renamed ZntAEf, is an ATPase with specificity for zinc and plays a role in dealing with several host defences, i.e. zinc overload, oxidative stress and lysozyme; it provides E. faecalis cells with the ability to survive inside macrophages. As these three host defence mechanisms are important at several sites in the host, i.e. inside macrophages and in saliva, this work suggested that ZntAEf constitutes a crucial E. faecalis defence mechanism that is likely to contribute to the ability of this bacterium to endure life inside its host.

  3. The Spx Regulator Modulates Stress Responses and Virulence in Enterococcus faecalis

    PubMed Central

    Kajfasz, Jessica K.; Mendoza, Jorge E.; Gaca, Anthony O.; Miller, James H.; Koselny, Kristy A.; Giambiagi-deMarval, Marcia; Wellington, Melanie; Abranches, Jacqueline

    2012-01-01

    The ability to cope with endogenous or host-generated reactive oxygen species is considered a key virulence attribute of the opportunistic pathogen Enterococcus faecalis, a leading cause of hospital-acquired infections. In this study, we used in silico and mutational analyses to identify and characterize the role of the Spx global regulator in oxidative stress tolerance and virulence in E. faecalis. While the Δspx strain grew as well as the wild-type strain under anaerobic conditions, the mutant strain exhibited impaired growth under aerobic conditions and was highly sensitive to oxidative stress agents. The spx mutant strain was also sensitive to a variety of other stressful conditions, including antibiotic stress and killing by the mouse-derived macrophage cell line J774. Using a murine model of foreign body-associated peritonitis, we demonstrated that the ability of the Δspx strain to colonize the peritoneum and disseminate in the bloodstream was significantly reduced compared to that of the parent strain. Transcriptional analysis revealed that a large number of known oxidative stress genes are under positive control by Spx. Collectively, our results show that Spx is a major stress gene regulator and is implicated in the pathophysiology of E. faecalis. The relationship of Spx to other oxidative stress regulators is also discussed. PMID:22508863

  4. Pheromone killing of multidrug-resistant Enterococcus faecalis V583 by native commensal strains

    PubMed Central

    Gilmore, Michael S.; Rauch, Marcus; Ramsey, Matthew M.; Himes, Paul R.; Varahan, Sriram; Manson, Janet M.; Lebreton, Francois; Hancock, Lynn Ernest

    2015-01-01

    Multidrug-resistant Enterococcus faecalis possess numerous mobile elements that encode virulence and antibiotic resistance traits as well as new metabolic pathways, often constituting over one-quarter of the genome. It was of interest to determine how this large accretion of mobile elements affects competitive growth in the gastrointestinal (GI) tract consortium. We unexpectedly observed that the prototype clinical isolate strain V583 was actively killed by GI tract flora, whereas commensal enterococci flourished. It was found that killing of V583 resulted from lethal cross-talk between accumulated mobile elements and that this cross-talk was induced by a heptapeptide pheromone produced by native E. faecalis present in the fecal consortium. These results highlight two important aspects of the evolution of multidrug-resistant enterococci: (i) the accretion of mobile elements in E. faecalis V583 renders it incompatible with commensal strains, and (ii) because of this incompatibility, multidrug-resistant strains sharing features found in V583 cannot coexist with commensal strains. The accumulation of mobile elements in hospital isolates of enterococci can include those that are inherently incompatible with native flora, highlighting the importance of maintaining commensal populations as means of preventing colonization and subsequent infection by multidrug-resistant strains. PMID:26039987

  5. Genome-based characterization of hospital-adapted Enterococcus faecalis lineages

    PubMed Central

    Raven, Kathy E.; Reuter, Sandra; Gouliouris, Theodore; Reynolds, Rosy; Russell, Julie E.; Brown, Nicholas M.; Török, M. Estée; Parkhill, Julian; Peacock, Sharon J.

    2016-01-01

    Vancomycin-resistant Enterococcus faecalis (VREfs) is an important nosocomial pathogen1,2. We undertook whole genome sequencing of E. faecalis associated with bloodstream infection in the UK and Ireland over more than a decade to determine the population structure and genetic associations with hospital adaptation. Three lineages predominated in the population, two of which (L1 and L2) were nationally distributed, and one (L3) geographically restricted. Genome comparison with a global collection identified that L1 and L3 were also present in the USA, but were genetically distinct. Over 90% of VREfs belonged to L1–L3, with resistance acquired and lost multiple times in L1 and L2, but only once followed by clonal expansion in L3. Putative virulence and antibiotic resistance genes were over-represented in L1, L2 and L3 isolates combined, versus the remainder. Each of the three main lineages contained a mixture of vancomycin-resistant and -susceptible E. faecalis (VSEfs), which has important implications for infection control and antibiotic stewardship. PMID:27213049

  6. Enterococcus faecalis reconfigures its gene regulatory network activation under copper exposure

    PubMed Central

    Latorre, Mauricio; Galloway-Peña, Jessica; Roh, Jung Hyeob; Budinich, Marko; Reyes-Jara, Angélica; Murray, Barbara E.; Maass, Alejandro; González, Mauricio

    2014-01-01

    A gene regulatory network was generated in the bacterium Enterococcus faecalis in order to understand how this organism can activate its expression under different copper concentrations. The topological evaluation of the network showed common patterns described in other organisms. Integrating microarray experiments allowed the identification of sub-networks activated under low (0.05 mM CuSO4) and high (0.5 mM CuSO4) copper concentrations. The analysis indicates the presence of specific functionally activated modules induced by copper, highlighting the regulons LysR, ArgR as global regulators and CopY, Fur and LexA as local regulators. Taking advantage of the fact that E. faecalis presented a homeostatic module isolated, we produced an in vivo intervention removing this system from the cell without affecting the connectivity of the global transcriptional network. This strategy led us to find that this bacterium can reconfigure its gene expression to maintain cellular homeostasis, activating new modules principally related to glucose metabolism and transcriptional processes. Finally, these results position E. faecalis as the organism having the most complete and controllable systemic model of copper homeostasis available to date. PMID:24382465

  7. Genome-based characterization of hospital-adapted Enterococcus faecalis lineages.

    PubMed

    Raven, Kathy E; Reuter, Sandra; Gouliouris, Theodore; Reynolds, Rosy; Russell, Julie E; Brown, Nicholas M; Török, M Estée; Parkhill, Julian; Peacock, Sharon J

    2016-01-01

    Vancomycin-resistant Enterococcus faecalis (VREfs) is an important nosocomial pathogen(1,2). We undertook whole genome sequencing of E. faecalis associated with bloodstream infection in the UK and Ireland over more than a decade to determine the population structure and genetic associations with hospital adaptation. Three lineages predominated in the population, two of which (L1 and L2) were nationally distributed, and one (L3) geographically restricted. Genome comparison with a global collection identified that L1 and L3 were also present in the USA, but were genetically distinct. Over 90% of VREfs belonged to L1-L3, with resistance acquired and lost multiple times in L1 and L2, but only once followed by clonal expansion in L3. Putative virulence and antibiotic resistance genes were over-represented in L1, L2 and L3 isolates combined, versus the remainder. Each of the three main lineages contained a mixture of vancomycin-resistant and -susceptible E. faecalis (VSEfs), which has important implications for infection control and antibiotic stewardship. PMID:27572164

  8. Identification of nor-β-lapachone derivatives as potential antibacterial compounds against Enterococcus faecalis clinical strain.

    PubMed

    Lourenço, André L; Abreu, Paula A; Leal, Bruno; da Silva Júnior, Eufrânio N; Pinto, Antonio V; Pinto, Maria do Carmo F R; Souza, Alessandra M T; Novais, Juliana S; Paiva, Marcela B; Cabral, Lucio M; Rodrigues, Carlos R; Ferreira, Vitor F; Castro, Helena C

    2011-02-01

    A broad-spectrum antibiotic therapy has led to medical complications and emergence of multiresistant bacteria including Enterococcus faecalis. In this study, we designed, synthesized, and evaluated the antibacterial activity of 13 nor-β-lapachone derivatives against a drug resistant E. faecalis strain. Two triazole substituted compounds (1e = 8 μg/ml and 1c = 16 μg/ml) and the non-substituted derivative (1a = 8 μg/ml) were promising compared to chloramphenicol (12 μg/ml), an antibiotic currently available in the market. We also performed a structure-activity relationship analysis using a molecular modeling approach that pointed the low HOMO energy values; HOMO density concentrated on the nor-β-lapachone ring, lipophilicity, solubility and number HBA as important stereoelectronic features for the antibacterial profile. In addition the triazole compounds presented low theoretical toxicity profile, and drug-score higher than commercial antibiotics also fulfilling the Lipinski "Rule of Five", which pointed them as promising candidates for further studies in infections caused by multiresistant E. faecalis hospital strains.

  9. The effect of Carvacrol on Enterococcus faecalis as a final irrigant

    PubMed Central

    Nosrat, Ali; Bolhari, Behnam; Sharifian, Mohammad Reza; Aligholi, Marziyeh; Mortazavi, Mahsa Sadat

    2009-01-01

    INTRODUCTION: Sodium hypochlorite (NaOCl) is an effective antimicrobial irrigant, however its toxic effects and deterrent odor are not ideal. Carvacrol is an edible plant extract with anti-inflammatory and anti-bacterial properties that is effective against Enterococcus (E) faecalis. The aim of this study was to evaluate Carvacrol's antibacterial efficacy against E. faecalis bacteria as a final irrigant. MATERIALS AND METHODS: Forty extracted single-rooted human teeth were utilized. After mechanical preparations, samples were randomly divided into three experimental (A, B and C) and two control groups. E. faecalis was cultured in both experimental and positive control groups. After bacterial counting in all canals, 5.25% NaOCl, 0.6% Carvacrol emulsion and MTAD were used as final irrigants in groups A, B and C respectively. Data were analyzed using Kruskal-Wallis and Mann-Whitney U tests. RESULTS: There was no meaningful difference in bacterial reduction between groups A and B; however, group C showed significantly lower efficacy compared to other groups (P<0.05). CONCLUSION: The 0.6% Carvacrol disinfects root canals effectively. It also has anti-inflammatory qualities and therefore may be an acceptable alternative for NaOCl. PMID:24003329

  10. Transcriptome profiling of TDC cluster deletion mutant of Enterococcus faecalis V583.

    PubMed

    Perez, Marta; Ladero, Victor; Del Rio, Beatriz; Redruello, Begoña; de Jong, Anne; Kuipers, Oscar P; Kok, Jan; Martin, M Cruz; Fernandez, Maria; Alvarez, Miguel A

    2016-09-01

    The species Enterococcus faecalis is able to catabolise the amino acid tyrosine into the biogenic amine tyramine by the tyrosine decarboxilase (TDC) pathway Ladero et al. (2012) [1]. The TDC cluster comprises four genes: tyrS, an aminoacyl-tRNA synthetase-like gene; tdcA, which encodes the tyrosine decarboxylase; tyrP, a tyrosine/tyramine exchanger gene and nhaC-2, which encodes an Na(+)/H(+) antiporter and whose role in the tyramine biosynthesis remains unknown [2]. In E. faecalis V583 the last three genes are co-transcribed as a single polycistronic mRNA forming the catabolic operon, while tyrS is transcribed independently of the catabolic genes as a monocistronic mRNA [2]. The catabolic operon is transcriptionally induced by tyrosine and acidic pH. On the opposite, the tyrS expression is repressed by tyrosine concentrations [2]. In this work we report the transcriptional profiling of the TDC cluster deletion mutant (E. faecalis V583 ΔTDC) [2] compared to the wild-type strain, both grown in M17 medium supplemented with tyrosine. The transcriptional profile data of TDC cluster-regulated genes were deposited in the Gene Expression Omnibus (GEO) database under accession no. GSE77864.

  11. Antimicrobial activity of tetraacetylethylenediamine-sodium perborate versus sodium hypochlorite against Enterococcus faecalis

    PubMed Central

    Shakouie, Sahar; Salem Milani, Amin; Eskandarnejad, Mahsa; Rahimi, Saeed; Froughreyhani, Mohammad; Galedar, Saeede; Ranjbar, Ehsan

    2016-01-01

    Background. This study evaluated the antimicrobial activity of Tetraacetylethylenediamine-sodium perborate (TAED-SP) in comparison to 2.5% and 5% sodium hypochlorite (NaOCl) against Enterococcus faecalis. Methods. A standard suspension of E. faecalis was inoculated on 60 plates containing Mueller-Hinton agar culture medium. Four sterile disks of Beckman filtration paper were placed on each plate. TAED-SP, 5% and 2.5% NaOCl were placed on three disks. Sterile physiologic saline was placed on the fourth disk as negative control. After 24-hour incubation, the diameter of the inhibition zone around the disks was measured using a transparent ruler. One-way Analysis of Variance (ANOVA) was used to compare the mean zone of microbial growth in the groups. P-values less than 0.05 were considered statistically significant. Results. There was a significant difference in the diameter of the inhibition zones between groups (P < 0.05). The Tukey post hoc test showed a higher diameter of the inhibitory zone with TAED-SP than that of 2.5% NaOCl. However, there were no significant differences between the inhibitory zones of TAED-SP and 5% NaOCl. Conclusion. TAED-SP and 5% NaOCl have similar antibacterial activity against E. faecalis; however, TAED-SP has a greater antibacterial effect compared to 2.5% NaOCl. PMID:27092214

  12. Draft Genome Sequence of the Bacteriocinogenic Strain Enterococcus faecalis DBH18, Isolated from Mallard Ducks (Anas platyrhynchos)

    PubMed Central

    Arbulu, Sara; Jimenez, Juan J.; Borrero, Juan; Sánchez, Jorge; Frantzen, Cyril; Herranz, Carmen; Nes, Ingolf F.; Cintas, Luis M.; Diep, Dzung B.

    2016-01-01

    Here, we report the draft genome sequence of Enterococcus faecalis DBH18, a bacteriocinogenic lactic acid bacterium (LAB) isolated from mallard ducks (Anas platyrhynchos). The assembly contains 2,836,724 bp, with a G+C content of 37.6%. The genome is predicted to contain 2,654 coding DNA sequences (CDSs) and 50 RNAs. PMID:27417838

  13. Draft Genome Sequence of the Bacteriocinogenic Strain Enterococcus faecalis DBH18, Isolated from Mallard Ducks (Anas platyrhynchos).

    PubMed

    Arbulu, Sara; Jimenez, Juan J; Borrero, Juan; Sánchez, Jorge; Frantzen, Cyril; Herranz, Carmen; Nes, Ingolf F; Cintas, Luis M; Diep, Dzung B; Hernández, Pablo E

    2016-01-01

    Here, we report the draft genome sequence of Enterococcus faecalis DBH18, a bacteriocinogenic lactic acid bacterium (LAB) isolated from mallard ducks (Anas platyrhynchos). The assembly contains 2,836,724 bp, with a G+C content of 37.6%. The genome is predicted to contain 2,654 coding DNA sequences (CDSs) and 50 RNAs. PMID:27417838

  14. The effect of different root canal medicaments on the elimination of Enterococcus faecalis ex vivo

    PubMed Central

    Dammaschke, Till; Jung, Nina; Harks, Inga; Schafer, Edgar

    2013-01-01

    Objective: The aim of this study was to evaluate the antimicrobial effect of chlorhexidine gel (CHX-G) 2%, chlorhexidine powder (CHX-P) 1%, povidone-iodine (PVP-I), polyhexanide and camphorated-and-mentholated chlorophenol (ChKM) ex vivo. Materials and Methods: For every medicament group 10 root segments (15 mm long) of extracted human teeth were prepared to ISO-size 45 and sterilized (n = 50). The root segments were then inoculated with Enterococcus faecalis and aerobically incubated at 37°C. After 1 week, ten root canals were filled with one of the medicaments, respectively and aerobically incubated at 37°C for another week. Ten teeth served as positive controls and were filled with sterile saline solution. After 7 days, the medicaments were inactivated and all root canals were instrumented to ISO-size 50. The obtained dentin samples were dispersed in Ringer solution followed by the preparation of serial dilutions. 10 μl per sample were applied to an agar plate and incubated at 37°C for 48 h. The colony forming units were counted and the reduction factors (RFs) were calculated and statistically analyzed. Results: Compared with the positive controls all medicaments exhibited an antibacterial effect against E. faecalis. The RFs for CHX-G, CHX-P and ChKM were significantly higher compared to PVP-I and polyhexanide (P < 0.05). In contrast to PVP-I and polyhexanide, CHX-G, CHX-P and ChKM were able to eliminate E. faecalis from all dentin samples. Conclusions: Within the limitations of this ex vivo investigation, 2% CHX-G and CHX-P were as effective as ChKM against E. faecalis. Thus, when choosing a root canal medicament the better biocompatibility of CHX compared with ChKM should be taken in consideration. PMID:24932119

  15. Antibacterial Activity of Diode Laser and Sodium Hypochlorite in Enterococcus Faecalis-Contaminated Root Canals

    PubMed Central

    Sohrabi, Khosrow; Sooratgar, Aidin; Zolfagharnasab, Kaveh; Kharazifard, Mohammad Javad; Afkhami, Farzaneh

    2016-01-01

    Introduction: The aim of the present in vitro study was to evaluate the disinfection ability of 980-nm diode laser in comparison with sodium hypochlorite (NaOCl) as a common root canal irrigant in canals infected with Enterococcus faecalis (E. faecalis). Methods and Materials: The root canals of 18 extracted single-rooted premolars were prepared by rotary system. After decoronation, the roots were autoclaved. One specimen was chosen for the negative control, and the remaining teeth were incubated with E. faecalis suspension for two weeks. Subsequently, one specimen was selected as the positive control and the remaining samples were divided into two groups (n=8). The samples of the first group were irrigated with 5.25% NaOCl and the second group were treated with a 980-nm diode laser. Microbial samples were taken from the root canals and bacterial cultivation was carried out. The average value and the standard deviation of colony-forming units (CFU) of each specimen were measured using descriptive statistics. The student’s t-test was used to compare the reduction in CFU in each group. The equality of variance of CFU was measured by the Levene’s test. Results: NaOCl resulted in 99.87% removal of the bacteria and showed significantly more antibacterial effect compared to the 980-nm diode laser which led to 96.56% bacterial reduction (P<0.05). Conclusion: Although 5.25% NaOCl seems to reduce E. faecalis more effectively, the diode laser also reduced the bacterial count. Therefore a 980-nm diode laser could be considered as a complementary disinfection method in root canal treatment. PMID:26843870

  16. Effect of chitosan-ethylenediamine tetraacetic acid on Enterococcus faecalis dentinal biofilm and smear layer removal

    PubMed Central

    Geethapriya, Nagarajan; Subbiya, Arunajatesan; Padmavathy, Kesavaram; Mahalakshmi, Krishnan; Vivekanandan, Paramasivam; Sukumaran, Virudhachalam Ganapathy

    2016-01-01

    Objective: The objective of the study was to evaluate the effectiveness of chitosan and chitosan-ethylenediamine tetraacetic acid (EDTA) (3:1,1:1,1:3) in comparison with 5.2% sodium hypochlorite (NaOCl) in disinfecting Enterococcus faecalis biofilm on root canal dentin and in the removal of smear layer with minimal erosion. Materials and Methods: Seventy single-rooted extracted human mandibular premolars (n = 70) were selected for the study. Forty tooth samples were biomechanically prepared, vertically sectioned, and sterilized by autoclaving. The tooth sections were artificially infected with E. faecalis (ATCC 29212 [n = 35] and clinical isolate [SBEF2, n = 35]) to form mature dentinal biofilm in vitro. The tooth samples were treated with the test solutions: chitosan and chitosan-EDTA (3:1, 1:1, 1:3), and the killing time was determined. The smear layer removal ability of the test solutions (Group A: chitosan-EDTA [1:1], Group B: EDTA, Group C: control) (n = 10 tooth/group) was assessed. Results: Chitosan and chitosan-EDTA (3:1, 1:1, 1:3) exhibited antibacterial activity against both the strains of E. faecalis. Chitosan and chitosan-EDTA caused 3 log reduction in the viable count of the sessile cells of E. faecalis at 15 min while 5.2% NaOCl exhibited 99.98% inhibition at 15 min. Chitosan-EDTA (1:1) was found to be effective in removing the smear layer and showed lesser erosion than EDTA at the coronal and middle portions. Conclusion: Chitosan-EDTA (1:1) is a potential root canal irrigant that performs a dual role – root canal disinfection and smear layer removal. PMID:27656070

  17. Global Regulation of Gene Expression by the MafR Protein of Enterococcus faecalis

    PubMed Central

    Ruiz-Cruz, Sofía; Espinosa, Manuel; Goldmann, Oliver; Bravo, Alicia

    2016-01-01

    Enterococcus faecalis is a natural inhabitant of the human gastrointestinal tract. However, as an opportunistic pathogen, it is able to colonize other host niches and cause life-threatening infections. Its adaptation to new environments involves global changes in gene expression. The EF3013 gene (here named mafR) of E. faecalis strain V583 encodes a protein (MafR, 482 residues) that has sequence similarity to global response regulators of the Mga/AtxA family. The enterococcal OG1RF genome also encodes the MafR protein (gene OG1RF_12293). In this work, we have identified the promoter of the mafR gene using several in vivo approaches. Moreover, we show that MafR influences positively the transcription of many genes on a genome-wide scale. The most significant target genes encode components of PTS-type membrane transporters, components of ABC-type membrane transporters, and proteins involved in the metabolism of carbon sources. Some of these genes were previously reported to be up-regulated during the growth of E. faecalis in blood and/or in human urine. Furthermore, we show that a mafR deletion mutant strain induces a significant lower degree of inflammation in the peritoneal cavity of mice, suggesting that enterococcal cells deficient in MafR are less virulent. Our work indicates that MafR is a global transcriptional regulator. It might facilitate the adaptation of E. faecalis to particular host niches and, therefore, contribute to its potential virulence. PMID:26793169

  18. Effect of chitosan-ethylenediamine tetraacetic acid on Enterococcus faecalis dentinal biofilm and smear layer removal

    PubMed Central

    Geethapriya, Nagarajan; Subbiya, Arunajatesan; Padmavathy, Kesavaram; Mahalakshmi, Krishnan; Vivekanandan, Paramasivam; Sukumaran, Virudhachalam Ganapathy

    2016-01-01

    Objective: The objective of the study was to evaluate the effectiveness of chitosan and chitosan-ethylenediamine tetraacetic acid (EDTA) (3:1,1:1,1:3) in comparison with 5.2% sodium hypochlorite (NaOCl) in disinfecting Enterococcus faecalis biofilm on root canal dentin and in the removal of smear layer with minimal erosion. Materials and Methods: Seventy single-rooted extracted human mandibular premolars (n = 70) were selected for the study. Forty tooth samples were biomechanically prepared, vertically sectioned, and sterilized by autoclaving. The tooth sections were artificially infected with E. faecalis (ATCC 29212 [n = 35] and clinical isolate [SBEF2, n = 35]) to form mature dentinal biofilm in vitro. The tooth samples were treated with the test solutions: chitosan and chitosan-EDTA (3:1, 1:1, 1:3), and the killing time was determined. The smear layer removal ability of the test solutions (Group A: chitosan-EDTA [1:1], Group B: EDTA, Group C: control) (n = 10 tooth/group) was assessed. Results: Chitosan and chitosan-EDTA (3:1, 1:1, 1:3) exhibited antibacterial activity against both the strains of E. faecalis. Chitosan and chitosan-EDTA caused 3 log reduction in the viable count of the sessile cells of E. faecalis at 15 min while 5.2% NaOCl exhibited 99.98% inhibition at 15 min. Chitosan-EDTA (1:1) was found to be effective in removing the smear layer and showed lesser erosion than EDTA at the coronal and middle portions. Conclusion: Chitosan-EDTA (1:1) is a potential root canal irrigant that performs a dual role – root canal disinfection and smear layer removal.

  19. Novel Structural Components Contribute to the High Thermal Stability of Acyl Carrier Protein from Enterococcus faecalis.

    PubMed

    Park, Young-Guen; Jung, Min-Cheol; Song, Heesang; Jeong, Ki-Woong; Bang, Eunjung; Hwang, Geum-Sook; Kim, Yangmee

    2016-01-22

    Enterococcus faecalis is a Gram-positive, commensal bacterium that lives in the gastrointestinal tracts of humans and other mammals. It causes severe infections because of high antibiotic resistance. E. faecalis can endure extremes of temperature and pH. Acyl carrier protein (ACP) is a key element in the biosynthesis of fatty acids responsible for acyl group shuttling and delivery. In this study, to understand the origin of high thermal stabilities of E. faecalis ACP (Ef-ACP), its solution structure was investigated for the first time. CD experiments showed that the melting temperature of Ef-ACP is 78.8 °C, which is much higher than that of Escherichia coli ACP (67.2 °C). The overall structure of Ef-ACP shows the common ACP folding pattern consisting of four α-helices (helix I (residues 3-17), helix II (residues 39-53), helix III (residues 60-64), and helix IV (residues 68-78)) connected by three loops. Unique Ef-ACP structural features include a hydrophobic interaction between Phe(45) in helix II and Phe(18) in the α1α2 loop and a hydrogen bonding between Ser(15) in helix I and Ile(20) in the α1α2 loop, resulting in its high thermal stability. Phe(45)-mediated hydrophobic packing may block acyl chain binding subpocket II entry. Furthermore, Ser(58) in the α2α3 loop in Ef-ACP, which usually constitutes a proline in other ACPs, exhibited slow conformational exchanges, resulting in the movement of the helix III outside the structure to accommodate a longer acyl chain in the acyl binding cavity. These results might provide insights into the development of antibiotics against pathogenic drug-resistant E. faecalis strains.

  20. The Heterodimeric ABC Transporter EfrCD Mediates Multidrug Efflux in Enterococcus faecalis

    PubMed Central

    Hürlimann, Lea M.; Corradi, Valentina; Hohl, Michael; Bloemberg, Guido V.; Tieleman, D. Peter

    2016-01-01

    Nosocomial infections with Enterococcus faecalis are an emerging health problem. However, drug efflux pumps contributing to intrinsic drug resistance are poorly studied in this Gram-positive pathogen. In this study, we functionally investigated seven heterodimeric ABC transporters of E. faecalis that are annotated as drug efflux pumps. Deletion of ef0789-ef0790 on the chromosome of E. faecalis resulted in increased susceptibility to daunorubicin, doxorubicin, ethidium, and Hoechst 33342, and the corresponding transporter was named EfrCD. Unexpectedly, the previously described heterodimeric multidrug ABC transporter EfrAB contributes marginally to drug efflux in the endogenous context of E. faecalis. In contrast, heterologous expression in Lactococcus lactis revealed that EfrAB, EfrCD, and the product of ef2226-ef2227 (EfrEF) mediate the efflux of fluorescent substrates and confer resistance to multiple dyes and drugs, including fluoroquinolones. Four of seven transporters failed to exhibit drug efflux activity for the set of drugs and dyes tested, even upon overexpression in L. lactis. Since all seven transporters were purified as heterodimers after overexpression in L. lactis, a lack of drug efflux activity is not attributed to poor expression or protein aggregation. Reconstitution of the purified multidrug transporters EfrAB, EfrCD, and EfrEF in proteoliposomes revealed functional coupling between ATP hydrolysis and drug binding. Our analysis creates an experimental basis for the accurate prediction of drug efflux transporters and indicates that many annotated multidrug efflux pumps might be incapable of drug transport and thus might fulfill other physiological functions in the cell. PMID:27381387

  1. Expression of Alzheimer-Type Neurofibrillary Epitopes in Primary Rat Cortical Neurons Following Infection with Enterococcus faecalis

    PubMed Central

    Underly, Robert; Song, Mee-Sook; Dunbar, Gary L.; Weaver, Charles L.

    2016-01-01

    The neurofibrillary tau pathology and amyloid deposits seen in Alzheimer’s disease (AD) also have been seen in bacteria-infected brains. However, few studies have examined the role of these bacteria in the generation of tau pathology. One suggested link between infection and AD is edentulism, the complete loss of teeth. Edentulism can result from chronic periodontal disease due to infection by Enterococcus faecalis. The current study assessed the ability to generate early Alzheimer-like neurofibrillary epitopes in primary rat cortical neurons through bacterial infection by E. faecalis. Seven-day old cultured neurons were infected with E. faecalis for 24 and 48 h. An upward molecular weight shift in tau by Western blotting (WB) and increased appearance of tau reactivity in cell bodies and degenerating neurites was found in the 48 h infection group for the antibody CP13 (phospho-Serine 202). A substantial increase in reactivity of Alz-50 was seen at 24 and 48 h after infection. Furthermore, extensive microtubule-associated protein 2 (MAP2) reactivity also was seen at 24 and 48 h post-infection. Our preliminary findings suggest a potential link between E. faecalis infection and intracellular changes that may help facilitate early AD-like neurofibrillary pathology. HighlightsEnterococcus faecalis used in the generation of AD neurofibrillary epitopes in rat.Infection increases Alz-50, phospho-Serine 202 tau, and MAP2 expression.Infection by Enterococcus may play a role in early Alzheimer neurofibrillary changes. PMID:26834627

  2. The sex pheromone system of Enterococcus faecalis. More than just a plasmid-collection mechanism?

    PubMed

    Wirth, R

    1994-06-01

    The sex pheromone system of Enterococcus faecalis was discovered by observing a clumping reaction of E. faecalis strains during conjugative transfer of plasmids. It was found that only a special type of E. faecalis plasmids, the so-called sex pheromone plasmids, are transferred via this mechanism. Various experiments, especially by the group of D. B. Clewell, led to the formulation of a model describing how the sex pheromone system works. Small linear peptides, the so-called sex pheromones, are excreted by strains not possessing the corresponding sex pheromone plasmid. Donor strains harboring the plasmid do not produce the corresponding sex pheromone; they react to the presence of the peptide by production of a plasmid-encoded adhesin, the so-called aggregation substance. This adhesin allows contact between the non-motile mating partners; after conjugative transfer of the plasmid, the former recipient possesses and replicates the new plasmid. Thereby the population of E. faecalis strains is shifted to a high percentage of donor strains. This is especially true because a donor strain will still excrete sex pheromones corresponding to plasmids it does not harbor; therefore, such a strain can also function as recipient for other sex pheromone plasmids it does not possess. Various aspects of this unique plasmid collection mechanism have been studied during the last few years. The data indicate that, with the exception of pAM373, all sex pheromone plasmids possess one DNA region which is highly similar to and codes for the adhesin. It is also becoming more and more clear that regulatory functions/proteins are not conserved between different sex pheromone plasmids. Induction of adhesin synthesis needs the action of a regulatory cascade composed of unique features; at the moment we are just beginning to understand this cascade. By sequencing the first structural gene for one of those adhesins, we realized that the aggregation substance might act also as an adhesin for

  3. Virulence Genes among Enterococcus faecalis and Enterococcus faecium Isolated from Coastal Beaches and Human and Nonhuman Sources in Southern California and Puerto Rico

    PubMed Central

    Talavera, Ginamary Negrón; Hernández, Luis A. Ríos; Ambrose, Richard F.; Jay, Jennifer A.

    2016-01-01

    Most Enterococcus faecalis and E. faecium are harmless to humans; however, strains harboring virulence genes, including esp, gelE, cylA, asa1, and hyl, have been associated with human infections. E. faecalis and E. faecium are present in beach waters worldwide, yet little is known about their virulence potential. Here, multiplex PCR was used to compare the distribution of virulence genes among E. faecalis and E. faecium isolated from beaches in Southern California and Puerto Rico to isolates from potential sources including humans, animals, birds, and plants. All five virulence genes were found in E. faecalis and E. faecium from beach water, mostly among E. faecalis. gelE was the most common among isolates from all source types. There was a lower incidence of asa1, esp, cylA, and hyl genes among isolates from beach water, sewage, septage, urban runoff, sea wrack, and eelgrass as compared to human isolates, indicating that virulent strains of E. faecalis and E. faecium may not be widely disseminated at beaches. A higher frequency of asa1 and esp among E. faecalis from dogs and of asa1 among birds (mostly seagull) suggests that further studies on the distribution and virulence potential of strains carrying these genes may be warranted. PMID:27144029

  4. Virulence Genes among Enterococcus faecalis and Enterococcus faecium Isolated from Coastal Beaches and Human and Nonhuman Sources in Southern California and Puerto Rico.

    PubMed

    Ferguson, Donna M; Talavera, Ginamary Negrón; Hernández, Luis A Ríos; Weisberg, Stephen B; Ambrose, Richard F; Jay, Jennifer A

    2016-01-01

    Most Enterococcus faecalis and E. faecium are harmless to humans; however, strains harboring virulence genes, including esp, gelE, cylA, asa1, and hyl, have been associated with human infections. E. faecalis and E. faecium are present in beach waters worldwide, yet little is known about their virulence potential. Here, multiplex PCR was used to compare the distribution of virulence genes among E. faecalis and E. faecium isolated from beaches in Southern California and Puerto Rico to isolates from potential sources including humans, animals, birds, and plants. All five virulence genes were found in E. faecalis and E. faecium from beach water, mostly among E. faecalis. gelE was the most common among isolates from all source types. There was a lower incidence of asa1, esp, cylA, and hyl genes among isolates from beach water, sewage, septage, urban runoff, sea wrack, and eelgrass as compared to human isolates, indicating that virulent strains of E. faecalis and E. faecium may not be widely disseminated at beaches. A higher frequency of asa1 and esp among E. faecalis from dogs and of asa1 among birds (mostly seagull) suggests that further studies on the distribution and virulence potential of strains carrying these genes may be warranted.

  5. Virulence Genes among Enterococcus faecalis and Enterococcus faecium Isolated from Coastal Beaches and Human and Nonhuman Sources in Southern California and Puerto Rico.

    PubMed

    Ferguson, Donna M; Talavera, Ginamary Negrón; Hernández, Luis A Ríos; Weisberg, Stephen B; Ambrose, Richard F; Jay, Jennifer A

    2016-01-01

    Most Enterococcus faecalis and E. faecium are harmless to humans; however, strains harboring virulence genes, including esp, gelE, cylA, asa1, and hyl, have been associated with human infections. E. faecalis and E. faecium are present in beach waters worldwide, yet little is known about their virulence potential. Here, multiplex PCR was used to compare the distribution of virulence genes among E. faecalis and E. faecium isolated from beaches in Southern California and Puerto Rico to isolates from potential sources including humans, animals, birds, and plants. All five virulence genes were found in E. faecalis and E. faecium from beach water, mostly among E. faecalis. gelE was the most common among isolates from all source types. There was a lower incidence of asa1, esp, cylA, and hyl genes among isolates from beach water, sewage, septage, urban runoff, sea wrack, and eelgrass as compared to human isolates, indicating that virulent strains of E. faecalis and E. faecium may not be widely disseminated at beaches. A higher frequency of asa1 and esp among E. faecalis from dogs and of asa1 among birds (mostly seagull) suggests that further studies on the distribution and virulence potential of strains carrying these genes may be warranted. PMID:27144029

  6. Interference in Pheromone-Responsive Conjugation of a High-Level Bacitracin Resistant Enterococcus faecalis Plasmid of Poultry Origin

    PubMed Central

    Tremblay, Cindy-Love; Archambault, Marie

    2013-01-01

    The current study reports on contact interference of a high-level bacitracin- resistant pheromone-responsive plasmid of Enterococcus faecalis strain 543 of poultry origin during conjugative transfer of bcr antimicrobial resistance genes using a polyclonal antiserum aggregation substance44–560 (AS). After induction with pheromones produced by the recipient strain E. faecalis JH2-2, clumping of the donor E. faecalis strain 543 was observed as well as high transfer frequencies of bcr in short time broth mating. Filter mating assays from donor strain E. faecalis 543 to the recipient strain E. faecalis JH2-2 revealed conjugative transfer of asa1 (AS), bcrRAB and traB (negative regulator pheromone response) genes. The presence of these genes in transconjugants was confirmed by antimicrobial susceptibility testing, PCR, Southern hybridization and sequencing. A significant reduction in formation of aggregates was observed when the polyclonal anti-AS44–560 was added in the pheromone-responsive conjugation experiments as compared to the induced state. Moreover, interference of anti-AS44–560 antibodies in pheromone-responsive conjugation was demonstrated by a reduction in horizontal transfer of asa1 and bcr genes between E. faecalis strain 543 and E. faecalis JH2-2. Reducing the pheromone-responsive conjugation of E. faecalis is of interest because of its clinical importance in the horizontal transfer of antimicrobial resistance. PMID:24030654

  7. Comparison of Enterococcus faecium and Enterococcus faecalis Strains Isolated from Water and Clinical Samples: Antimicrobial Susceptibility and Genetic Relationships

    PubMed Central

    Castillo-Rojas, Gonzalo; Mazari-Hiríart, Marisa; Ponce de León, Sergio; Amieva-Fernández, Rosa I.; Agis-Juárez, Raúl A.; Huebner, Johannes; López-Vidal, Yolanda

    2013-01-01

    Enterococci are part of the normal intestinal flora in a large number of mammals, and these microbes are currently used as indicators of fecal contamination in water and food for human consumption. These organisms are considered one of the primary causes of nosocomial and environmental infections due to their ability to survive in the environment and to their intrinsic resistance to antimicrobials. The aims of this study were to determine the biochemical patterns and antimicrobial susceptibilities of Enterococcus faecalis and E. faecium isolates from clinical samples and from water (groundwater, water from the Xochimilco wetland, and treated water from the Mexico City Metropolitan Area) and to determine the genetic relationships among these isolates. A total of 121 enterococcus strains were studied; 31 and 90 strains were isolated from clinical samples and water (groundwater, water from the Xochimilco wetland, and water for agricultural irrigation), respectively. Identification to the species level was performed using a multiplex PCR assay, and antimicrobial profiles were obtained using a commercial kit. Twenty-eight strains were analyzed by pulsed-field gel electrophoresis (PFGE). E. faecium strains isolated from water showed an atypical biochemical pattern. The clinical isolates showed higher resistance to antibiotics than those from water. Both the enterococci isolated from humans, and those isolated from water showed high genetic diversity according to the PFGE analysis, although some strains seemed to be closely related. In conclusion, enterococci isolated from humans and water are genetically different. However, water represents a potential route of transmission to the community and a source of antimicrobial resistance genes that may be readily transmitted to other, different bacterial species. PMID:23560050

  8. Comparison of Enterococcus faecium and Enterococcus faecalis Strains isolated from water and clinical samples: antimicrobial susceptibility and genetic relationships.

    PubMed

    Castillo-Rojas, Gonzalo; Mazari-Hiríart, Marisa; Ponce de León, Sergio; Amieva-Fernández, Rosa I; Agis-Juárez, Raúl A; Huebner, Johannes; López-Vidal, Yolanda

    2013-01-01

    Enterococci are part of the normal intestinal flora in a large number of mammals, and these microbes are currently used as indicators of fecal contamination in water and food for human consumption. These organisms are considered one of the primary causes of nosocomial and environmental infections due to their ability to survive in the environment and to their intrinsic resistance to antimicrobials. The aims of this study were to determine the biochemical patterns and antimicrobial susceptibilities of Enterococcus faecalis and E. faecium isolates from clinical samples and from water (groundwater, water from the Xochimilco wetland, and treated water from the Mexico City Metropolitan Area) and to determine the genetic relationships among these isolates. A total of 121 enterococcus strains were studied; 31 and 90 strains were isolated from clinical samples and water (groundwater, water from the Xochimilco wetland, and water for agricultural irrigation), respectively. Identification to the species level was performed using a multiplex PCR assay, and antimicrobial profiles were obtained using a commercial kit. Twenty-eight strains were analyzed by pulsed-field gel electrophoresis (PFGE). E. faecium strains isolated from water showed an atypical biochemical pattern. The clinical isolates showed higher resistance to antibiotics than those from water. Both the enterococci isolated from humans, and those isolated from water showed high genetic diversity according to the PFGE analysis, although some strains seemed to be closely related. In conclusion, enterococci isolated from humans and water are genetically different. However, water represents a potential route of transmission to the community and a source of antimicrobial resistance genes that may be readily transmitted to other, different bacterial species.

  9. Inductive effects of environmental concentration of atrazine on Escherichia coli and Enterococcus faecalis.

    PubMed

    Koutsotoli, A D; Dimou, D S; Alamanos, Y P; Maipa, V E

    2005-01-01

    Atrazine solutions (0.1, 1, 10 and 100 microg/L) inoculated with Escherichia coli and Enterococcus faecalis under natural conditions significantly increased (p < or = 0.05) the population levels of both test bacteria; it indicates the ability of bacterial cells to degrade atrazine and to use the original compound or its degradation products as nutrient(s). In some cases, alterations in the morphology of the colonies were also observed on selective solid media. Biochemical differentiation was also found and, on the other hand, a loss of culturability was recorded; this suggests that bacteria have entered in a viable but nonculturable state. A re-appearance of the colonies occurred after inoculation on tryptone-soy agar with atrazine. PMID:16408845

  10. Treatment of enterococcus faecalis bacteria by a helium atmospheric cold plasma brush with oxygen addition

    NASA Astrophysics Data System (ADS)

    Chen, Wei; Huang, Jun; Du, Ning; Liu, Xiao-Di; Wang, Xing-Quan; Lv, Guo-Hua; Zhang, Guo-Ping; Guo, Li-Hong; Yang, Si-Ze

    2012-07-01

    An atmospheric cold plasma brush suitable for large area and low-temperature plasma-based sterilization is designed. Results demonstrate that the He/O2 plasma more effectively kills Enterococcus faecalis than the pure He plasma. In addition, the sterilization efficiency values of the He/O2 plasma depend on the oxygen fraction in Helium gas. The atmospheric cold plasma brush using a proper ratio of He/O2 (2.5%) reaches the optimum sterilization efficiency. After plasma treatment, the cell structure and morphology changes can be observed by the scanning electron microscopy. Optical emission measurements indicate that reactive species such as O and OH play a significant role in the sterilization process.

  11. Treatment of enterococcus faecalis bacteria by a helium atmospheric cold plasma brush with oxygen addition

    SciTech Connect

    Chen Wei; Huang Jun; Wang Xingquan; Lv Guohua; Zhang Guoping; Du Ning; Liu Xiaodi; Guo Lihong; Yang Size

    2012-07-01

    An atmospheric cold plasma brush suitable for large area and low-temperature plasma-based sterilization is designed. Results demonstrate that the He/O{sub 2} plasma more effectively kills Enterococcus faecalis than the pure He plasma. In addition, the sterilization efficiency values of the He/O{sub 2} plasma depend on the oxygen fraction in Helium gas. The atmospheric cold plasma brush using a proper ratio of He/O{sub 2} (2.5%) reaches the optimum sterilization efficiency. After plasma treatment, the cell structure and morphology changes can be observed by the scanning electron microscopy. Optical emission measurements indicate that reactive species such as O and OH play a significant role in the sterilization process.

  12. P087, a lactococcal phage with a morphogenesis module similar to an Enterococcus faecalis prophage.

    PubMed

    Villion, Manuela; Chopin, Marie-Christine; Deveau, Hélène; Ehrlich, S Dusko; Moineau, Sylvain; Chopin, Alain

    2009-05-25

    The virulent lactococcal phage P087 was isolated from a dairy environment in 1978. This phage was then recognized as the reference member for one of the ten phage groups currently known to infect Lactococcus lactis strains. The double-stranded DNA genome of this Siphoviridae phage is composed of 60,074 bp and is circularly permuted. Five tRNA and 88 orfs were found within an uncommon genome architecture. Eleven structural proteins were also identified through SDS-PAGE and LC-MS/MS analyses. Of note, 11 translated orfs from the structural module of phage P087 have identities to gene products found in a prophage located in the genome of Enterococcus faecalis V583. The alignment of both genomic sequences suggests that DNA exchanges could occur between these two phages which are infecting low G+C bacteria found in similar ecological niches.

  13. Effect of freezing on photoreactivation of Escherichia coli and Enterococcus faecalis.

    PubMed

    Williams, A; Gao, W; Leung, K T

    2012-06-01

    The effect of freezing on photoreactivation of two strains of Escherichia coli (ATCC strain 25922 and O157:H7 strain 961019) and two strains of Enterococcus faecalis (strain ATCC 51299, vancomycin-resistant and strain ATCC 29212, vancomycin-sensitive) following ultraviolet irradiation were examined. The level of log photoreactivation of the freezing treated test organisms (frozen at -7, -15, or -30 degrees C then thawed at room temperature prior to ultraviolet irradiation) was compared with that of the samples that had not been frozen. Freezing had obvious impact on the response of the test organisms to visible light following ultraviolet irradiation. Significantly lower levels of photoreactivation were observed in the freezing treated cells. The effect of freezing on the ability of the test microbes to photoreactivate seems to be strain and species dependent. Overall, the experimental results suggest that less photoreactivation could be expected if freezing is used as a treatment method prior to ultraviolet disinfection.

  14. Cloning of an Enterococcus faecalis endocarditis antigen: homology with adhesins from some oral streptococci.

    PubMed Central

    Lowe, A M; Lambert, P A; Smith, A W

    1995-01-01

    Serum from a patient with Enterococcus faecalis endocarditis was used to identify the gene efaA cloned in Lambda ZapII in Escherichia coli. Nucleotide sequence analysis revealed a 924-bp open reading frame encoding a protein with a predicted molecular weight of 34,768. The amino acid sequence of EfaA shows 55 to 60% homology to a group of streptococcal proteins, FimA from Streptococcus parasanguis, SsaB from Streptococcus sanguis, ScaA from Streptococcus gordonii, and PsaA from Streptococcus pneumoniae. Members of this group have been shown to be adhesins, and we hypothesize that EfaA may function as an adhesin in endocarditis. PMID:7822045

  15. Effects of Enterococcus faecalis CECT 7121 on Cryptosporidium parvum infection in mice.

    PubMed

    Del Coco, Valeria F; Sparo, Mónica D; Sidoti, Alicia; Santín, Mónica; Basualdo, Juan Angel; Córdoba, María Alejandra

    2016-08-01

    Cryptosporidium is an opportunistic protozoan parasite of humans and animals worldwide and causes diarrheal disease that is typically self-limiting in immunocompetent hosts but often life threatening to immunocompromised individuals. However, there is a lack of completely efficient therapy available. Probiotics have attracted the attention as potential antiparasite compounds against protozoa involved in intestinal infections. This study investigated the effects of administration of probiotic Enterococcus faecalis CECT 7121 on Cryptosporidium parvum infection in immunosuppressed mice. Effects on C. parvum infection at the intestinal mucosa were studied and scored at each portion of the gut. It was demonstrated that Ef CECT 7121 interfered with C. parvum infection when both probiotic and parasite were present in the same intestinal location suggesting that Ef CECT 7121 supplementation can alleviate the negative effects of C. parvum infection. PMID:27193238

  16. Antimicrobial activity of herbal medicines (tulsi extract, neem extract) and chlorhexidine against Enterococcus faecalis in Endodontics: An in vitro study

    PubMed Central

    Chandrappa, Pradeep Muttagadur; Dupper, Akash; Tripathi, Pragya; Arroju, Ramakrishna; Sharma, Preeti; Sulochana, Konthoujam

    2015-01-01

    Background: Successful endodontic treatment depends on effective disinfection and complete sealing of root canal. Various medicaments are advised for disinfecting root canal, such as herbal and non-herbal medicaments. This study was done to assess the antimicrobial activity of herbal medicines (neem extract, tulsi extract) and chlorhexidine against Enterococcus faecalis in Endodontics. Materials and Methods: Agar diffusion method was used to evaluate the antimicrobial action of different medicines. Sixty samples were segregated into four groups with 15 samples in each: Group I: chlorhexidine 2%, Group II: neem extract, Group III: tulsi extract, and Group IV: distilled water. The inhibition zones against E. faecalis were recorded and statistically assessed using one-way analysis of variance (ANOVA) test (P < 0.05). Results: Significant antibacterial effect against E. faecalis was observed with chlorhexidine followed by neem extract and tulsi extract. Conclusion: Herbal medicines seemed to be effective against E. faecalis compared to 2% chlorhexidine gluconate. PMID:26942123

  17. The Natural Surfactant Glycerol Monolaurate Significantly Reduces Development of Staphylococcus aureus and Enterococcus faecalis Biofilms

    PubMed Central

    Hess, Donavon J.; Henry-Stanley, Michelle J.

    2015-01-01

    Abstract Background: Bacterial biofilms are involved in a large proportion of clinical infections, including device-related infections. Unfortunately, biofilm-associated bacteria are typically less susceptible to antibiotics, and infected devices must often be removed. On the basis of a recent observation that lipid-rich biofilm matrix material is present in early biofilm formation and may protect a population of bacteria from interacting with ordinarily diffusible small molecules, we hypothesized that surfactants may be useful in preventing biofilm development. Methods: Experimental Staphylococcus aureus or Enterococcus faecalis biofilms were cultivated on surgical suture suspended in a growth medium supplemented with the natural surfactant glycerol monolaurate (GML) or with a component molecule, lauric acid. After 16 h incubation, the numbers of viable biofilm-associated bacteria were measured by standard microbiologic techniques and biofilm biomass was measured using the colorimetric crystal violet assay. Results: Both GML and lauric acid were effective in inhibiting biofilm development as measured by decreased numbers of viable biofilm-associated bacteria as well as decreased biofilm biomass. Compared with lauric acid on a molar basis, GML represented a more effective inhibitor of biofilms formed by either S. aureus or E. faecalis. Conclusions: Because the natural surfactant GML inhibited biofilm development, resulting data were consistent with the hypothesis that lipids may play an important role in biofilm growth, implying that interfering with lipid formation may help control development of clinically relevant biofilms. PMID:26110557

  18. Biodegradation of C.I. Reactive Red 195 by Enterococcus faecalis strain YZ66.

    PubMed

    Mate, Madhuri Sahasrabudhe; Pathade, Girish

    2012-03-01

    Synthetic dyes are extensively used in textile dyeing, paper, printing, colour photography, pharmaceutics, cosmetics and other industries. Among these, azodyes represents the largest and most versatile class of synthetic dyes. As high as 50% of the dyes are released into the environment during manufacture and usage. Traditional methods of treatment are found to be expensive and have operational problems. Biological decolourization has been investigated as a method to transform, degrade or mineralize azo dyes. In the present studies bacteria from soil from dye waste area, dye waste, sewage and dung were subjected to acclimatization with C.I. Reactive Red 195 an azo dye, in the basal nutrient media. The most promising bacterial isolate was used for further dye degradation studies. The 16s rRNA gene sequencing and biochemical characteristics revealed the isolated organism as Enterococcus faecalis strain YZ66. The strain showed 99.5% decolourization of the selected dye (Reactive Red 195-50 mg/l) within one and half hour in static anoxic condition. The optimum pH and temperature for the decolourization was 5.0 and 40°C respectively. The biodegradation was monitored by UV-Vis, FTIR, TLC and HPLC. The final products were characterized by Gas chromatography and Mass Spectrophotometry. Toxicity study demonstrated no toxicity of the biodegradation product. The results suggest that the isolated organism E. faecalis strain YZ 66 can be used as a useful tool to treat waste water containing reactive dyes.

  19. Tn5381, a conjugative transposon identifiable as a circular form in Enterococcus faecalis.

    PubMed Central

    Rice, L B; Marshall, S H; Carias, L L

    1992-01-01

    We have identified two 19-kb conjugative transposons (Tn5381 and Tn5383) in separate strains of multiply resistant Enterococcus faecalis. These transposons confer resistance to tetracycline and minocycline via a tetM gene, are capable of both chromosomal and plasmid integration in a Rec- environment, and transfer between strains in the absence of detectable plasmid DNA at frequencies ranging from < 1 x 10(-9) to 2 x 10(-5) per donor CFU, depending on the donor strain and the growth conditions. Hybridization studies indicate that these transposons are closely related to Tn916. We have identified bands of ca. 19 kb on agarose gel separations of alkaline lysis preparations from E. faecalis strains containing chromosomal copies of Tn5381, which we have confirmed to be a circularized form of this transposon. This phenomenon has previously been observed only when Tn916 has been cloned in Escherichia coli. Overnight growth of donor strains in the presence of subinhibitory concentrations of tetracycline results in an approximately 10-fold increase in transfer frequency of Tn5381 into enterococcal recipients and an increase in the amount of the circular form of Tn5381 as detectable by hybridization. These results suggest that Tn5381 is a Tn916-related conjugative transposon for which the appearance of a circular form and the conjugative-transfer frequency are regulated by a mechanism(s) affected by the presence of tetracycline in the growth medium. Images PMID:1331026

  20. Eradication of Intracanal Enterococcus Faecalis Biofilm by Passive Ultrasonic Irrigation and RinsEndo System

    PubMed Central

    Toljan, Ivana; Bago, Ivona; Anić, Ivica

    2016-01-01

    Aim The aim of the study was to compare the antimicrobial efficacy of three irrigation techniques after the use of standardized volume of NaOCl and with standardized time and irrigation. Methodology Forty-eight single rooted teeth were inoculated with an Enterococcus faecalis suspension for 24 h. The remaining six canals served as negative controls. The 36 root canals were randomly distributed into three experimental groups; group 1, conventional syringe irrigation; group 2, automated-dynamic irrigation (RinsEndo); group 3, passive ultrasonic irrigation (PUI). In the first protocol, the standardized volume of 3% NaOCl (20 mL) was used and in the second protocol, and standardized irrigation time (45 seconds) was used. Samples from root canals were cultured and the colony-forming units (CFUs) were counted. Results When the volume of the irrigant was standardized, RinsEndo was more effective than PUI (p<0.01). When the irrigation time was standardized, there were no significant differences between any irrigation techniques (p>0.05). The RinsEndo group had the highest percentage of minimal counts of E. faecalis CFUs. Conclusions RinsEndo was more effective than PUI only when the volume of the irrigant was standardized. However, the RinsEndo provided higher bacterial reduction in both protocols when using the least amount of the irrigant and providing longer contact time. PMID:27688422

  1. Overexpression, crystallization and preliminary X-ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis

    SciTech Connect

    Kang, Ji Yong; Lee, Hyung Ho; Yoon, Hye Jin; Kim, Hyoun Sook; Suh, Se Won

    2006-11-01

    Phosphopantetheine adenylyltransferase from En. faecalis was crystallized and X-ray diffraction data were collected to 2.70 Å resolution. Phosphopantetheine adenylyltransferase, an essential enzyme in the coenzyme A biosynthetic pathway, catalyzes the reversible transfer of an adenylyl group from ATP to 4′-phosphopantetheine, yielding 3′-dephospho-CoA and pyrophosphate. Enterococcus faecalis PPAT has been overexpressed in Escherichia coli as a fusion with a C-terminal purification tag and crystallized at 297 K using a reservoir solution consisting of 0.1 M sodium HEPES pH 7.5, 0.8 M sodium dihydrogen phosphate and 0.8 M potassium dihydrogen phosphate. X-ray diffraction data were collected to 2.70 Å at 100 K. The crystals belong to the primitive tetragonal space group P4{sub 1} (or P4{sub 3}), with unit-cell parameters a = b = 160.81, c = 225.68 Å. Four copies of the hexameric molecule are likely to be present in the asymmetric unit, giving a crystal volume per protein weight (V{sub M}) of 3.08 Å{sup 3} Da{sup −1} and a solvent content of 60.1%.

  2. Bactericial effect of a non-thermal plasma needle against Enterococcus faecalis biofilms

    NASA Astrophysics Data System (ADS)

    Jiang, Chunqi; Schaudinn, C.; Jaramillo, D. E.; Sedghizadeh, P. P.; Webster, P.; Costerton, J. W.

    2011-10-01

    Up to 3 cm long submillimeter-in-scale plasma needle was generated in ambient atmosphere for root canal disinfection. Powered with 1-2 kHz, multi-kilovolt nanosecond electric pulses, this He/(1%)O2 plasma jet consists of ionization fronts propagating at speeds of the order of 107 cm/s. Plasma treatment of Enterococcus faecalis biofilms on hydroxyapatite (HA) discs for 5 min resulted in severe damage of the bacterial cells and sterilized HA surfaces of more than 3 mm in diameter, observed by the scanning electron microscopy. With a curing dielectric microtube placed 1 cm or less below the nozzle, the plasma jet entered even at a sharp angle and followed the curvature of the tube, and reached the bottom of the tube. The bactericidal effect of the plasma needle against E. faecalis biofilm grown on the inner surfaces of the tube was demonstrated. However, the bactericidal effect weakens or diminishes for the bacteria grown deeper in the tube, indicating improvement of the plasma treatment scheme is needed. Mechanisms of the plasma bactericidal effects are discussed. Supported by the National Institute of Dental and Craniofacial Research and the Air Force Office of Scientific Research.

  3. Structural Studies on Cytosolic Domain of Magnesium Transporter MgtE from Enterococcus faecalis

    SciTech Connect

    Ragumani, S.; Sauder, J; Burley, S; Swaminathan, S

    2009-01-01

    Magnesium (Mg{sup 2+}) is an essential element for growth and maintenance of living cells. It acts as a cofactor for many enzymes and is also essential for stability of the plasma membrane. There are two distinct classes of magnesium transporters identified in bacteria that convey Mg{sup 2+} from periplasm to cytoplasm [ATPase-dependent (MgtA and MgtB) and constitutively active (CorA and MgtE)]. Previously published work on Mg{sup 2+} transporters yielded structures of full length MgtE from Thermus thermophilus, determined at 3.5 {angstrom} resolution, and its cytoplasmic domain with and without bond Mg{sup 2+} determined at 2.3 and 3.9 {angstrom} resolution, respectively. Here, they report the crystal structure of the Mg{sup 2+} bound form of the cytosolic portion of MgtE (residues 6-262) from Enterococcus faecalis at 2.2 {angstrom} resolution. The present structure and magnesium bound cytosolic domain structure from T. thermophilus (PDB ID: 2YVY) are structurally similar. Three magnesium binding sites are common to both MgtE full length and the present structure. Their work revealed an additional Mg{sup 2+} binding site in the E. faecalis structure. In this report, they discuss the functional significance of Mg{sup 2+} binding sites in the cytosolic domains of MgtE transporters.

  4. Eradication of Intracanal Enterococcus Faecalis Biofilm by Passive Ultrasonic Irrigation and RinsEndo System

    PubMed Central

    Toljan, Ivana; Bago, Ivona; Anić, Ivica

    2016-01-01

    Aim The aim of the study was to compare the antimicrobial efficacy of three irrigation techniques after the use of standardized volume of NaOCl and with standardized time and irrigation. Methodology Forty-eight single rooted teeth were inoculated with an Enterococcus faecalis suspension for 24 h. The remaining six canals served as negative controls. The 36 root canals were randomly distributed into three experimental groups; group 1, conventional syringe irrigation; group 2, automated-dynamic irrigation (RinsEndo); group 3, passive ultrasonic irrigation (PUI). In the first protocol, the standardized volume of 3% NaOCl (20 mL) was used and in the second protocol, and standardized irrigation time (45 seconds) was used. Samples from root canals were cultured and the colony-forming units (CFUs) were counted. Results When the volume of the irrigant was standardized, RinsEndo was more effective than PUI (p<0.01). When the irrigation time was standardized, there were no significant differences between any irrigation techniques (p>0.05). The RinsEndo group had the highest percentage of minimal counts of E. faecalis CFUs. Conclusions RinsEndo was more effective than PUI only when the volume of the irrigant was standardized. However, the RinsEndo provided higher bacterial reduction in both protocols when using the least amount of the irrigant and providing longer contact time.

  5. The Intraperitoneal Transcriptome of the Opportunistic Pathogen Enterococcus faecalis in Mice

    PubMed Central

    Muller, Cécile; Cacaci, Margherita; Sauvageot, Nicolas; Sanguinetti, Maurizio; Rattei, Thomas; Eder, Thomas; Giard, Jean-Christophe; Kalinowski, Jörn; Hain, Torsten; Hartke, Axel

    2015-01-01

    Enterococcus faecalis is a Gram-positive lactic acid intestinal opportunistic bacterium with virulence potential. For a better understanding of the adapation of this bacterium to the host conditions, we performed a transcriptome analysis of bacteria isolated from an infection site (mouse peritonitis) by RNA-sequencing. We identified a total of 211 genes with significantly higher transcript levels and 157 repressed genes. Our in vivo gene expression database reflects well the infection process since genes encoding important virulence factors like cytolysin, gelatinase or aggregation substance as well as stress response proteins, are significantly induced. Genes encoding metabolic activities are the second most abundant in vivo induced genes demonstrating that the bacteria are metabolically active and adapt to the special nutrient conditions of the host. α- and β- glucosides seem to be important substrates for E. faecalis inside the host. Compared to laboratory conditions, the flux through the upper part of glycolysis seems to be reduced and more carbon may enter the pentose phosphate pathway. This may reflect the need of the bacteria under infection conditions to produce more reducing power for biosynthesis. Another important substrate is certainly glycerol since both pathways of glycerol catabolism are strongly induced. Strongly in vivo induced genes should be important for the infection process. This assumption has been verified in a virulence test using well characterized mutants affected in glycerol metabolism. This showed indeed that mutants unable to metabolize this sugar alcohol are affected in organ colonisation in a mouse model. PMID:25978463

  6. The Presence and Origin of Enterococcus faecalis in Cabo Rojo, Puerto Rico

    NASA Astrophysics Data System (ADS)

    Zachman, A. J.; Sturm, P.; Viqueira Ríos, R.

    2015-12-01

    Currently, a watershed management plan is being developed for Cabo Rojo region in Southwest Puerto Rico. This project fills in major gaps for water quality data on the Rio Viejo, a tributary on the Guanajibio River. The Rio Viejo flows through the town of Cabo Rojo, a town of 51,245 people. The project has identified 5 sites along the river to track bacterial loads. In the tropics, Enterococcus faecalis is an important indicator for fecal contamination in surface waters as it does not reproduce as quickly soils as E. coli. A combination of EPA 1600 and 9230B from Standard Methods for the Examination of Water and Wastewater for identification of E. faecalis were utilized. The assay is a four step procedure that identifies the four criteria of bacteria in the group D Streptococcus system. The criteria require that the bacteria are Gram-positive cocci and Esculin-positive. There also must be growth in Brain Heart Infusion Broth at 35C and 45C as well as growth in Brain Heart Infusion broth + 6.5% NaCl. Further research will be conducted at North Carolina State University to ascertain the vertebrate species that is the source of the contamination through the use of qPCR.

  7. [Emergence of linezolid-resistant Enterococcus faecalis strains from two inpatients in a pediatric ward].

    PubMed

    Nihonyanagi, Shin; Adachi, Yuzuru; Onuki, Tomoyo; Nakazaki, Nobuhiko; Hirata, Yasuyosi; Fujiki, Kuniko; Takayama, Yoko; Kanoh, Yuhsaku; Bandoh, Yuki; Dantsuji, Yurika; Hanaki, Hideaki; Sunakawa, Keisuke

    2012-09-01

    We report herein on the isolation of three linezolid-resistant Enterococcus faecalis strains in 2011 from two pediatric inpatients at Kitasato University Hospital, Japan. Three linezolid resistant strains were isolated from two patients who shared the same room of a pediatric inpatient ward. Two linezolid resistant strains were isolated from patient A who had been treated with a total of 17,600mg of linezolid during 60 days of hospitalization (strains 1 and 2). The linezolid resistant E. faecalis persisted through the time that the patient had been discharged from the hospital. Another linezolid resistant strain was isolated from patient B who had no history of linezolid administration. The resistant strain in patient B phased out spontaneously. The minimum inhibitory concentration of linezolid in these strains ranged from 8.0 to 16.0 microg/mL. PCR amplification of the chromosomal gene encoding domain V of the 23S rRNA and subsequent nucleotide sequencing revealed that all the strains had at least one G2576T mutation. The pulse-field-gel electrophoretograms of the DNA treated with the SmaI restriction enzyme showed an identical profile suggesting that they were derived from a single resistant strain. These results suggested that the resistant strain occurred in patient A and was transmitted to patient B within the inpatient ward. PMID:23198574

  8. Effect of high-intensity focused ultrasound on Enterococcus faecalis planktonic suspensions and biofilms.

    PubMed

    Iqbal, Kulsum; Ohl, Siew-Wan; Khoo, Boo-Cheong; Neo, Jennifer; Fawzy, Amr S

    2013-05-01

    In this study, the effect of high-intensity focused ultrasound (HIFU) on Enterococcus faecalis on both planktonic suspensions and biofilms was investigated. E. faecalis persist in secondary dental infections as biofilms. Glass-bottom Petri dishes with biofilms were centered at the focal point of the HIFU wave generated by a 250-kHz transducer. Specimens were subjected to HIFU exposure at different periods of 30, 60 and 120 s. The viable bacteria, removal effect and bacterial viability of biofilms attached to the Petri dish surface were studied by colony-forming units (CFUs), scanning electron microscopy and confocal microscopy, respectively. The removal and bactericidal effects of HIFU are dependent on the exposure time. A significant reduction in biofilm thickness and CFU was found with the increase in HIFU exposure. The removal or bactericidal effect of HIFU was more significant starting from 60 s of exposure. This study highlighted the potential application of HIFU as a novel method for root canal disinfection. PMID:23453374

  9. Identification and Characterization of a Bacitracin Resistance Network in Enterococcus faecalis

    PubMed Central

    Fang, Chong; Shaaly, Aishath; Leslie, David J.; Weimar, Marion R.; Kalamorz, Falk; Carne, Alan; Cook, Gregory M.

    2014-01-01

    Resistance of Enterococcus faecalis against antimicrobial peptides, both of host origin and produced by other bacteria of the gut microflora, is likely to be an important factor in the bacterium's success as an intestinal commensal. The aim of this study was to identify proteins with a role in resistance against the model antimicrobial peptide bacitracin. Proteome analysis of bacitracin-treated and untreated cells showed that bacitracin stress induced the expression of cell wall-biosynthetic proteins and caused metabolic rearrangements. Among the proteins with increased production, an ATP-binding cassette (ABC) transporter with similarity to known peptide antibiotic resistance systems was identified and shown to mediate resistance against bacitracin. Expression of the transporter was dependent on a two-component regulatory system and a second ABC transporter, which were identified by genome analysis. Both resistance and the regulatory pathway could be functionally transferred to Bacillus subtilis, proving the function and sufficiency of these components for bacitracin resistance. Our data therefore show that the two ABC transporters and the two-component system form a resistance network against antimicrobial peptides in E. faecalis, where one transporter acts as the sensor that activates the TCS to induce production of the second transporter, which mediates the actual resistance. PMID:24342648

  10. In vitro evaluation of the antimicrobial efficacy of chitosan and other endodontic irrigants against Enterococcus faecalis.

    PubMed

    Shenoi, Pratima R; Morey, Elakshi S; Makade, Chetana S; Gunwal, Mohit K; Khode, Rajiv T; Wanmali, Sunay S

    2016-01-01

    The success of endodontic treatment is directly enhanced by elimination of microorganisms in infected root canals. Recently, chitosan, a natural, nontoxic biopolymer, has been introduced as an irrigant that has the capacity to remove the smear layer. The antimicrobial properties of chitosan as an endodontic irrigant have not yet been explored. The purpose of this study was to compare the antimicrobial efficacy of BioPure MTAD, 0.2% chitosan, 1% chitosan, 2% chlorhexidine gluconate, and 3% sodium hypochlorite (NaOCl) against Enterococcus faecalis, which is frequently isolated from persistent root canal infections. The agar well diffusion method was used to measure the antimicrobial activities of these irrigants. Saline was used as a negative control. The order of effectiveness was determined by the measurement of inhibition zones. Data were analyzed using 1-way analysis of variance and the Duncan multiple range test. BioPure MTAD had a significantly larger mean inhibition zone against E faecalis than the other irrigants (P < 0.001). Although 0.2% chitosan did not show any inhibition zones, 1% chitosan was as effective as 3% NaOCl (P = 0.352), and both irrigants showed significantly greater effectivity than 2% chlorhexidine (P < 0.001). Thus, 1% chitosan can be an effective natural antimicrobial substitute for synthetic irrigants. PMID:27599284

  11. Involvement of Enterococcus faecalis Small RNAs in Stress Response and Virulence

    PubMed Central

    Michaux, Charlotte; Hartke, Axel; Martini, Cecilia; Reiss, Swantje; Albrecht, Dirk; Budin-Verneuil, Aurélie; Sanguinetti, Maurizio; Engelmann, Susanne; Hain, Torsten; Verneuil, Nicolas

    2014-01-01

    Candidate small RNAs (sRNAs) have recently been identified in Enterococcus faecalis, a Gram-positive opportunistic pathogen, and six of these candidate sRNAs with unknown functions were selected for a functional study. Deletion mutants and complemented strains were constructed, and their virulence was tested. We were unable to obtain the ef0869-0870 mutant, likely due to an essential role, and the ef0820-0821 sRNA seemed not to be involved in virulence. In contrast, the mutant lacking ef0408-0409 sRNA, homologous to the RNAII component of the toxin-antitoxin system, appeared more virulent and more able to colonize mouse organs. The three other mutants showed reduced virulence. In addition, we checked the responses of these mutant strains to several stresses encountered in the gastrointestinal tract or during the infection process. In parallel, the activities of the sRNA promoters were measured using transcriptional fusion constructions. To attempt to identify the regulons of these candidate sRNAs, proteomics profiles of the mutant strains were compared with that of the wild type. This showed that the selected sRNAs controlled the expression of proteins involved in diverse cellular processes and the stress response. The combined data highlight the roles of certain candidate sRNAs in the adaptation of E. faecalis to environmental changes and in the complex transition process from a commensal to a pathogen. PMID:24914223

  12. Human salivary proteins with affinity to lipoteichoic acid of Enterococcus faecalis.

    PubMed

    Baik, Jung Eun; Choe, Hyuk-Il; Hong, Sun Woong; Kang, Seok-Seong; Ahn, Ki Bum; Cho, Kun; Yun, Cheol-Heui; Han, Seung Hyun

    2016-09-01

    Enterococcus faecalis is associated with refractory apical periodontitis and its lipoteichoic acid (Ef.LTA) is considered as a major virulence factor. Although the binding proteins of Ef.LTA may play an important role for mediating infection and immunity in the oral cavity, little is known about Ef.LTA-binding proteins (Ef.LTA-BPs) in saliva. In this study, we identified salivary Ef.LTA-BPs with biotinylated Ef.LTA (Ef.LTA-biotin) through mass spectrometry. The biotinylation of Ef.LTA was confirmed by binding capacity with streptavidin-FITC on CHO/CD14/TLR2 cells. The biological activity of Ef.LTA-biotin was determined based on the induction of nitric oxide and macrophage inflammatory protein-1α in a macrophage cell-line, RAW 264.7. To identify salivary Ef.LTA-BPs, the Ef.LTA-biotin was mixed with a pool of human saliva obtained from nine healthy subjects followed by precipitation with a streptavidin-coated bead. Ef.LTA-BPs were then separated with 12% SDS-PAGE and subjected to the mass spectrometry. Six human salivary Ef.LTA-BPs including short palate lung and nasal epithelium carcinoma-associated protein 2, zymogen granule protein 16 homolog B, hemoglobin subunit α and β, apolipoprotein A-I, and lipocalin-1 were identified with statistical significance (P<0.05). Ef.LTA-BPs were validated with lipocalin-1 using pull-down assay. Hemoglobin inhibited the biofilm formation of E. faecalis whereas lipocalin-1 did not show such effect. Collectively, the identified Ef.LTA-BPs could provide clues for our understanding of the pathogenesis of E. faecalis and host immunity in oral cavity. PMID:27474971

  13. Pilin and Sortase Residues Critical for Endocarditis- and Biofilm-Associated Pilus Biogenesis in Enterococcus faecalis

    PubMed Central

    Nielsen, Hailyn V.; Flores-Mireles, Ana L.; Kau, Andrew L.; Kline, Kimberly A.; Pinkner, Jerome S.; Neiers, Fabrice; Normark, Staffan; Henriques-Normark, Birgitta

    2013-01-01

    Enterococci commonly cause hospital-acquired infections, such as infective endocarditis and catheter-associated urinary tract infections. In animal models of these infections, a long hairlike extracellular protein fiber known as the endocarditis- and biofilm-associated (Ebp) pilus is an important virulence factor for Enterococcus faecalis. For Ebp and other sortase-assembled pili, the pilus-associated sortases are essential for fiber formation as they create covalent isopeptide bonds between the sortase recognition motif and the pilin-like motif of the pilus subunits. However, the molecular requirements governing the incorporation of the three pilus subunits (EbpA, EbpB, and EbpC) have not been investigated in E. faecalis. Here, we show that a Lys residue within the pilin-like motif of the EbpC subunit was necessary for EbpC polymerization. However, incorporation of EbpA into the pilus fiber only required its sortase recognition motif (LPXTG), while incorporation of EbpB only required its pilin-like motif. Only the sortase recognition motif would be required for incorporation of the pilus tip subunit, while incorporation of the base subunit would only require the pilin recognition motif. Thus, these data support a model with EbpA at the tip and EbpB at the base of an EbpC polymer. In addition, the housekeeping sortase, SrtA, was found to process EbpB and its predicted catalytic Cys residue was required for efficient cell wall anchoring of mature Ebp pili. Thus, we have defined molecular interactions involved in fiber polymerization, minor subunit organization, and pilus subcellular compartmentalization in the E. faecalis Ebp pilus system. These studies advance our understanding of unique molecular mechanisms of sortase-assembled pilus biogenesis. PMID:23913319

  14. Abundance and diversity of plasmid-associated genes among clinical isolates of Enterococcus faecalis.

    PubMed

    Wardal, Ewa; Gawryszewska, Iwona; Hryniewicz, Waleria; Sadowy, Ewa

    2013-11-01

    Enterococcus faecalis, a normal compound of the human intestinal microbiome, plays an important role in hospital-acquired infections. Plasmids make a significant contribution to the acquisition of the novel traits such as antimicrobial resistance and virulence by this pathogen. The study investigated the plasmid content and the diversity of plasmid-associated genes in a group of 152 hospital isolates of E. faecalis. The majority of plasmids visualized by pulsed-field gel electrophoresis of S1 nuclease-digested DNA fell into the range of 50-100 kb. PCR-based screening allowed detection of genes of the rep1(pIP501), rep2(pRE25), rep4(pMBB1), rep6(pS86), rep7(pT181), rep8(pAM373), rep9(pAD1/pTEF2/pCF10), rep10(pIM13) and rep13(pC194) families in 29 different combinations. The par and ω-ε-ζ plasmid stabilization systems were ubiquitous (45 isolates, 29.6% and 88 isolates, 57.9%, respectively), while the axe-txe system was not found. The asa1 gene homologues encoding aggregation substance characteristic for the pAD1 and related group of pheromone-responsive plasmids were present in 106 isolates. A variety of sequence variants, including novel ones, of genes associated with pheromone-responsive plasmids, such as rep8(pAM373), rep9(pAD1/pTEF2/pCF10), par, and asa1 were observed. In conclusion, there is a big and only partially characterized pool of diverse plasmids in clinical E. faecalis.

  15. Antibacterial Efficacy of Super-Oxidized Water on Enterococcus faecalis Biofilms in Root Canal

    PubMed Central

    Zan, Recai; Alacam, Tayfun; Hubbezoglu, Ihsan; Tunc, Tutku; Sumer, Zeynep; Alici, Oguzhan

    2016-01-01

    Background The success of endodontic treatment depends on a few crucial factors. One of these factors is the complete chemomechanic preparation of root canal against various bacteria. In particular, the effect of resistant bacteria may cause intense pain with flare-up and formation of periapical lesions. Therefore, the strong effect of irrigants plays an important role in terms of the complete elimination of these bacteria to achieve long-term successful treatment. Objectives The aim of this study was to investigate the antibacterial effects of super-oxidized water (SPO) in root canals infected with Enterococcus faecalis biofilms. Methods One hundred twenty single-root, premolar teeth were selected. Initially, the teeth were prepared and then disinfected. E. faecalis were inoculated and kept at 37°C for 24 hours in the root canals. The re-inoculation procedure was repeated on the first, fourth, seventh, and tenth days. The infected root canals were divided into one negative (saline) and one positive (sodium hypochlorite) control group and four experimental groups (super-oxidized water: 1, 2, 3, or 5 minutes) (n = 20). Paper points were placed in the root canals to control and evaluate the biofilm formation. Biofilms were counted on blood agar plates, and data was evaluated and statistically analyzed using one-way ANOVA and Tukey’s test. Results Although sodium hypochlorite (NaOCl) showed no statistically significant difference when compared with three and five minutes of SPO irrigation (P > 0.05), NaOCl showed statistically significant differences among all other groups (P < 0.05). Conclusions Super-oxidized water indicated a remarkable and similar bactericidal effect to that of traditional NaOCl against E. faecalis biofilms. In terms of successful endodontic treatment approaches, super-oxidized water may be used as an effective irrigation solution in clinics. PMID:27800142

  16. Antibacterial Efficacy of Calcium Hypochlorite with Vibringe Sonic Irrigation System on Enterococcus faecalis: An In Vitro Study.

    PubMed

    Dumani, Aysin; Guvenmez, Hatice Korkmaz; Yilmaz, Sehnaz; Yoldas, Oguz; Kurklu, Zeliha Gonca Bek

    2016-01-01

    Aim. The purpose of this study was to compare the in vitro efficacy of calcium hypochlorite (Ca[OCl]2) and sodium hypochlorite (NaOCl) associated with sonic (Vibringe) irrigation system in root canals which were contaminated with Enterococcus faecalis. Material and Methods. The root canals of 84 single-rooted premolars were enlarged up to a file 40, autoclaved, inoculated with Enterococcus faecalis, and incubated for 21 days. The samples were divided into 7 groups according to the irrigation protocol: G0: no treatment; G1: distilled water; G2: 2.5% NaOCl; G3: 2.5% Ca(OCl)2; G4: distilled water with sonic activation; G5: 2.5% NaOCl with sonic activation; and G6: 2.5% Ca(OCl)2 with sonic activation. Before and after decontamination procedures microbiological samples were collected and the colony-forming units were counted and the percentages of reduction were calculated. Results. Distilled water with syringe irrigation and sonic activation groups demonstrated poor antibacterial effect on Enterococcus faecalis compared to other experimental groups (p < 0.05). There was no statistically significant difference between syringe and sonic irrigation systems with Ca(OCl)2 and NaOCl. Conclusion. The antimicrobial property of Ca(OCl)2 has been investigated and compared with that of NaOCl. Both conventional syringe irrigation and sonic irrigation were found effective at removing E. faecalis from the root canal of extracted human teeth.

  17. Antibacterial Efficacy of Calcium Hypochlorite with Vibringe Sonic Irrigation System on Enterococcus faecalis: An In Vitro Study

    PubMed Central

    Dumani, Aysin; Guvenmez, Hatice Korkmaz; Yilmaz, Sehnaz; Yoldas, Oguz; Kurklu, Zeliha Gonca Bek

    2016-01-01

    Aim. The purpose of this study was to compare the in vitro efficacy of calcium hypochlorite (Ca[OCl]2) and sodium hypochlorite (NaOCl) associated with sonic (Vibringe) irrigation system in root canals which were contaminated with Enterococcus faecalis. Material and Methods. The root canals of 84 single-rooted premolars were enlarged up to a file 40, autoclaved, inoculated with Enterococcus faecalis, and incubated for 21 days. The samples were divided into 7 groups according to the irrigation protocol: G0: no treatment; G1: distilled water; G2: 2.5% NaOCl; G3: 2.5% Ca(OCl)2; G4: distilled water with sonic activation; G5: 2.5% NaOCl with sonic activation; and G6: 2.5% Ca(OCl)2 with sonic activation. Before and after decontamination procedures microbiological samples were collected and the colony-forming units were counted and the percentages of reduction were calculated. Results. Distilled water with syringe irrigation and sonic activation groups demonstrated poor antibacterial effect on Enterococcus faecalis compared to other experimental groups (p < 0.05). There was no statistically significant difference between syringe and sonic irrigation systems with Ca(OCl)2 and NaOCl. Conclusion. The antimicrobial property of Ca(OCl)2 has been investigated and compared with that of NaOCl. Both conventional syringe irrigation and sonic irrigation were found effective at removing E. faecalis from the root canal of extracted human teeth. PMID:27218106

  18. Screening of In Vivo Activated Genes in Enterococcus faecalis during Insect and Mouse Infections and Growth in Urine

    PubMed Central

    Hanin, Aurelie; Sava, Irina; Bao, YinYin; Huebner, Johannes; Hartke, Axel; Auffray, Yanick; Sauvageot, Nicolas

    2010-01-01

    Enterococcus faecalis is part of the commensal microbiota of humans and its main habitat is the gastrointestinal tract. Although harmless in healthy individuals, E. faecalis has emerged as a major cause of nosocomial infections. In order to better understand the transformation of a harmless commensal into a life-threatening pathogen, we developed a Recombination-based In Vivo Expression Technology for E. faecalis. Two R-IVET systems with different levels of sensitivity have been constructed in a E. faecalis V583 derivative strain and tested in the insect model Galleria mellonella, during growth in urine, in a mouse bacteremia and in a mouse peritonitis model. Our combined results led to the identification of 81 in vivo activated genes. Among them, the ef_3196/7 operon was shown to be strongly induced in the insect host model. Deletion of this operonic structure demonstrated that this two-component system was essential to the E. faecalis pathogenic potential in Galleria. Gene ef_0377, induced in insect and mammalian models, has also been further analyzed and it has been demonstrated that this ankyrin-encoding gene was also involved in E. faecalis virulence. Thus these R-IVET screenings led to the identification of new E. faecalis factors implied in in vivo persistence and pathogenic potential of this opportunistic pathogen. PMID:20686694

  19. Bacteriocin protein BacL1 of Enterococcus faecalis is a peptidoglycan D-isoglutamyl-L-lysine endopeptidase.

    PubMed

    Kurushima, Jun; Hayashi, Ikue; Sugai, Motoyuki; Tomita, Haruyoshi

    2013-12-27

    Enterococcus faecalis strains are commensal bacteria in humans and other animals, and they are also the causative agent of opportunistic infectious diseases. Bacteriocin 41 (Bac41) is produced by certain E. faecalis clinical isolates, and it is active against other E. faecalis strains. Our genetic analyses demonstrated that the extracellular products of the bacL1 and bacA genes, which are encoded in the Bac41 operon, coordinately express the bacteriocin activity against E. faecalis. In this study, we investigated the molecular functions of the BacL1 and BacA proteins. Immunoblotting and N-terminal amino acid sequence analysis revealed that BacL1 and BacA are secreted without any processing. The coincidental treatment with the recombinant BacL1 and BacA showed complete bacteriocin activity against E. faecalis, but neither BacL1 nor BacA protein alone showed the bacteriocin activity. Interestingly, BacL1 alone demonstrated substantial degrading activity against the cell wall fraction of E. faecalis in the absence of BacA. Furthermore, MALDI-TOF MS analysis revealed that BacL1 has a peptidoglycan D-isoglutamyl-L-lysine endopeptidase activity via a NlpC/P60 homology domain. These results collectively suggest that BacL1 serves as a peptidoglycan hydrolase and, when BacA is present, results in the lysis of viable E. faecalis cells.

  20. In vivo broiler experiments to assess anti-Campylobacter jejuni activity of a live Enterococcus faecalis strain.

    PubMed

    Robyn, J; Rasschaert, G; Hermans, D; Pasmans, F; Heyndrickx, M

    2013-01-01

    Bacterial gastroenteritis caused by thermotolerant Campylobacter species, mainly Campylobacter jejuni, has been the most reported zoonotic disease in many developed countries in recent years. Reducing Campylobacter shedding on the farm could result in a reduction of the number of campylobacteriosis cases. In 2 independent broiler seeder experiments, in which broiler chickens were orally inoculated with 2 amounts of Enterococcus faecalis MB 5259, we established whether a live E. faecalis strain was capable of reducing cecal Campylobacter colonization in broiler chickens. In previous in vitro experiments it has been demonstrated that this E. faecalis MB 5259 displays anti-Campylobacter activity. The effect of pH and bile salts on E. faecalis MB 5259 showed that growth and survival of E. faecalis MB 5259 can be impaired during passage through the gastrointestinal tract of broiler chickens. Despite these results E. faecalis MB 5259 was capable of colonizing the broiler ceca. Contrary to the in vitro experiments, in which E. faecalis MB 5259 inhibited C. jejuni MB 4185 growth, no inhibition was observed in the in vivo experiments independent of the inoculum size. PMID:23243257

  1. Prevalence of Virulence Factors and Vancomycin-resistant Genes among Enterococcus faecalis and E. faecium Isolated from Clinical Specimens

    PubMed Central

    NASAJ, Mona; MOUSAVI, Seyed Masoud; HOSSEINI, Seyed Mostafa; ARABESTANI, Mohammad Reza

    2016-01-01

    Background: The aim of this study was to determine the occurrence of virulence determinants and vancomycin-resistant genes among Enterococcus faecalis and E. faecium obtained from various clinical sources. Methods: The study was performed on the 280 enterococcal isolated from clinical specimens in Hamadan hospitals, western Iran in 2012–14. Antibiotic susceptibility testing was performed using disk diffusion and Minimal Inhibitory Concentration (MIC) methods. The presence of vancomycin-resistant genes and virulence genes was investigated using PCR. Results: Totally 280 enterococcal isolates were identified as follows: E. faecalis (62.5%), E. faecium (24%) and Enterococcus spp (13.5%). The results of antibiotic susceptibility testing showed that resistance rates to vancomycin and teicoplanin in E. faecalis and E. faecium isolates were 5% and 73%, respectively. Of Sixty vancomycin-resistant Enterococci strains, fifty-one isolates were identified as E. faecium (VREfm) and nine as E. faecalis (VREfs). Prevalence of esp, hyl, and asa1 genes were determined as 82%, 71.6%, and 100%, respectively in E. faecium strains; and 78%, 56/6%, and 97%, respectively in E. faecalis strains. Conclusion: The increased frequency of VREF, as seen with rapid rise in the number of vanA isolates should be considered in infection control practices. PMID:27648425

  2. High-level aminoglycoside resistance in Enterococcus faecalis and Enterococcus faecium causing invasive infection: Twelve-year surveillance in the Minami Ibaraki Area.

    PubMed

    Osuka, Hanako; Nakajima, Jun; Oishi, Tsuyoshi; Funayama, Yasunori; Ebihara, Tsugio; Ishikawa, Hiroichi; Saito, Kazuto; Koganemaru, Hiroshi; Hitomi, Shigemi

    2016-01-01

    We examined prevalence of high-level aminoglycoside resistance (HLAR) in Enterococcus faecalis and Enterococcus faecium causing invasive infection in the Minami Ibaraki Area. Ten strains of both species each, recovered from the blood or the cerebrospinal fluid between 2003 and 2014, were randomly selected every year. High-level resistance to gentamicin (HLR-GM) and streptomycin (HLR-SM) was detected in 34% (41 of 120 strains) and 18% (21) of E. faecalis and 9% (11) and 39% (48) of E. faecium, respectively. In comparisons of the proportions among three four-year periods, HLR-SM among E. faecium was significantly lower in the 2011-2014 period. All strains with HLR-GM were positive for the aac(6')-Ie-aph(2″)-Ia gene. The ant(6')-Ia gene was detected in all with HLR-SM except for one E. faecalis strain. The present study showed that prevalence of HLR-GM among E. faecalis and E. faecium causing invasive infection in this area was nearly equivalent to that described in previous studies in Japan and that proportions of strains with HLAR did not vary during the study period except for that of HLR-SM among E. faecium.

  3. The mazEF toxin-antitoxin system as an attractive target in clinical isolates of Enterococcus faecium and Enterococcus faecalis.

    PubMed

    Soheili, Sara; Ghafourian, Sobhan; Sekawi, Zamberi; Neela, Vasantha Kumari; Sadeghifard, Nourkhoda; Taherikalani, Morovat; Khosravi, Afra; Ramli, Ramliza; Hamat, Rukman Awang

    2015-01-01

    The toxin-antitoxin (TA) system is a regulatory system where two sets of genes encode the toxin and its corresponding antitoxin. In this study, the prevalence of TA systems in independently isolated clinical isolates of Enterococcus faecium and Enterococcus faecalis was determined, the dominant TA system was identified, different virulence genes in E. faecium and E. faecalis were surveyed, the level of expression of the virulence and TA genes in normal and stress conditions was determined, and finally their associations with the TA genes were defined. Remarkably, the analysis demonstrated higBA and mazEF in all clinical isolates, and their locations were on chromosomes and plasmids, respectively. On the other hand, a quantitative analysis of TA and virulence genes revealed that the expression level in both genes is different under normal and stress conditions. The results obtained by anti-mazF peptide nucleic acids demonstrated that the expression level of virulence genes had decreased. These findings demonstrate an association between TA systems and virulence factors. The mazEF on the plasmids and the higBA TA genes on the chromosomes of all E. faecium and E. faecalis strains were dominant. Additionally, there was a decrease in the expression of virulence genes in the presence of anti-mazF peptide nucleic acids. Therefore, it is suggested that mazEF TA systems are potent and sensitive targets in all E. faecium and E. faecalis strains.

  4. Biochemical characterization of an anti-Candida factor produced by Enterococcus faecalis

    PubMed Central

    2012-01-01

    Background Because Candida albicans is resistant to several antifungal antibiotics, there is a need to identify other less toxic natural products, particularly antimicrobial proteins, peptides or bacteriocin like inhibitory substances. An attempt has been made to purify and characterise an anti-Candida compound produced by Enterococcus faecalis. Results An anti-Candida protein (ACP) produced by E. faecalis active against 8 C. albicans strains was characterised and partially purified. The ACP showed a broad-spectrum activity against multidrug resistant C. albicans MTCC 183, MTCC 7315, MTCC 3958, NCIM 3557, NCIM 3471 and DI. It was completely inactivated by treatment with proteinase K and partially by pronase E. The ACP retained biological stability after heat-treatment at 90°C for 20 min, maintained activity over a pH range 6–10, and remained active after treatment with α-amylase, lipase, organic solvents, and detergents. The antimicrobial activity of the E. faecalis strain was found exclusively in the extracellular filtrate produced in the late logarithmic growth phase. The highest activity (1600 AU mL-1) against C. albicans MTCC 183 was recorded at 48 h of incubation, and activity decreased thereafter. The peptide showed very low haemagglutination and haemolytic activities against human red blood cells. The antimicrobial substance was purified by salt-fractionation and chromatography. Partially purified ACP had a molecular weight of approximately 43 KDa in Tricine-PAGE analysis. The 12 amino acid N terminal sequence was obtained by Edman degradation. The peptide was de novo sequenced by ESI-MS, and the deduced combined sequence when compared to other bacteriocins and antimicrobial peptide had no significant sequence similarity. Conclusions The inhibitory activity of the test strain is due to the synthesis of an antimicrobial protein. To our knowledge, this is the first report on the isolation of a promising non-haemolytic anti-Candida protein from E

  5. Complete Genome Sequence of Enterococcus faecalis Strain P8-1 Isolated from Wild Magellanic Penguin (Spheniscus magellanicus) Feces on the South Coast of Brazil

    PubMed Central

    Prichula, Janira; Campos, Fabricio Souza; Pereira, Rebeca Inhoque; Cardoso, Leonardo Almansa; Wachholz, Guilherme Raffo; Pieta, Luiza; Mariot, Roberta Fogliatto; de Moura, Tiane Martin; Tavares, Maurício; d’Azevedo, Pedro Alves; Frazzon, Ana Paula Guedes

    2016-01-01

    Enterococcus faecalis strains have a ubiquitous nature that allows them to survive in different niches. Studies involving enterococci isolated from marine animals are scarce. Therefore, in this study, we report the complete genome sequence of E. faecalis strain P8-1 isolated from feces of a Magellanic penguin on the south coast of Brazil. PMID:26769928

  6. Complete Genome Sequence of Enterococcus faecalis Strain P8-1 Isolated from Wild Magellanic Penguin (Spheniscus magellanicus) Feces on the South Coast of Brazil.

    PubMed

    Prichula, Janira; Campos, Fabricio Souza; Pereira, Rebeca Inhoque; Cardoso, Leonardo Almansa; Wachholz, Guilherme Raffo; Pieta, Luiza; Mariot, Roberta Fogliatto; de Moura, Tiane Martin; Tavares, Maurício; d'Azevedo, Pedro Alves; Frazzon, Jeverson; Frazzon, Ana Paula Guedes

    2016-01-01

    Enterococcus faecalis strains have a ubiquitous nature that allows them to survive in different niches. Studies involving enterococci isolated from marine animals are scarce. Therefore, in this study, we report the complete genome sequence of E. faecalis strain P8-1 isolated from feces of a Magellanic penguin on the south coast of Brazil. PMID:26769928

  7. Substantivity of Ag-Ca-Si mesoporous nanoparticles on dentin and its ability to inhibit Enterococcus faecalis.

    PubMed

    Fan, Wei; Wu, Yujie; Ma, Tengjiao; Li, Yanyun; Fan, Bing

    2016-01-01

    The main purpose of this study was to investigate the substantivity of Ag-Ca-Si mesoporous nanoparticles (Ag-MCSNs) on dentin and its residual antibacterial effects against Enterococcus faecalis. Ag-MCSNs were fabricated and characterized, ion release profile and pH were tested, and the ability to inhibit planktonic E. faecalis as well as the cytotoxicity was evaluated. Dentin slices were medicated with Ca(OH)2 paste, 2 % chlorhexidine gel and Ag-MCSNs paste for 7 days and then irrigated. Dentin slices were then immersed in E. faecalis suspension for 6 days and then transferred to fresh brain heart infusion solution. The optical density value within 10 h after immersing and transferring were measured and compared among groups. Results indicated that Ag-MCSNs showed high pH, sustained Ag(+)-Ca(2+)-SiO3 (2-) ion release, and high substantivity on dentin. The Ag-MCSNs exhibited strong antibacterial effects against planktonic E. faecalis and much better residual inhibition effects against E. faecalis growth on dentin than Ca(OH)2 paste (P < 0.05). The Ag-MCSNs showed excellent antibacterial ability against E. faecalis and high substantivity on dentin, which might be developed to a new effective intra-canal medicament for human teeth.

  8. Effect of the probiotic Enterococcus faecium NCIMB10415 on cell numbers of total Enterococcus spp., E. faecium and E. faecalis in the intestine of piglets.

    PubMed

    Vahjen, W; Taras, D; Simon, O

    2007-03-01

    Sows and their piglets were fed a diet supplemented with or without the probiotic E. faecium NCIMB10415 (also known as SF68). Piglets were sacrificed 14, 28, 35 and 56 days after birth and DNA from intestinal segments was extracted and purified. A real time PCR assay was used to distinguish Enterococcus spp. (16s rDNA based), E. faecium (Efaafm gene), E. faecalis (Efaafs gene) as well as the probiotic strain (unique plasmid sequence). Extracts of autoclaved sow feces inoculated with E. faecium and E. faecalis cultures were used to calibrate real time PCR results. The probiotic strain was detected in 14 day old suckling piglets before the piglets had access to the starter diet. In piglets of the probiotic group, probiotic E. faecium cell counts were always a significant proportion of total E. faecium cells in stomach digesta (4-20%), however only a small fraction of the total Enterococcus spp. cell number on day 14 and 28 in all intestinal segments (0.1-0.7%). Compared to control samples, the probiotic E. faecium strain significantly (p < or = 0.05) decreased the amount of total Enterococcus spp. and E. faecalis cells in the colon of 14 day old suckling piglets as well as in jejunum and colon samples one week after weaning. E. faecium cell counts were not modified on any sampling day or intestinal segment. This study showed that the presence of probiotic E. faecium NCIMB10415 coincided with reduced total E. faecalis, but not total E. faecium cell numbers in the intestine of piglets. In view of unchanged cell numbers and ratios in sow feces, modifications must have taken place within the intestine of suckling piglets.

  9. Cell Wall Chemical Composition of Enterococcus faecalis in the Viable but Nonculturable State

    PubMed Central

    Signoretto, Caterina; del Mar Lleò, Maria; Tafi, Maria Carla; Canepari, Pietro

    2000-01-01

    The viable but nonculturable (VBNC) state is a survival mechanism adopted by many bacteria (including those of medical interest) when exposed to adverse environmental conditions. In this state bacteria lose the ability to grow in bacteriological media but maintain viability and pathogenicity and sometimes are able to revert to regular division upon restoration of normal growth conditions. The aim of this work was to analyze the biochemical composition of the cell wall of Enterococcus faecalis in the VBNC state in comparison with exponentially growing and stationary cells. VBNC enterococcal cells appeared as slightly elongated and were endowed with a wall more resistant to mechanical disruption than dividing cells. Analysis of the peptidoglycan chemical composition showed an increase in total cross-linking, which rose from 39% in growing cells to 48% in VBNC cells. This increase was detected in oligomers of a higher order than dimers, such as trimers (24% increase), tetramers (37% increase), pentamers (65% increase), and higher oligomers (95% increase). Changes were also observed in penicillin binding proteins (PBPs), the enzymes involved in the terminal stages of peptidoglycan assembly, with PBPs 5 and 1 being prevalent, and in autolytic enzymes, with a threefold increase in the activity of latent muramidase-1 in E. faecalis in the VBNC state. Accessory wall polymers such as teichoic acid and lipoteichoic acid proved unchanged and doubled in quantity, respectively, in VBNC cells in comparison to dividing cells. It is suggested that all these changes in the cell wall of VBNC enterococci are specific to this particular physiological state. This may provide indirect confirmation of the viability of these cells. PMID:10788366

  10. Drosophila host model reveals new enterococcus faecalis quorum-sensing associated virulence factors.

    PubMed

    Teixeira, Neuza; Varahan, Sriram; Gorman, Matthew J; Palmer, Kelli L; Zaidman-Remy, Anna; Yokohata, Ryoji; Nakayama, Jiro; Hancock, Lynn E; Jacinto, António; Gilmore, Michael S; de Fátima Silva Lopes, Maria

    2013-01-01

    Enterococcus faecalis V583 is a vancomycin-resistant clinical isolate which belongs to the hospital-adapted clade, CC2. This strain harbours several factors that have been associated with virulence, including the fsr quorum-sensing regulatory system that is known to control the expression of GelE and SprE proteases. To discriminate between genes directly regulated by Fsr, and those indirectly regulated as the result of protease expression or activity, we compared gene expression in isogenic mutants of V583 variously defective in either Fsr quorum sensing or protease expression. Quorum sensing was artificially induced by addition of the quorum signal, GBAP, exogenously in a controlled manner. The Fsr regulon was found to be restricted to five genes, gelE, sprE, ef1097, ef1351 and ef1352. Twelve additional genes were found to be dependent on the presence of GBAP-induced proteases. Induction of GelE and SprE by GBAP via Fsr resulted in accumulation of mRNA encoding lrgAB, and this induction was found to be lytRS dependent. Drosophila infection was used to discern varying levels of toxicity stemming from mutations in the fsr quorum regulatory system and the genes that it regulates, highlighting the contribution of LrgAB and bacteriocin EF1097 to infection toxicity. A contribution of SprE to infection toxicity was also detected. This work brought to light new players in E. faecalis success as a pathogen and paves the way for future studies on host tolerance mechanisms to infections caused by this important nosocomial pathogen.

  11. Reduction of Enterococcus faecalis in curved root canals after various sizes and tapers of canal preparation

    PubMed Central

    Moshari, Amir Abbas; Akhlaghi, Nahid Mohammadzadeh; Rahimifard, Nahid; Darmiani, Soheila

    2015-01-01

    Aims: The aim of this study was to evaluate the reduction of Enterococcus faecalis in curved root canals after various sizes and tapers of the canal preparation. Materials and Methods: Mandibular first molars (n = 103) with curved mesiobuccal canals were divided into one control (n = 5) and 7 experimental (n = 14) groups, were inoculated with E. faecalis (ATTC 29212) and prepared with the following RaCe files (FKG Dentaire) as master apical file: Groups: 25.04, 25.06, 30.04, 30.06, 35.04, 35.06 and 40.06. All the experimental groups were irrigated with 2 mL of 1% sodium hypochlorite during instrumentation and finally rinsed with 17% ethylenediaminetetraacetic acid (EDTA) (2 mL) followed by 5.25% NaOCl (2 mL) and sterile distilled water. Colony counting was performed after incubation. Statistical Analysis Used: Resulting data were analyzed using one-way ANOVA and Tukey's post-hoc test, (P < 0.05). Results and Conclusions: All the experimental groups showed significant bacterial reduction (P < 0.001). Although the greater the size/taper or both led to more decreased amount of bacteria, differences between the groups with the identical size and different tapers, and among the groups with the same taper and different sizes were not significant. Based on this study, 25.04 along with using 2 mL of 1% NaOCl during instrumentation, and using 17% EDTA and 5.25% NaOCl as final rinse successively after the termination of preparation, can effectively reduce intra-canal bacteria and preserve root structure. PMID:26180416

  12. Fine-Tuned Transcriptional Regulation of Malate Operons in Enterococcus faecalis

    PubMed Central

    Mortera, Pablo; Espariz, Martín; Suárez, Cristian; Repizo, Guillermo; Deutscher, Josef; Alarcón, Sergio; Blancato, Víctor

    2012-01-01

    In Enterococcus faecalis, the mae locus is constituted by two putative divergent operons, maePE and maeKR. The first operon encodes a putative H+/malate symporter (MaeP) and a malic enzyme (MaeE) previously shown to be essential for malate utilization in this bacterium. The maeKR operon encodes two putative proteins with significant similarity to two-component systems involved in sensing malate and activating its assimilation in bacteria. Our transcriptional and genetic assays showed that maePE and maeKR are induced in response to malate by the response regulator MaeR. In addition, we observed that both operons were partially repressed in the presence of glucose. Accordingly, the cometabolism of this sugar and malate was detected. The binding of the complex formed by CcpA and its corepressor P-Ser-HPr to a cre site located in the mae region was demonstrated in vitro and explains the carbon catabolite repression (CCR) observed for the maePE operon. However, our results also provide evidence for a CcpA-independent CCR mechanism regulating the expression of both operons. Finally, a biomass increment of 40 or 75% was observed compared to the biomass of cells grown only on glucose or malate, respectively. Cells cometabolizing both carbon sources exhibit a higher rate of glucose consumption and a lower rate of malate utilization. The growth improvement achieved by E. faecalis during glucose-malate cometabolism might explain why this microorganism employs different regulatory systems to tightly control the assimilation of both carbon sources. PMID:22247139

  13. Atomic force microscopy visualization of injuries in Enterococcus faecalis surface caused by Er,Cr:YSGG and diode lasers

    PubMed Central

    López-Jiménez, Lidia; Viñas, Miguel; Vinuesa, Teresa

    2015-01-01

    Aim: To visualize by Atomic Force Microscopy the alterations induced on Enterococcus. faecalis surface after treatment with 2 types of laser: Erbium chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser and Diode laser. Material and Methods: Bacterial suspensions from overnight cultures of E. faecalis were irradiated during 30 seconds with the laser-lights at 1 W and 2 W of power, leaving one untreated sample as control. Surface alterations on treated E. faecalis were visualized by Atomic Force Microscopy (AFM) and its surface roughness determined. Results: AFM imaging showed that at high potency of laser both cell morphology and surface roughness resulted altered, and that several cell lysis signs were easily visualized. Surface roughness clearly increase after the treatment with Er,Cr:YSGG at 2W of power, while the other treatments gave similar values of surface roughness. The effect of lasers on bacterial surfaces visualized by AFM revealed drastic alterations. Conclusions: AFM is a good tool to evaluate surface injuries after laser treatment; and could constitute a measure of antimicrobial effect that can complete data obtained by determination of microbial viability. Key words:Atomic force microscopy, Er,Cr:YSGG laser, diode laser, Enterococcus faecalis, surface roughness. PMID:25475770

  14. The Transcriptome of the Nosocomial Pathogen Enterococcus faecalis V583 Reveals Adaptive Responses to Growth in Blood

    PubMed Central

    Vebø, Heidi C.; Snipen, Lars; Nes, Ingolf F.; Brede, Dag A.

    2009-01-01

    Background Enterococcus faecalis plays a dual role in human ecology, predominantly existing as a commensal in the alimentary canal, but also as an opportunistic pathogen that frequently causes nosocomial infections like bacteremia. A number of virulence factors that contribute to the pathogenic potential of E. faecalis have been established. However, the process in which E. faecalis gains access to the bloodstream and establishes a persistent infection is not well understood. Methodology/Principal Findings To enhance our understanding of how this commensal bacterium adapts during a bloodstream infection and to examine the interplay between genes we designed an in vitro experiment using genome-wide microarrays to investigate what effects the presence of and growth in blood have on the transcriptome of E. faecalis strain V583. We showed that growth in both 2xYT supplemented with 10% blood and in 100% blood had a great impact on the transcription of many genes in the V583 genome. We identified several immediate changes signifying cellular processes that might contribute to adaptation and growth in blood. These include modulation of membrane fatty acid composition, oxidative and lytic stress protection, acquisition of new available substrates, transport functions including heme/iron transporters and genes associated with virulence in E. faecalis. Conclusions/Significance The results presented here reveal that cultivation of E. faecalis in blood in vitro has a profound impact on its transcriptome, which includes a number of virulence traits. Observed regulation of genes and pathways revealed new insight into physiological features and metabolic capacities which enable E. faecalis to adapt and grow in blood. A number of the regulated genes might potentially be useful candidates for development of new therapeutic approaches for treatment of E. faecalis infections. PMID:19888459

  15. Partial purification and characterization of bacteriocin produced by Enterococcus faecalis DU10 and its probiotic attributes.

    PubMed

    Perumal, Venkatesh; Repally, Ayyanna; Dasari, Ankaiah; Venkatesan, Arul

    2016-10-01

    A novel bacteriocin produced by avian duck isolated lactic acid bacterium Enterococcus faecalis DU10 was isolated. This bacteriocin showed a broad spectrum of antibacterial activity against important food-borne pathogens and was purified by size exclusion chromatography followed by reverse-phase high-performance liquid chromatography in a C-18 column. Tricine-SDS PAGE revealed the presence of a band with an estimated molecular mass of 6.3 kDa. The zymogram clearly linked the antimicrobial activity with this band. This result was further confirmed by mass-assisted laser desorption ionization time-of-flight mass spectrometry, since a sharp peak corresponding to 6.313 kDa was detected and the functional groups were revealed by Fourier transform infrared spectroscopy. Bacteriocin DU10 activity was found sensitive to proteinase-K and pepsin and partially affected by trypsin and α-chymotrypsin. The activity of bacteriocin DU10 was partially resistant to heat treatments ranging from 30 to 90°C for 30 min. It also withstood a treatment at 121°C for 10 min. Cytotoxicity of bacteriocin DU10 by methyl-thiazolyl-diphenyl-tetrazolium bromide assay showed that the viability of HT-29 and HeLa cells decreased 60 ± 0.7% and 43 ± 4.8%, respectively, in the presence of 3,200 AU/mL of bacteriocin. The strain withstood 0.3% w/v of bile oxgall and pH 2 affected the bacterial growth between 2 and 4 hr of incubation. Adhesion properties examined with HT-29 cell line showed 69.85% initial population of strain E. faecalis DU10, which was found to be strongly adhered to this cell line. These results conclude bacteriocin DU10 may be used as a potential biopreservative and E. faecalis DU10 may be used as a potential probiont to control Salmonella infections. PMID:26786752

  16. Partial purification and characterization of bacteriocin produced by Enterococcus faecalis DU10 and its probiotic attributes.

    PubMed

    Perumal, Venkatesh; Repally, Ayyanna; Dasari, Ankaiah; Venkatesan, Arul

    2016-10-01

    A novel bacteriocin produced by avian duck isolated lactic acid bacterium Enterococcus faecalis DU10 was isolated. This bacteriocin showed a broad spectrum of antibacterial activity against important food-borne pathogens and was purified by size exclusion chromatography followed by reverse-phase high-performance liquid chromatography in a C-18 column. Tricine-SDS PAGE revealed the presence of a band with an estimated molecular mass of 6.3 kDa. The zymogram clearly linked the antimicrobial activity with this band. This result was further confirmed by mass-assisted laser desorption ionization time-of-flight mass spectrometry, since a sharp peak corresponding to 6.313 kDa was detected and the functional groups were revealed by Fourier transform infrared spectroscopy. Bacteriocin DU10 activity was found sensitive to proteinase-K and pepsin and partially affected by trypsin and α-chymotrypsin. The activity of bacteriocin DU10 was partially resistant to heat treatments ranging from 30 to 90°C for 30 min. It also withstood a treatment at 121°C for 10 min. Cytotoxicity of bacteriocin DU10 by methyl-thiazolyl-diphenyl-tetrazolium bromide assay showed that the viability of HT-29 and HeLa cells decreased 60 ± 0.7% and 43 ± 4.8%, respectively, in the presence of 3,200 AU/mL of bacteriocin. The strain withstood 0.3% w/v of bile oxgall and pH 2 affected the bacterial growth between 2 and 4 hr of incubation. Adhesion properties examined with HT-29 cell line showed 69.85% initial population of strain E. faecalis DU10, which was found to be strongly adhered to this cell line. These results conclude bacteriocin DU10 may be used as a potential biopreservative and E. faecalis DU10 may be used as a potential probiont to control Salmonella infections.

  17. Molecular Epidemiologic Analysis of Enterococcus faecalis Isolates in Cuba by Multilocus Sequence Typing

    PubMed Central

    Kobayashi, Nobumichi; Nagashima, Shigeo

    2009-01-01

    We carried out the first study of Enterococcus faecalis clinical isolates in Cuba by multilocus sequence typing linking the molecular typing data with the presence of virulence determinants and the antibiotic resistance genes. A total of 23 E. faecalis isolates recovered from several clinic sources and geographic areas of Cuba during a period between 2000 and 2005 were typed by multilocus sequence typing. Thirteen sequence types (STs) including five novel STs were identified, and the ST 64 (clonal complex [CC] 8), ST 6 (CC2), ST 21(CC21), and ST 16 (CC58) were found in more than one strain. Sixty-seven percent of STs corresponded to STs reported previously in Spain, Poland, and The Netherlands, and other STs (ST115, ST64, ST6, and ST40) were genetically close to those detected in the United States. Prevalence of both antimicrobial resistance genes [aac(6′)-aph(2″), aph(3′), ant(6), ant(3″)(9), aph(2″)-Id, aph(2″)-Ic, erm(B), erm(A), erm(C), mef(A), tet(M), and tet(L)] and virulence genes (agg, gelE, cylA, esp, ccf, and efaAfs) were examined by polymerase chain reaction. Aminoglycoside resistance genes aac(6′)-Ie-aph(2″)-Ia, aph(3′), ant(6), ant(3″)(9) were more frequently detected in ST6, ST16, ST23, ST64, and ST115. The multidrug resistance was distributed to all STs detected, except for ST117 and singleton ST225. The presence of cyl gene was specifically linked to the ST64 and ST16. Presence of the esp, gel, and agg genes was not specific to any particular ST. This research provided the first insight into the population structure of E. faecalis in Cuba, that is, most Cuban strains were related to European strains, whereas others to U.S. strains. The CC2, CC21, and CC8, three of the biggest CCs in the world, were evidently circulating in Cuba, associated with multidrug resistance and virulence traits. PMID:19857135

  18. CRISPR-Cas and Restriction-Modification Act Additively against Conjugative Antibiotic Resistance Plasmid Transfer in Enterococcus faecalis.

    PubMed

    Price, Valerie J; Huo, Wenwen; Sharifi, Ardalan; Palmer, Kelli L

    2016-01-01

    Enterococcus faecalis is an opportunistic pathogen and a leading cause of nosocomial infections. Conjugative pheromone-responsive plasmids are narrow-host-range mobile genetic elements (MGEs) that are rapid disseminators of antibiotic resistance in the faecalis species. Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas and restriction-modification confer acquired and innate immunity, respectively, against MGE acquisition in bacteria. Most multidrug-resistant E. faecalis isolates lack CRISPR-Cas and possess an orphan locus lacking cas genes, CRISPR2, that is of unknown function. Little is known about restriction-modification defense in E. faecalis. Here, we explore the hypothesis that multidrug-resistant E. faecalis strains are immunocompromised. We assessed MGE acquisition by E. faecalis T11, a strain closely related to the multidrug-resistant hospital isolate V583 but which lacks the ~620 kb of horizontally acquired genome content that characterizes V583. T11 possesses the E. faecalis CRISPR3-cas locus and a predicted restriction-modification system, neither of which occurs in V583. We demonstrate that CRISPR-Cas and restriction-modification together confer a 4-log reduction in acquisition of the pheromone-responsive plasmid pAM714 in biofilm matings. Additionally, we show that the orphan CRISPR2 locus is functional for genome defense against another pheromone-responsive plasmid, pCF10, only in the presence of cas9 derived from the E. faecalis CRISPR1-cas locus, which most multidrug-resistant E. faecalis isolates lack. Overall, our work demonstrated that the loss of only two loci led to a dramatic reduction in genome defense against a clinically relevant MGE, highlighting the critical importance of the E. faecalis accessory genome in modulating horizontal gene transfer. Our results rationalize the development of antimicrobial strategies that capitalize upon the immunocompromised status of multidrug-resistant E. faecalis. IMPORTANCE

  19. CRISPR-Cas and Restriction-Modification Act Additively against Conjugative Antibiotic Resistance Plasmid Transfer in Enterococcus faecalis.

    PubMed

    Price, Valerie J; Huo, Wenwen; Sharifi, Ardalan; Palmer, Kelli L

    2016-01-01

    Enterococcus faecalis is an opportunistic pathogen and a leading cause of nosocomial infections. Conjugative pheromone-responsive plasmids are narrow-host-range mobile genetic elements (MGEs) that are rapid disseminators of antibiotic resistance in the faecalis species. Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas and restriction-modification confer acquired and innate immunity, respectively, against MGE acquisition in bacteria. Most multidrug-resistant E. faecalis isolates lack CRISPR-Cas and possess an orphan locus lacking cas genes, CRISPR2, that is of unknown function. Little is known about restriction-modification defense in E. faecalis. Here, we explore the hypothesis that multidrug-resistant E. faecalis strains are immunocompromised. We assessed MGE acquisition by E. faecalis T11, a strain closely related to the multidrug-resistant hospital isolate V583 but which lacks the ~620 kb of horizontally acquired genome content that characterizes V583. T11 possesses the E. faecalis CRISPR3-cas locus and a predicted restriction-modification system, neither of which occurs in V583. We demonstrate that CRISPR-Cas and restriction-modification together confer a 4-log reduction in acquisition of the pheromone-responsive plasmid pAM714 in biofilm matings. Additionally, we show that the orphan CRISPR2 locus is functional for genome defense against another pheromone-responsive plasmid, pCF10, only in the presence of cas9 derived from the E. faecalis CRISPR1-cas locus, which most multidrug-resistant E. faecalis isolates lack. Overall, our work demonstrated that the loss of only two loci led to a dramatic reduction in genome defense against a clinically relevant MGE, highlighting the critical importance of the E. faecalis accessory genome in modulating horizontal gene transfer. Our results rationalize the development of antimicrobial strategies that capitalize upon the immunocompromised status of multidrug-resistant E. faecalis. IMPORTANCE

  20. CRISPR-Cas and Restriction-Modification Act Additively against Conjugative Antibiotic Resistance Plasmid Transfer in Enterococcus faecalis

    PubMed Central

    Price, Valerie J.; Huo, Wenwen; Sharifi, Ardalan

    2016-01-01

    ABSTRACT Enterococcus faecalis is an opportunistic pathogen and a leading cause of nosocomial infections. Conjugative pheromone-responsive plasmids are narrow-host-range mobile genetic elements (MGEs) that are rapid disseminators of antibiotic resistance in the faecalis species. Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas and restriction-modification confer acquired and innate immunity, respectively, against MGE acquisition in bacteria. Most multidrug-resistant E. faecalis isolates lack CRISPR-Cas and possess an orphan locus lacking cas genes, CRISPR2, that is of unknown function. Little is known about restriction-modification defense in E. faecalis. Here, we explore the hypothesis that multidrug-resistant E. faecalis strains are immunocompromised. We assessed MGE acquisition by E. faecalis T11, a strain closely related to the multidrug-resistant hospital isolate V583 but which lacks the ~620 kb of horizontally acquired genome content that characterizes V583. T11 possesses the E. faecalis CRISPR3-cas locus and a predicted restriction-modification system, neither of which occurs in V583. We demonstrate that CRISPR-Cas and restriction-modification together confer a 4-log reduction in acquisition of the pheromone-responsive plasmid pAM714 in biofilm matings. Additionally, we show that the orphan CRISPR2 locus is functional for genome defense against another pheromone-responsive plasmid, pCF10, only in the presence of cas9 derived from the E. faecalis CRISPR1-cas locus, which most multidrug-resistant E. faecalis isolates lack. Overall, our work demonstrated that the loss of only two loci led to a dramatic reduction in genome defense against a clinically relevant MGE, highlighting the critical importance of the E. faecalis accessory genome in modulating horizontal gene transfer. Our results rationalize the development of antimicrobial strategies that capitalize upon the immunocompromised status of multidrug-resistant E. faecalis

  1. Biofilms of Enterococcus faecalis and Enterococcus faecium isolated from the processing of ricotta and the control of these pathogens through cleaning and sanitization procedures.

    PubMed

    da Silva Fernandes, Meg; Kabuki, Dirce Yorika; Kuaye, Arnaldo Yoshiteru

    2015-05-01

    The biofilm formation of Enterococcus faecalis and Enterococcus faecium isolated from the processing of ricotta on stainless steel coupons was evaluated, and the effect of cleaning and sanitization procedures in the control of these biofilms was determined. The formation of biofilms was observed while varying the incubation temperature (7, 25 and 39°C) and time (0, 1, 2, 4, 6 and 8 days). At 7°C, the counts of E. faecalis and E. faecium were below 2 log10 CFU/cm(2). For the temperatures of 25 and 39°C, after 1 day, the counts of E. faecalis and E. faecium were 5.75 and 6.07 log10 CFU/cm(2), respectively, which is characteristic of biofilm formation. The tested sanitation procedures a) acid-anionic tensioactive cleaning, b) anionic tensioactive cleaning+sanitizer and c) acid-anionic tensioactive cleaning+sanitizer were effective in removing the biofilms, reducing the counts to levels below 0.4 log10 CFU/cm(2). The sanitizer biguanide was the least effective, and peracetic acid was the most effective. These studies revealed the ability of enterococci to form biofilms and the importance of the cleaning step and the type of sanitizer used in sanitation processes for the effective removal of biofilms.

  2. Biofilms of Enterococcus faecalis and Enterococcus faecium isolated from the processing of ricotta and the control of these pathogens through cleaning and sanitization procedures.

    PubMed

    da Silva Fernandes, Meg; Kabuki, Dirce Yorika; Kuaye, Arnaldo Yoshiteru

    2015-05-01

    The biofilm formation of Enterococcus faecalis and Enterococcus faecium isolated from the processing of ricotta on stainless steel coupons was evaluated, and the effect of cleaning and sanitization procedures in the control of these biofilms was determined. The formation of biofilms was observed while varying the incubation temperature (7, 25 and 39°C) and time (0, 1, 2, 4, 6 and 8 days). At 7°C, the counts of E. faecalis and E. faecium were below 2 log10 CFU/cm(2). For the temperatures of 25 and 39°C, after 1 day, the counts of E. faecalis and E. faecium were 5.75 and 6.07 log10 CFU/cm(2), respectively, which is characteristic of biofilm formation. The tested sanitation procedures a) acid-anionic tensioactive cleaning, b) anionic tensioactive cleaning+sanitizer and c) acid-anionic tensioactive cleaning+sanitizer were effective in removing the biofilms, reducing the counts to levels below 0.4 log10 CFU/cm(2). The sanitizer biguanide was the least effective, and peracetic acid was the most effective. These studies revealed the ability of enterococci to form biofilms and the importance of the cleaning step and the type of sanitizer used in sanitation processes for the effective removal of biofilms. PMID:25702883

  3. Heat-Killed Enterococcus faecalis EF-2001 Ameliorates Atopic Dermatitis in a Murine Model.

    PubMed

    Choi, Eun-Ju; Iwasa, Masahiro; Han, Kwon-Il; Kim, Wan-Jae; Tang, Yujiao; Hwang, Young Joung; Chae, Jeong Ryong; Han, Weon Cheol; Shin, Yu-Su; Kim, Eun-Kyung

    2016-03-01

    Recent reports have shown the immunomodulatory effect of heat-killed lactic acid bacteria. Atopic dermatitis (AD) is an allergic skin disease, caused by immune dysregulation among other factors. The aim of this study was to assess the effect of heat-killed Enterococcus faecalis EF-2001 (EF-2001) on AD. We established an in vivo AD model by repeated local exposure of Dermatophagoides farinae extract (DFE; house dust mite extract) and 2,4-dinitrochlorobenzene (DNCB) to the ears of mice. After oral administration of EF-2001 for four weeks, the epidermal and dermal ear thickness, mast cell infiltration, and serum immunoglobulin levels were measured. In addition, the gene expression levels of pathogenic cytokines in the ears, lymph nodes, and splenocytes were assayed. EF-2001 attenuated AD symptoms based on the ear thickness, histopathological analysis, and serum immunoglobulin levels. Moreover, EF-2001 decreased the DFE/DNCB-induced expression of various pathogenic cytokines in the ears, lymph nodes, and splenocytes. These results suggest that EF-2001 has therapeutic potential in the treatment of AD owing to its immunomodulatory effects. PMID:26959058

  4. Heat-Killed Enterococcus faecalis EF-2001 Ameliorates Atopic Dermatitis in a Murine Model

    PubMed Central

    Choi, Eun-Ju; Iwasa, Masahiro; Han, Kwon-Il; Kim, Wan-Jae; Tang, Yujiao; Hwang, Young Joung; Chae, Jeong Ryong; Han, Weon Cheol; Shin, Yu-Su; Kim, Eun-Kyung

    2016-01-01

    Recent reports have shown the immunomodulatory effect of heat-killed lactic acid bacteria. Atopic dermatitis (AD) is an allergic skin disease, caused by immune dysregulation among other factors. The aim of this study was to assess the effect of heat-killed Enterococcus faecalis EF-2001 (EF-2001) on AD. We established an in vivo AD model by repeated local exposure of Dermatophagoides farinae extract (DFE; house dust mite extract) and 2,4-dinitrochlorobenzene (DNCB) to the ears of mice. After oral administration of EF-2001 for four weeks, the epidermal and dermal ear thickness, mast cell infiltration, and serum immunoglobulin levels were measured. In addition, the gene expression levels of pathogenic cytokines in the ears, lymph nodes, and splenocytes were assayed. EF-2001 attenuated AD symptoms based on the ear thickness, histopathological analysis, and serum immunoglobulin levels. Moreover, EF-2001 decreased the DFE/DNCB-induced expression of various pathogenic cytokines in the ears, lymph nodes, and splenocytes. These results suggest that EF-2001 has therapeutic potential in the treatment of AD owing to its immunomodulatory effects. PMID:26959058

  5. Insidious Onset of Tetraparesis due to Cervical Epidural Abscess from Enterococcus faecalis.

    PubMed

    Soultanis, Konstantinos Chr; Sakellariou, Vasileios I; Starantzis, Konstantinos A; Stavropoulos, Nikolaos A; Papagelopoulos, Panayiotis J

    2013-01-01

    We report a case of cervical epidural abscess from Enterococcus faecalis, which caused an insidious onset of tetraparesis. This 53-year-old female with a history of diabetes mellitus and chronic renal failure under hemodialysis presented with pain and progressive weakness of upper and lower extremities without fever. Although a recent MRI she did at the beginning of symptoms showed no significant pathologies, except for a cervical disc herniation and adjacent spinal degeneration, and stenosis that confused the diagnostic procedure, newer imaging with CT and MRI, which was performed due to progression of tetraparesis, revealed the formation of a cervical epidural abscess. Surgical drainage was done after a complete infection workup. The patient showed immediate neurological improvement after surgery. She received antibiotics intravenously for 3 weeks and orally for another 6 weeks. The patient was free from complications 24 months after surgery. A high index of suspicion is most important in making a rapid and correct diagnosis of spinal epidural abscess. The classic clinical triad (fever, local pain, and neurologic deficits) is not sensitive enough for early detection. Continuous clinical, laboratory, and imaging monitoring are of paramount importance. Early diagnosis and surgical intervention could optimize the final functional outcome. PMID:23573096

  6. The polyamine N-acetyltransferase-like enzyme PmvE plays a role in the virulence of Enterococcus faecalis.

    PubMed

    Martini, Cecilia; Michaux, Charlotte; Bugli, Francesca; Arcovito, Alessandro; Iavarone, Federica; Cacaci, Margherita; Paroni Sterbini, Francesco; Hartke, Axel; Sauvageot, Nicolas; Sanguinetti, Maurizio; Posteraro, Brunella; Giard, Jean-Christophe

    2015-01-01

    We previously showed that the mutant strain of Enterococcus faecalis lacking the transcriptional regulator SlyA is more virulent than the parental strain. We hypothesized that this phenotype was due to overexpression of the second gene of the slyA operon, ef_3001, renamed pmvE (for polyamine metabolism and virulence of E. faecalis). PmvE shares strong homologies with N(1)-spermidine/spermine acetyltransferase enzymes involved in the metabolism of polyamines. In this study, we used an E. faecalis strain carrying the recombinant plasmid pMSP3535-pmvE (V19/p3535-pmvE), which allows the induction of pmvE by addition of nisin. Thereby, we showed that the overexpression of PmvE increased the virulence of E. faecalis in the Galleria mellonella infection model, as well as the persistence within peritoneal macrophages. We were also able to show a direct interaction between the His-tagged recombinant PmvE (rPmvE) protein and putrescine by the surface plasmon resonance (SPR) technique on a Biacore instrument. Moreover, biochemical assays showed that PmvE possesses an N-acetyltransferase activity toward polyamine substrates. Our results suggest that PmvE contributes to the virulence of E. faecalis, likely through its involvement in the polyamine metabolism. PMID:25385793

  7. An in vitro study on the effects of nisin on the antibacterial activities of 18 antibiotics against Enterococcus faecalis.

    PubMed

    Tong, Zhongchun; Zhang, Yuejiao; Ling, Junqi; Ma, Jinglei; Huang, Lijia; Zhang, Luodan

    2014-01-01

    Enterococcus faecalis rank among the leading causes of nosocomial infections worldwide and possesses both intrinsic and acquired resistance to a variety of antibiotics. Development of new antibiotics is limited, and pathogens continually generate new antibiotic resistance. Many researchers aim to identify strategies to effectively kill this drug-resistant pathogen. Here, we evaluated the effect of the antimicrobial peptide nisin on the antibacterial activities of 18 antibiotics against E. faecalis. The MIC and MBC results showed that the antibacterial activities of 18 antibiotics against E. faecalis OG1RF, ATCC 29212, and strain E were significantly improved in the presence of 200 U/ml nisin. Statistically significant differences were observed between the results with and without 200 U/ml nisin at the same concentrations of penicillin or chloramphenicol (p<0.05). The checkerboard assay showed that the combination of nisin and penicillin or chloramphenicol had a synergetic effect against the three tested E. faecalis strains. The transmission electron microscope images showed that E. faecalis was not obviously destroyed by penicillin or chloramphenicol alone but was severely disrupted by either antibiotic in combination with nisin. Furthermore, assessing biofilms by a confocal laser scanning microscope showed that penicillin, ciprofloxacin, and chloramphenicol all showed stronger antibiofilm actions in combination with nisin than when these antibiotics were administered alone. Therefore, nisin can significantly improve the antibacterial and antibiofilm activities of many antibiotics, and certain antibiotics in combination with nisin have considerable potential for use as inhibitors of this drug-resistant pathogen.

  8. The effect of sodium hypochlorite on Enterococcus faecalis when grown on dentine as a single- and multi-species biofilm.

    PubMed

    Yap, Benlee; Zilm, Peter S; Briggs, Nancy; Rogers, Anthony H; Cathro, Peter C

    2014-12-01

    Enterococcus faecalis is often involved in the aetiology of apical periodontitis after endodontic treatment. This project aimed to establish, on dentine in vitro, a multi-species biofilm containing E. faecalis, and to determine if the organism had an increased resistance to sodium hypochlorite compared with an axenic biofilm. Biofilms were established on dentine discs in flow cells with either E. faecalis alone (axenic) or together with Fusobacterium nucleatum and Streptococcus sanguinis. Following treatment with either 0.9% sodium hypochlorite or saline, the viability of E. faecalis was determined by serial plating and qualitative analysis was performed by scanning electron microscopy and confocal laser scanning microscopy. Viable counts indicated that 0.9% NaOCl is highly effective against E. faecalis grown alone and as part of a multi-species biofilm (P = 0.0005 and P = 0.001, respectively). No significant difference in its survival in the two biofilm types was found (P = 0.8276).

  9. Antibacterial and residual antimicrobial activities against Enterococcus faecalis biofilm: A comparison between EDTA, chlorhexidine, cetrimide, MTAD and QMix

    NASA Astrophysics Data System (ADS)

    Zhang, Rui; Chen, Min; Lu, Yan; Guo, Xiangjun; Qiao, Feng; Wu, Ligeng

    2015-08-01

    We compared the antibacterial and residual antimicrobial activities of five root canal irrigants (17% EDTA,2% chlorhexidine,0.2% cetrimide, MTAD, and QMix) in a model of Enterococcus faecalis biofilm formation. Sixty dentin blocks with 3-week E. faecalis biofilm were divided into six equal groups and flushed with irrigant for 2 min or left untreated. A blank control group was also established. Antibacterial activities of the irrigants were evaluated by counting colony forming units. To test residual antimicrobial activities, 280 dentin blocks were divided into seven equal groups and flushed with irrigant for 2 min or left untreated and then incubated with E. faecalis suspension for 48 h, or used as a blank. No bacteria were observed in the blank control group. The number of viable E. faecalis was significantly fewer in the irrigant-treated groups compared with the untreated control (P < 0.05). Among the five irrigants, QMix had the strongest antibacterial activity. Residual antimicrobial activities of CHX were significantly higher at 12 h, 24 h and 36 h compared to untreated control (P < 0.05). All five root canal irrigants were effective to some extent against E. faecalis, but QMix and CHX had the strongest, and CHX the longest (up to 36 h), antimicrobial activity.

  10. Effects of storage temperature on tyramine production by Enterococcus faecalis R612Z1 in water-boiled salted ducks.

    PubMed

    Liu, Fang; Du, Lihui; Wu, Haihong; Wang, Daoying; Zhu, Yongzhi; Geng, Zhiming; Zhang, Muhan; Xu, Weimin

    2014-10-01

    Tyramine production by Enterococcus faecalis R612Z1 in water-boiled salted ducks was evaluated during storage at different temperatures. The results showed that E. faecalis R612Z1 could produce tyramine in meat samples when the storage temperature was no less than 4°C. The E. faecalis R612Z1 counts of the meat samples reached 10(8) CFU/g on day 7 at 4°C and on day 4 at 10°C. However, the tyramine content of the meat samples stored at 10°C increased to 23.73 μg/g (on day 10), which was greater than the level in the samples stored at 4°C (7.56 μg/g). Reverse transcription quantitative PCR detection of the expression level of the tyrDC gene in E. faecalis R612Z1 in the meat samples revealed no significant changes at different storage temperatures. Thus, the changes in tyramine production of E. faecalis R612Z1 may be due to the different enzymatic activities at different storage temperatures.

  11. Antimicrobial activity of some essential oils against oral multidrug-resistant Enterococcus faecalis in both planktonic and biofilm state

    PubMed Central

    Benbelaïd, Fethi; Khadir, Abdelmounaïm; Abdoune, Mohamed Amine; Bendahou, Mourad; Muselli, Alain; Costa, Jean

    2014-01-01

    Objective To evaluate some essential oils in treatment of intractable oral infections, principally caused by biofilm of multidrug-resistant Enterococcus faecalis (E. faecalis), such as persistent endodontic infections in which their treatment exhibits a real challenge for dentists. Methods Ten chemically analyzed essential oils by gas chromatography-mass spectrometry were evaluated for antimicrobial activity against sensitive and resistant clinical strains of E. faecalis in both planktonic and biofilm state using two methods, disk diffusion and broth micro-dilution. Results Studied essential oils showed a good antimicrobial activity and high ability in E. faecalis biofilm eradication, whether for sensitive or multidrug-resistant strains, especially those of Origanum glandulosum and Thymbra capitata with interesting minimum inhibitory concentration, biofilm inhibitory concentration, and biofilm eradication concentration values which doesn't exceed 0.063%, 0.75%, and 1.5%, respectively. Conclusions Findings of this study indicate that essential oils extracted from aromatic plants can be used in treatment of intractable oral infections, especially caused by biofilm of multidrug-resistant E. faecalis. PMID:25182948

  12. An in vitro study on the effects of nisin on the antibacterial activities of 18 antibiotics against Enterococcus faecalis.

    PubMed

    Tong, Zhongchun; Zhang, Yuejiao; Ling, Junqi; Ma, Jinglei; Huang, Lijia; Zhang, Luodan

    2014-01-01

    Enterococcus faecalis rank among the leading causes of nosocomial infections worldwide and possesses both intrinsic and acquired resistance to a variety of antibiotics. Development of new antibiotics is limited, and pathogens continually generate new antibiotic resistance. Many researchers aim to identify strategies to effectively kill this drug-resistant pathogen. Here, we evaluated the effect of the antimicrobial peptide nisin on the antibacterial activities of 18 antibiotics against E. faecalis. The MIC and MBC results showed that the antibacterial activities of 18 antibiotics against E. faecalis OG1RF, ATCC 29212, and strain E were significantly improved in the presence of 200 U/ml nisin. Statistically significant differences were observed between the results with and without 200 U/ml nisin at the same concentrations of penicillin or chloramphenicol (p<0.05). The checkerboard assay showed that the combination of nisin and penicillin or chloramphenicol had a synergetic effect against the three tested E. faecalis strains. The transmission electron microscope images showed that E. faecalis was not obviously destroyed by penicillin or chloramphenicol alone but was severely disrupted by either antibiotic in combination with nisin. Furthermore, assessing biofilms by a confocal laser scanning microscope showed that penicillin, ciprofloxacin, and chloramphenicol all showed stronger antibiofilm actions in combination with nisin than when these antibiotics were administered alone. Therefore, nisin can significantly improve the antibacterial and antibiofilm activities of many antibiotics, and certain antibiotics in combination with nisin have considerable potential for use as inhibitors of this drug-resistant pathogen. PMID:24586598

  13. Biofilm formation on polystyrene under different temperatures by antibiotic resistant Enterococcus faecalis and Enterococcus faecium isolated from food.

    PubMed

    Marinho, A R; Martins, P D; Ditmer, E M; d'Azevedo, P A; Frazzon, J; Van Der Sand, S T; Frazzon, A P G

    2013-01-01

    The ability of antibiotic resistant E. faecalis and E. faecium isolated from food to form biofilm at different temperatures in the absence or presence of 0.75% glucose was evaluated. A synergistic effect on biofilm at 10 °C, 28 °C, 37 °C and 45 °C and glucose was observed for E. faecalis and E. faecium.

  14. Transcriptome Analysis of Enterococcus faecalis during Mammalian Infection Shows Cells Undergo Adaptation and Exist in a Stringent Response State

    PubMed Central

    Frank, Kristi L.; Colomer-Winter, Cristina; Grindle, Suzanne M.; Lemos, José A.; Schlievert, Patrick M.; Dunny, Gary M.

    2014-01-01

    As both a commensal and a major cause of healthcare-associated infections in humans, Enterococcus faecalis is a remarkably adaptable organism. We investigated how E. faecalis adapts in a mammalian host as a pathogen by characterizing changes in the transcriptome during infection in a rabbit model of subdermal abscess formation using transcriptional microarrays. The microarray experiments detected 222 and 291 differentially regulated genes in E. faecalis OG1RF at two and eight hours after subdermal chamber inoculation, respectively. The profile of significantly regulated genes at two hours post-inoculation included genes involved in stress response, metabolism, nutrient acquisition, and cell surface components, suggesting genome-wide adaptation to growth in an altered environment. At eight hours post-inoculation, 88% of the differentially expressed genes were down-regulated and matched a transcriptional profile consistent with a (p)ppGpp-mediated stringent response. Subsequent subdermal abscess infections with E. faecalis mutants lacking the (p)ppGpp synthetase/hydrolase RSH, the small synthetase RelQ, or both enzymes, suggest that intracellular (p)ppGpp levels, but not stringent response activation, influence persistence in the model. The ability of cells to synthesize (p)ppGpp was also found to be important for growth in human serum and whole blood. The data presented in this report provide the first genome-wide insights on E. faecalis in vivo gene expression and regulation measured by transcriptional profiling during infection in a mammalian host and show that (p)ppGpp levels affect viability of E. faecalis in multiple conditions relevant to mammalian infection. The subdermal abscess model can serve as a novel experimental system for studying the E. faecalis stringent response in the context of the mammalian immune system. PMID:25545155

  15. Horizontal transfer of antibiotic resistance from Enterococcus faecium of fermented meat origin to clinical isolates of E. faecium and Enterococcus faecalis.

    PubMed

    Jahan, Musarrat; Zhanel, George G; Sparling, Richard; Holley, Richard A

    2015-04-16

    Enterococcus species are part of the normal intestinal flora of a large number of mammals including humans and consequently, they can be used as indicators of faecal contamination in food and water for human consumption. Their presence in large numbers in foods may indicate a lapse in sanitation and their ability to serve as a genetic reservoir of transferable antibiotic resistance is of concern. In the present study, Enterococcus spp., isolated from commercially fermented meat and human clinical specimen were studied to determine genetic relationships. SmaI pulsed-field gel electrophoresis (PFGE) patterns exhibited genomic heterogeneity within and between both groups of isolates. However, in spite of this heterogeneity there were still substantial phenotypic similarities which suggested that food might be a potential vehicle for distribution of resistant bacteria among humans. In vitro conjugation experiments demonstrated transfer of the tetracycline resistant determinant, tet(M), from Enterococcus faecium S27 isolated from fermented sausage to clinical isolates of both E. faecium and Enterococcus faecalis. The streptomycin resistance of E. faecium S27 was also transferred to a clinical strain, E. faecalis 82916, which was confirmed by the presence of the streptomycin resistance gene, aadA, in the donor and transconjugant strains. Since the aadA gene is associated with a class 1 integron, results also suggested that resistance transfer might have occurred via an integron. It appears this is the first identification of a class 1 integron in E. faecium isolated from food. The importance of food enterococci as a reservoir of antibiotic resistance genes and the potential for their genetic transfer to human strains following consumption of uncooked or undercooked contaminated meat is underlined by this work.

  16. Native Microbial Colonization of Drosophila melanogaster and Its Use as a Model of Enterococcus faecalis Pathogenesis▿ †

    PubMed Central

    Cox, Christopher R.; Gilmore, Michael S.

    2007-01-01

    Enterococci are commensal organisms of the gastrointestinal (GI) tracts of a broad range of mammalian and insect hosts, but they are also leading causes of nosocomial infection. Little is known about the ecological role of enterococci in the GI tract consortia. To develop a tractable model for studying the roles of these organisms as commensals and pathogens, we characterized the Drosophila melanogaster microflora and examined the occurrence of enterococci in the gastrointestinal consortium of Drosophila. In a survey of laboratory-reared Drosophila and wild-captured flies, we found that Drosophila was naturally colonized by representatives of five bacterial phyla. Among these organisms were several species of enterococci, including Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinaraum, and Enterococcus durans, as well as a previously detected but uncultured Enterococcus species. Drosophila could be cured of enterococcal carriage by antibiotic treatment and could be reassociated with laboratory strains. High-level colonization by a well-characterized strain expressing the enterococcal cytolysin was found to be detrimental to Drosophila compared to the effect of an isogenic, noncytolytic control. The anatomical distribution of enterococci in the Drosophila GI tract was determined by immunohistochemical staining of thin sections of naturally colonized and reassociated flies. PMID:17220307

  17. Biofilm formation on polystyrene under different temperatures by antibiotic resistant Enterococcus faecalis and Enterococcus faecium isolated from food

    PubMed Central

    Marinho, A.R.; Martins, P.D.; Ditmer, E.M.; d’Azevedo, P.A.; Frazzon, J.; Van Der Sand, S.T.; Frazzon, A.P.G.

    2013-01-01

    The ability of antibiotic resistant E. faecalis and E. faecium isolated from food to form biofilm at different temperatures in the absence or presence of 0.75% glucose was evaluated. A synergistic effect on biofilm at 10 °C, 28 °C, 37 °C and 45 °C and glucose was observed for E. faecalis and E. faecium. PMID:24294231

  18. Cyclooxygenase-2 Generates the Endogenous Mutagen trans-4-Hydroxy-2-nonenal in Enterococcus faecalis-infected Macrophages

    PubMed Central

    Wang, Xingmin; Allen, Toby D.; Yang, Yonghong; Moore, Danny R.; Huycke, Mark M.

    2013-01-01

    Infection of macrophages by the human intestinal commensal Enterococcus faecalis generates DNA damage and chromosomal instability in mammalian cells through bystander effects. These effects are characterized by clastogenesis and damage to mitotic spindles in target cells and are mediated, in part, by trans-4-hydroxy-2-nonenal (4-HNE). In this study we investigated the role of cyclooxygenase (COX) and lipoxygenase (LOX) in producing this reactive aldehyde using E. faecalis-infected macrophages and interleukin-10 knockout mice colonized with this commensal. 4-HNE production by E. faecalis-infected macrophages was significantly reduced by COX and LOX inhibitors. The infection of macrophages led to decreased Cox1 and Alox5 expression while COX-2 and 4-HNE increased. Silencing Alox5 and Cox1 with gene-specific siRNAs had no effect on 4-HNE production. In contrast, silencing Cox2 significantly decreased 4-HNE production by E. faecalis-infected macrophages. Depleting intracellular glutathione increased 4-HNE production by these cells. Next, to confirm COX-2 as a source for 4-HNE, we assayed the products generated by recombinant human COX-2 and found 4-HNE in a concentration-dependent manner using arachidonic acid as a substrate. Finally, tissue macrophages in colon biopsies from interleukin-10 knockout mice colonized with E. faecalis were positive for COX-2 by immunohistochemical staining. This was associated with increased staining for 4-HNE-protein adducts in surrounding stroma. These data show that E. faecalis, a human intestinal commensal, can trigger macrophages to produce 4-HNE through COX-2. Importantly, it reinforces the concept of COX-2 as a procarcinogenic enzyme capable of damaging DNA in target cells through bystander effects that contribute to colorectal carcinogenesis. PMID:23321929

  19. Immunostimulatory Effects Triggered by Enterococcus faecalis CECT7121 Probiotic Strain Involve Activation of Dendritic Cells and Interferon-Gamma Production

    PubMed Central

    Molina, Matías Alejandro; Díaz, Ailén Magalí; Hesse, Christina; Ginter, Wiebke; Gentilini, María Virginia; Nuñez, Guillermo Gabriel; Canellada, Andrea Mercedes; Sparwasser, Tim; Berod, Luciana; Castro, Marisa Silvia; Manghi, Marcela Alejandra

    2015-01-01

    Probiotics can modulate the immune system, conferring beneficial effects on the host. Understanding how these microorganisms contribute to improve the health status is still a challenge. Previously, we have demonstrated that Enterococcus faecalis CECT7121 implants itself and persists in the murine gastrointestinal tract, and enhances and skews the profile of cytokines towards the Th1 phenotype in several biological models. Given the importance of dendritic cells (DCs) in the orchestration of immunity, the aim of this work was to elucidate the influence of E. faecalis CECT7121 on DCs and the outcome of the immune responses. In this work we show that E. faecalis CECT7121 induces a strong dose-dependent activation of DCs and secretion of high levels of IL-12, IL-6, TNFα, and IL-10. This stimulation is dependent on TLR signaling, and skews the activation of T cells towards the production of IFNγ. The influence of this activation in the establishment of Th responses in vivo shows the accumulation of specific IFNγ-producing cells. Our findings indicate that the activation exerted by E. faecalis CECT7121 on DCs and its consequence on the cellular adaptive immune response may have broad therapeutic implications in immunomodulation. PMID:25978357

  20. C-Terminal WxL Domain Mediates Cell Wall Binding in Enterococcus faecalis and Other Gram-Positive Bacteria▿

    PubMed Central

    Brinster, Sophie; Furlan, Sylviane; Serror, Pascale

    2007-01-01

    Analysis of the genome sequence of Enterococcus faecalis clinical isolate V583 revealed novel genes encoding surface proteins. Twenty-seven of these proteins, annotated as having unknown functions, possess a putative N-terminal signal peptide and a conserved C-terminal region characterized by a novel conserved domain designated WxL. Proteins having similar characteristics were also detected in other low-G+C-content gram-positive bacteria. We hypothesized that the WxL region might be a determinant of bacterial cell location. This hypothesis was tested by generating protein fusions between the C-terminal regions of two WxL proteins in E. faecalis and a nuclease reporter protein. We demonstrated that the C-terminal regions of both proteins conferred a cell surface localization to the reporter fusions in E. faecalis. This localization was eliminated by introducing specific deletions into the domains. Interestingly, exogenously added protein fusions displayed binding to whole cells of various gram-positive bacteria. We also showed that the peptidoglycan was a binding ligand for WxL domain attachment to the cell surface and that neither proteins nor carbohydrates were necessary for binding. Based on our findings, we propose that the WxL region is a novel cell wall binding domain in E. faecalis and other gram-positive bacteria. PMID:16963569

  1. Antimicrobial effect of Lippia sidoides and thymol on Enterococcus faecalis biofilm of the bacterium isolated from root canals.

    PubMed

    Veras, H N H; Rodrigues, F F G; Botelho, M A; Menezes, I R A; Coutinho, H D M; da Costa, J G M

    2014-01-01

    The species Lippia sidoides Cham. (Verbenaceae) is utilized in popular medicine as a local antiseptic on the skin and mucosal tissues. Enterococcus faecalis is the bacterium isolated from root canals of teeth with persistent periapical lesions and has the ability to form biofilm, where it is responsible for the failure of endodontic treatments. Essential oil of L. sidoides (EOLS) and its major component, thymol, were evaluated for reducing the CFU in biofilms of E. faecalis in vitro. The essential oil was obtained by hydrodistillation and examined with respect to the chemical composition, by gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis has led to the identification of thymol (84.9%) and p-cymene (5.33%). EOLS and thymol reduced CFU in biofilms of E. faecalis in vitro (time of maturation, 72 h), with an exposure time of 30 and 60 min at concentrations of 2.5 and 10%. There was no statistical difference in effect between EOLS and thymol, demontrating that this phenolic monoterpene was the possible compound responsible for the antimicrobial activity of EOLS. This study provides a basis for the possible utilization of EOLS as an adjuvant in the treatment of root canals that show colonization by E. faecalis. PMID:24683344

  2. Immunostimulatory Effects Triggered by Enterococcus faecalis CECT7121 Probiotic Strain Involve Activation of Dendritic Cells and Interferon-Gamma Production.

    PubMed

    Molina, Matías Alejandro; Díaz, Ailén Magalí; Hesse, Christina; Ginter, Wiebke; Gentilini, María Virginia; Nuñez, Guillermo Gabriel; Canellada, Andrea Mercedes; Sparwasser, Tim; Berod, Luciana; Castro, Marisa Silvia; Manghi, Marcela Alejandra

    2015-01-01

    Probiotics can modulate the immune system, conferring beneficial effects on the host. Understanding how these microorganisms contribute to improve the health status is still a challenge. Previously, we have demonstrated that Enterococcus faecalis CECT7121 implants itself and persists in the murine gastrointestinal tract, and enhances and skews the profile of cytokines towards the Th1 phenotype in several biological models. Given the importance of dendritic cells (DCs) in the orchestration of immunity, the aim of this work was to elucidate the influence of E. faecalis CECT7121 on DCs and the outcome of the immune responses. In this work we show that E. faecalis CECT7121 induces a strong dose-dependent activation of DCs and secretion of high levels of IL-12, IL-6, TNFα, and IL-10. This stimulation is dependent on TLR signaling, and skews the activation of T cells towards the production of IFNγ. The influence of this activation in the establishment of Th responses in vivo shows the accumulation of specific IFNγ-producing cells. Our findings indicate that the activation exerted by E. faecalis CECT7121 on DCs and its consequence on the cellular adaptive immune response may have broad therapeutic implications in immunomodulation.

  3. IS256 abolishes gelatinase activity and biofilm formation in a mutant of the nosocomial pathogen Enterococcus faecalis V583.

    PubMed

    Perez, Marta; Calles-Enríquez, Marina; del Rio, Beatriz; Ladero, Victor; Martín, María Cruz; Fernández, María; Alvarez, Miguel A

    2015-07-01

    Enterococcus faecalis is one of the most controversial species of lactic acid bacteria. Some strains are used as probiotics, while others are associated with severe and life-threatening nosocomial infections. Their pathogenicity depends on the acquisition of multidrug resistance and virulence factors. Gelatinase, which is required in the first steps of biofilm formation, is an important virulence determinant involved in E. faecalis pathogenesis, including endocarditis and peritonitis. The gene that codes for gelatinase (gelE) is controlled by the Fsr quorum-sensing system, whose encoding genes (fsrA, fsrB, fsrC, and fsrD) are located immediately upstream of gelE. The integration of a DNA fragment into the fsr locus of a derived mutant of E. faecalis V583 suppressed the gelatinase activity and prevented biofilm formation. Sequence analysis indicated the presence of IS256 integrated into the fsrC gene at nucleotide position 321. Interestingly, IS256 is also associated with biofilm formation in Staphylococcus epidermidis and Staphylococcus aureus. This is the first description of an insertion sequence that prevents biofilm formation in E. faecalis.

  4. Immunostimulatory Effects Triggered by Enterococcus faecalis CECT7121 Probiotic Strain Involve Activation of Dendritic Cells and Interferon-Gamma Production.

    PubMed

    Molina, Matías Alejandro; Díaz, Ailén Magalí; Hesse, Christina; Ginter, Wiebke; Gentilini, María Virginia; Nuñez, Guillermo Gabriel; Canellada, Andrea Mercedes; Sparwasser, Tim; Berod, Luciana; Castro, Marisa Silvia; Manghi, Marcela Alejandra

    2015-01-01

    Probiotics can modulate the immune system, conferring beneficial effects on the host. Understanding how these microorganisms contribute to improve the health status is still a challenge. Previously, we have demonstrated that Enterococcus faecalis CECT7121 implants itself and persists in the murine gastrointestinal tract, and enhances and skews the profile of cytokines towards the Th1 phenotype in several biological models. Given the importance of dendritic cells (DCs) in the orchestration of immunity, the aim of this work was to elucidate the influence of E. faecalis CECT7121 on DCs and the outcome of the immune responses. In this work we show that E. faecalis CECT7121 induces a strong dose-dependent activation of DCs and secretion of high levels of IL-12, IL-6, TNFα, and IL-10. This stimulation is dependent on TLR signaling, and skews the activation of T cells towards the production of IFNγ. The influence of this activation in the establishment of Th responses in vivo shows the accumulation of specific IFNγ-producing cells. Our findings indicate that the activation exerted by E. faecalis CECT7121 on DCs and its consequence on the cellular adaptive immune response may have broad therapeutic implications in immunomodulation. PMID:25978357

  5. Comparative evaluation of antimicrobial efficacy of QMix™ 2 in 1, sodium hypochlorite, and chlorhexidine against Enterococcus faecalis and Candida albicans

    PubMed Central

    Elakanti, Soujanya; Cherukuri, Gayathri; Rao, Venkateswara G; Chandrasekhar, Veeramachaneni; Rao, Anitha S; Tummala, Muralidhar

    2015-01-01

    Aim/Objective: The aim of this study is to compare the antimicrobial efficacy of QMix™ 2 in 1, sodium hypochlorite (NaOCl), and chlorhexidine (CHX) against Enterococcus faecalis and Candida albicans. Materials and Methods: Eighty freshly extracted, single-rooted human mandibular premolar teeth were instrumented and autoclaved. Samples were divided into two groups of 40 teeth each based on the type of microorganism used. Group I was inoculated with E. faecalis and Group II with C. albicans and incubated for 3 days. Each group was subdivided into four subgroups based on the type of irrigant used. Group IA, IIA, 5.25% NaOCl; Group IB, IIB, 2% CHX; Group IC, IIC, QMix™ 2 in 1; and Group ID, IID, 0.9% saline (the control group). Ten microliters of the sample from each canal was taken and was placed on Brain Heart Infusion agar and Sabouraud dextrose agar. The plates were incubated at 37°C for 24 h and colony forming units (CFUs) that were grown were counted. Data was analyzed with analysis of variance (ANOVA) followed by post-hoc Games-Howell test. Results: The greatest antimicrobial effects were observed in samples treated with QMix™ 2 in 1 (P < 0.001). No statistical significant difference was found between 5.25% NaOCl and 2% CHX (P > 0.001) against E. faecalis and C. albicans. Conclusion: QMix™ 2 in 1 demonstrated significant antimicrobial efficacy against E. faecalis and C. albicans. PMID:25829691

  6. Spread of an Enterococcus faecalis sequence type 6 (CC2) clone in patients undergoing selective decontamination of the digestive tract.

    PubMed

    Muruzábal-Lecumberri, Izaskun; Girbau, Cecilia; Canut, Andrés; Alonso, Rodrigo; Fernández-Astorga, Aurora

    2015-03-01

    Enterococcus faecalis (E. faecalis) is a common cause of nosocomial infection in immunocompromised patients. The presence and dissemination of high-risk clonal complexes, such as CC2, is an ongoing problem in hospitals. The aim of this work was to characterize 24 E. faecalis isolates from ICU patients undergoing selective decontamination of the digestive tract (SDD) by phenotypical (antimicrobial susceptibility) and genotypical (presence of virulence genes, RAPD-PCR and MLST) methods. Our results showed high prevalence of the ST6 E. faecalis clone (91.6%), especially adapted to the hospital environment, with a multidrug resistance pattern and a multitude of putative virulence genes. In addition, ST179 (4.2%) and ST191 (4.2%) were detected. By RAPD-PCR analysis, the 22 isolates identified as ST6 showed six different DNA patterns, while the two remaining isolates, ST179 and ST191, showed two additional profiles. CC2 is a known clonal complex with high adaptability to hospital environment and worldwide distribution. The high prevalence of the ST6 clone in the studied population could be related to the presence of gentamicin in the SDD mixture since most strains were gentamicin resistant. Consequently, strict surveillance should be applied for rapid detection and control of this clone to prevent future spread outside the ICU.

  7. Enterococcus faecalis inhibits superantigen toxic shock syndrome toxin-1-induced interleukin-8 from human vaginal epithelial cells through tetramic acids.

    PubMed

    Brosnahan, Amanda J; Merriman, Joseph A; Salgado-Pabón, Wilmara; Ford, Bradley; Schlievert, Patrick M

    2013-01-01

    The vaginal mucosa can be colonized by many bacteria including commensal organisms and potential pathogens, such as Staphylococcus aureus. Some strains of S. aureus produce the superantigen toxic shock syndrome toxin-1, which can penetrate the vaginal epithelium to cause toxic shock syndrome. We have observed that a female was mono-colonized with Enterococcus faecalis vaginally as tested in aerobic culture, even upon repeated culture for six months, suggesting this organism was negatively influencing colonization by other bacteria. In recent studies, we demonstrated an "outside-in" mechanism of cytokine signaling and consequent inflammation that facilitates the ability of potential pathogens to initiate infection from mucosal surfaces. Thus, we hypothesized that this strain of E. faecalis may make anti-inflammatory factors which block disease progression of more pathogenic organisms. E. faecalis MN1 inhibited interleukin-8 production from human vaginal epithelial cells in response to the vaginal pathogens Candida albicans, Gardnerella vaginalis, and Neisseria gonorrhoeae, as well as to toxic shock syndrome toxin-1. We further demonstrated that this organism secretes two tetramic acid compounds which appear responsible for inhibition of interleukin-8 production, as well as inhibition of T cell proliferation due to toxic shock syndrome toxin-1. Microbicides that include anti-inflammatory molecules, such as these tetramic acid compounds naturally produced by E. faecalis MN1, may be useful in prevention of diseases that develop from vaginal infections. PMID:23613823

  8. The Two Functional Enoyl-Acyl Carrier Protein Reductases of Enterococcus faecalis Do Not Mediate Triclosan Resistance

    PubMed Central

    Zhu, Lei; Bi, Hongkai; Ma, Jincheng; Hu, Zhe; Zhang, Wenbin; Cronan, John E.; Wang, Haihong

    2013-01-01

    ABSTRACT Enoyl-acyl carrier protein (enoyl-ACP) reductase catalyzes the last step of the elongation cycle in the synthesis of bacterial fatty acids. The Enterococcus faecalis genome contains two genes annotated as enoyl-ACP reductases, a FabI-type enoyl-ACP reductase and a FabK-type enoyl-ACP reductase. We report that expression of either of the two proteins restores growth of an Escherichia coli fabI temperature-sensitive mutant strain under nonpermissive conditions. In vitro assays demonstrated that both proteins support fatty acid synthesis and are active with substrates of all fatty acid chain lengths. Although expression of E. faecalis fabK confers to E. coli high levels of resistance to the antimicrobial triclosan, deletion of fabK from the E. faecalis genome showed that FabK does not play a detectable role in the inherent triclosan resistance of E. faecalis. Indeed, FabK seems to play only a minor role in modulating fatty acid composition. Strains carrying a deletion of fabK grow normally without fatty acid supplementation, whereas fabI deletion mutants make only traces of fatty acids and are unsaturated fatty acid auxotrophs. PMID:24085780

  9. Antibacterial efficacy of Mangifera indica L. kernel and Ocimum sanctum L. leaves against Enterococcus faecalis dentinal biofilm

    PubMed Central

    Subbiya, Arunajatesan; Mahalakshmi, Krishnan; Pushpangadan, Sivan; Padmavathy, Kesavaram; Vivekanandan, Paramasivam; Sukumaran, Vridhachalam Ganapathy

    2013-01-01

    Introduction: The Enterococcus faecalis biofilm in the root canal makes it difficult to be eradicated by the conventional irrigants with no toxicity to the tissues. Hence, plant products with least side effects are explored for their use as irrigants in the root canal therapy. Aim: To evaluate and compare the antibacterial efficacy of Mangifera indica L. kernel (mango kernel) and Ocimum sanctum L. leaves (tulsi) extracts with conventional irrigants (5% sodium hypochlorite (NaOCl) and 2% chlorhexidine) against E. faecalis dentinal biofilm. Materials and Methods: Agar diffusion and broth microdilution assay was performed with the herbal extracts and conventional irrigants (2% chlorhexidine and 5% NaOCl) against E. faecalis planktonic cells. The assay was extended onto 3 week E. faecalis dentinal biofilm. Results: Significant reduction of colony forming units (CFU)/mL was observed for the herbal groups and the antibacterial activity of the herbal groups was at par with 5% NaOCl. Conclusions: The antibacterial activity of these herbal extracts is found to be comparable with that of conventional irrigants both on the biofilm and planktonic counterparts. PMID:24082577

  10. Antimicrobial Effect of Lippia sidoides and Thymol on Enterococcus faecalis Biofilm of the Bacterium Isolated from Root Canals

    PubMed Central

    Veras, H. N. H.; Rodrigues, F. F. G.; Botelho, M. A.; Menezes, I. R. A.; Coutinho, H. D. M.; da Costa, J. G. M.

    2014-01-01

    The species Lippia sidoides Cham. (Verbenaceae) is utilized in popular medicine as a local antiseptic on the skin and mucosal tissues. Enterococcus faecalis is the bacterium isolated from root canals of teeth with persistent periapical lesions and has the ability to form biofilm, where it is responsible for the failure of endodontic treatments. Essential oil of L. sidoides (EOLS) and its major component, thymol, were evaluated for reducing the CFU in biofilms of E. faecalis in vitro. The essential oil was obtained by hydrodistillation and examined with respect to the chemical composition, by gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis has led to the identification of thymol (84.9%) and p-cymene (5.33%). EOLS and thymol reduced CFU in biofilms of E. faecalis in vitro (time of maturation, 72 h), with an exposure time of 30 and 60 min at concentrations of 2.5 and 10%. There was no statistical difference in effect between EOLS and thymol, demonstrating that this phenolic monoterpene was the possible compound responsible for the antimicrobial activity of EOLS. This study provides a basis for the possible utilization of EOLS as an adjuvant in the treatment of root canals that show colonization by E. faecalis. PMID:24683344

  11. Whole-genome mapping of 5′ RNA ends in bacteria by tagged sequencing: a comprehensive view in Enterococcus faecalis

    PubMed Central

    Innocenti, Nicolas; Golumbeanu, Monica; Fouquier d'Hérouël, Aymeric; Lacoux, Caroline; Bonnin, Rémy A.; Kennedy, Sean P.; Wessner, Françoise; Serror, Pascale; Bouloc, Philippe; Repoila, Francis; Aurell, Erik

    2015-01-01

    Enterococcus faecalis is the third cause of nosocomial infections. To obtain the first snapshot of transcriptional organizations in this bacterium, we used a modified RNA-seq approach enabling to discriminate primary from processed 5′ RNA ends. We also validated our approach by confirming known features in Escherichia coli. We mapped 559 transcription start sites (TSSs) and 352 processing sites (PSSs) in E. faecalis. A blind motif search retrieved canonical features of SigA- and SigN-dependent promoters preceding transcription start sites mapped. We discovered 85 novel putative regulatory RNAs, small- and antisense RNAs, and 72 transcriptional antisense organizations. Presented data constitute a significant insight into bacterial RNA landscapes and a step toward the inference of regulatory processes at transcriptional and post-transcriptional levels in a comprehensive manner. PMID:25737579

  12. Characterization of a bacteriocin produced by Enterococcus faecalis N1-33 and its application as a food preservative.

    PubMed

    Hata, Tomomi; Alemu, Melaku; Kobayashi, Miho; Suzuki, Chise; Nitisinprasert, Sunee; Ohmomo, Sadahiro

    2009-03-01

    A bacteriocin-producing strain, N1-33, isolated from fermented bamboo shoot was identified as Enterococcus faecalis. The pH-adjusted culture supernatant of this strain consisted of several peptides with bacteriocin activity, and the supernatant inhibited the growth of pathogenic bacteria such as Listeria monocytogenes. The major peptide with bacteriocin activity was purified, and the first 39 amino acid residues of the bacteriocin were found to be identical to enterocin MR10A produced by E. faecalis MRR10-3. Addition of the pH-adjusted and concentrated culture supernatant of strain N1-33 caused a marked reduction in the growth of Bacillus cereus in custard cream and L. monocytogenes in pickled cucumber. These results suggest the potential use of the bacteriocin produced by strain N1-33 in food biopreservation.

  13. Detection of Identical Isolates of Enterococcus faecalis from the Blood and Oral Mucosa in a Patient with Infective Endocarditis.

    PubMed

    Okui, Akemi; Soga, Yoshihiko; Kokeguchi, Susumu; Nose, Motoko; Yamanaka, Reiko; Kusano, Nobuchika; Morita, Manabu

    2015-01-01

    The detection of infective endocarditis (IE) of oral origin has been previously discussed. However, there are few reports confirming this infection using molecular biological techniques. We herein describe the case of a 67-year-old man who developed IE. Blood culture samples and strains obtained from the gingival and buccal mucosa showed 100% identity to Enterococcus faecalis JCM 5803 on sequencing of 16S rRNA gene fragments. A random amplification of polymorphic DNA (RAPD) analysis showed the same pattern for these samples, thus confirming the identity of E. faecalis isolates in the blood and oral mucosa. Our observations provide novel information regarding the level of identity between IE pathogens and oral bacteria.

  14. Detection of Identical Isolates of Enterococcus faecalis from the Blood and Oral Mucosa in a Patient with Infective Endocarditis.

    PubMed

    Okui, Akemi; Soga, Yoshihiko; Kokeguchi, Susumu; Nose, Motoko; Yamanaka, Reiko; Kusano, Nobuchika; Morita, Manabu

    2015-01-01

    The detection of infective endocarditis (IE) of oral origin has been previously discussed. However, there are few reports confirming this infection using molecular biological techniques. We herein describe the case of a 67-year-old man who developed IE. Blood culture samples and strains obtained from the gingival and buccal mucosa showed 100% identity to Enterococcus faecalis JCM 5803 on sequencing of 16S rRNA gene fragments. A random amplification of polymorphic DNA (RAPD) analysis showed the same pattern for these samples, thus confirming the identity of E. faecalis isolates in the blood and oral mucosa. Our observations provide novel information regarding the level of identity between IE pathogens and oral bacteria. PMID:26179542

  15. Ex situ study of Enterococcus faecalis survival in the recreational waters of the southern coast of the Caspian Sea

    PubMed Central

    Irankhah, Sahar; Soudi, Mohammad Reza; Gharavi, Sara

    2016-01-01

    Background and Objectives: The US Environmental Protection Agency has suggested faecal enterococci as the primary bacterial indicators. Of more importance is their direct correlation with swimmer-associated gastroenteritis in recreation water quality monitoring. In contrast to other seawater bodies with 3.5% salinity, the recreational waters in the southern coast of the Caspian Sea possess its own salinity (about 1% w/v) and thus require further investigations to determine the capacity of Enterococcus faecalis as the sole primary microbial index in this unique aquatic environment. Materials and Methods: The survey of the presence and survival of E. faecalis as a microbial index in the recreational waters of the southern Caspian Sea was carried out using a microcosm as an experimental model. The concentration of E. faecalis cells in samples of seawater were estimated by a standard membrane filtration method using m-Enterococcus agar as the selective culture medium. As the current standard culture-based methods are not reliable enough for the detection of non-growing, damaged and under-tension bacteria, PCR was used to identify the possible VBNC form of the bacterium after disappearance of the culturable cells. Results and Conclusion: A continuous decline in the number of culturable E. faecalis cells resulted in apparent elimination of the bacteria from seawater in a defined period. Detection of intact DNA was possible in the following 60 days. The salinity of about 1% and the self-purification properties of the Caspian Sea make the conditions feasible for the use of this microorganism as a measure of water quality throughout the region. The results confirmed the presence of damaged bacterial cells, namely VBNC forms, indicating the necessity of examining of the sea water samples by using molecular approaches or repair procedures. PMID:27307975

  16. Involvement of pyruvate dehydrogenase in product formation in pyruvate-limited anaerobic chemostat cultures of Enterococcus faecalis NCTC 775.

    PubMed

    Snoep, J L; Teixeira de Mattos, M J; Postma, P W; Neijssel, O M

    1990-01-01

    Enterococcus faecalis NCTC 775 was grown anaerobically in chemostat culture with pyruvate as the energy source. At low culture pH values, high in vivo and in vitro activities were found for both pyruvate dehydrogenase and lactate dehydrogenase. At high culture pH values the carbon flux was shifted towards pyruvate formate lyase. Some mechanisms possibly involved in this metabolic switch are discussed. In particular attention is paid to the NADH/NAD ratio (redox potential) and the fructose-1,6-bisphosphate-dependent lactate dehydrogenase activity as possible regulatory factors.

  17. Escherichia coli and Enterococcus faecalis are able to incorporate and enhance a pre-formed Gardnerella vaginalis biofilm.

    PubMed

    Castro, Joana; Machado, Daniela; Cerca, Nuno

    2016-04-01

    Gardnerella vaginalis is the most frequent microorganism found in bacterial vaginosis (BV), while Escherichia coli and Enterococcus faecalis are amongst the most frequent pathogens found in urinary tract infections (UTIs). This study aimed to evaluate possible interactions between UTIs pathogens and G. vaginalis using an in vitro dual-species biofilm model. Our results showed that dual-species biofilms reached significantly higher bacterial concentration than monospecies biofilms. Moreover, visualization of dual-populations species in the biofilms, using the epifluorescence microscopy, revealed that all of the urogenital pathogens coexisted with G. vaginalis. In conclusion, our work demonstrates that uropathogens can incorporate into mature BV biofilms. PMID:26782142

  18. Escherichia coli and Enterococcus faecalis are able to incorporate and enhance a pre-formed Gardnerella vaginalis biofilm.

    PubMed

    Castro, Joana; Machado, Daniela; Cerca, Nuno

    2016-04-01

    Gardnerella vaginalis is the most frequent microorganism found in bacterial vaginosis (BV), while Escherichia coli and Enterococcus faecalis are amongst the most frequent pathogens found in urinary tract infections (UTIs). This study aimed to evaluate possible interactions between UTIs pathogens and G. vaginalis using an in vitro dual-species biofilm model. Our results showed that dual-species biofilms reached significantly higher bacterial concentration than monospecies biofilms. Moreover, visualization of dual-populations species in the biofilms, using the epifluorescence microscopy, revealed that all of the urogenital pathogens coexisted with G. vaginalis. In conclusion, our work demonstrates that uropathogens can incorporate into mature BV biofilms.

  19. Antibiotic resistance genes and virulence factors in Enterococcus faecium and Enterococcus faecalis from diseased farm animals: pigs, cattle and poultry.

    PubMed

    Seputiene, V; Bogdaite, A; Ruzauskas, M; Suziedeliene, E

    2012-01-01

    Eighty enterococcal isolates (E. faecium, n = 38, E. faecalis, n = 42) from diseased farm animals (swine, cattle, poultry) in Lithuania have been studied for the prevalence of antibiotic resistance and for resistance and virulence genetic determinants. 86% of E. faecium and 71% of E. faecalis isolates were multidrug resistant (resistant to three or more unrelated antibiotics). Resistance to aminoglycosides, tetracycline and erythromycin was found most frequently in both species (61%, 69%) and was linked to aph(3')-IIIa, aac(6')-Ie-aph(2")-Ia, ant(6)-Ia (aminoglycoside resistance), tetM, tetL (tetracycline resistance), ermA, ermB (erythromycin resistance) gene combinations, which were supplemented with chloramphenicol resistance genes catA7, catA8 (E. faecalis) and catA9 (E. faecium). All E. faecalis isolates harboured genes coding for virulence factors agg, esp, fsr gelE alone or in combinations with the high prevalence of esp gene in isolates from cattle (63%) and pigs (79%). The origin-dependent incidence of agg gene variants prgB and asp1 was observed. The results indicate the existence of a large pool of potentially virulent and multidrug resistant E. faecalis in diseased farm animals posing risk to humans.

  20. Heterologous expression and characterization of tyrosine decarboxylase from Enterococcus faecalis R612Z1 and Enterococcus faecium R615Z1.

    PubMed

    Liu, Fang; Xu, Wenjuan; Du, Lihui; Wang, Daoying; Zhu, Yongzhi; Geng, Zhiming; Zhang, Muhan; Xu, Weimin

    2014-04-01

    Tyrosine decarboxylase (TDC) is responsible for tyramine production and can catalyze phenylalanine to produce β-phenylethylamine. Enterococcus strains are a group of bacteria predominantly producing tyramine and β-phenylethylamine in water-boiled salted duck. In this study, the heterologous expression and characterization of two TDCs from Enterococcus faecalis R612Z1 (612TDC) and Enterococcus faecium R615Z1 (615TDC) were studied. The recombinant putative proteins of 612TDC and 615TDC were heterologously expressed in Escherichia coli. 612TDC is a 620-amino-acid protein with a molecular mass of 70.0 kDa, whereas 615TDC is a 625-amino-acid protein with a molecular mass of 70.3 kDa. Both 612TDC and 615TDC showed an optimum temperature of 25 °C for the tyrosine and phenylalanine substrates. However, 612TDC revealed maximal activity at pH 5.5, whereas 615TDC revealed maximal activity at pH 6.0. Kinetic studies showed that 612TDC and 615TDC exhibited higher specificity for tyrosine than for phenylalanine. The catalysis abilities of both 612TDC and 615TDC for phenylalanine were restrained significantly with the increase in NaCl concentration, but this was not the case for tyrosine. This study revealed that the enzyme properties of the purified recombinant 612TDC and 615TDC were similar, although their amino acid sequences had 84% identity. PMID:24680070

  1. Comparative Assessment of Antimicrobial Efficacy of Different Antibiotic Coated Gutta-Percha Cones on Enterococcus faecalis An Invitro Study

    PubMed Central

    Karibasappa, Gundabaktha Nagappa; Dodamani, Arun Suresh; Vishwakarma, Prashanth Kumar; Mali, Gaurao Vasant

    2016-01-01

    Introduction The major goal of endodontic treatment is to eliminate bacteria from the root canals and prevent re-infection. Enterococcus faecalis (E. faecalis) has been attributed to be the most common organism for the endodontic treatment failures. The choice of endodontic material that have high antimicrobial efficacy can help in decreasing/avoiding growth of micro-organisms and facilitate the success rate of treatment. Aim The present study was designed with an aim to evaluate the antimicrobial efficacy of antibiotic coated gutta-percha cones on E. faecalis. Materials and Methods The present study was an invitro experimental study, conducted at Department of Public Health Densitry and Department of Microbiology. Gutta-percha cones were coated with different medicaments like Zinc Oxide-Eugenol cement (ZOE i.e. Group A), ZOE plus Amoxicillin-Clavulanic acid combination (Group B), ZOE plus Amoxicillin (Group C), ZOE plus Ofloxacin-Ornidazole combination (Group D). Agar plates were inoculated with E. faecalis and antibiotic coated gutta-percha cones along with conventional gutta-percha cones (coated only with ZOE) were placed in those agar plates. After 24hours incubation; diameter of zone of inhibition around gutta-percha stick was considered to assess the antimicrobial activity. Results were statistically analysed with analysis of variance (ANOVA) followed by Tukey post-hoc test for group-wise comparisons. Results Mean diameter of zone of inhibition (in mm) obtained for Group A, Group B, Group C and Group D were 5±0.03, 26.6±0.05, 21.5±0.04 and 15.8±0.03 respectively. The difference in values of different antibiotics was statistically significant. The p-value < 0.001 was considered statistically significant. Conclusion Group B was most effective against E.faecalis compared to other combinations used which increase the success rate of endodontic treatment as compared to conventional gutta-percha cones. PMID:27790583

  2. The antibacterial activity of sodium hypochlorite and chlorhexidine against Enterococcus faecalis: A review on agar diffusion and direct contact methods

    PubMed Central

    Luddin, Norhayati; Ahmed, Hany Mohamed Aly

    2013-01-01

    Complete debridement and disinfection of the root canal system are fundamental requirements for successful endodontic treatment. Despite the morphological challenges of the internal root anatomy, root canal irrigants play an important role in the optimization of the root canal preparation, which is essentially a chemo-mechanical procedure. Enterococcus faecalis is one of the most resistant microorganisms that dominants the microbial ecosystem of persistent periradicular lesions in retreatment cases. For that reason, many in vitro and in vivo studies evaluated and compared the antibacterial activity of sodium hypochlorite and chlorhexidine at varying concentrations using different experimental models against this microorganism. However, many controversies with regard to the ideal irrigant and concentration do in fact exist. Hence, this review aims to discuss the antibacterial activity of these two main root canal irrigants against Enterococcus faecalis using the agar diffusion and direct contact methods and the possible modulating factors responsible for inconsistent findings among different studies. In addition, the disinfection potential of both chemical agents on gutta percha and Resilon cones are also discussed. The source of this review was conducted through an electronic literature search using PubMed database from December 1997 until December 2011, which analyze the related laboratory investigations of both irrigants, published in major endodontic journals. PMID:23349569

  3. Adaptation to NaCl Reduces the Susceptibility of Enterococcus faecalis to Melaleuca alternifolia (Tea Tree) Oil.

    PubMed

    Lim, Ee Lin; Hammer, Katherine Ann

    2015-10-01

    This study investigated the hypothesis that the salt adaptation response of Enterococcus faecalis alters susceptibility to tea tree oil (TTO). Six E. faecalis isolates were adapted to 6.5 % NaCl, and then exposed to TTO in phosphate-buffered saline (PBS). One isolate was also exposed to TTO in Brain Heart Infusion Broth (BHIB). The viability of salt-adapted and non-adapted control cells was determined at 0, 45 and 90 min and compared. MICs for several antibiotics and TTO were also determined by E test and broth microdilution, respectively. Results showed that susceptibility to TTO in PBS was significantly reduced after salt adaptation for five isolates (83 %) (P < 0.05). Mean differences between salt-adapted and non-adapted cell counts were 2.51 log at 45 min and 2.13 log at 90 min. However, when E. faecalis ATCC 19433 was exposed to TTO in BHIB, no significant differences were seen. In conclusion, salt adaptation resulted in reduced susceptibility to TTO in PBS for the majority of isolates, indicating that cross protection had occurred. This effect was absent in BHIB, suggesting that the uptake of compatible solutes from the growth medium protected non-adapted cells from TTO. Whether this has implications for the clinical effectiveness of TTO remains to be determined.

  4. Biochemical and Structural Basis for Inhibition of Enterococcus faecalis Hydroxymethylglutaryl-CoA Synthase, mvaS, by Hymeglusin

    SciTech Connect

    Skaff, D. Andrew; Ramyar, Kasra X.; McWhorter, William J.; Barta, Michael L.; Geisbrecht, Brian V.; Miziorko, Henry M.

    2012-07-25

    Hymeglusin (1233A, F244, L-659-699) is established as a specific {beta}-lactone inhibitor of eukaryotic hydroxymethylglutaryl-CoA synthase (HMGCS). Inhibition results from formation of a thioester adduct to the active site cysteine. In contrast, the effects of hymeglusin on bacterial HMG-CoA synthase, mvaS, have been minimally characterized. Hymeglusin blocks growth of Enterococcus faecalis. After removal of the inhibitor from culture media, a growth curve inflection point at 3.1 h is observed (vs 0.7 h for the uninhibited control). Upon hymeglusin inactivation of purified E. faecalis mvaS, the thioester adduct is more stable than that measured for human HMGCS. Hydroxylamine cleaves the thioester adduct; substantial enzyme activity is restored at a rate that is 8-fold faster for human HMGCS than for mvaS. Structural results explain these differences in enzyme-inhibitor thioester adduct stability and solvent accessibility. The E. faecalis mvaS-hymeglusin cocrystal structure (1.95 {angstrom}) reveals virtually complete occlusion of the bound inhibitor in a narrow tunnel that is largely sequestered from bulk solvent. In contrast, eukaryotic (Brassica juncea) HMGCS binds hymeglusin in a more solvent-exposed cavity.

  5. The fibronectin-binding protein EfbA contributes to pathogenesis and protects against infective endocarditis caused by Enterococcus faecalis.

    PubMed

    Singh, Kavindra V; La Rosa, Sabina Leanti; Somarajan, Sudha R; Roh, Jung Hyeob; Murray, Barbara E

    2015-12-01

    EfbA is a PavA-like fibronectin adhesin of Enterococcus faecalis previously shown to be important in experimental urinary tract infection. Here, we expressed and purified the E. faecalis OG1RF EfbA and confirmed that this protein binds with high affinity to immobilized fibronectin, collagen I, and collagen V. We constructed an efbA deletion mutant and demonstrated that its virulence was significantly attenuated (P < 0.0006) versus the wild type in a mixed inoculum rat endocarditis model. Furthermore, efbA deletion resulted in diminished ability to bind fibronectin (P < 0.0001) and reduced biofilm (P < 0.001). Reintroduction of efbA into the original chromosomal location restored virulence, adherence to fibronectin, and biofilm formation to wild-type levels. Finally, vaccination of rats with purified recombinant EfbA protein protected against OG1RF endocarditis (P = 0.008 versus control). Taken together, our results demonstrate that EfbA is an important factor involved in E. faecalis endocarditis and that rEfbA immunization is effective in preventing such infection, likely by interfering with bacterial adherence.

  6. PCR and electrospray ionization mass spectrometry for detection of persistent enterococcus faecalis in cerebrospinal fluid following treatment of postoperative ventriculitis.

    PubMed

    Farrell, John J; Tsung, Andrew J; Flier, Lisa; Martinez, Derek L; Beam, Sarah B; Chen, Clifford; Lowery, Kristin S; Sampath, Rangarajan; Bonomo, Robert A

    2013-10-01

    We describe the use of PCR and electrospray ionization followed by mass spectrometry (PCR/ESI-MS) to evaluate "culture-negative" cerebrospinal fluid (CSF) from a 67-year-old man who developed postoperative bacterial ventriculitis following a suboccipital craniotomy for resection of an ependymoma in the 4th ventricle. CSF samples were obtained on seven occasions, beginning in the operating room at the time of insertion of a right ventriculoperitoneal shunt (VPS) and continuing until his death, 6 weeks later. During the course of the illness, two initial CSF specimens taken before the initiation of antimicrobial treatment were notable for growth of Enterococcus faecalis. Once antimicrobial treatment was initiated, all CSF cultures were negative. PCR/ESI-MS detected genetic evidence of E. faecalis in all CSF samples, but the level of detection (LOD) decreased once antimicrobial treatment was initiated. When our patient returned with symptoms of meningitis 3 days after the completion of antibiotic treatment, CSF cultures remained negative, but PCR/ESI-MS again found genetic evidence for E. faecalis at levels comparable to the pretreatment levels seen initially. This unique case and these findings suggest that determination of CSF LOD by PCR/ESI-MS may be a very sensitive indicator of persistent infection in patients on antibiotic therapy for complex CNS infections and may have relevance for treatment duration and assessment of persistent or recurrent infection at the completion of therapy.

  7. The Fibronectin-Binding Protein EfbA Contributes to Pathogenesis and Protects against Infective Endocarditis Caused by Enterococcus faecalis

    PubMed Central

    Singh, Kavindra V.; La Rosa, Sabina Leanti; Somarajan, Sudha R.; Roh, Jung Hyeob

    2015-01-01

    EfbA is a PavA-like fibronectin adhesin of Enterococcus faecalis previously shown to be important in experimental urinary tract infection. Here, we expressed and purified the E. faecalis OG1RF EfbA and confirmed that this protein binds with high affinity to immobilized fibronectin, collagen I, and collagen V. We constructed an efbA deletion mutant and demonstrated that its virulence was significantly attenuated (P < 0.0006) versus the wild type in a mixed inoculum rat endocarditis model. Furthermore, efbA deletion resulted in diminished ability to bind fibronectin (P < 0.0001) and reduced biofilm (P < 0.001). Reintroduction of efbA into the original chromosomal location restored virulence, adherence to fibronectin, and biofilm formation to wild-type levels. Finally, vaccination of rats with purified recombinant EfbA protein protected against OG1RF endocarditis (P = 0.008 versus control). Taken together, our results demonstrate that EfbA is an important factor involved in E. faecalis endocarditis and that rEfbA immunization is effective in preventing such infection, likely by interfering with bacterial adherence. PMID:26351286

  8. Inhibitory effect of gels loaded with a low concentration of antibiotics against biofilm formation by Enterococcus faecalis and Porphyromonas gingivalis.

    PubMed

    A Algarni, Amnah; H Yassen, Ghaeth; L Gregory, Richard

    2015-09-01

    We explored longitudinally the inhibitory effect of gels loaded with 1 mg/mL modified triple antibiotic paste (MTAP) or double antibiotic paste (DAP) against biofilm formation by Enterococcus faecalis and Porphyromonas gingivalis. Methylcellulose-based antibiotic gels of MTAP (ciprofloxacin, metronidazole and clindamycin) and DAP (ciprofloxacin and metronidazole) were prepared at a concentration of 1 mg/mL. Individually cultured E. faecalis and P. gingivalis bacterial suspensions were treated with MTAP, DAP, or placebo (vehicle only) gels at different dilutions and allowed to grow in 96-well microtiter plates. Untreated bacterial suspensions served as a negative control. Crystal violet assays were used to evaluate biofilm formation after 48 h. The ability of the gels to inhibit biofilm formation was determined immediately, and at 1 month and 3 months after the gels had been prepared. Data were analyzed using a mixed-model ANOVA. The MTAP and DAP gels significantly reduced biofilm formation by both bacterial species at all time points, regardless of the tested dilution. No-significant differences in biofilm-inhibitory effects between MTAP and DAP gels were observed at the majority of the tested dilutions through various time points. Gels loaded with 1 mg/mL MTAP and DAP demonstrated a significant antibiofilm effect against E.faecalis and P. gingivalis. PMID:26369485

  9. Adaptation to NaCl Reduces the Susceptibility of Enterococcus faecalis to Melaleuca alternifolia (Tea Tree) Oil.

    PubMed

    Lim, Ee Lin; Hammer, Katherine Ann

    2015-10-01

    This study investigated the hypothesis that the salt adaptation response of Enterococcus faecalis alters susceptibility to tea tree oil (TTO). Six E. faecalis isolates were adapted to 6.5 % NaCl, and then exposed to TTO in phosphate-buffered saline (PBS). One isolate was also exposed to TTO in Brain Heart Infusion Broth (BHIB). The viability of salt-adapted and non-adapted control cells was determined at 0, 45 and 90 min and compared. MICs for several antibiotics and TTO were also determined by E test and broth microdilution, respectively. Results showed that susceptibility to TTO in PBS was significantly reduced after salt adaptation for five isolates (83 %) (P < 0.05). Mean differences between salt-adapted and non-adapted cell counts were 2.51 log at 45 min and 2.13 log at 90 min. However, when E. faecalis ATCC 19433 was exposed to TTO in BHIB, no significant differences were seen. In conclusion, salt adaptation resulted in reduced susceptibility to TTO in PBS for the majority of isolates, indicating that cross protection had occurred. This effect was absent in BHIB, suggesting that the uptake of compatible solutes from the growth medium protected non-adapted cells from TTO. Whether this has implications for the clinical effectiveness of TTO remains to be determined. PMID:26159776

  10. ANTIMICROBIAL ACTIVITY OF SODIUM HYPOCHLORITE ASSOCIATED WITH INTRACANAL MEDICATION FOR Candida albicans AND Enterococcus faecalis INOCULATED IN ROOT CANALS

    PubMed Central

    Valera, Marcia Carneiro; da Silva, Katy Costa Godinho; Maekawa, Lilian Eiko; Carvalho, Cláudio Antonio Talge; Koga-Ito, Cristiane Yumi; Camargo, Carlos Henrique Ribeiro; Silva e Lima, Raphael

    2009-01-01

    Objective: The purpose of this study was to evaluate the action of sodium hypochlorite (NaOCl) associated with an intracanal medication against Candida albicans and Enterococcus faecalis inoculated in root canals. Material and Methods: Thirty-six human single-rooted teeth with single root canals were used. The canals were contaminated with C. albicans and E. faecalis for 21 days and were then instrumented with 1% NaOCl. The roots were divided into 3 groups (n=12) according to the intracanal medication applied: calcium hydroxide paste, 2% chlorhexidine (CHX) gel, and 2% CHX gel associated with calcium hydroxide. The following collections were made from the root canals: a) initial sample (IS): 21 days after contamination (control), b) S1: after instrumentation, c) S2: 14 days after intracanal medication placement; S3: 7 days after intracanal medication removal. The results were analyzed statistically by the Kruskal-Wallis test at 5% significance level. Results and Conclusions: Both 1% NaOCl irrigation and the intracanal medications were effective in eliminating E. faecalis and C. albicans inoculated in root canals. PMID:20027425

  11. Spread of Enterococcal Surface Protein in Antibiotic Resistant Entero-coccus faecium and Enterococcus faecalis isolates from Urinary Tract Infections

    PubMed Central

    Kafil, Hossein S; Mobarez, Ashraf M

    2015-01-01

    Enterococci rank among leading cause of nosocomial bacteremia and urinary tract infection in hospital and community acquired infections. Several traits that may contribute to enhanced virulence have been identified in Enterococci. Extracellular surface protein (Esp) is a virulence factor that contributes in biofilm formation and resistance to environmental stresses. In this study we aimed to determine occurrence of esp in E. faecium and E. faecalis isolates isolated from urinary tract infections and to investigate whether there is any correlation between presence of esp and antibiotic resistance. One hundred and sixty six isolates were collected from patients with UTI and after identification by biochemical and PCR, antibiotic resistances were examined. The presence of esp was investigated by primer-specific PCR. 43.3% of isolates identified as E. faecium and 56.7% as E. faecalis. The esp gene was found in 76.1% of E. faecium isolates and 77.9% of E. faecalis isolate. There were significant correlation between esp positive E. faecium and resistance to Vancomycin (p<0.01), also in E.faecalis we found correlation between esp positive and resistance to Ampicillin, Chloramphenicol and Tetracycline (p<0.01, p<0.01, p<0.01 respectively). Occurrence of esp in our isolates from urinary tract infection was high that indicates importance of this gene in urinary tract infections and shows importance of ability to forming biofilm and hydrophobicity of surface of Enterococci for causing urinary infection by Enterococci. Also, our finding showed significant correlation between resistance to antibiotics and presence of esp in Enterococci. PMID:26161154

  12. Mortality in Kittens Is Associated with a Shift in Ileum Mucosa-Associated Enterococci from Enterococcus hirae to Biofilm-Forming Enterococcus faecalis and Adherent Escherichia coli

    PubMed Central

    Ghosh, Anuradha; Borst, Luke; Stauffer, Stephen H.; Suyemoto, Mitsu; Moisan, Peter; Zurek, Ludek

    2013-01-01

    Approximately 15% of foster kittens die before 8 weeks of age, with most of these kittens demonstrating clinical signs or postmortem evidence of enteritis. While a specific cause of enteritis is not determined in most cases, these kittens are often empirically administered probiotics that contain enterococci. The enterococci are members of the commensal intestinal microbiota but also can function as opportunistic pathogens. Given the complicated role of enterococci in health and disease, it would be valuable to better understand what constitutes a “healthy” enterococcal community in these kittens and how this microbiota is impacted by severe illness. In this study, we characterized the ileum mucosa-associated enterococcal community of 50 apparently healthy and 50 terminally ill foster kittens. In healthy kittens, Enterococcus hirae was the most common species of ileum mucosa-associated enterococci and was often observed to adhere extensively to the small intestinal epithelium. These E. hirae isolates generally lacked virulence traits. In contrast, non-E. hirae enterococci, notably Enterococcus faecalis, were more commonly isolated from the ileum mucosa of kittens with terminal illness. Isolates of E. faecalis had numerous virulence traits and multiple antimicrobial resistances. Moreover, the attachment of Escherichia coli to the intestinal epithelium was significantly associated with terminal illness and was not observed in any kitten with adherent E. hirae. These findings identify a significant difference in the species of enterococci cultured from the ileum mucosa of kittens with terminal illness compared to the species cultured from healthy kittens. In contrast to prior case studies that associated enteroadherent E. hirae with diarrhea in young animals, these controlled studies identified E. hirae as more often isolated from healthy kittens and adherence of E. hirae as more common and extensive in healthy kittens than in sick kittens. PMID:23966487

  13. A Comparison between Antibacterial Activity of Propolis and Aloe vera on Enterococcus faecalis (an In Vitro Study).

    PubMed

    Ehsani, Maryam; Amin Marashi, Mahmood; Zabihi, Ebrahim; Issazadeh, Maryam; Khafri, Soraya

    2013-01-01

    Removing the bacteria, including Enterococcus faecalis, from the root canal is one of the important aims in endodontic treatment.We aimed to compare the antibacterial activity of Chlorhexidine with two natural drugs. The antibacterial activities of three different propolis extracts (alcohol concentrations: 0, 15, 40%) and Aloe vera gel on E. faecalis were compared using three methods: disk diffusion, microdilution and direct contact test. In addition to the above bacterium, the Aloe vera gel effect on Staphylococcus aureus and Streptococcus mutans was evaluated. Disk diffusion test revealed that propolis ethanolic extracts (the alcohol concentration of 15 and 40%) and Aloe vera gel have antibacterial activities but aqueous extract of propolis did not show any effect in this test. The MICs for propolis ethanolic extracts, Aloe vera gel and aqueous extract of propolis (0% alcohol) were 313 µg/ml, 750 µg/ml, 2250 µg/ml, and ≥ 500 µg/ml respectively, much higher than the Chlorhexidine one. In direct contact test, contrary to Aloe vera, all three propolis extracts showed antibacterial effects on E. faecalis. The Aloe vera gel also showed significant antibacterial effect on S.aureus and S.mutans. The hydroalcoholic extracts of propolis and Aloe vera gel had antibacterial effects on E. faecalis, however, propolis is more potent than Aloe vera. The antibacterial effect of Aloe vera on S. aureus and S. mutans is low (MIC ≥ 2250 µg/ml). Appropriate concentrations of alcoholic extracts of propolis and some fractions of Aloe vera gel might be good choices for disinfecting the root canal in endodontic treatments.

  14. The in Vitro Antibacterial Efficacy of Persian Green Tea Extract as an Intracanal Irrigant on Enterococcus faecalis Biofilm

    PubMed Central

    Ramezanali, Fatemeh; Samimi, Shiva; Kharazifard, Mohammadjavad; Afkhami, Farzaneh

    2016-01-01

    Introduction: The aim of this study was to compare the antibacterial effect of Persian green tea extract (GTE) and 2.5% sodium hypochlorite (NaOCl) against Enterococcus faecalis (E. faecalis) as an intracanal irrigant. Methods and Materials: Thirty freshly extracted teeth were instrumented and sectioned into mesial and distal segments. The specimens were put into wells containing 2 mL of E. faecalis-containing medium. After 3 weeks, the specimens were removed and divided randomly into three groups (n=20). Each group was exposed to 3 mL of different irrigants for 3 min. Groups 1, 2 and 3 were irrigated with GTE, 2.5% NaOCl and normal saline, respectively. Biofilm formed in the middle third of the root canal was carved by sterile scalpel and cultured in Mueller-Hinton medium. Number of colony forming units (CFU) was counted on each plate. In addition, antimicrobial activity of the irrigants was evaluated by the agar disc diffusion test. The diameter of inhibition zone (IZ) around each irrigant was evaluated. The Kruskal-Wallis and Dunn tests were used to analysis the data. Results: While in NaOCl group no bacterial colonies were observed, the mean number of E. faecalis in GTE and control groups were 275±74 CFU/mL (P<0.001) and 119×108±11×108 (P<0.001), respectively. The mean of IZ in NaOCl and GTE groups were 24.35±0.78 and 6.9±0.87 mm, in order of appearance (P<0.001). Zone of inhibition was not observed around the control group (P<0.001). Conclusion: This research highlighted the potential role of plant extracts in antimicrobial root canal irrigation protocol. PMID:27790260

  15. Bacillus subtilis as a platform for molecular characterisation of regulatory mechanisms of Enterococcus faecalis resistance against cell wall antibiotics.

    PubMed

    Fang, Chong; Stiegeler, Emanuel; Cook, Gregory M; Mascher, Thorsten; Gebhard, Susanne

    2014-01-01

    To combat antibiotic resistance of Enterococcus faecalis, a better understanding of the molecular mechanisms, particularly of antibiotic detection, signal transduction and gene regulation is needed. Because molecular studies in this bacterium can be challenging, we aimed at exploiting the genetically highly tractable Gram-positive model organism Bacillus subtilis as a heterologous host. Two fundamentally different regulators of E. faecalis resistance against cell wall antibiotics, the bacitracin sensor BcrR and the vancomycin-sensing two-component system VanSB-VanRB, were produced in B. subtilis and their functions were monitored using target promoters fused to reporter genes (lacZ and luxABCDE). The bacitracin resistance system BcrR-BcrAB of E. faecalis was fully functional in B. subtilis, both regarding regulation of bcrAB expression and resistance mediated by the transporter BcrAB. Removal of intrinsic bacitracin resistance of B. subtilis increased the sensitivity of the system. The lacZ and luxABCDE reporters were found to both offer sensitive detection of promoter induction on solid media, which is useful for screening of large mutant libraries. The VanSB-VanRB system displayed a gradual dose-response behaviour to vancomycin, but only when produced at low levels in the cell. Taken together, our data show that B. subtilis is a well-suited host for the molecular characterization of regulatory systems controlling resistance against cell wall active compounds in E. faecalis. Importantly, B. subtilis facilitates the careful adjustment of expression levels and genetic background required for full functionality of the introduced regulators.

  16. Influence of the length of remaining root canal filling and post space preparation on the coronal leakage of Enterococcus faecalis

    PubMed Central

    Mozini, Alexandra Conca Alves; Vansan, Luis P.; Sousa Neto, Manoel D.; Pietro, Rosimeire

    2009-01-01

    This study evaluated the sealing ability of different lengths of remaining root canal filling and post space preparation against coronal leakage of Enterococcus faecalis. Forty-one roots of maxillary incisors were biomechanically prepared, maintaining standardized canal diameter at the middle and coronal thirds. The roots were autoclaved and all subsequent steps were undertaken in a laminar flow chamber. The canals of 33 roots were obturated with AH Plus sealer and gutta-percha. The root canal fillings were reduced to 3 predetermined lengths (n=11): G1=6 mm, G2=4 mm and G3=2 mm. The remaining roots served as positive and negative controls. Bacterial leakage test apparatuses were fabricated with the roots attached to Eppendorf tubes keeping 2 mm of apex submerged in BHI in glass flasks. The specimens received an E. faecalis inoculum of 1 x 107 cfu/mL every 3 days and were observed for bacterial leakage daily during 60 days. Data were submitted to ANOVA, Tukey’s test and Fisher’s test. At 60 days, G1 (6 mm) and G2 (4 mm) presented statistically similar results (p>0.05) (54.4% of specimens with bacterial leakage) and both groups differed significantly (p<0.01) from G3 (2 mm), which presented 100% of specimens with E. faecalis leakage. It may be concluded that the shortest endodontic obturation remnant leaked considerably more than the other lengths, although none of the tested conditions avoids coronal leakage of E. faecalis. PMID:24031339

  17. Enterococcus faecalis strains from food, environmental, and clinical origin produce ACE-inhibitory peptides and other bioactive peptides during growth in bovine skim milk.

    PubMed

    Gútiez, Loreto; Gómez-Sala, Beatriz; Recio, Isidra; del Campo, Rosa; Cintas, Luis M; Herranz, Carmen; Hernández, Pablo E

    2013-08-16

    Enterococcus faecalis isolates from food and environmental origin were evaluated for their angiotensin-converting enzyme (ACE)-inhibitory activity (ACE-IA) after growth in bovine skim milk (BSM). Most (90% active) but not all (10% inactive) E. faecalis strains produced BSM-derived hydrolysates with high ACE-IA. Known ACE-inhibitory peptides (ACE-IP) and an antioxidant peptide were identified in the E. faecalis hydrolysates by reversed-phase high-performance liquid chromatography-tandem mass spectrometry (RP-HPLC-MS/MS). Antimicrobial activity against Pediococcus damnosus CECT4797 and Listeria ivanovii CECT913 was also observed in the E. faecalis hydrolysates. The incidence of virulence factors in the E. faecalis strains with ACE-IA and producers of ACE-IP was variable but less virulence factors were observed in the food and environmental strains than in the clinical reference strains. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) based analysis demonstrated that food and environmental E. faecalis strains were genetically different from those of clinical origin. When evaluated, most E. faecalis strains of clinical origin also originated BSM-derived hydrolysates with high ACE-IA due to the production of ACE-IP. Accordingly, the results of this work suggest that most E. faecalis strains of food, environmental and clinical origin produce BSM-derived bioactive peptides with human health connotations and potential biotechnological applications.

  18. Molecular characterization of resistance, virulence and clonality in vancomycin-resistant Enterococcus faecium and Enterococcus faecalis: A hospital-based study in Beijing, China.

    PubMed

    Yang, Jing-xian; Li, Tong; Ning, Yong-zhong; Shao, Dong-hua; Liu, Jing; Wang, Shu-qin; Liang, Guo-wei

    2015-07-01

    The incidence of vancomycin-resistant enterococcus (VRE) in China is increasing, the molecular epidemiology of VRE in China is only partly known. This study was conducted to assess the molecular characterization of resistance, virulence and clonality of 69 vancomycin-resistant Enterococcus faecium (VREfm) and seven vancomycin-resistant Enterococcus faecalis (VREfs) isolates obtained from a Chinese hospital between July 2011 and July 2013. The glycopeptide resistance genes (VanA and VanB) were screened by multiplex PCR. The presence of five putative virulence genes (esp, gelE, asa1, hyl and cylA) were evaluated by another multiplex PCR. Multilocus sequence typing (MLST) scheme was used to assess the clonality. All 76 VRE isolates exhibited VanA phenotype and harbored VanA gene. Esp was the only gene detected both in VREfm and VREfs strains, accounting for 89.9% and 42.9%, respectively. The hyl gene was merely positive in 27.5% of VREfm strains. MLST analysis demonstrated three STs (ST6, ST4 and ST470) in VREfs and twelve STs (ST78, ST571, ST17, ST564, ST389, ST18, ST547, ST341, ST414, ST343, ST262 and ST203) in VREfm, which were all designated as CC17 by eBURST algorithm. An outbreak of VREfm belonging to ST571 was found to happen within the neurology ward in this hospital. To our knowledge, this is the first report of ST6 (CC2) VREfs strains in China and the first outbreak report of VREfm strains belonging to ST571 around the world. Our data could offer important information for understanding the molecular features of VRE in China.

  19. Increased pheromone cCF10 expression in Enterococcus faecalis biofilm formed by isolates from renal transplant patients.

    PubMed

    Jarzembowski, Tomasz; Daca, Agnieszka; Bryl, Ewa; Wiśniewska, Katarzyna; Gołębiewska, Justyna; Dębska-Ślizień, Alicja; Rutkowski, Bolesław; Witkowski, Jacek

    2012-12-01

    Renal transplant recipients are at a high risk of developing infectious complications even caused by commensal bacteria. This is because of various physiological non-immunological, and immunological protective mechanisms are not fully efficient in RTx patients. Therefore, rapid and precise diagnostic tools are essential in this particular group of patients. We aimed to develop simple and sensitive protocol Flow-Fish for the study of gene expression in enterococci and to compare expression of genes involved in virulence regulation in biofilm and planktonic form of Enterococcus faecalis. Proper optimization of the method was demonstrated with analysis of dehydrogenase gene expression. According to expectation reduction of the dehydrogenase gene expression was observed in biofilm. Furthermore, expression of studied gene was higher in clinical than in commensal strains. We have also found that in contrast to dehydrogenase gene, pheromone cCF10 gene expression increasing then clinical strains formed biofilm.

  20. The influence of different peritoneal dialysis fluids on the in vitro activity of ampicillin, daptomycin, and linezolid against Enterococcus faecalis.

    PubMed

    Kussmann, M; Schuster, L; Zeitlinger, M; Pichler, P; Reznicek, G; Wiesholzer, M; Burgmann, H; Poeppl, W

    2015-11-01

    Intraperitoneal administration of antibiotics is recommended for the treatment of peritoneal dialysis-related peritonitis. However, little data are available on a possible interference between peritoneal dialysis fluids and the activity of antimicrobial agents. Thus, the present in vitro study set out to investigate the influence of different peritoneal dialysis fluids on the antimicrobial activity of ampicillin, linezolid, and daptomycin against Enterococcus faecalis. Time-kill curves in four different peritoneal dialysis fluids were performed over 24 h with four different concentrations (1 × MIC, 4 × MIC, 8 × MIC, 30 × MIC) of each antibiotic evaluated. Cation-adjusted Mueller-Hinton broth was used as the comparator solution. All four peritoneal dialysis fluids evaluated had a bacteriostatic effect on the growth of Enterococcus faecalis. Compared to the cation-adjusted Mueller-Hinton broth comparator solution, the antimicrobial activity of all antibiotics tested was reduced. For ampicillin and linezolid, no activity was found in any peritoneal dialysis fluid, regardless of the concentration. Daptomycin demonstrated dose-dependent activity in all peritoneal dialysis fluids. Bactericidal activity was observed at the highest concentrations evaluated in Dianeal® PDG4 and Extraneal®, but not in concentrations lower than 30 × MIC and not in Nutrineal® PD4 and Physioneal® 40. The antimicrobial activity of ampicillin and linezolid is limited in peritoneal dialysis fluids in vitro. Daptomycin is highly effective in peritoneal dialysis fluids and might, thus, serve as an important treatment option in peritoneal dialysis-related peritonitis. Further studies are needed to evaluate the clinical impact of the present findings.

  1. Inactivation of Staphylococcus aureus and Enterococcus faecalis by a direct-current, cold atmospheric-pressure air plasma microjet☆

    PubMed Central

    Tian, Ye; Sun, Peng; Wu, Haiyan; Bai, Na; Wang, Ruixue; Zhu, Weidong; Zhang, Jue; Liu, Fuxiang

    2010-01-01

    Objective A direct-current, cold atmospheric-pressure air plasma microjet (PMJ) was performed to inactivate Staphylococcus aureus (S. aureus) and Enterococcus faecalis (E. faecalis) in air. The process of sterilization and morphology of bacteria was observed. We wish to know the possible inactivation mechanisms of PMJ and explore a potential application in dental and other temperature sensitive treatment. Methods In this study, we employed a direct current, atmospheric pressure, cold air PMJ to inactivate bacterias. Scanning electron microscopy was employed to evaluate the morphology of S. aureus and showed rupture of cell walls after the plasma treatment and Optical emission spectrum (OES) were used to understand the possible inactivation mechanisms of PMJ. Results The inactivation rates could reach 100% in 5 min. When the distance between the exit nozzle of the PMJ device and Petri dish was extended from 1 cm to 3 cm, effective inactivation was also observed with a similar inactivation curve. Conclusion The inactivation of bacteria is attributed to the abundant reactive oxygen and nitrogen species, as well as ultroviolet radiation in the plasma. Different life spans and defensibilities of these killing agents may hold the key to understanding the different inactivation curves at different treatment distances. PMID:23554639

  2. Inefficient translation renders the Enterococcus faecalis fabK enoyl-acyl carrier protein reductase phenotypically cryptic.

    PubMed

    Bi, Hongkai; Zhu, Lei; Wang, Haihong; Cronan, John E

    2014-01-01

    Enoyl-acyl carrier protein (ACP) reductase catalyzes the last step of the bacterial fatty acid elongation cycle. Enterococcus faecalis is unusual in that it encodes two unrelated enoyl-ACP reductases, FabI and FabK. We recently reported that deletion of the gene encoding FabI results in an unsaturated fatty acid (UFA) auxotroph despite the presence of fabK, a gene encoding a second fully functional enoyl-ACP reductase. By process of elimination, our prior report argued that poor expression was the reason that fabK failed to functionally replace FabI. We now report that FabK is indeed poorly expressed and that the expression defect is at the level of translation rather than transcription. We isolated four spontaneous mutants that allowed growth of the E. faecalis ΔfabI strain on fatty acid-free medium. Each mutational lesion (single base substitution or deletion) extended the fabK ribosome binding site. Inactivation of fabK blocked growth, indicating that the mutations acted only on fabK rather than a downstream gene. The mutations activated fabK translation to levels that supported fatty acid synthesis and hence cell growth. Furthermore, site-directed and random mutagenesis experiments showed that point mutations that resulted in increased complementarity to the 3' end of the 16S rRNA increased FabK translation to levels sufficient to support growth, whereas mutations that decreased complementarity blocked fabK translation.

  3. Antibacterial Activity of Synthetic Peptides Derived from Lactoferricin against Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 29212

    PubMed Central

    León-Calvijo, María A.; Leal-Castro, Aura L.; Almanzar-Reina, Giovanni A.; Rosas-Pérez, Jaiver E.; García-Castañeda, Javier E.; Rivera-Monroy, Zuly J.

    2015-01-01

    Peptides derived from human and bovine lactoferricin were designed, synthesized, purified, and characterized using RP-HPLC and MALDI-TOF-MS. Specific changes in the sequences were designed as (i) the incorporation of unnatural amino acids in the sequence, the (ii) reduction or (iii) elongation of the peptide chain length, and (iv) synthesis of molecules with different number of branches containing the same sequence. For each peptide, the antibacterial activity against Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 29212 was evaluated. Our results showed that Peptides I.2 (RWQWRWQWR) and I.4 ((RRWQWR)4K2Ahx2C2) exhibit bigger or similar activity against E. coli (MIC 4–33 μM) and E. faecalis (MIC 10–33 μM) when they were compared with lactoferricin protein (LF) and some of its derivate peptides as II.1 (FKCRRWQWRMKKLGA) and IV.1 (FKCRRWQWRMKKLGAPSITCVRRAE). It should be pointed out that Peptides I.2 and I.4, containing the RWQWR motif, are short and easy to synthesize; our results demonstrate that it is possible to design and obtain synthetic peptides that exhibit enhanced antibacterial activity using a methodology that is fast and low-cost and that allows obtaining products with a high degree of purity and high yield. PMID:25815317

  4. The effects of solution chemistry on the sticking efficiencies of viable Enterococcus faecalis: An atomic force microscopy and modeling study

    NASA Astrophysics Data System (ADS)

    Cail, Tracy L.; Hochella, Michael F.

    2005-06-01

    Atomic force microscopy (AFM) and Derjaguin-Landau-Verwey-Overbeek (DLVO) theory in combination with the interaction force boundary layer (IFBL) model have been used to empirically and theoretically calculate sticking efficiencies (α) of Enterococcus faecalis cells against a silica glass surface. Sticking efficiencies were calculated in solutions of varying pH and ionic strength and related to maximum distances of transport through a hypothetical soil block using colloid filtration theory. AFM measurements show that the repulsive and attractive forces between E. faecalis cells and a glass surface are a function of ionic strength but are less sensitive to changes in solution pH. Zeta (ζ)-potential measurements of the cells and glass surfaces correlate with these trends. Calculated DLVO energy profiles predict much greater sensitivity to changing solution chemistry. Sticking efficiencies derived from AFM measurements range from 9.6 × 10 -17 to 1 in solutions of low ionic strength (IS) and from 2.6 × 10 -33 to 1 at higher IS. Corresponding α values determined from DLVO theory are essentially zero in all tested solutions. Sticking efficiencies calculated in this study are smaller than values determined from column and field studies in similar systems; however, α derived from AFM data and the IFBL model more closely represent field data than do values calculated from DLVO energy values. A comparison with different methods of calculating α suggests that reversible adhesion may be significant in column-scale transport studies.

  5. Decolorization and detoxification of sulfonated toxic diazo dye C.I. Direct Red 81 by Enterococcus faecalis YZ 66.

    PubMed

    Sahasrabudhe, Madhuri M; Saratale, Rijuta G; Saratale, Ganesh D; Pathade, Girish R

    2014-01-01

    Isolated Enterococcus faecalis YZ 66 strain shows ability to decolorize various industrial dyes among which, it showed complete decolorization and degradation of toxic, sulfonated recalcitrant diazo dye Direct Red 81 (50 mg/L) within 1.5 h of incubation under static anoxic condition. The optimum pH and temperature for decolorization was 7.0 and 40°C, respectively. Significant induction in the activity of intracellular oxidoreductive enzymes suggested its involvement in the decolorization of Direct Red 81. The biodegradation of Direct Red 81 was monitored by UV-Visible, FT-IR spectroscopy and HPLC. The final products were characterized by GC-MS and possible pathway of the degradation of the dye was proposed. The phytotoxicity assay (with respect to plants Sorghum vulgare and Phaseolus mungo) revealed that the degradation of Direct Red 81 produced nontoxic metabolites. Finally E. faecalis was employed to decolorize actual industrial effluent showing decolorization (in terms of ADMI value) with moderate COD and BOD reduction. Moreover the result increases the applicability of the strain for the treatment of industrial wastewaters containing dye pollutants. PMID:25649265

  6. Decolorization and detoxification of sulfonated toxic diazo dye C.I. Direct Red 81 by Enterococcus faecalis YZ 66.

    PubMed

    Sahasrabudhe, Madhuri M; Saratale, Rijuta G; Saratale, Ganesh D; Pathade, Girish R

    2014-01-01

    Isolated Enterococcus faecalis YZ 66 strain shows ability to decolorize various industrial dyes among which, it showed complete decolorization and degradation of toxic, sulfonated recalcitrant diazo dye Direct Red 81 (50 mg/L) within 1.5 h of incubation under static anoxic condition. The optimum pH and temperature for decolorization was 7.0 and 40°C, respectively. Significant induction in the activity of intracellular oxidoreductive enzymes suggested its involvement in the decolorization of Direct Red 81. The biodegradation of Direct Red 81 was monitored by UV-Visible, FT-IR spectroscopy and HPLC. The final products were characterized by GC-MS and possible pathway of the degradation of the dye was proposed. The phytotoxicity assay (with respect to plants Sorghum vulgare and Phaseolus mungo) revealed that the degradation of Direct Red 81 produced nontoxic metabolites. Finally E. faecalis was employed to decolorize actual industrial effluent showing decolorization (in terms of ADMI value) with moderate COD and BOD reduction. Moreover the result increases the applicability of the strain for the treatment of industrial wastewaters containing dye pollutants.

  7. Protection of mice from oral Candidiasis by heat-killed enterococcus faecalis, possibly through its direct binding to Candida albicans.

    PubMed

    Ishijima, Sanae A; Hayama, Kazumi; Ninomiya, Kentaro; Iwasa, Masahiro; Yamazaki, Masatoshi; Abe, Shigeru

    2014-01-01

    To develop a new therapy against oral candidiasis, a commensal microorganism, Enterococcus faecalis was tested for its ability to modulate Candida growth in vitro and its therapeutic activities against a murine model in vivo. Addition of heat-killed E. faecalis strain EF2001 (EF2001) isolated from healthy human feces to the culture of C. albicans strain TIMM1768 inhibited adherence of the latter to a microtiter plate in a dose dependent manner and Candida cells surrounded by EF2001 were increased. To examine the protective activities of EF2001 in vivo, heat-killed EF2001 was applied orally before and after inoculation of Candida to the tongue of mice previously immunosuppressed. Two days after inoculation this inoculation, both the symptom score and CFU from swabbed-tongue were significantly reduced in the EF2001-treated animals. Histological analysis indicated that EF2001 may potentiate the accumulation of polymorphnuclear cells near a Candida-infected region. These results suggest that oral administration of EF2001 has protective activity against oral candidiasis and that the in vivo activity may be reflected by direct interaction between EF2001 and Candida cells in vitro and the potentiation of an immunostimulatory effect of EF2001.

  8. Development of an intracanal mature Enterococcus faecalis biofilm and its susceptibility to some antimicrobial intracanal medications; an in vitro study

    PubMed Central

    Saber, Shehab El-Din Mohamed; El-Hady, Soha A.

    2012-01-01

    Objectives: To develop a mature biofilm of Enterococcus faecalis inside the root canal system and to test its susceptibility to some antimicrobial medications in vitro. Methods: Single rooted premolars were mechanically enlarged, sterilized, and then infected with a clinical isolate of E. faecalis. Biofilm formation and maturation was monitored using SEM. Biofilm bacteria were exposed to Amoxicillin+clavulanate, Ciprofloxacin, Clindamycin, Doxycycline, and calcium hydroxide as intracanal medications for 1 week. Finally bacterial samples were collected, and colony-forming units were enumerated. Results: SEM examination confirmed the formation of a mature biofilm at the end of the incubation period. All the chemotherapeutic agents used were significantly better than Calcium hydroxide in elimination of biofilm bacteria. The antimicrobial effect of Amoxicillin + clavulanate, Ciprofloxacin and Clindamycin was significantly better than Doxycycline (P=.05). However the difference in the antimicrobial effectiveness among them was statistically non-significant (P=.05). Conclusions: The method used for bacterial biofilm development and maturation is reliable and can be used to assess the anti bacterial potential of endodontic materials. Also, the local application of antibacterial agents can be beneficial in resistant cases of apical periodontitis but only after careful culture and sensitivity testing to choose the appropriate agent for the existing flora. PMID:22229006

  9. Identification of vancomycin interaction with Enterococcus faecalis within 30 min of interaction time using Raman spectroscopy.

    PubMed

    Assmann, Cora; Kirchhoff, Johanna; Beleites, Claudia; Hey, Jessica; Kostudis, Sophia; Pfister, Wolfgang; Schlattmann, Peter; Popp, Jürgen; Neugebauer, Ute

    2015-11-01

    Vancomycin is an important glycopeptide antibiotic which is used to treat serious infections caused by Gram-positive bacteria. However, during the last years, a tremendous rise in vancomycin resistances, especially among Enterococci, was reported, making fast diagnostic methods inevitable. In this contribution, we apply Raman spectroscopy to systematically characterize vancomycin-enterococci interactions over a time span of 90 min using a sensitive Enterococcus faecalis strain and two different vancomycin concentrations above the minimal inhibitory concentration (MIC). Successful action of the drug on the pathogen could be observed already after 30 min of interaction time. Characteristic spectral changes are visualized with the help of multivariate statistical analysis (linear discriminant analysis and partial least squares regressions). Those changes were employed to train a statistical model to predict vancomycin treatment based on the Raman spectra. The robustness of the model was tested using data recorded by an independent operator. Classification accuracies of >90 % were obtained for vancomycin concentrations in the lower range of a typical trough serum concentration recommended for most patients during appropriate vancomycin therapy. Characterization of drug-pathogen interactions by means of label-free spectroscopic methods, such as Raman spectroscopy, can provide the knowledge base for innovative and fast susceptibility tests which could speed up microbiological analysis as well as finding applications in novel antibiotic screenings assays. Graphical Abstract E. faecalis is incubated with vancomycin and characterized by means of Raman spectroscopy after different time points. Characteristic spectral changes reveal efficient vancomycin-enterococci-interaction.

  10. Penicillin-resistant, ampicillin-susceptible Enterococcus faecalis of hospital origin: pbp4 gene polymorphism and genetic diversity.

    PubMed

    Conceição, Natália; da Silva, Lucas Emanuel Pinheiro; Darini, Ana Lúcia da Costa; Pitondo-Silva, André; de Oliveira, Adriana Gonçalves

    2014-12-01

    Despite the spread of penicillin-resistant, ampicillin-susceptible Enterococcus faecalis (PRASEF) isolates in diverse countries, the mechanisms leading to this unusual resistance phenotype have not yet been investigated. The aim of this study was to evaluate whether polymorphism in the pbp4 gene is associated with penicillin resistance in PRASEF isolates and to determine their genetic diversity. E. faecalis isolates were recovered from different clinical specimens of hospitalized patients from February 2006 to June 2010. The β-lactam minimal inhibitory concentrations (MICs) were determined by E-test®. The PCR-amplified pbp4 gene was sequenced with an automated sequencer. The genetic diversities of the isolates were established by PFGE (pulsed-field gel electrophoresis) and MLST (multilocus sequencing typing). Seventeen non-producing β-lactamase PRASEF and 10 penicillin-susceptible, ampicillin-susceptible E. faecalis (PSASEF) strains were analyzed. A single-amino-acid substitution (Asp-573→Glu) in the penicillin-binding domain was significantly found in all PRASEF isolates by sequencing of the pbp4 gene but not in the penicillin-susceptible isolates. In contrast to the PSASEF isolates, a majority of the PRASEFs had similar PFGE profiles. Six representative PRASEF isolates were resolved by MLST into ST9 and ST524 and belong to the globally dispersed clonal complex 9 (CC9). In conclusion, it appears quite likely that the amino acid alteration (Asp-573→Glu) found in the PBP4 of the Brazilian PRASEF isolates may account for their reduced susceptibility to penicillin, although other resistance mechanisms remain to be investigated.

  11. Uses and limitations of green fluorescent protein as a viability marker in Enterococcus faecalis: An observational investigation.

    PubMed

    Hoogenkamp, Michel A; Crielaard, Wim; Krom, Bastiaan P

    2015-08-01

    Enterococci are capable of producing biofilms that are notoriously difficult to treat and remove, for instance in root canal infections. The tenacious nature of these organisms makes screening of known and novel antimicrobial compounds necessary. While traditionally growth and fluorescence-based screening methods have proven useful, these methods have their limitations when applied to enterococci (e.g. time consuming, no kinetic data, diffusion properties of the fluorescent dyes). The aim of this study was to develop and validate a GFP-based high-throughput screening system to assess the bactericidal activity of a broad range of antimicrobial agents on Enterococcus faecalis and its biofilms. The effect of antimicrobial compounds on cell viability and GFP fluorescence of enterococcal planktonic and biofilm cells was determined using colony forming unit counts, fluorescence spectrophotometry and real-time imaging devices. There was a linear correlation between cell viability and GFP fluorescence. The intensity of the GFP signal was effected by the extracellular pH. For a range of antimicrobials however, there was no correlation between these two parameters. In contrast, for oxidizing agents such as sodium hypochlorite, the antimicrobial of choice for root canal disinfection, there was a correlation between loss of fluorescence and loss of viability. To conclude, the use of a GFP-based system to monitor the antimicrobial activity of compounds on E. faecalis is possible despite significant limitations. This approach is useful for analysis of susceptibility to oxidizing agents. Using real-time measuring devices to follow GFP fluorescence it should be possible to investigate the mode of action and rate of diffusion of oxidizing agents in E. faecalis biofilm.

  12. Protective effect of Enterococcus faecalis DAPTO 512 on the intestinal tract and gut mucosa: milk allergy application.

    PubMed

    Belkaaloul, K; Haertlé, T; Chobert, J M; Merah, R; Taibi, K; Saad El Hachemi, H A; Hemch, S; Amier, L; Chekroun, A; Saidi, D; Kheroua, O

    2015-01-01

    The allergenicity of β-lactoglobulin (β-Lg) was studied by using Ussing chamber in a murine model of β-Lg allergy supplemented with hydrolysates obtained after fermentation of milk for 48 h at 37 (°)C with Enterococcus faecalis DAPTO 512, isolated from cow milk and identified by 16S rDNA sequence analysis. Balb/c mice were sensitised intraperitoneally with β-Lg. Three groups of mice were formed: group 1, composed of naive mice used as control received only NaCl; group 2, positive control composed of mice sensitised intraperitoneally with β-Lg; group 3, formed by mice which were given hydrolysates of 48 h then sensitised with β-Lg. After 48 h of fermentation β-casein and β-Lg were degraded by E. faecalis DAPTO 512. β-Lg immunisation was associated with strong IgG and IgE production in case of positive controls and a significant increase in short current circuit (Isc) and high conductance (G) responses were observed. The control and the hydrolysate groups showed a significant decrease in the production of IgG and IgE anti β-Lg compared to the positive control. The allergenic potential of β-Lg was markedly reduced in the group that received hydrolysates (Isc and G remained unchanged after intestine challenge with β-Lg). The histological scrutiny showed villi atrophy, lymphocyte hyperplasia and a significant chorion detachment in the positive control group. In the group administered with hydrolysates of fermented milk, inflammatory signs were lower, the villi were long and thin and lymphocytes were less dense. The results showed that feeding of milk fermented with E. faecalis DAPTO 512 during 18 days prior to β-Lg allergy induction exerts a protecting effect on the murine intestine and induces a significant decrease in the β-Lg allergenicity.

  13. Activities of fosfomycin and rifampin on planktonic and adherent Enterococcus faecalis strains in an experimental foreign-body infection model.

    PubMed

    Oliva, Alessandra; Furustrand Tafin, Ulrika; Maiolo, Elena Maryka; Jeddari, Safaa; Bétrisey, Bertrand; Trampuz, Andrej

    2014-01-01

    Enterococcal implant-associated infections are difficult to treat because antibiotics generally lack activity against enterococcal biofilms. We investigated fosfomycin, rifampin, and their combinations against planktonic and adherent Enterococcus faecalis (ATCC 19433) in vitro and in a foreign-body infection model. The MIC/MBClog values were 32/>512 μg/ml for fosfomycin, 4/>64 μg/ml for rifampin, 1/2 μg/ml for ampicillin, 2/>256 μg/ml for linezolid, 16/32 μg/ml for gentamicin, 1/>64 μg/ml for vancomycin, and 1/5 μg/ml for daptomycin. In time-kill studies, fosfomycin was bactericidal at 8× and 16× MIC, but regrowth of resistant strains occurred after 24 h. With the exception of gentamicin, no complete inhibition of growth-related heat production was observed with other antimicrobials on early (3 h) or mature (24 h) biofilms. In the animal model, fosfomycin alone or in combination with daptomycin reduced planktonic counts by ≈4 log10 CFU/ml below the levels before treatment. Fosfomycin cleared planktonic bacteria from 74% of cage fluids (i.e., no growth in aspirated fluid) and eradicated biofilm bacteria from 43% of cages (i.e., no growth from removed cages). In combination with gentamicin, fosfomycin cleared 77% and cured 58% of cages; in combination with vancomycin, fosfomycin cleared 33% and cured 18% of cages; in combination with daptomycin, fosfomycin cleared 75% and cured 17% of cages. Rifampin showed no activity on planktonic or adherent E. faecalis, whereas in combination with daptomycin it cured 17% and with fosfomycin it cured 25% of cages. Emergence of fosfomycin resistance was not observed in vivo. In conclusion, fosfomycin showed activity against planktonic and adherent E. faecalis. Its role against enterococcal biofilms should be further investigated, especially in combination with rifampin and/or daptomycin treatment.

  14. Structural elucidation of SrtA enzyme in Enterococcus faecalis: an emphasis on screening of potential inhibitors against the biofilm formation.

    PubMed

    Selvaraj, Chandrabose; Sivakamavalli, Jeyachandran; Vaseeharan, Baskaralingam; Singh, Poonam; Singh, Sanjeev Kumar

    2014-07-01

    Enterococcus faecalis is a pathogenic Gram-positive bacterium, which mainly infects humans through urinary tract infections. SrtA is an essential enzyme for survival of E. faecalis, and inhibition of this particular enzyme will reduce the virulence of biofilm formation. It is proved to be associated with the microbial surface protein embedded signal transduction mechanism and promising as a suitable anti-microbial drug target for E. faecalis. The present work gives an inclusive description of SrtA isolated from E. faecalis through computational and experimental methodologies. For exploring the mechanism of SrtA and to screen potential leads against E. faecalis, we have generated three-dimensional models through homology modeling. The 3D model showed conformational stability over time, confirming the quality of the starting 3D model. Large scale 100 ns molecular dynamics simulations show the intramolecular changes occurring in SrtA, and multiple conformations of structure based screening elucidate potential leads against this pathogen. Experimental results showed that the screened compounds are active showing anti-microbial and anti-biofilm activity, as SrtA is known to play an important role in E. faecalis biofilm formation. Experimental results also suggest that SrtA specific screened compounds have better anti-biofilm activity than the available inhibitors. Therefore, we believe that development of these compounds would be an impetus to design the novel chief SrtA inhibitors against E. faecalis.

  15. Development and Use of an Efficient System for Random mariner Transposon Mutagenesis To Identify Novel Genetic Determinants of Biofilm Formation in the Core Enterococcus faecalis Genome▿ †

    PubMed Central

    Kristich, Christopher J.; Nguyen, Vy T.; Le, Thinh; Barnes, Aaron M. T.; Grindle, Suzanne; Dunny, Gary M.

    2008-01-01

    Enterococcus faecalis is a gram-positive commensal bacterium of the gastrointestinal tract and an important opportunistic pathogen. Despite the increasing clinical significance of the enterococci, most of the genetic analysis of these organisms has focused on mobile genetic elements, and existing tools for manipulation and analysis of the core E. faecalis chromosome are limited. We are interested in a comprehensive analysis of the genetic determinants for biofilm formation encoded within the core E. faecalis genome. To identify such determinants, we developed a substantially improved system for transposon mutagenesis in E. faecalis based on a mini-mariner transposable element. Mutagenesis of wild-type E. faecalis with this element yielded predominantly mutants carrying a single copy of the transposable element, and insertions were distributed around the entire chromosome in an apparently random fashion. We constructed a library of E. faecalis transposon insertion mutants and screened this library to identify mutants exhibiting a defect in biofilm formation. Biofilm-defective mutants were found to carry transposon insertions both in genes that were previously known to play a role in biofilm formation and in new genes lacking any known function; for several genes identified in the screen, complementation analysis confirmed a direct role in biofilm formation. These results provide significant new information about the genetics of enterococcal biofilm formation and demonstrate the general utility of our transposon system for functional genomic analysis of E. faecalis. PMID:18408066

  16. Structural elucidation of SrtA enzyme in Enterococcus faecalis: an emphasis on screening of potential inhibitors against the biofilm formation.

    PubMed

    Selvaraj, Chandrabose; Sivakamavalli, Jeyachandran; Vaseeharan, Baskaralingam; Singh, Poonam; Singh, Sanjeev Kumar

    2014-07-01

    Enterococcus faecalis is a pathogenic Gram-positive bacterium, which mainly infects humans through urinary tract infections. SrtA is an essential enzyme for survival of E. faecalis, and inhibition of this particular enzyme will reduce the virulence of biofilm formation. It is proved to be associated with the microbial surface protein embedded signal transduction mechanism and promising as a suitable anti-microbial drug target for E. faecalis. The present work gives an inclusive description of SrtA isolated from E. faecalis through computational and experimental methodologies. For exploring the mechanism of SrtA and to screen potential leads against E. faecalis, we have generated three-dimensional models through homology modeling. The 3D model showed conformational stability over time, confirming the quality of the starting 3D model. Large scale 100 ns molecular dynamics simulations show the intramolecular changes occurring in SrtA, and multiple conformations of structure based screening elucidate potential leads against this pathogen. Experimental results showed that the screened compounds are active showing anti-microbial and anti-biofilm activity, as SrtA is known to play an important role in E. faecalis biofilm formation. Experimental results also suggest that SrtA specific screened compounds have better anti-biofilm activity than the available inhibitors. Therefore, we believe that development of these compounds would be an impetus to design the novel chief SrtA inhibitors against E. faecalis. PMID:24718729

  17. Behavior of Listeria monocytogenes in a multi-species biofilm with Enterococcus faecalis and Enterococcus faecium and control through sanitation procedures.

    PubMed

    da Silva Fernandes, Meg; Kabuki, Dirce Yorika; Kuaye, Arnaldo Yoshiteru

    2015-05-01

    The formation of mono-species biofilm (Listeria monocytogenes) and multi-species biofilms (Enterococcus faecium, Enterococcus faecalis, and L. monocytogenes) was evaluated. In addition, the effectiveness of sanitation procedures for the control of the multi-species biofilm also was evaluated. The biofilms were grown on stainless steel coupons at various incubation temperatures (7, 25 and 39°C) and contact times (0, 1, 2, 4, 6 and 8 days). In all tests, at 7°C, the microbial counts were below 0.4 log CFU/cm(2) and not characteristic of biofilms. In mono-species biofilm, the counts of L. monocytogenes after 8 days of contact were 4.1 and 2.8 log CFU/cm(2) at 25 and 39°C, respectively. In the multi-species biofilms, Enterococcus spp. were present at counts of 8 log CFU/cm(2) at 25 and 39°C after 8 days of contact. However, the L. monocytogenes in multi-species biofilms was significantly affected by the presence of Enterococcus spp. and by temperature. At 25°C, the growth of L. monocytogenes biofilms was favored in multi-species cultures, with counts above 6 log CFU/cm(2) after 8 days of contact. In contrast, at 39°C, a negative effect was observed for L. monocytogenes biofilm growth in mixed cultures, with a significant reduction in counts over time and values below 0.4 log CFU/cm(2) starting at day 4. Anionic tensioactive cleaning complemented with another procedure (acid cleaning, disinfection or acid cleaning+disinfection) eliminated the multi-species biofilms under all conditions tested (counts of all micro-organisms<0.4 log CFU/cm(2)). Peracetic acid was the most effective disinfectant, eliminating the multi-species biofilms under all tested conditions (counts of the all microorganisms <0.4 log CFU/cm(2)). In contrast, biguanide was the least effective disinfectant, failing to eliminate biofilms under all the test conditions.

  18. Behavior of Listeria monocytogenes in a multi-species biofilm with Enterococcus faecalis and Enterococcus faecium and control through sanitation procedures.

    PubMed

    da Silva Fernandes, Meg; Kabuki, Dirce Yorika; Kuaye, Arnaldo Yoshiteru

    2015-05-01

    The formation of mono-species biofilm (Listeria monocytogenes) and multi-species biofilms (Enterococcus faecium, Enterococcus faecalis, and L. monocytogenes) was evaluated. In addition, the effectiveness of sanitation procedures for the control of the multi-species biofilm also was evaluated. The biofilms were grown on stainless steel coupons at various incubation temperatures (7, 25 and 39°C) and contact times (0, 1, 2, 4, 6 and 8 days). In all tests, at 7°C, the microbial counts were below 0.4 log CFU/cm(2) and not characteristic of biofilms. In mono-species biofilm, the counts of L. monocytogenes after 8 days of contact were 4.1 and 2.8 log CFU/cm(2) at 25 and 39°C, respectively. In the multi-species biofilms, Enterococcus spp. were present at counts of 8 log CFU/cm(2) at 25 and 39°C after 8 days of contact. However, the L. monocytogenes in multi-species biofilms was significantly affected by the presence of Enterococcus spp. and by temperature. At 25°C, the growth of L. monocytogenes biofilms was favored in multi-species cultures, with counts above 6 log CFU/cm(2) after 8 days of contact. In contrast, at 39°C, a negative effect was observed for L. monocytogenes biofilm growth in mixed cultures, with a significant reduction in counts over time and values below 0.4 log CFU/cm(2) starting at day 4. Anionic tensioactive cleaning complemented with another procedure (acid cleaning, disinfection or acid cleaning+disinfection) eliminated the multi-species biofilms under all conditions tested (counts of all micro-organisms<0.4 log CFU/cm(2)). Peracetic acid was the most effective disinfectant, eliminating the multi-species biofilms under all tested conditions (counts of the all microorganisms <0.4 log CFU/cm(2)). In contrast, biguanide was the least effective disinfectant, failing to eliminate biofilms under all the test conditions. PMID:25655573

  19. Surface-Associated Lipoproteins Link Enterococcus faecalis Virulence to Colitogenic Activity in IL-10-Deficient Mice Independent of Their Expression Levels.

    PubMed

    Ocvirk, Soeren; Sava, Irina G; Lengfelder, Isabella; Lagkouvardos, Ilias; Steck, Natalie; Roh, Jung H; Tchaptchet, Sandrine; Bao, Yinyin; Hansen, Jonathan J; Huebner, Johannes; Carroll, Ian M; Murray, Barbara E; Sartor, R Balfour; Haller, Dirk

    2015-06-01

    The commensal Enterococcus faecalis is among the most common causes of nosocomial infections. Recent findings regarding increased abundance of enterococci in the intestinal microbiota of patients with inflammatory bowel diseases and induction of colitis in IL-10-deficient (IL-10-/-) mice put a new perspective on the contribution of E. faecalis to chronic intestinal inflammation. Based on the expression of virulence-related genes in the inflammatory milieu of IL-10-/- mice using RNA-sequencing analysis, we characterized the colitogenic role of two bacterial structures that substantially impact on E. faecalis virulence by different mechanisms: the enterococcal polysaccharide antigen and cell surface-associated lipoproteins. Germ-free wild type and IL-10-/- mice were monoassociated with E. faecalis wild type OG1RF or the respective isogenic mutants for 16 weeks. Intestinal tissue and mesenteric lymph nodes (MLN) were collected to characterize tissue pathology, loss of intestinal barrier function, bacterial adhesion to intestinal epithelium and immune cell activation. Bone marrow-derived dendritic cells (BMDC) were stimulated with bacterial lysates and E. faecalis virulence was additionally investigated in three invertebrate models. Colitogenic activity of wild type E. faecalis (OG1RF score: 7.2±1.2) in monoassociated IL-10-/- mice was partially impaired in E. faecalis lacking enterococcal polysaccharide antigen (ΔepaB score: 4.7±2.3; p<0.05) and was almost completely abrogated in E. faecalis deficient for lipoproteins (Δlgt score: 2.3±2.3; p<0.0001). Consistently both E. faecalis mutants showed significantly impaired virulence in Galleria mellonella and Caenorhabditis elegans. Loss of E-cadherin in the epithelium was shown for all bacterial strains in inflamed IL-10-/- but not wild type mice. Inactivation of epaB in E. faecalis reduced microcolony and biofilm formation in vitro, altered bacterial adhesion to intestinal epithelium of germ-free Manduca sexta larvae

  20. Surface-Associated Lipoproteins Link Enterococcus faecalis Virulence to Colitogenic Activity in IL-10-Deficient Mice Independent of Their Expression Levels.

    PubMed

    Ocvirk, Soeren; Sava, Irina G; Lengfelder, Isabella; Lagkouvardos, Ilias; Steck, Natalie; Roh, Jung H; Tchaptchet, Sandrine; Bao, Yinyin; Hansen, Jonathan J; Huebner, Johannes; Carroll, Ian M; Murray, Barbara E; Sartor, R Balfour; Haller, Dirk

    2015-06-01

    The commensal Enterococcus faecalis is among the most common causes of nosocomial infections. Recent findings regarding increased abundance of enterococci in the intestinal microbiota of patients with inflammatory bowel diseases and induction of colitis in IL-10-deficient (IL-10-/-) mice put a new perspective on the contribution of E. faecalis to chronic intestinal inflammation. Based on the expression of virulence-related genes in the inflammatory milieu of IL-10-/- mice using RNA-sequencing analysis, we characterized the colitogenic role of two bacterial structures that substantially impact on E. faecalis virulence by different mechanisms: the enterococcal polysaccharide antigen and cell surface-associated lipoproteins. Germ-free wild type and IL-10-/- mice were monoassociated with E. faecalis wild type OG1RF or the respective isogenic mutants for 16 weeks. Intestinal tissue and mesenteric lymph nodes (MLN) were collected to characterize tissue pathology, loss of intestinal barrier function, bacterial adhesion to intestinal epithelium and immune cell activation. Bone marrow-derived dendritic cells (BMDC) were stimulated with bacterial lysates and E. faecalis virulence was additionally investigated in three invertebrate models. Colitogenic activity of wild type E. faecalis (OG1RF score: 7.2±1.2) in monoassociated IL-10-/- mice was partially impaired in E. faecalis lacking enterococcal polysaccharide antigen (ΔepaB score: 4.7±2.3; p<0.05) and was almost completely abrogated in E. faecalis deficient for lipoproteins (Δlgt score: 2.3±2.3; p<0.0001). Consistently both E. faecalis mutants showed significantly impaired virulence in Galleria mellonella and Caenorhabditis elegans. Loss of E-cadherin in the epithelium was shown for all bacterial strains in inflamed IL-10-/- but not wild type mice. Inactivation of epaB in E. faecalis reduced microcolony and biofilm formation in vitro, altered bacterial adhesion to intestinal epithelium of germ-free Manduca sexta larvae

  1. Surface-Associated Lipoproteins Link Enterococcus faecalis Virulence to Colitogenic Activity in IL-10-Deficient Mice Independent of Their Expression Levels

    PubMed Central

    Lengfelder, Isabella; Lagkouvardos, Ilias; Steck, Natalie; Roh, Jung H.; Tchaptchet, Sandrine; Bao, Yinyin; Hansen, Jonathan J.; Huebner, Johannes; Carroll, Ian M.; Murray, Barbara E.; Sartor, R. Balfour; Haller, Dirk

    2015-01-01

    The commensal Enterococcus faecalis is among the most common causes of nosocomial infections. Recent findings regarding increased abundance of enterococci in the intestinal microbiota of patients with inflammatory bowel diseases and induction of colitis in IL-10-deficient (IL-10-/-) mice put a new perspective on the contribution of E. faecalis to chronic intestinal inflammation. Based on the expression of virulence-related genes in the inflammatory milieu of IL-10-/- mice using RNA-sequencing analysis, we characterized the colitogenic role of two bacterial structures that substantially impact on E. faecalis virulence by different mechanisms: the enterococcal polysaccharide antigen and cell surface-associated lipoproteins. Germ-free wild type and IL-10-/- mice were monoassociated with E. faecalis wild type OG1RF or the respective isogenic mutants for 16 weeks. Intestinal tissue and mesenteric lymph nodes (MLN) were collected to characterize tissue pathology, loss of intestinal barrier function, bacterial adhesion to intestinal epithelium and immune cell activation. Bone marrow-derived dendritic cells (BMDC) were stimulated with bacterial lysates and E. faecalis virulence was additionally investigated in three invertebrate models. Colitogenic activity of wild type E. faecalis (OG1RF score: 7.2±1.2) in monoassociated IL-10-/- mice was partially impaired in E. faecalis lacking enterococcal polysaccharide antigen (ΔepaB score: 4.7±2.3; p<0.05) and was almost completely abrogated in E. faecalis deficient for lipoproteins (Δlgt score: 2.3±2.3; p<0.0001). Consistently both E. faecalis mutants showed significantly impaired virulence in Galleria mellonella and Caenorhabditis elegans. Loss of E-cadherin in the epithelium was shown for all bacterial strains in inflamed IL-10-/- but not wild type mice. Inactivation of epaB in E. faecalis reduced microcolony and biofilm formation in vitro, altered bacterial adhesion to intestinal epithelium of germ-free Manduca sexta larvae

  2. Detection of opsonic antibodies against Enterococcus faecalis cell wall carbohydrates in immune globulin preparations.

    PubMed

    Hufnagel, M; Sixel, K; Hammer, F; Kropec, A; Sava, I G; Theilacker, C; Berner, R; Huebner, J

    2014-08-01

    Three different commercially available polyvalent immune globulins (IG) were investigated for the existence of antibodies against cell wall carbohydrates of four different E. faecalis serotypes (using a cell wall carbohydrate-enzyme-linked immunosorbent assay), and whether these antibodies mediated opsonic killing (using an opsonic-killing assay). All three IG preparations contained antibodies against all four serotypes (CPS-A to CPS-D). However, only one of the three IG preparations showed opsonic killing against all four serotypes. Average killing was higher against serotypes A and B (72 and 79 %, respectively) than against serotypes C and D (30 and 37 %, respectively). Such IG preparations could play a role as an adjuvant therapeutic option in life-threatening infections with E. faecalis, particularly when resistant strains are involved.

  3. Bacteriocin protein BacL1 of Enterococcus faecalis targets cell division loci and specifically recognizes L-Ala2-cross-bridged peptidoglycan.

    PubMed

    Kurushima, Jun; Nakane, Daisuke; Nishizaka, Takayuki; Tomita, Haruyoshi

    2015-01-01

    Bacteriocin 41 (Bac41) is produced from clinical isolates of Enterococcus faecalis and consists of two extracellular proteins, BacL1 and BacA. We previously reported that BacL1 protein (595 amino acids, 64.5 kDa) is a bacteriolytic peptidoglycan D-isoglutamyl-L-lysine endopeptidase that induces cell lysis of E. faecalis when an accessory factor, BacA, is copresent. However, the target of BacL1 remains unknown. In this study, we investigated the targeting specificity of BacL1. Fluorescence microscopy analysis using fluorescent dye-conjugated recombinant protein demonstrated that BacL1 specifically localized at the cell division-associated site, including the equatorial ring, division septum, and nascent cell wall, on the cell surface of target E. faecalis cells. This specific targeting was dependent on the triple repeat of the SH3 domain located in the region from amino acid 329 to 590 of BacL1. Repression of cell growth due to the stationary state of the growth phase or to treatment with bacteriostatic antibiotics rescued bacteria from the bacteriolytic activity of BacL1 and BacA. The static growth state also abolished the binding and targeting of BacL1 to the cell division-associated site. Furthermore, the targeting of BacL1 was detectable among Gram-positive bacteria with an L-Ala-L-Ala-cross-bridging peptidoglycan, including E. faecalis, Streptococcus pyogenes, or Streptococcus pneumoniae, but not among bacteria with alternate peptidoglycan structures, such as Enterococcus faecium, Enterococcus hirae, Staphylococcus aureus, or Listeria monocytogenes. These data suggest that BacL1 specifically targets the L-Ala-L-Ala-cross-bridged peptidoglycan and potentially lyses the E. faecalis cells during cell division.

  4. Partial Purification and Characterization of the Mode of Action of Enterocin S37: A Bacteriocin Produced by Enterococcus faecalis S37 Isolated from Poultry Feces

    PubMed Central

    Belguesmia, Y.; Choiset, Y.; Prévost, H.; Dalgalarrondo, M.; Chobert, J.-M.; Drider, D.

    2010-01-01

    The aim of this research was to purify and characterize the mode of action of enterocin S37, a bacteriocin produced by Enterococcus faecalis S37, a strain recently isolated from the chicken feces. Enterocin S37 has a molecular weight comprised between 4 and 5 kDa. It remained active after 1 h at 80oC and at pH values ranging from 4.0 to 9.0. Furthermore, cell-free supernatant of Enterococcus faecalis S37 and purified enterocin S37 were active against Gram-positive bacteria including Listeria monocytogenes EGDe, L. innocua F, Enterococcus faecalis JH2-2, and Lactobacillus brevis F145. The purification of enterocin S37 was performed by ammonium sulfate precipitation followed up by hydrophobic-interaction chromatography procedures. Treatment of enterocin S37 with proteinase K, α-chymotrypsin, and papain confirmed its proteinaceous nature, while its treatment with lysozyme and lipase resulted in no alteration of activity. Enterocin S37 is hydrophobic, anti-Listeria and likely acting by depletion of intracellular K+ ions upon action on KATP channels. This study contributed to gain more insights into the mode of action of enterocins. PMID:20811593

  5. Biosorption and biodegradation of Acid Orange 7 by Enterococcus faecalis strain ZL: optimization by response surface methodological approach.

    PubMed

    Lim, Chi Kim; Bay, Hui Han; Aris, Azmi; Abdul Majid, Zaiton; Ibrahim, Zaharah

    2013-07-01

    Reactive dyes account for one of the major sources of dye wastes in textile effluent. In this study, decolorization of the monoazo dye, Acid Orange 7 (AO7) by the Enterococcus faecalis strain ZL that isolated from a palm oil mill effluent treatment plant has been investigated. Decolorization efficiency of azo dye is greatly affected by the types of nutrients and the size of inoculum used. In this work, one-factor-at-a-time (method and response surface methodology (RSM) was applied to optimize these operational factors and also to study the combined interaction between them. Analysis of AO7 decolorization was done using Fourier transform infrared (FTIR) spectroscopy, desorption study, UV-Vis spectral analysis, field emission scanning electron microscopy (FESEM), and high performance liquid chromatography (HPLC). The optimum condition via RSM for the color removal of AO7 was found to be as follows: yeast extract, 0.1% w/v, glycerol concentration of 0.1% v/v, and inoculum density of 2.5% v/v at initial dye concentration of 100 mg/L at 37 °C. Decolorization efficiency of 98% was achieved in only 5 h. The kinetic of AO7 decolorization was found to be first order with respect to dye concentration with a k value of 0.87/h. FTIR, desorption study, UV-Vis spectral analysis, FESEM, and HPLC findings indicated that the decolorization of AO7 was mainly due to the biosorption as well as biodegradation of the bacterial cells. In addition, HPLC analyses also showed the formation of sulfanilic acid as a possible degradation product of AO7 under facultative anaerobic condition. This study explored the ability of E. faecalis strain ZL in decolorizing AO7 by biosorption as well as biodegradation process.

  6. Pyruvate Formate-Lyase Is Essential for Fumarate-Independent Anaerobic Glycerol Utilization in the Enterococcus faecalis Strain W11

    PubMed Central

    Ikegami, Yuki

    2014-01-01

    Although anaerobic glycerol metabolism in Enterococcus faecalis requires exogenous fumarate for NADH oxidation, E. faecalis strain W11 can metabolize glycerol in the absence of oxygen without exogenous fumarate. In this study, metabolic end product analyses and reporter assays probing the expression of enzymes involved in pyruvate metabolism were performed to investigate this fumarate-independent anaerobic metabolism of glycerol in W11. Under aerobic conditions, the metabolic end products of W11 cultured with glycerol were similar to those of W11 cultured with glucose. However, when W11 was cultured anaerobically, most of the glucose was converted to l-lactate, but glycerol was converted to ethanol and formate. During anaerobic culture with glycerol, the expression of the l-lactate dehydrogenase and pyruvate dehydrogenase E1αβ genes in W11 was downregulated, whereas the expression of the pyruvate formate-lyase (Pfl) and aldehyde/alcohol dehydrogenase genes was upregulated. These changes in the expression levels caused the change in the composition of end products. A pflB gene disruptant (Δpfl mutant) of W11 could barely utilize glycerol under anaerobic conditions, but the growth of the Δpfl mutant cultured with either glucose or dihydroxyacetone (DHA) under anaerobic conditions was the same as that of W11. Glucose metabolism and DHA generates one NADH molecule per pyruvate molecule, whereas glycerol metabolism in the dehydrogenation pathway generates two NADH molecules per pyruvate molecule. These findings demonstrate that NADH generated from anaerobic glycerol metabolism in the absence of fumarate is oxidized through the Pfl-ethanol fermentation pathway. Thus, Pfl is essential to avoid the accumulation of excess NADH during fumarate-independent anaerobic glycerol metabolism. PMID:24769696

  7. Pyruvate formate-lyase is essential for fumarate-independent anaerobic glycerol utilization in the Enterococcus faecalis strain W11.

    PubMed

    Doi, Yuki; Ikegami, Yuki

    2014-07-01

    Although anaerobic glycerol metabolism in Enterococcus faecalis requires exogenous fumarate for NADH oxidation, E. faecalis strain W11 can metabolize glycerol in the absence of oxygen without exogenous fumarate. In this study, metabolic end product analyses and reporter assays probing the expression of enzymes involved in pyruvate metabolism were performed to investigate this fumarate-independent anaerobic metabolism of glycerol in W11. Under aerobic conditions, the metabolic end products of W11 cultured with glycerol were similar to those of W11 cultured with glucose. However, when W11 was cultured anaerobically, most of the glucose was converted to l-lactate, but glycerol was converted to ethanol and formate. During anaerobic culture with glycerol, the expression of the l-lactate dehydrogenase and pyruvate dehydrogenase E1αβ genes in W11 was downregulated, whereas the expression of the pyruvate formate-lyase (Pfl) and aldehyde/alcohol dehydrogenase genes was upregulated. These changes in the expression levels caused the change in the composition of end products. A pflB gene disruptant (Δpfl mutant) of W11 could barely utilize glycerol under anaerobic conditions, but the growth of the Δpfl mutant cultured with either glucose or dihydroxyacetone (DHA) under anaerobic conditions was the same as that of W11. Glucose metabolism and DHA generates one NADH molecule per pyruvate molecule, whereas glycerol metabolism in the dehydrogenation pathway generates two NADH molecules per pyruvate molecule. These findings demonstrate that NADH generated from anaerobic glycerol metabolism in the absence of fumarate is oxidized through the Pfl-ethanol fermentation pathway. Thus, Pfl is essential to avoid the accumulation of excess NADH during fumarate-independent anaerobic glycerol metabolism. PMID:24769696

  8. The structure and competitive substrate inhibition of dihydrofolate reductase from Enterococcus faecalis reveal restrictions to cofactor docking.

    PubMed

    Bourne, Christina R; Wakeham, Nancy; Webb, Nicole; Nammalwar, Baskar; Bunce, Richard A; Berlin, K Darrell; Barrow, William W

    2014-02-25

    We are addressing bacterial resistance to antibiotics by repurposing a well-established classic antimicrobial target, the dihydrofolate reductase (DHFR) enzyme. In this work, we have focused on Enterococcus faecalis, a nosocomial pathogen that frequently harbors antibiotic resistance determinants leading to complicated and difficult-to-treat infections. An inhibitor series with a hydrophobic dihydrophthalazine heterocycle was designed from the anti-folate trimethoprim. We have examined the potency of this inhibitor series based on inhibition of DHFR enzyme activity and bacterial growth, including in the presence of the exogenous product analogue folinic acid. The resulting preferences were rationalized using a cocrystal structure of the DHFR from this organism with a propyl-bearing series member (RAB-propyl). In a companion apo structure, we identify four buried waters that act as placeholders for a conserved hydrogen-bonding network to the substrate and indicate an important role in protein stability during catalytic cycling. In these structures, the nicotinamide of the nicotinamide adenine dinucleotide phosphate cofactor is visualized outside of its binding pocket, which is exacerbated by RAB-propyl binding. Finally, homology models of the TMP(R) sequences dfrK and dfrF were constructed. While the dfrK-encoded protein shows clear sequence changes that would be detrimental to inhibitor binding, the dfrF-encoded protein model suggests the protein would be relatively unstable. These data suggest a utility for anti-DHFR compounds for treating infections arising from E. faecalis. They also highlight a role for water in stabilizing the DHFR substrate pocket and for competitive substrate inhibitors that may gain advantages in potency by the perturbation of cofactor dynamics.

  9. Enterococcus faecalis lipoteichoic acid suppresses Aggregatibacter actinomycetemcomitans lipopolysaccharide-induced IL-8 expression in human periodontal ligament cells.

    PubMed

    Im, Jintaek; Baik, Jung Eun; Kim, Kyoung Whun; Kang, Seok-Seong; Jeon, Jun Ho; Park, Ok-Jin; Kim, Hyun Young; Kum, Kee-Yeon; Yun, Cheol-Heui; Han, Seung Hyun

    2015-08-01

    Periodontitis is caused by multi-bacterial infection and Aggregatibacter actinomycetemcomitans and Enterococcus faecalis are closely associated with inflammatory periodontal diseases. Although lipopolysaccharide (LPS) of A. actinomycetemcomitans (Aa.LPS) and lipoteichoic acid of E. faecalis (Ef.LTA) are considered to be major virulence factors evoking inflammatory responses, their combinatorial effect on the induction of chemokines has not been investigated. In this study, we investigated the interaction between Aa.LPS and Ef.LTA on IL-8 expression in human periodontal ligament (PDL) cells. Aa.LPS, but not Ef.LTA, substantially induced IL-8 expression at the protein and mRNA levels. Interestingly, Ef.LTA suppressed Aa.LPS-induced IL-8 expression without affecting the binding of Aa.LPS to Toll-like receptor (TLR) 4. Ef.LTA reduced Aa.LPS-induced phosphorylation of mitogen-activated protein kinases, including ERK, JNK and p38 kinase. Furthermore, Ef.LTA inhibited the Aa.LPS-induced transcriptional activities of the activating protein 1, CCAAT/enhancer-binding protein and nuclear factor-kappa B transcription factors, all of which are known to regulate IL-8 gene expression. Ef.LTA augmented the expression of IL-1 receptor-associated kinase-M (IRAK-M), a negative regulator of TLR intracellular signaling pathways, in the presence of Aa.LPS at both the mRNA and protein levels. Small interfering RNA silencing IRAK-M reversed the attenuation of Aa.LPS-induced IL-8 expression by Ef.LTA. Collectively, these results suggest that Ef.LTA down-regulates Aa.LPS-induced IL-8 expression in human PDL cells through up-regulation of the negative regulator IRAK-M.

  10. Unraveling antimicrobial resistance genes and phenotype patterns among Enterococcus faecalis isolated from retail chicken products in Japan.

    PubMed

    Hidano, Arata; Yamamoto, Takehisa; Hayama, Yoko; Muroga, Norihiko; Kobayashi, Sota; Nishida, Takeshi; Tsutsui, Toshiyuki

    2015-01-01

    Multidrug-resistant enterococci are considered crucial drivers for the dissemination of antimicrobial resistance determinants within and beyond a genus. These organisms may pass numerous resistance determinants to other harmful pathogens, whose multiple resistances would cause adverse consequences. Therefore, an understanding of the coexistence epidemiology of resistance genes is critical, but such information remains limited. In this study, our first objective was to determine the prevalence of principal resistance phenotypes and genes among Enterococcus faecalis isolated from retail chicken domestic products collected throughout Japan. Subsequent analysis of these data by using an additive Bayesian network (ABN) model revealed the co-appearance patterns of resistance genes and identified the associations between resistance genes and phenotypes. The common phenotypes observed among E. faecalis isolated from the domestic products were the resistances to oxytetracycline (58.4%), dihydrostreptomycin (50.4%), and erythromycin (37.2%), and the gene tet(L) was detected in 46.0% of the isolates. The ABN model identified statistically significant associations between tet(L) and erm(B), tet(L) and ant(6)-Ia, ant(6)-Ia and aph(3')-IIIa, and aph(3')-IIIa and erm(B), which indicated that a multiple-resistance profile of tetracycline, erythromycin, streptomycin, and kanamycin is systematic rather than random. Conversely, the presence of tet(O) was only negatively associated with that of erm(B) and tet(M), which suggested that in the presence of tet(O), the aforementioned multiple resistance is unlikely to be observed. Such heterogeneity in linkages among genes that confer the same phenotypic resistance highlights the importance of incorporating genetic information when investigating the risk factors for the spread of resistance. The epidemiological factors that underlie the persistence of systematic multiple-resistance patterns warrant further investigations with appropriate

  11. Unraveling Antimicrobial Resistance Genes and Phenotype Patterns among Enterococcus faecalis Isolated from Retail Chicken Products in Japan

    PubMed Central

    Hidano, Arata; Yamamoto, Takehisa; Hayama, Yoko; Muroga, Norihiko; Kobayashi, Sota; Nishida, Takeshi; Tsutsui, Toshiyuki

    2015-01-01

    Multidrug-resistant enterococci are considered crucial drivers for the dissemination of antimicrobial resistance determinants within and beyond a genus. These organisms may pass numerous resistance determinants to other harmful pathogens, whose multiple resistances would cause adverse consequences. Therefore, an understanding of the coexistence epidemiology of resistance genes is critical, but such information remains limited. In this study, our first objective was to determine the prevalence of principal resistance phenotypes and genes among Enterococcus faecalis isolated from retail chicken domestic products collected throughout Japan. Subsequent analysis of these data by using an additive Bayesian network (ABN) model revealed the co-appearance patterns of resistance genes and identified the associations between resistance genes and phenotypes. The common phenotypes observed among E. faecalis isolated from the domestic products were the resistances to oxytetracycline (58.4%), dihydrostreptomycin (50.4%), and erythromycin (37.2%), and the gene tet(L) was detected in 46.0% of the isolates. The ABN model identified statistically significant associations between tet(L) and erm(B), tet(L) and ant(6)-Ia, ant(6)-Ia and aph(3’)-IIIa, and aph(3’)-IIIa and erm(B), which indicated that a multiple-resistance profile of tetracycline, erythromycin, streptomycin, and kanamycin is systematic rather than random. Conversely, the presence of tet(O) was only negatively associated with that of erm(B) and tet(M), which suggested that in the presence of tet(O), the aforementioned multiple resistance is unlikely to be observed. Such heterogeneity in linkages among genes that confer the same phenotypic resistance highlights the importance of incorporating genetic information when investigating the risk factors for the spread of resistance. The epidemiological factors that underlie the persistence of systematic multiple-resistance patterns warrant further investigations with

  12. L-Lactic acid production from glycerol coupled with acetic acid metabolism by Enterococcus faecalis without carbon loss.

    PubMed

    Murakami, Nao; Oba, Mana; Iwamoto, Mariko; Tashiro, Yukihiro; Noguchi, Takuya; Bonkohara, Kaori; Abdel-Rahman, Mohamed Ali; Zendo, Takeshi; Shimoda, Mitsuya; Sakai, Kenji; Sonomoto, Kenji

    2016-01-01

    Glycerol is a by-product in the biodiesel production process and considered as one of the prospective carbon sources for microbial fermentation including lactic acid fermentation, which has received considerable interest due to its potential application. Enterococcus faecalis isolated in our laboratory produced optically pure L-lactic acid from glycerol in the presence of acetic acid. Gas chromatography-mass spectrometry analysis using [1, 2-(13)C2] acetic acid proved that the E. faecalis strain QU 11 was capable of converting acetic acid to ethanol during lactic acid fermentation of glycerol. This indicated that strain QU 11 restored the redox balance by oxidizing excess NADH though acetic acid metabolism, during ethanol production, which resulted in lactic acid production from glycerol. The effects of pH control and substrate concentration on lactic acid fermentation were also investigated. Glycerol and acetic acid concentrations of 30 g/L and 10 g/L, respectively, were expected to be appropriate for lactic acid fermentation of glycerol by strain QU 11 at a pH of 6.5. Furthermore, fed-batch fermentation with 30 g/L glycerol and 10 g/L acetic acid wholly exhibited the best performance including lactic acid production (55.3 g/L), lactic acid yield (0.991 mol-lactic acid/mol-glycerol), total yield [1.08 mol-(lactic acid and ethanol)]/mol-(glycerol and acetic acid)], and total carbon yield [1.06 C-mol-(lactic acid and ethanol)/C-mol-(glycerol and acetic acid)] of lactic acid and ethanol. In summary, the strain QU 11 successfully produced lactic acid from glycerol with acetic acid metabolism, and an efficient fermentation system was established without carbon loss.

  13. Comparison of the Antimicrobial Efficacy of Two Antibiotics Sparfloxacin and Augmentin as Experimental Root Canal Irrigating Solutions against Enterococcus faecalis - An Invitro Study

    PubMed Central

    Venigalla, Bhuvan Shome; Surakanti, Jayaprada Reddy; Thumu, Jayaprakash; Chennamaneni, Krishna Chaitanya; Kalluru, Rama S.

    2016-01-01

    Introduction One of the main goals of endodontic treatment is root canal disinfection and to prevent subsequent chances of reinfection. Adjuvant to instrumentation, root canal irrigants are required to eliminate the bacteria found on the root canal walls and lateral canals within the dentinal tubules. Aim To measure and compare the antibacterial efficacy of two antibiotics as experimental root canal irrigating solutions against Enterococcus faecalis (E. faecalis). Materials and Methods Fifteen Brain Heart Infusion agar plates were inoculated with Enterococcus faecalis-American Type Culture Collection (ATCC) 29212. 5 micrograms (mcg) Sparfloxacin discs, 30mcg Augmentin discs, and sterile paper test discs saturated with 2% Chlorhexidine (CHX), 3% Sodium Hypochlorite (NaOCl) and 5% NaOCl solutions were placed on agar plates. Sodium Chloride 0.9% (NaCl) paper discs were used as controls. Fifteen plates were incubated aerobically at 37°C. Results were expressed as per the terms of the diameter of the inhibition zone. Results Results suggested a statistically significant difference in the zones of inhibition between five irrigating solutions (p < 0.001). Conclusion Although, zones of inhibition were found in all the groups, 5mcg Sparfloxacin and 30mcg Augmentin showed maximum antimicrobial activity against E.faecalis. PMID:27135003

  14. Molecular detection of virulence factors among food and clinical Enterococcus faecalis strains in South Brazil.

    PubMed

    Medeiros, A W; Pereira, R I; Oliveira, D V; Martins, P D; d'Azevedo, P A; Van der Sand, S; Frazzon, J; Frazzon, A P G

    2014-01-01

    The present report aimed to perform a molecular epidemiological survey by investigating the presence of virulence factors in E. faecalis isolated from different human clinical (n = 57) and food samples (n = 55) in Porto Alegre, Brazil, collected from 2006 to 2009. In addition, the ability to form biofilm in vitro on polystyrene and the β-haemolytic and gelatinase activities were determined. Clinical strains presented a higher prevalence of aggregation substance (agg), enterococcal surface protein (esp) and cytolysin (cylA) genes when compared with food isolates. The esp gene was found only in clinical strains. On the other hand, the gelatinase (gelE) and adherence factor (ace) genes had similar prevalence among the strains, showing the widespread occurrence of these virulence factors among food and clinical E. faecalis strains in South Brazil. More than three virulence factor genes were detected in 77.2% and 18.2% of clinical and food strains, respectively. Gelatinase and β-haemolysin activities were not associated with the presence of gelE and cylA genes. The ability to produce biofilm was detected in 100% of clinical and 94.6% of food isolates, and clinical strains were more able to form biofilm than the food isolates (Student's t-test, p < 0.01). Results from the statistical analysis showed significant associations between strong biofilm formation and ace (p = 0.015) and gelE (p = 0.007) genes in clinical strains. In conclusion, our data indicate that E. faecalis strains isolated from clinical and food samples possess distinctive patterns of virulence factors, with a larger number of genes that encode virulence factors detected in clinical strains.

  15. The Effect of Addition of an EPS Degrading Enzyme with and without Detergent to 2% Chlorhexidine on Disruption of Enterococcus faecalis Biofilm: A Confocal Laser Scanning Microscopic Study

    PubMed Central

    Nagendrababu, Venkateshbabu; John, Aby; Deivanayagam, Kandaswamy

    2015-01-01

    Background Enterococcus faecalis is one of the most commonly occurring organisms retrieved from root canal treated teeth that show refractory apical periodontitis. Though it is well known that the ability of E. faecalis to form a matrix-encased biofilm contributes to its pathogenicity, the role of extracellular dextran and DNA in biofilm formation and its effect on the susceptibility of the biofilm to chlorhexidine remains poorly understood. It was hypothesized that the addition of an Extracellular Polymeric Substance (EPS) degrading enzyme along with a detergent to chlorhexidine may increase the susceptibility of the E. faecalis biofilm. Aim To evaluate the sensitivity of Enterococcus faecalis biofilms treated with DNase enzyme and their susceptibility to 2% chlorhexidine used alone or in conjunction with a detergent in a dentin disinfection model and examine under confocal laser scanning microscopy (CLSM). Materials and Methods Semi cylindrical shaped dentin specimens were infected with E. faecalis and incubated for 24 hours. Following incubation, the infected dentin specimens were exposed for 3 minutes to the four disinfecting solutions and grouped accordingly. {Group I- Sterile saline, Group II- 2% Chlorhexidine (CHX), Group III– Dnase1 Enzyme + 2% CHX, Group IV- DNase1 Enzyme + 2% CHX & Tween 80. Bacterial viability was then assessed by staining the specimens and examining under CLSM to analyse the proportion of dead and live bacteria within the dentinal tubules. Results The Groups II, III and IV showed statistically significant (p<0.05) percentage of dead bacteria compared to the control (Group I). However there was no significant difference in the killing effectiveness within the experimental groups (II-IV) at (p<0.05). Conclusion EPS degrading enzyme (DNase I) disrupts the biofilm and increases the susceptibility of E.faecalis when exposed to 2% Chlorhexidine and the use of a surfactant with this combination significantly contributes to improving the

  16. Effects of intracanal irrigant MTAD Combined with nisin at sub-minimum inhibitory concentration levels on Enterococcus faecalis growth and the expression of pathogenic genes.

    PubMed

    Tong, Zhongchun; Huang, Lijia; Ling, Junqi; Mao, Xueli; Ning, Yang; Deng, Dongmei

    2014-01-01

    Exposure to antibiotics is considered to be the major driver in the selection of antibiotic-resistant bacteria and may induce diverse biological responses in bacteria. MTAD is a common intracanal irrigant, but its bactericidal activity remains to be improved. Previous studies have indicated that the antimicrobial peptide nisin can significantly improve the bactericidal activity of MTAD against Enterococcus faecalis. However, the effects of MTAD and its modification at sub-minimum inhibitory concentration (sub-MIC) levels on Enterococcus faecalis growth and the expression of pathogenic genes still need to be explored. In this study, the results of post-antibiotic effects (PAE) and post-antibiotic sub-MIC effects (PASME) showed that MTADN (nisin in combination with MTAD) had the best post-antibiotic effect. E. faecalis after challenge with MTAD was less sensitive to alkaline solutions compared with MTAN (nisin in place of doxycycline in MTAD) and MTADN. E. faecalis induced with sub-MIC of MTAD generated resistance to the higher concentration, but induction of E. faecalis with MTAN did not cause resistance to higher concentrations. Furthermore, real-time polymerase chain reaction (RT-PCR) showed that the stress caused by sub-MIC exposure to MTAD, MTAN, or MTADN resulted in up- or down-regulation of nine stress genes and four virulence-associated genes in E. faecalis and resulted in different stress states. These findings suggested that nisin improved the post-antibacterial effect of MTAD at sub-MIC levels and has considerable potential for use as a modification of MTAD.

  17. Effects of Intracanal Irrigant MTAD Combined with Nisin at Sub-Minimum Inhibitory Concentration Levels on Enterococcus faecalis Growth and the Expression of Pathogenic Genes

    PubMed Central

    Ling, Junqi; Mao, Xueli; Ning, Yang; Deng, Dongmei

    2014-01-01

    Exposure to antibiotics is considered to be the major driver in the selection of antibiotic-resistant bacteria and may induce diverse biological responses in bacteria. MTAD is a common intracanal irrigant, but its bactericidal activity remains to be improved. Previous studies have indicated that the antimicrobial peptide nisin can significantly improve the bactericidal activity of MTAD against Enterococcus faecalis. However, the effects of MTAD and its modification at sub-minimum inhibitory concentration (sub-MIC) levels on Enterococcus faecalis growth and the expression of pathogenic genes still need to be explored. In this study, the results of post-antibiotic effects (PAE) and post-antibiotic sub-MIC effects (PASME) showed that MTADN (nisin in combination with MTAD) had the best post-antibiotic effect. E. faecalis after challenge with MTAD was less sensitive to alkaline solutions compared with MTAN (nisin in place of doxycycline in MTAD) and MTADN. E. faecalis induced with sub-MIC of MTAD generated resistance to the higher concentration, but induction of E. faecalis with MTAN did not cause resistance to higher concentrations. Furthermore, real-time polymerase chain reaction (RT-PCR) showed that the stress caused by sub-MIC exposure to MTAD, MTAN, or MTADN resulted in up- or down-regulation of nine stress genes and four virulence-associated genes in E. faecalis and resulted in different stress states. These findings suggested that nisin improved the post-antibacterial effect of MTAD at sub-MIC levels and has considerable potential for use as a modification of MTAD. PMID:24603760

  18. Molecular characterization of a widespread, pathogenic, and antibiotic resistance-receptive Enterococcus faecalis lineage and dissemination of its putative pathogenicity island.

    PubMed

    Nallapareddy, Sreedhar R; Wenxiang, Huang; Weinstock, George M; Murray, Barbara E

    2005-08-01

    Enterococcus faecalis, a common cause of endocarditis and known for its capacity to transfer antibiotic resistance to other pathogens, has recently emerged as an important, multidrug-resistant nosocomial pathogen. However, knowledge of its lineages and the potential of particular clones of this species to disseminate and cause disease is limited. Using a nine-gene multilocus sequence typing (MLST) scheme, we identified an evolving and widespread clonal complex of E. faecalis that has caused outbreaks and life-threatening infections. Moreover, this unusual clonal complex was found to contain isolates of unexpected relatedness, including the first known U.S. vancomycin-resistant enterococcus (E. faecalis strain V583), the first known penicillinase-producing (Bla(+)) E. faecalis isolate, and the previously described widespread clone of penicillinase producers, a trait found in <0.1% of E. faecalis isolates. All members of this clonal cluster (designated as BVE for Bla(+) Van(r) endocarditis) were found to contain a previously described putative pathogenicity island (PAI). Further analysis of this PAI demonstrated its dissemination worldwide, albeit with considerable variability, confirmed its association with clinical isolates, and found a common insertion site in different clonal lineages. PAI deletions, MLST, and the uncommon resistances were used to predict the evolution of the BVE clonal cluster. The finding of a virulent and highly successful clonal complex of E. faecalis with different members resistant to the primary therapies of choice, ampicillin and vancomycin, has important implications for the evolution of virulence and successful lineages and for public health monitoring and control.

  19. Effects of intracanal irrigant MTAD Combined with nisin at sub-minimum inhibitory concentration levels on Enterococcus faecalis growth and the expression of pathogenic genes.

    PubMed

    Tong, Zhongchun; Huang, Lijia; Ling, Junqi; Mao, Xueli; Ning, Yang; Deng, Dongmei

    2014-01-01

    Exposure to antibiotics is considered to be the major driver in the selection of antibiotic-resistant bacteria and may induce diverse biological responses in bacteria. MTAD is a common intracanal irrigant, but its bactericidal activity remains to be improved. Previous studies have indicated that the antimicrobial peptide nisin can significantly improve the bactericidal activity of MTAD against Enterococcus faecalis. However, the effects of MTAD and its modification at sub-minimum inhibitory concentration (sub-MIC) levels on Enterococcus faecalis growth and the expression of pathogenic genes still need to be explored. In this study, the results of post-antibiotic effects (PAE) and post-antibiotic sub-MIC effects (PASME) showed that MTADN (nisin in combination with MTAD) had the best post-antibiotic effect. E. faecalis after challenge with MTAD was less sensitive to alkaline solutions compared with MTAN (nisin in place of doxycycline in MTAD) and MTADN. E. faecalis induced with sub-MIC of MTAD generated resistance to the higher concentration, but induction of E. faecalis with MTAN did not cause resistance to higher concentrations. Furthermore, real-time polymerase chain reaction (RT-PCR) showed that the stress caused by sub-MIC exposure to MTAD, MTAN, or MTADN resulted in up- or down-regulation of nine stress genes and four virulence-associated genes in E. faecalis and resulted in different stress states. These findings suggested that nisin improved the post-antibacterial effect of MTAD at sub-MIC levels and has considerable potential for use as a modification of MTAD. PMID:24603760

  20. Comparative evaluation of effect of different irrigation solutions against Enterococcus faecalis: A polymerase chain reaction-based study

    PubMed Central

    Seelan, R. Gnana; Kumar, Arvind; Jonathan, R.; Maheswari, Uma; Raja, Jacob; Chelliah, P.

    2015-01-01

    Enterococcus faecalis is the most isolated or detected species from oral infections including marginal periodontitis, infected root canals, periradicular abscesses and also detected in cases of failed endodontic therapy. To prevent endodontic treatment failure irrigation is mandatory for the effective removal of smear layer, pulp tissue, and microorganisms. Cultivation and other traditional identification methods have been demonstrated to have several limitations when it comes to microbiological identification. Polymerase chain reaction was selected because it has an added advantage over traditional microbiological methods. Materials and Methods: Twenty single rooted premolars were taken were taken stored in 0.1% thymol solution at 4°C decoronated to obtain 12 mm length, teeth were autoclaved at 121°C, canals were instrumented up to 35k file (International Organization for Standardization). The samples were randomly divided into three groups Group I - 5.25% sodium hypochlorite (NaOCL) and 17% ethylenediaminetetraacetic acid (EDTA), Group - II 5.25% NaOCL and 2% chlorhexidine (CHX), Group III - 5.25% NaOCL and 17% EDTA and 2% CHX. Results: The results showed that Group III which is 5% NaOCl followed by 17% EDTA and followed by 2% CHX showed maximum antimicrobial activity in all the three different time intervals. PMID:26538921

  1. VirB8-like protein TraH is crucial for DNA transfer in Enterococcus faecalis

    PubMed Central

    Fercher, Christian; Probst, Ines; Kohler, Verena; Goessweiner-Mohr, Nikolaus; Arends, Karsten; Grohmann, Elisabeth; Zangger, Klaus; Meyer, N. Helge; Keller, Walter

    2016-01-01

    Untreatable bacterial infections caused by a perpetual increase of antibiotic resistant strains represent a serious threat to human healthcare in the 21st century. Conjugative DNA transfer is the most important mechanism for antibiotic resistance and virulence gene dissemination among bacteria and is mediated by a protein complex, known as type IV secretion system (T4SS). The core of the T4SS is a multiprotein complex that spans the bacterial envelope as a channel for macromolecular secretion. We report the NMR structure and functional characterization of the transfer protein TraH encoded by the conjugative Gram-positive broad-host range plasmid pIP501. The structure exhibits a striking similarity to VirB8 proteins of Gram-negative secretion systems where they play an essential role in the scaffold of the secretion machinery. Considering TraM as the first VirB8-like protein discovered in pIP501, TraH represents the second protein affiliated with this family in the respective transfer operon. A markerless traH deletion in pIP501 resulted in a total loss of transfer in Enterococcus faecalis as compared with the pIP501 wild type (wt) plasmid, demonstrating that TraH is essential for pIP501 mediated conjugation. Moreover, oligomerization state and topology of TraH in the native membrane were determined providing insights in molecular organization of a Gram-positive T4SS. PMID:27103580

  2. Library Screen Identifies Enterococcus faecalis CcpA, the Catabolite Control Protein A, as an Effector of Ace, a Collagen Adhesion Protein Linked to Virulence

    PubMed Central

    Gao, Peng; Pinkston, Kenneth L.; Bourgogne, Agathe; Cruz, Melissa R.; Garsin, Danielle A.; Murray, Barbara E.

    2013-01-01

    The Enterococcus faecalis cell wall-anchored protein Ace is an important virulence factor involved in cell adhesion and infection. Expression of Ace on the cell surface is affected by many factors, including stage of growth, culture temperature, and environmental components, such as serum, urine, and collagen. However, the mechanisms that regulate or modulate Ace display are not well understood. With interest in identifying genes associated with Ace expression, we utilized a whole-cell enzyme-linked immunosorbent assay (ELISA)-based screening method to identify mutants from a transposon insertion mutant library which exhibited distinct Ace surface expression profiles. We identified a ccpA insertion mutant which showed significantly decreased levels of Ace surface expression at early growth phase versus those of wild-type OG1RF. Confirmation of the observation was achieved through flow cytometry and complementation analysis. Compared to the wild type, the E. faecalis ccpA mutant had an impaired ability to adhere to collagen when grown to early exponential phase, consistent with the lack of Ace expression in the early growth phase. As a key component of carbon catabolite regulation, CcpA has been previously reported to play a critical role in regulating expression of proteins involved in E. faecalis carbohydrate uptake and utilization. Our discovery is the first to associate CcpA with the production of a major E. faecalis virulence factor, providing new insights into the regulation of E. faecalis pathogenesis. PMID:23974022

  3. Library screen identifies Enterococcus faecalis CcpA, the catabolite control protein A, as an effector of Ace, a collagen adhesion protein linked to virulence.

    PubMed

    Gao, Peng; Pinkston, Kenneth L; Bourgogne, Agathe; Cruz, Melissa R; Garsin, Danielle A; Murray, Barbara E; Harvey, Barrett R

    2013-10-01

    The Enterococcus faecalis cell wall-anchored protein Ace is an important virulence factor involved in cell adhesion and infection. Expression of Ace on the cell surface is affected by many factors, including stage of growth, culture temperature, and environmental components, such as serum, urine, and collagen. However, the mechanisms that regulate or modulate Ace display are not well understood. With interest in identifying genes associated with Ace expression, we utilized a whole-cell enzyme-linked immunosorbent assay (ELISA)-based screening method to identify mutants from a transposon insertion mutant library which exhibited distinct Ace surface expression profiles. We identified a ccpA insertion mutant which showed significantly decreased levels of Ace surface expression at early growth phase versus those of wild-type OG1RF. Confirmation of the observation was achieved through flow cytometry and complementation analysis. Compared to the wild type, the E. faecalis ccpA mutant had an impaired ability to adhere to collagen when grown to early exponential phase, consistent with the lack of Ace expression in the early growth phase. As a key component of carbon catabolite regulation, CcpA has been previously reported to play a critical role in regulating expression of proteins involved in E. faecalis carbohydrate uptake and utilization. Our discovery is the first to associate CcpA with the production of a major E. faecalis virulence factor, providing new insights into the regulation of E. faecalis pathogenesis.

  4. Production of lactic acid using a new homofermentative Enterococcus faecalis isolate.

    PubMed

    Subramanian, Mohan Raj; Talluri, Suvarna; Christopher, Lew P

    2015-03-01

    Lactic acid is an intermediate-volume specialty chemical for a wide range of food and industrial applications such as pharmaceuticals, cosmetics and chemical syntheses. Although lactic acid production has been well documented, improved production parameters that lead to reduced production costs are always of interest in industrial developments. In this study, we describe the production of lactic acid at high concentration, yield and volumetric productivity utilizing a novel homofermentative, facultative anaerobe Enterococcus faecalis CBRD01. The highest concentration of 182 g lactic acid l(-1) was achieved after 38 h of fed-batch fermentation on glucose. The bacterial isolate utilized only 2-13% of carbon for its growth and energy metabolism, while 87-98% of carbon was converted to lactic acid at an overall volumetric productivity of 5 g l(-1)  h(-1). At 13 h of fermentation, the volumetric productivity of lactate production reached 10.3 g l(-1)  h(-1), which is the highest ever reported for microbial production of lactic acid. The lactic acid produced was of high purity as formation of other metabolites was less than 0.1%. The present investigation demonstrates a new opportunity for enhanced production of lactic acid with potential for reduced purification costs. PMID:24894833

  5. Production of lactic acid using a new homofermentative Enterococcus faecalis isolate.

    PubMed

    Subramanian, Mohan Raj; Talluri, Suvarna; Christopher, Lew P

    2015-03-01

    Lactic acid is an intermediate-volume specialty chemical for a wide range of food and industrial applications such as pharmaceuticals, cosmetics and chemical syntheses. Although lactic acid production has been well documented, improved production parameters that lead to reduced production costs are always of interest in industrial developments. In this study, we describe the production of lactic acid at high concentration, yield and volumetric productivity utilizing a novel homofermentative, facultative anaerobe Enterococcus faecalis CBRD01. The highest concentration of 182 g lactic acid l(-1) was achieved after 38 h of fed-batch fermentation on glucose. The bacterial isolate utilized only 2-13% of carbon for its growth and energy metabolism, while 87-98% of carbon was converted to lactic acid at an overall volumetric productivity of 5 g l(-1)  h(-1). At 13 h of fermentation, the volumetric productivity of lactate production reached 10.3 g l(-1)  h(-1), which is the highest ever reported for microbial production of lactic acid. The lactic acid produced was of high purity as formation of other metabolites was less than 0.1%. The present investigation demonstrates a new opportunity for enhanced production of lactic acid with potential for reduced purification costs.

  6. Genetic modifications to temperate Enterococcus faecalis phage Ef11 that abolish the establishment of lysogeny and sensitivity to repressor, and increase host range and productivity of lytic infection.

    PubMed

    Zhang, H; Fouts, D E; DePew, J; Stevens, R H

    2013-06-01

    Ef11 is a temperate bacteriophage originally isolated by induction from a lysogenic Enterococcus faecalis strain recovered from an infected root canal, and the Ef11 prophage is widely disseminated among strains of E. faecalis. Because E. faecalis has emerged as a significant opportunistic human pathogen, we were interested in examining the genes and regulatory sequences predicted to be critical in the establishment/maintenance of lysogeny by Ef11 as a first step in the construction of the genome of a virulent, highly lytic phage that could be used in treating serious E. faecalis infections. Passage of Ef11 in E. faecalis JH2-2 yielded a variant that produced large, extensively spreading plaques in lawns of indicator cells, and elevated phage titres in broth cultures. Genetic analysis of the cloned virus producing the large plaques revealed that the variant was a recombinant between Ef11 and a defective FL1C-like prophage located in the E. faecalis JH2-2 chromosome. The recombinant possessed five ORFs of the defective FL1C-like prophage in place of six ORFs of the Ef11 genome. Deletion of the putative lysogeny gene module (ORFs 31-36) and replacement of the putative cro promoter from the recombinant phage genome with a nisin-inducible promoter resulted in no loss of virus infectivity. The genetic construct incorporating all the aforementioned Ef11 genomic modifications resulted in the generation of a variant that was incapable of lysogeny and insensitive to repressor, rendering it virulent and highly lytic, with a notably extended host range. PMID:23579685

  7. A propidium monoazide-quantitative PCR method for the detection and quantification of viable Enterococcus faecalis in large-volume samples of marine waters.

    PubMed

    Salam, Khaled W; El-Fadel, Mutasem; Barbour, Elie K; Saikaly, Pascal E

    2014-10-01

    The development of rapid detection assays of cell viability is essential for monitoring the microbiological quality of water systems. Coupling propidium monoazide with quantitative PCR (PMA-qPCR) has been successfully applied in different studies for the detection and quantification of viable cells in small-volume samples (0.25-1.00 mL), but it has not been evaluated sufficiently in marine environments or in large-volume samples. In this study, we successfully integrated blue light-emitting diodes for photoactivating PMA and membrane filtration into the PMA-qPCR assay for the rapid detection and quantification of viable Enterococcus faecalis cells in 10-mL samples of marine waters. The assay was optimized in phosphate-buffered saline and seawater, reducing the qPCR signal of heat-killed E. faecalis cells by 4 log10 and 3 log10 units, respectively. Results suggest that high total dissolved solid concentration (32 g/L) in seawater can reduce PMA activity. Optimal PMA-qPCR standard curves with a 6-log dynamic range and detection limit of 10(2) cells/mL were generated for quantifying viable E. faecalis cells in marine waters. The developed assay was compared with the standard membrane filter (MF) method by quantifying viable E. faecalis cells in seawater samples exposed to solar radiation. The results of the developed PMA-qPCR assay did not match that of the standard MF method. This difference in the results reflects the different physiological states of E. faecalis cells in seawater. In conclusion, the developed assay is a rapid (∼5 h) method for the quantification of viable E. faecalis cells in marine recreational waters, which should be further improved and tested in different seawater settings.

  8. Functional Analysis of the Citrate Activator CitO from Enterococcus faecalis Implicates a Divalent Metal in Ligand Binding

    PubMed Central

    Blancato, Víctor S.; Pagliai, Fernando A.; Magni, Christian; Gonzalez, Claudio F.; Lorca, Graciela L.

    2016-01-01

    The regulator of citrate metabolism, CitO, from Enterococcus faecalis belongs to the FCD family within the GntR superfamily. In the presence of citrate, CitO binds to cis-acting sequences located upstream of the cit promoters inducing the expression of genes involved in citrate utilization. The quantification of the molecular binding affinities, performed by isothermal titration calorimetry (ITC), indicated that CitO has a high affinity for citrate (KD = 1.2 ± 0.2 μM), while it did not recognize other metabolic intermediates. Based on a structural model of CitO where a putative small molecule and a metal binding site were identified, it was hypothesized that the metal ion is required for citrate binding. In agreement with this model, citrate binding to CitO sharply decreased when the protein was incubated with EDTA. This effect was reverted by the addition of Ni2+, and Zn2+ to a lesser extent. Structure-based site-directed mutagenesis was conducted and it was found that changes to alanine in residues Arg97 and His191 resulted in decreased binding affinities for citrate, as determined by EMSA and ITC. Further assays using lacZ fusions confirmed that these residues in CitO are involved in sensing citrate in vivo. These results indicate that the molecular modifications induced by a ligand and a metal binding in the C-terminal domain of CitO are required for optimal DNA binding activity, and consequently, transcriptional activation. PMID:26903980

  9. Comparative antimicrobial efficacy of herbal alternatives (Emblica officinalis, Psidium guajava), MTAD, and 2.5% sodium hypochlorite against Enterococcus faecalis: An in vitro study

    PubMed Central

    Dubey, Sandeep

    2016-01-01

    Aim The objective of this study was to evaluate the antimicrobial efficacy of herbal alternatives (Emblica officinalis, Psidium guajava), BioPure MTAD, and 2.5% sodium hypochlorite against Enterococcus faecalis. Materials and method The testing of the antimicrobial efficacy of selected medicaments against E. faecalis was done by the agar disk-diffusion method. Whatman paper discs of 6 mm diameter were prepared and soaked with the test solution. These discs were then placed onto the previously seeded agar Petri plates. Later, these plates were incubated for 48 h at 37 °C under the appropriate gaseous conditions in a CO2 incubator. A zone of inhibition was recorded in millimeter for each plate and the results were analyzed statistically. Result MTAD was found to be superior in its antibacterial abilities against E. faecalis compared with the other irrigants used. All the other tested irrigants showed significant zone of inhibition. Conclusions BioPure MTAD offers better antibacterial efficacy than NaOCl. E. officinalis and P. guajava are effective antibacterial agents against E. faecalis and can be used to reduce root canal microflora and root canal failures. PMID:26937369

  10. Expression of the agmatine deiminase pathway in Enterococcus faecalis is activated by the AguR regulator and repressed by CcpA and PTS(Man) systems.

    PubMed

    Suárez, Cristian; Espariz, Martín; Blancato, Víctor S; Magni, Christian

    2013-01-01

    Although the agmatine deiminase system (AgDI) has been investigated in Enterococcus faecalis, little information is available with respect to its gene regulation. In this study we demonstrate that the presence of exogenous agmatine induces the expression of agu genes in this bacterium. In contrast to the homologous and extensively characterized AgDI system of S. mutants, the aguBDAC operon in E. faecalis is not induced in response to low pH. In spite of this, agmatine catabolism in this bacterium contributes by neutralizing the external medium while enhancing bacterial growth. Our results indicate that carbon catabolic repression (CCR) operates on the AgDI system via a mechanism that involves interaction of CcpA and P-Ser-HPr with a cre site found in an unusual position considering the aguB promoter (55 nt upstream the +1 position). In addition, we found that components of the mannose phosphotransferase (PTS(Man)) system also contributed to CCR in E. faecalis since a complete relief of the PTS-sugars repressive effect was observed only in a PTS(Man) and CcpA double defective strain. Our gene context analysis revealed that aguR is present in oral and gastrointestinal microorganisms. Thus, regulation of the aguBDAC operon in E. faecalis seems to have evolved to obtain energy and resist low pH conditions in order to persist and colonize gastrointestinal niches.

  11. Use of Enterococcus faecalis and Bacillus atrophaeus as surrogates to establish and maintain laboratory proficiency for concentration of water samples using ultrafiltration.

    PubMed

    Mapp, Latisha; Klonicki, Patricia; Takundwa, Prisca; Hill, Vincent R; Schneeberger, Chandra; Knee, Jackie; Raynor, Malik; Hwang, Nina; Chambers, Yildiz; Miller, Kenneth; Pope, Misty

    2015-11-01

    The U.S. Environmental Protection Agency's (EPA) Water Laboratory Alliance (WLA) currently uses ultrafiltration (UF) for concentration of biosafety level 3 (BSL-3) agents from large volumes (up to 100-L) of drinking water prior to analysis. Most UF procedures require comprehensive training and practice to achieve and maintain proficiency. As a result, there was a critical need to develop quality control (QC) criteria. Because select agents are difficult to work with and pose a significant safety hazard, QC criteria were developed using surrogates, including Enterococcus faecalis and Bacillus atrophaeus. This article presents the results from the QC criteria development study and results from a subsequent demonstration exercise in which E. faecalis was used to evaluate proficiency using UF to concentrate large volume drinking water samples. Based on preliminary testing EPA Method 1600 and Standard Methods 9218, for E. faecalis and B. atrophaeus respectively, were selected for use during the QC criteria development study. The QC criteria established for Method 1600 were used to assess laboratory performance during the demonstration exercise. Based on the results of the QC criteria study E. faecalis and B. atrophaeus can be used effectively to demonstrate and maintain proficiency using ultrafiltration.

  12. Biofilm forming capacity of Enterococcus faecalis on Gutta-percha points treated with four disinfectants using confocal scanning laser microscope: An in vitro study

    PubMed Central

    Ravi Chandra, Polavarapu Venkata; Kumar, Vemisetty Hari; Reddy, Surakanti Jayaprada; Kiran, Dandolu Ram; Krishna, Muppala Nagendra; Kumar, Golla Vinay

    2015-01-01

    Background: The aim of this study was to evaluate and compare the in vitro biofilm forming capacity of Enterococcus faecalis on Gutta-percha points disinfected with four disinfectants. Materials and Methods: A total of 50 Gutta-percha points used in this study were divided into four test groups based on disinfectant (5.25% sodium hypochlorite, 2% chlorhexidine gluconate, 20% neem, 13% benzalkonium chloride [BAK]), and one control group. The Gutta-percha points were initially treated with corresponding disinfectants followed by anaerobic incubation in Brain Heart Infusion broth suspended with human serum and E. faecalis strain for 14 days. After incubation, these Gutta-percha points were stained with Acridine Orange (Sigma – Aldrich Co., St. Louis, MO, USA) and 0.5 mm thick cross section samples were prepared. The biofilm thickness of E. faecalis was analyzed quantitatively using a confocal scanning laser microscope. Results statistically analyzed using analysis of variance. P < 0.05 was considered to be significant. Results: Confocal scanning laser microscope showed reduced amount of E. faecalis biofilm on Gutta-percha points treated with BAK and sodium hypochlorite. Post-hoc (least square differences) test revealed that there is no statistically significant difference between BAK and sodium hypochlorite groups (P > 0.05). Conclusion: This study illustrates that the Gutta-percha points disinfected with sodium hypochlorite and BAK showed minimal biofilm growth on its surface. PMID:26288622

  13. Multilocus sequence typing scheme for Enterococcus faecalis reveals hospital-adapted genetic complexes in a background of high rates of recombination.

    PubMed

    Ruiz-Garbajosa, Patricia; Bonten, Marc J M; Robinson, D Ashley; Top, Janetta; Nallapareddy, Sreedhar R; Torres, Carmen; Coque, Teresa M; Cantón, Rafael; Baquero, Fernando; Murray, Barbara E; del Campo, Rosa; Willems, Rob J L

    2006-06-01

    A multilocus sequence typing (MLST) scheme based on seven housekeeping genes was used to investigate the epidemiology and population structure of Enterococcus faecalis. MLST of 110 isolates from different sources and geographic locations revealed 55 different sequence types that grouped into four major clonal complexes (CC2, CC9, CC10, and CC21) by use of eBURST. Two of these clonal complexes, CC2 and CC9, are particularly fit in the hospital environment, as CC2 includes the previously described BVE clonal complex identified by an alternative MLST scheme and CC9 includes exclusively isolates from hospitalized patients. Identical alleles were found in genetically diverse isolates with no linkage disequilibrium, while the different MLST loci gave incongruent phylogenetic trees. This demonstrates that recombination is an important mechanism driving genetic variation in E. faecalis and suggests an epidemic population structure for E. faecalis. Our novel MLST scheme provides an excellent tool for investigating local and short-term epidemiology as well as global epidemiology, population structure, and genetic evolution of E. faecalis.

  14. Expression of the Agmatine Deiminase Pathway in Enterococcus faecalis Is Activated by the AguR Regulator and Repressed by CcpA and PTSMan Systems

    PubMed Central

    Blancato, Víctor S.; Magni, Christian

    2013-01-01

    Although the agmatine deiminase system (AgDI) has been investigated in Enterococcus faecalis, little information is available with respect to its gene regulation. In this study we demonstrate that the presence of exogenous agmatine induces the expression of agu genes in this bacterium. In contrast to the homologous and extensively characterized AgDI system of S. mutants, the aguBDAC operon in E. faecalis is not induced in response to low pH. In spite of this, agmatine catabolism in this bacterium contributes by neutralizing the external medium while enhancing bacterial growth. Our results indicate that carbon catabolic repression (CCR) operates on the AgDI system via a mechanism that involves interaction of CcpA and P-Ser-HPr with a cre site found in an unusual position considering the aguB promoter (55 nt upstream the +1 position). In addition, we found that components of the mannose phosphotransferase (PTSMan) system also contributed to CCR in E. faecalis since a complete relief of the PTS-sugars repressive effect was observed only in a PTSMan and CcpA double defective strain. Our gene context analysis revealed that aguR is present in oral and gastrointestinal microorganisms. Thus, regulation of the aguBDAC operon in E. faecalis seems to have evolved to obtain energy and resist low pH conditions in order to persist and colonize gastrointestinal niches. PMID:24155893

  15. Killing of Enterococcus faecalis by MTAD and chlorhexidine digluconate with or without cetrimide in the presence or absence of dentine powder or BSA.

    PubMed

    Portenier, Isabelle; Waltimo, Tuomas; Ørstavik, Dag; Haapasalo, Markus

    2006-02-01

    The antibacterial efficacy of irrigating solutions and local disinfectants used in endodontics appears poorer in vivo than in vitro. One explanation may be inactivation by compounds present in the root canal. MTAD (a mixture of tetracycline isomer, acid, and detergent) is a new root canal irrigation solution with antibacterial activity. The aim of this study was to investigate the antibacterial activity of MTAD and chlorhexidine towards two strains of Enterococcus faecalis and the inhibitory effects of dentine and bovine serum albumin on the antibacterial activity. Survival of bacteria exposed to the medicaments in the presence or absence of inhibitors was monitored in an in vitro model. Full concentration (100%) MTAD and 0.2% chlorhexidine rapidly killed both strains. Combining chlorhexidine with cetrimide further reduced the time required for killing. The presence of dentine or BSA caused a marked delay in killing by both medicaments. The two E. faecalis strains tested showed minor differences in their susceptibility to the disinfectants.

  16. Detection and characterization of a ST6 clone of vanB2-Enterococcus faecalis from three different hospitals in Spain.

    PubMed

    López, M; Rezusta, A; Seral, C; Aspiroz, C; Marne, C; Aldea, M J; Ferrer, I; Revillo, M J; Castillo, F J; Torres, C

    2012-03-01

    Thirteen vancomycin-resistant and teicoplanin-susceptible Enterococcus faecalis isolates were recovered from unrelated patients in three Spanish hospitals from November 2009 to December 2010. All isolates carried the vanB2 gene, showed indistinguishable or closely-related PFGE patterns and were ascribed to the sequence type ST6 (included into the high-risk clonal-complex CC2). They showed a multiresistance phenotype (erythromycin, tetracycline, ciprofloxacin and high-level-resistance to streptomycin, gentamicin and kanamycin) and harboured the aac(6')-aph(2"), ant(6)-Ia, and tet(M)+/-tet(L) genes. All isolates produced gelatinase and harboured the gelE gene, but not the esp or hyl genes. The inclusion of the vanB2 gene into the Tn5382 transposon was demonstrated in one isolate. Clonal dissemination of vanB2-containing the E. faecalis strain is demonstrated.

  17. [Submucosal bacterial abscesses of the ascending colon and liver associated with portal and superior mesenteric vein thrombosis due to Enterococcus faecalis infection: a case report].

    PubMed

    Norimura, Daisuke; Takeshima, Fuminao; Satou, Yoshiaki; Nakagoe, Tohru; Ohnita, Ken; Isomoto, Hajime; Nakao, Kazuhiko

    2014-06-01

    A 72-year-old woman with diabetes mellitus was admitted with fever and general fatigue. Blood biochemistry showed elevated hepatic and biliary enzyme levels, abdominal computed tomography showed multiple liver abscesses with portal and superior mesenteric vein thrombosis, and total colonoscopy revealed a submucosal bacterial abscess in the ascending colon. The abscesses were determined to be associated with Enterococcus faecalis infection. The patient was treated conservatively with antibiotics (meropenem) and anticoagulants (warfarin), which led to a gradual amelioration of symptoms and resolution of thrombosis.

  18. Inactivation of a 25.5 µm Enterococcus faecalis biofilm by a room-temperature, battery-operated, handheld air plasma jet

    NASA Astrophysics Data System (ADS)

    Pei, X.; Lu, X.; Liu, J.; Liu, D.; Yang, Y.; Ostrikov, K.; Chu, Paul K.; Pan, Y.

    2012-04-01

    Effective biofilm inactivation using a handheld, mobile plasma jet powered by a 12 V dc battery and operated in open air without any external gas supply is reported. This cold, room-temperature plasma is produced in self-repetitive nanosecond discharges with current pulses of ˜100 ns duration, current peak amplitude of ˜6 mA and repetition rate of ˜20 kHz. It is shown that the reactive plasma species penetrate to the bottom layer of a 25.5 µm-thick Enterococcus faecalis biofilm and produce a strong bactericidal effect. This is the thickest reported biofilm inactivated using room-temperature air plasmas.

  19. Identification of Enterococcus faecalis bacteria resistant to heavy metals and antibiotics in surface waters of the Mololoa River in Tepic, Nayarit, Mexico.

    PubMed

    Mondragón, Verónica Alejandra; Llamas-Pérez, Dámaris F; González-Guzmán, Gladis E; Márquez-González, Antonio R; Padilla-Noriega, Roberto; Durán-Avelar, Ma de Jesús; Franco, Bernardo

    2011-12-01

    Heavy metal and antibiotic resistance have been shown to have a strong correlation in nature, and their inter-relation is an important subject of study. We report an analysis of surface waters of the Mololoa River in the municipality of Tepic, state of Nayarit, Mexico. This river has two distinctive sources of contamination: sewage waters and trash confinements. Our findings demonstrate a correlation between the river flow pattern and resistance to heavy metals or to heavy metals and antibiotics in isolated bacteria of the genus Enterococcus, specifically Enterococcus faecalis. The Mololoa River provides a model to study the relationship between water flow and generation of biodiversity, and more importantly, it constitutes a model for studying genetic diversity of bacteria affecting human health.

  20. Evaluation of the Enterococcus faecalis Biofilm-Associated Virulence Factors AhrC and Eep in Rat Foreign Body Osteomyelitis and In Vitro Biofilm-Associated Antimicrobial Resistance

    PubMed Central

    Frank, Kristi L.; Vergidis, Paschalis; Brinkman, Cassandra L.; Greenwood Quaintance, Kerryl E.; Barnes, Aaron M. T.; Mandrekar, Jayawant N.; Schlievert, Patrick M.; Dunny, Gary M.; Patel, Robin

    2015-01-01

    Enterococcus faecalis can cause healthcare-associated biofilm infections, including those of orthopedic devices. Treatment of enterococcal prosthetic joint infection is difficult, in part, due to biofilm-associated antimicrobial resistance. We previously showed that the E. faecalis OG1RF genes ahrC and eep are in vitro biofilm determinants and virulence factors in animal models of endocarditis and catheter-associated urinary tract infection. In this study, we evaluated the role of these genes in a rat acute foreign body osteomyelitis model and in in vitro biofilm-associated antimicrobial resistance. Osteomyelitis was established for one week following the implantation of stainless steel orthopedic wires inoculated with E. faecalis strains OG1RF, ΩahrC, and ∆eep into the proximal tibiae of rats. The median bacterial loads recovered from bones and wires did not differ significantly between the strains at multiple inoculum concentrations. We hypothesize that factors present at the infection site that affect biofilm formation, such as the presence or absence of shear force, may account for the differences in attenuation in the various animal models we have used to study the ΩahrC and ∆eep strains. No differences among the three strains were observed in the planktonic and biofilm antimicrobial susceptibilities to ampicillin, vancomycin, daptomycin, linezolid, and tetracycline. These findings suggest that neither ahrC nor eep directly contribute to E. faecalis biofilm-associated antimicrobial resistance. Notably, the experimental evidence that the biofilm attachment mutant ΩahrC displays biofilm-associated antimicrobial resistance suggests that surface colonization alone is sufficient for E. faecalis cells to acquire the biofilm antimicrobial resistance phenotype. PMID:26076451

  1. Antimicrobial activity of enterocins from Enterococcus faecalis SL-5 against Propionibacterium acnes, the causative agent in acne vulgaris, and its therapeutic effect.

    PubMed

    Kang, Bong Seon; Seo, Jae-Gu; Lee, Gwa-Su; Kim, Jung-Hwa; Kim, Sei Yeon; Han, Ye Won; Kang, Hoon; Kim, Hyung Ok; Rhee, Ji Hwan; Chung, Myung-Jun; Park, Young Min

    2009-02-01

    A lactic acid bacterial strain was isolated from human fecal specimen and identified as Enterococcus faecalis SL-5. The isolated strain showed antimicrobial activity against Gram-positive pathogens assayed, especially the highest activity against Propionibacterium acnes. The antimicrobial substance was purified and verified as a bacteriocin (named ESL5) of E. faecalis SL-5 by activity-staining using P. acnes as an indicator. N-terminal sequence of ESL5 was determined (MGAIAKLVAK) and sequence analysis revealed that it is almost identical to the some of enterocins including L50A/B of E. faecium L50 and MR10A/B of E. faecalis MRR 10-3. From the sequencing data of L50A/B structural genes, the nucleotide sequence showed 100% identity with that of the MR10A/B structural genes, implying that ESL5 is an equivalent of enterocin MR10. Meanwhile, we also tested the therapeutic effect of anti-P. acnes activity in patients with mild to moderate acne because of its pathogenic role to acne vulgaris. For this purpose, a concentrated powder of CBT SL-5 was prepared using cell-free culture supernatant (CFCS) of E. faecalis SL-5 and included in a lotion for application in the patients. The study showed that CBT SL-5 lotion significantly reduced the inflammatory lesions like pustules compared to the placebo lotion. Therefore our results indicate that the anti-P. acnes activity produced by E. faecalis SL-5 has potential role to the treatment of acne as an alternative to topical antibiotics.

  2. Evaluation of the Enterococcus faecalis Biofilm-Associated Virulence Factors AhrC and Eep in Rat Foreign Body Osteomyelitis and In Vitro Biofilm-Associated Antimicrobial Resistance.

    PubMed

    Frank, Kristi L; Vergidis, Paschalis; Brinkman, Cassandra L; Greenwood Quaintance, Kerryl E; Barnes, Aaron M T; Mandrekar, Jayawant N; Schlievert, Patrick M; Dunny, Gary M; Patel, Robin

    2015-01-01

    Enterococcus faecalis can cause healthcare-associated biofilm infections, including those of orthopedic devices. Treatment of enterococcal prosthetic joint infection is difficult, in part, due to biofilm-associated antimicrobial resistance. We previously showed that the E. faecalis OG1RF genes ahrC and eep are in vitro biofilm determinants and virulence factors in animal models of endocarditis and catheter-associated urinary tract infection. In this study, we evaluated the role of these genes in a rat acute foreign body osteomyelitis model and in in vitro biofilm-associated antimicrobial resistance. Osteomyelitis was established for one week following the implantation of stainless steel orthopedic wires inoculated with E. faecalis strains OG1RF, ΩahrC, and ∆eep into the proximal tibiae of rats. The median bacterial loads recovered from bones and wires did not differ significantly between the strains at multiple inoculum concentrations. We hypothesize that factors present at the infection site that affect biofilm formation, such as the presence or absence of shear force, may account for the differences in attenuation in the various animal models we have used to study the ΩahrC and ∆eep strains. No differences among the three strains were observed in the planktonic and biofilm antimicrobial susceptibilities to ampicillin, vancomycin, daptomycin, linezolid, and tetracycline. These findings suggest that neither ahrC nor eep directly contribute to E. faecalis biofilm-associated antimicrobial resistance. Notably, the experimental evidence that the biofilm attachment mutant ΩahrC displays biofilm-associated antimicrobial resistance suggests that surface colonization alone is sufficient for E. faecalis cells to acquire the biofilm antimicrobial resistance phenotype.

  3. Identification of Enterococcus faecium and Enterococcus faecalis as vanC-type Vancomycin-Resistant Enterococci (VRE) from sewage and river water in the provincial city of Miyazaki, Japan.

    PubMed

    Nishiyama, Masateru; Iguchi, Atsushi; Suzuki, Yoshihiro

    2015-01-01

    As a first step for assessing the risk to human health posed by vancomycin-resistant enterococci (VRE) in the aquatic environment, we screened sewage and urban river water samples from Miyazaki, Japan for VRE. Because vancomycin-resistant organisms are not as prevalent in sewage and river water as vancomycin-susceptible organisms, the samples were screened by minimum inhibitory concentration test using the vancomycin-supplemented membrane-Enterococcus indoxyl-β-d-glucoside (mEI) agar. The isolates, presumed to be enterococci, were identified using 16S rRNA sequencing analysis. The percentages of VRE isolates screened using 4 μg mL(-1) vancomycin-supplemented mEI agar from sewage and urban river water samples were 12% and 24%, respectively. The vancomycin-resistant genes vanC1 and vanC2/3 were detected in the isolates from both samples by PCR analysis. All enterococci isolates containing vanC1, which is a specific gene for vanC-type of VRE, were identified as Enterococcus casseliflavus/gallinarum. Further, 92% enterococci isolates containing vanC2/3 were identified as E. casseliflavus/gallinarum, the remaining isolates containing vanC2/3 were E. faecium (4%) and E. faecalis (4%). Thereafter, the distribution of E. faecium and E. faecalis, which are the major types of enterococci in humans containing vanC2/3, was observed in the water samples collected.

  4. An in Vitro Evaluation of Antimicrobial Efficacy of 2% Chlorhexidine Gel, Propolis and Calcium Hydroxide Against Enterococcus faecalis in Human Root Dentin

    PubMed Central

    T S, Ashwini; Patil, Chetan R

    2014-01-01

    Aim: To evaluate in vitro the antimicrobial efficacy of 2% Chlorhexidine gel, Propolis and Calcium hydroxide against Enterococcus faecalis in human root dentin. Methodology: One hundred and twenty human extracted anterior teeth were decoronated below CEJ and the apical part of root was removed to obtain 6mm of middle of the root. GG no 3 was used to standardize the internal diameter of root canal. Dentin blocks were infected with E faecalis for 21 d. They were assigned into four groups (n = 30).Group 1, Saline (negative control); Group 2, Propolis; Group 3, 2% CHX; Group 4, Calcium hydroxide, At the end of 1, 3, and 5 days an assessment of microbial cells was carried out at a depth of 400 μm and colony counts were calculated.The data were analysed statistically with one-way analysis of variance followed by Scheffe multiple comparison test (p < 0.05). Results: The number of colony-forming units was significantly lower in all experimental groups compared to the control group – Saline. 2% Chlorhexidinegluconate produced better antimicrobial efficacy (100%) on day 1, 3 and 5. Propolis (66.37%) had greater antimicrobial activity than Calcium hydroxide (50.89%) on day 1 but there was no significant difference in their antimicrobial activities on day 3 and day 5. Conclusion: 2% Chlorhexidine gel showed the maximum antimicrobial activity against E faecalis and Calcium hydroxide the least. Propolis can be used as an effective alternative intracanal medicament. PMID:25584319

  5. Enterococcus faecalis endophthalmitis as a metastatic complication of hemodialysis vascular access-related sepsis: A case report and review of the literature.

    PubMed

    Sahin, Osman Zikrullah; Kara, Ekrem; Belice, Tahir; Ayaz, Teslime; Baydur Sahin, Serap; Ozturk, Cinar; Yildirim, Safak; Metin, Yavuz; Sahutoglu, Tuncay

    2016-07-01

    Catheter and/or arteriovenous (A-V) graft-related bacteremia is an important cause of morbidity and mortality among hemodialysis (HD) patients. Endocarditis, septic arthritis, epidural abscess, septic embolism, and osteomyelitis are the most common complications of catheter and/or A-V graft-related bacteremia; however, endogenous endophthalmitis is rarely seen. To the best of our knowledge, Enterococcus faecalis is the first case report in this population. We hereby report a case of endogenous endophthalmitis caused by E. faecalis as a complication of catheter and/or A-V graft-related bacteremia in a diabetic patient, who was undergoing HD for 5 years. We also discuss the etiology, clinical features, and outcomes of endogenous endophthalmitis in HD patients with a brief review of the literature. Although broad-spectrum parenteral (intravenous and intravitreal) antibiotics were used for 4 weeks, evisceration of the left eye could not be avoided. Endogenous endophthalmitis is a rare but rapidly blinding complication of catheter and/or A-V graft-related bacteremia in HD patients. It can develop as a result of silent catheter and/or A-V graft infections, which may lead to recurrent bacteremia. E. faecalis should be considered as a pathogen in this population who had recent history of catheter or A-V graft procedure. PMID:26346615

  6. Effects of pH, temperature and NaCl concentration on the growth kinetics, proteolytic activity and biogenic amine production of Enterococcus faecalis.

    PubMed

    Gardin, F; Martuscelli, M; Caruso, M C; Galgano, F; Crudele, M A; Favati, F; Guerzoni, M E; Suzzi, G

    2001-02-28

    In this work, the combined effects of temperature, pH and NaCl concentration on the growth dynamics of Enterococcus faecalis EF37, its proteolytic activity and its production of biogenic amines have been studied. The effects of the selected variables have been analysed using a Central Composite Design. The production of biogenic amines, under the adopted conditions, was found to be mainly dependent on the extent of growth of E. faecalis. Its proteolytic activity was not a limiting factor for the final amine production, because in the system studied (skim milk) an excess of precursors was guaranteed. Quantitatively, the most important biogenic amine produced was 2-phenylethylamine but substantial amounts of tyramine were detected in all the samples. This work confirms that the main biological feature influencing the biogenic amine formation is the extent of growth of microorganisms, like E. faecalis, characterised by decarboxylase activity. In the traditional and artisanal cheeses produced using raw milk, enterococci usually reach levels of 10(7) cells/g. With this perspective, it is important that the presence of biogenic amines due to the activities of these microorganisms is maintained within safe levels, without affecting the positive effects of enterococci on the final organoleptic characteristics of the cheese.

  7. CRISPRs of Enterococcus faecalis and E. hirae isolates from pig feces have species-specific repeats but share some common spacer sequences.

    PubMed

    Katyal, Isha; Chaban, Bonnie; Ng, Beata; Hill, Janet E

    2013-07-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) are currently a topic of interest in microbiology due to their role as a prokaryotic immune system. Investigations of CRISPR distribution and characterization to date have focused on pathogenic bacteria, while less is known about CRISPR in commensal bacteria, where they may have a significant role in the ecology of the microbiota of humans and other animals, and act as a recorder of interactions between bacteria and viruses. A combination of PCR and sequencing was used to determine prevalence and distribution of CRISPR arrays in Enterococcus faecalis and Enterococcus hirae isolates from the feces of healthy pigs. Both type II CRISPR-Cas and Orphan CRISPR (without Cas genes) were detected in the 195 isolates examined. CRISPR-Cas was detected in 52 (46/88) and 42 % (45/107) E. faecalis and E. hirae isolates, respectively. The prevalence of Orphan CRISPR arrays was higher in E. faecalis isolates (95 %, 84/88) compared with E. hirae isolates (49 %, 53/107). Species-specific repeat sequences were identified in Orphan CRISPR arrays, and 42 unique spacer sequences were identified. Only two spacers matched previously characterized pig virome sequences, and many were apparently derived from chromosomal sequences of enterococci. Surprisingly, 17 (40 %) of the spacers were detected in both species. Shared spacer sequences are evidence of a lack of species specificity in the agents and mechanisms responsible for integration of spacers, and the abundance of spacer sequences corresponding to bacterial chromosomal sequences reflects interspecific interactions within the intestinal microbiota.

  8. Influence of various herbal irrigants as a final rinse on the adherence of Enterococcus faecalis by fluorescence confocal laser scanning microscope

    PubMed Central

    Rosaline, Hannah; Kandaswamy, D; Gogulnath, D; Rubin, MI

    2013-01-01

    Aim: The aim of this study was to assess the antibacterial efficacy of three different herbal irrigants against Enterococcus faecalis. Materials and Methods: Single rooted teeth were extracted due to orthodontic and periodontal reasons. The teeth were then inoculated with E. faecalis. The teeth were randomly divided into three experimental groups and two control groups of six samples each. Group 1 specimens were treated with 5.2% sodium hypochlorite (NaOCL) for 30 min followed by 5 mmol/L Ethylenediaminetetraacetic acid (EDTA) for 5 min and saline as final irrigant. Group 2 specimens were treated with and 5.2% NaOCl for 30 min as final irrigant. Group 3 were treated with Morinda citrifolia (MC) for 30 min as final irrigant. Group 4 were treated with Azadiracta indica (AI) as final irrigant. Group 5 were treated with green tea (GT) for 30 min as final irrigant. The dentin specimens were carefully spread onto a microscope slide and stained with BacLight and examined in a confocal laser scanning microscope set to monitor fluorescein isothiocyanate and propidium iodide. A total of nine fields were examined for each treatment and the bacteria presented were counted. Statistical Analysis: Using the one-way ANOVA with multiple comparison, significantly less bacteria were found adhering to the samples treated with Neem followed by NaOCL, GT, MC, Saline. Results: AI treatment produced the maximum reduction in adherence of E. faecalis to dentin (9.30%) followed by NaOCl (12.50%), GT (27.30%), MC (44.20%) and saline (86.70%). Conclusion: Neem is effective in preventing adhesion of E. faecalis to dentin. PMID:23956540

  9. Effectiveness of a new method of disinfecting the root canal, using Er, Cr:YSGG laser to kill Enterococcus faecalis in an infected tooth model.

    PubMed

    Licata, M E; Albanese, A; Campisi, G; Geraci, D M; Russo, R; Gallina, G

    2015-02-01

    Some lasers have demonstrated to provide effective disinfection when used as adjunctive device to the conventional treatment. The aim of this in vitro study was to determine the effectiveness of the erbium, chromium:yttrium scandium gallium garnet (Er, Cr:YSGG) laser by measuring its bactericidal effect inside the root canal experimentally colonized with Enterococcus faecalis. The laser was tested at different irradiation times (30 and 60 s) and energy of impulses (75 and 25 mJ). A total of 52 single-rooted extracted human teeth were endodontically prepared with rotary instrumentation. All were sterilized and inoculated with a suspension of E. faecalis (105 bacteria/ml). The teeth were randomized into three treatment (group 1, group 2, and group 3) and one control groups. In all groups, teeth were chemically irrigated with 5.25% sodium hypochlorite and 17% ethylenediaminetetraacetic acid. Groups 1 and 2 were also irradiated at 30 and 60 s, respectively, with an Er, Cr:YSGG laser at 75 mJ. Teeth of group 3 were treated with laser for 60 s at 25 mJ. Samples were processed to detect the presence of E. faecalis. For all groups, a bactericidal effect was observed. The use of laser at 75 mJ with an irradiation time of 30 and 60 s eliminated a percentage of 92.3 and 100% of E. faecalis, respectively. In the control group, a reduction of 92.3% was observed. Lower percentage of reduction (46.1%) was obtained in teeth treated with laser at 25 mJ for 60 s. No statistical differences were observed between the groups (P = 0.543, Fisher's exact test). The results indicated a bactericidal effect of Er, Cr:YSGG laser irradiation at the settings used in this study. The highest bactericidal effect of this laser was observed at 60 s of irradiation time, using an energy pulse of 75 mJ.

  10. First isolation of oleate-dependent Enterococcus faecalis small-colony variants from the umbilical exudate of a paediatric patient with omphalitis.

    PubMed

    Kubota, Noriko; Kuzumoto, Kei; Hidaka, Eiko; Yoshizawa, Katsumi; Yumoto, Kayoko; Oana, Kozue; Ogiso, Yoshifumi; Nakamura, Tomohiko; Kawakami, Yoshiyuki

    2013-12-01

    An oleate-dependent Enterococcus faecalis isolate representing small-colony variants (SCVs) was isolated from the umbilical exudate of a 31-month-old Japanese male patient in Nagano Children's Hospital, Azumino, Japan. The patient had been suffering from recurrent omphalitis since early infancy. The initial E. faecalis SCV isolate formed small colonies on sheep blood agar plates and tiny colonies on chocolate and modified Drigalski agar, although no visible growth was observed in HK-semi solid medium after 48 h incubation in ambient air. Moreover, the SCV isolate, the colonial morphology of which was reminiscent of Streptococcus species, could not be identified using the MicroScan WalkAway-40 and API 20 Strep systems, both of which yielded profile numbers that did not correspond to any bacterial species, probably as a result of insufficient growth of the isolate. The SCV isolate was subsequently identified as E. faecalis based on its morphological, cultural and biochemical properties, and this was confirmed by sequencing the 16S rRNA gene of the organism. Investigations revealed that the addition of oleate, an unsaturated fatty acid, enabled the isolate to grow on every medium with normal-sized colony morphology. Although it has long been known that long-chain fatty acids, especially unsaturated oleic acid, have a major inhibitory effect on the growth of a variety of microorganisms, including not only mycobacteria but also streptococci, this is, to the best of our knowledge, the first clinical isolation of an oleate-dependent E. faecalis SCV isolate. In addition, oleic acid might be considered to affect the cell membrane permeability of carbohydrates or antimicrobial agents such as β-lactams.

  11. Activity of daptomycin or linezolid in combination with rifampin or gentamicin against biofilm-forming Enterococcus faecalis or E. faecium in an in vitro pharmacodynamic model using simulated endocardial vegetations and an in vivo survival assay using Galleria mellonella larvae.

    PubMed

    Luther, Megan K; Arvanitis, Marios; Mylonakis, Eleftherios; LaPlante, Kerry L

    2014-08-01

    Enterococci are the third most frequent cause of infective endocarditis. A high-inoculum stationary-phase in vitro pharmacodynamic model with simulated endocardial vegetations was used to simulate the human pharmacokinetics of daptomycin at 6 or 10 mg/kg of body weight/day or linezolid at 600 mg every 12 h (q12h), alone or in combination with gentamicin at 1.3 mg/kg q12h or rifampin at 300 mg q8h or 900 mg q24h. Biofilm-forming, vancomycin-susceptible Enterococcus faecalis and vancomycin-resistant Enterococcus faecium (vancomycin-resistant enterococcus [VRE]) strains were tested. At 24, 48, and 72 h, all daptomycin-containing regimens demonstrated significantly more activity (decline in CFU/g) than any linezolid-containing regimen against biofilm-forming E. faecalis. The addition of gentamicin to daptomycin (at 6 or 10 mg/kg) in the first 24 h significantly improved bactericidal activity. In contrast, the addition of rifampin delayed the bactericidal activity of daptomycin against E. faecalis, and the addition of rifampin antagonized the activities of all regimens against VRE at 24 h. Also, against VRE, the addition of gentamicin to linezolid at 72 h improved activity and was bactericidal. Rifampin significantly antagonized the activity of linezolid against VRE at 72 h. In in vivo Galleria mellonella survival assays, linezolid and daptomycin improved survival. Daptomycin at 10 mg/kg improved survival significantly over that with linezolid against E. faecalis. The addition of gentamicin improved the efficacy of daptomycin against E. faecalis and those of linezolid and daptomycin against VRE. We conclude that in enterococcal infection models, daptomycin has more activity than linezolid alone. Against biofilm-forming E. faecalis, the addition of gentamicin in the first 24 h causes the most rapid decline in CFU/g. Of interest, the addition of rifampin decreased the activity of daptomycin against both E. faecalis and VRE.

  12. Prevalence and risk factors of early fecal carriage of Enterococcus faecalis and Staphylococcus spp and their antimicrobial resistant patterns among healthy neonates born in a hospital setting in central Saudi Arabia

    PubMed Central

    El-Kersh, Talat A.; Marie, Mohammed A.; Al-Sheikh, Yazeed A.; Al-Agamy, Mohamed H.; Al-Bloushy, Ahmad A.

    2016-01-01

    Objectives: To investigate the prevalence, antibiotic resistant profiles, and risk factors of early fecal carriage of Enterococcus faecalis (E. faecalis) and staphylococci among 150 healthy Saudi neonates born in a hospital setting in central Saudi Arabia. Methods: This prospective study was conducted in Al-Bukayriyah General Hospital, Qassim, Saudi Arabia, between June 2012 and January 2013. The E. faecalis and Staphylococcus spp. isolates were identified manually, and Vitek2 system was used for identity confirmation at the species level and minimum inhibitory concentration-susceptibility testing. Results: Enterococcus faecalis (n=73) and Staphylococcus spp. (n=18) were recovered. Unlike staphylococci, E. faecalis colonization did not significantly vary from day one up to 7 days of life, regardless of the type of feeding, but it was relatively higher among vaginally versus cesarean delivery. Both Staphylococcus epidermidis (S. epidermidis) and Staphylococcus aureus carriage increase as the body weight increases, and this difference was significant (p=0.025) for S. epidermidis. High-level resistance in Gentamycin among E. faecalis isolates was 25% and 11% to Streptomycin. Thirty percent of S. epidermidis were resistant to oxacillin and exhibited multidrug-resistant (MDR) patterns of 5 resistant markers, which were also observed among 2/5 (40%) of Methicillin-resistant Staphylococcus aureus isolates. Conclusion: Enterococcus faecalis did not significantly vary in relation to type of delivery, age up to 7 days, and type of feeding. The neonatal fecal carriage of MDR isolates should be considered as a crucial reservoir to the further spread of antimicrobial resistance genes among hospitals, cross infections, and the community. PMID:26905350

  13. Crystallization and preliminary crystallographic analysis of an Enterococcus faecalis repressor protein, CylR2, involved in regulating cytolysin production through quorum-sensing

    SciTech Connect

    Ni, Shuisong; McAteer, Kathleen; Bussiere, Dirksen E.; Kennedy, Michael A.

    2004-06-01

    CylR2 is one of the two regulatory proteins associated with the quorum-sensing-dependent synthesis of cytolysin for the common pathogen Enterococcus faecalis. The protein was expressed with a C-terminal 6-histidine tag and purified to homogeneity with a cobalt affinity column followed by another size exclusion column. Both native and SeMet proteins were crystallized. A complete X-ray diffraction data set from the native crystal was collected to 2.3 resolution. The crystal was tetragonal, belonging to space group P41/43, with unit-cell dimensions a=b=66.2 , c=40.9 and a=b=g=90. The asymmetric unit contained two molecules of CylR2.

  14. Structure of Peptide Sex Pheromone Receptor PrgX and PrgX/Pheromone Complexes and Regulation of Conjugation in Enterococcus faecalis

    SciTech Connect

    Shi,K.; Brown, C.; Gu, Z.; Kozlowicz, B.; Dunny, G.; Ohlendorf, D.; Earhart, C.

    2005-01-01

    Many bacterial activities, including expression of virulence factors, horizontal genetic transfer, and production of antibiotics, are controlled by intercellular signaling using small molecules. To date, understanding of the molecular mechanisms of peptide-mediated cell-cell signaling has been limited by a dearth of published information about the molecular structures of the signaling components. Here, we present the molecular structure of PrgX, a DNA- and peptide-binding protein that regulates expression of the conjugative transfer genes of the Enterococcus faecalis plasmid pCF10 in response to an intercellular peptide pheromone signal. Comparison of the structures of PrgX and the PrgX/pheromone complex suggests that pheromone binding destabilizes PrgX tetramers, opening a 70-bp pCF10 DNA loop required for conjugation repression.

  15. Relative expression of genes involved in the resistance/sensitivity of Enterococcus faecalis JH2-2 to recombinant divercin RV41.

    PubMed

    Calvez, Ségolène; Prevost, H; Drider, Djamel

    2008-10-01

    The relative expression of mptABCD operon, glpQ, pde, rpoN, mptR and gap-1 was studied by reverse transcription combined with the real time polymerase chain reaction to understand the role of each gene in resistance/sensitivity of Enterococcus faecalis to class IIa bacteriocins such as recombinant divercin V41 (DvnRV41). Comparative critical threshold methods in presence or absence of DvnRV41 were then used to determine the level of expression of each gene cited above. In the presence of DvnRV41, the rpoN and glpQ genes were down-regulated, mptR, mptC, gap-1 and pde genes were up-regulated, whilst expression of mptB and mptD genes remained unmodified.

  16. Evaluation of the Antibacterial Efficacy of Azadirachta Indica, Commiphora Myrrha, Glycyrrhiza Glabra Against Enterococcus Faecalis using Real Time PCR

    PubMed Central

    Anand, Suresh; Rajan, Mathan; Venkateshbabu, Nagendrababu; Kandaswamy, Deivanayagam; Shravya, Yarramreddy; Rajeswari, Kalaiselvam

    2016-01-01

    Aim: To compare the antibacterial efficacy of Azadirachta indica (Neem), Commiphora myrrha (Myrrh), Glycyrrhiza glabra (Liquorice) with 2% Chlorhexidine (CHX) against E. faecalis by using Real Time PCR Materials and Methods: A total of fifty teeth specimens (n=50) were inoculated with E. faecalis for 21 days. Specimens were divided into five groups (Group 1: Myrrh, Group 2: Neem, Group 3: Liquorice, Group 4: 2% CHX and Group 5: Saline (negative control)). The intracanal medicaments were packed inside the tooth. After 5 days, the remaining microbial load was determined by using real time PCR Results: Threshold cycle (Ct) values of Myrrh extract, Neem extract, Liquorice Extract, 2% CHX and saline were found to be 30.94, 23.85, 21.38, 30.93 and 17.8 respectively Conclusion: Myrrh extract showed inhibition of E.faecalis equal to that of 2% CHX followed by Neem, Liquorice and Saline PMID:27386000

  17. Effectiveness of KTP laser versus 980 nm diode laser to kill Enterococcus faecalis in biofilms developed in experimentally infected root canals.

    PubMed

    Romeo, Umberto; Palaia, Gaspare; Nardo, Alessia; Tenore, Gianluca; Telesca, Vito; Kornblit, Roly; Del Vecchio, Alessandro; Frioni, Alessandra; Valenti, Piera; Berlutti, Francesca

    2015-04-01

    This study aimed to evaluate the antibacterial action of KTP (potassium-titanyl-phosphate) laser irradiations (compared with 980 nm diode laser), associated with conventional endodontic procedures, on Enterococcus faecalis biofilms. Fifty-six dental roots with single canals were prepared with Ni-Ti rotary instruments, autoclaved, inoculated with an E. faecalis suspension and incubated for 72 h. They were randomly allocated to control and treatment groups. Laser parameters were as follows: power 2.5 W, Ton 35 ms, Toff 50 ms (KTP laser); power 2.5 W, Ton 30 ms, Toff 30 ms (980 nm diode laser). To evaluate the residual bacterial load, BioTimer Assay was employed. The chemo-mechanical treatment together with laser irradiations (KTP and 980 nm diode lasers) achieved a considerable reduction of bacterial load (higher than 96% and 93%, respectively). Regarding both laser systems, comparisons with conventional endodontic procedures (mortality rate of about 67%) revealed statistically highly significant differences (P ≤ 0.01). This study confirms that laser systems can provide an additional aid in endodontic disinfection.

  18. An in vitro evaluation of the antimicrobial efficacy of Curcuma longa, Tachyspermum ammi, chlorhexidine gluconate, and calcium hydroxide on Enterococcus faecalis

    PubMed Central

    Kumar, Hemanshi

    2013-01-01

    Aim and Objectives: This in vitro study was designed to comparatively evaluate the antimicrobial efficacy of Curcuma longa (turmeric – T1-10%, T2-20%); Tachyspermum ammi (ajwain – A1-10%, A2-20%); chlorhexidine (CHX) gluconate gel (hexigel – 1%); and calcium hydroxide (10%) as intracanal medicaments against Enterococcus faecalis. Materials and Methods: Agar plates were prepared using brain-heart infusion (BHI) agar. Cultures of E. faecalis were grown in BHI broth at 37°C. Well diffusion method was used to derive results. Plates were inoculated for 72 h at 37°C and microbial zones of inhibition were recorded. Statistical analysis was performed with repeated measures analysis of variance. Results: C. longa (T2-20%) and CHX gluconate gel (hexigel – CHX-1%) showed larger zones of microbial inhibition than C. longa (T1-10%) that were statistically significant (P < 0.05) and were highly significant when compared to T. ammi (ajwain – A1 and A2) and calcium hydroxide. Conclusion: C. longa can be used as intracanal medicament in endodontic failure cases. PMID:23716967

  19. L-Lactic acid production by combined utilization of agricultural bioresources as renewable and economical substrates through batch and repeated-batch fermentation of Enterococcus faecalis RKY1.

    PubMed

    Reddy, Lebaka Veeranjaneya; Kim, Young-Min; Yun, Jong-Sun; Ryu, Hwa-Won; Wee, Young-Jung

    2016-06-01

    Enterococcus faecalis RKY1 was used to produce l-lactic acid from hydrol, soybean curd residues (SCR), and malt. Hydrol was efficiently metabolized to l-lactic acid with optical purity of >97.5%, though hydrol contained mixed sugars such as glucose, maltose, maltotriose, and maltodextrin. Combined utilization of hydrol, SCR, and malt was enough to sustain lactic acid fermentation by E. faecalis RKY1. In order to reduce the amount of nitrogen sources and product inhibition, cell-recycle repeated-batch fermentation was employed, where a high cell mass (26.3g/L) was obtained. Lactic acid productivity was improved by removal of lactic acid from fermentation broth by membrane filtration and by linearly increased cell density. When the total of 10 repeated-batch fermentations were carried out using 100g/L hydrol, 150g/L SCR hydrolyzate, and 20g/L malt hydrolyzate as the main nutrients, lactic acid productivity was increased significantly from 3.20g/L/h to 6.37g/L/h.

  20. Role of EfrAB efflux pump in biocide tolerance and antibiotic resistance of Enterococcus faecalis and Enterococcus faecium isolated from traditional fermented foods and the effect of EDTA as EfrAB inhibitor.

    PubMed

    Lavilla Lerma, Leyre; Benomar, Nabil; Valenzuela, Antonio Sánchez; Casado Muñoz, María del Carmen; Gálvez, Antonio; Abriouel, Hikmate

    2014-12-01

    Enterococcus faecalis and Enterococcus faecium isolated from various traditional fermented foods of both animal and vegetable origins have shown multidrug resistance to several antibiotics and tolerance to biocides. Reduced susceptibility was intra and inter-species dependent and was due to specific and unspecific mechanisms such as efflux pumps. EfrAB, a heterodimeric ABC transporter efflux pump, was detected in 100% of multidrug resistant (MDR) E. faecalis strains and only in 12% of MDR E. faecium strains. EfrAB expression was induced by half of minimum inhibitory concentration (MIC) of gentamicin, streptomycin and chloramphenicol. However, expression of efrA and efrB genes was highly dependent on the strain tested and on the antimicrobial used. Our results indicated that 3 mM EDTA highly reduced the MICs of almost all drugs tested. Nevertheless, the higher reductions (>8 folds) were obtained with gentamicin, streptomycin, chlorhexidine and triclosan. Reductions of MICs were correlated with down-regulation of EfrAB expression (10-140 folds) in all three MDR enterococci strains. This is the first report describing the role of EfrAB in the efflux of antibiotics and biocides which reflect also the importance of EfrAB in multidrug resistance in enterococci. EDTA used at low concentration as food preservative could be one of the best choices to prevent spread of multidrug resistant enterococci throughout food chain by decreasing EfrAB expression. EfrAB could be an attractive target not only in enterococci present in food matrix but also those causing infections as well by using EDTA as therapeutic agent in combination with low doses of antibiotics.

  1. Nosocomial infection caused by vancomycin-susceptible multidrug-resistant Enterococcus faecalis over a long period in a university hospital in Japan.

    PubMed

    Kudo, Michiaki; Nomura, Takahiro; Yomoda, Sachie; Tanimoto, Koichi; Tomita, Haruyoshi

    2014-11-01

    Compared with other developed countries, vancomycin-resistant enterococci (VRE) are not widespread in clinical environments in Japan. There have been no VRE outbreaks and only a few VRE strains have sporadically been isolated in our university hospital in Gunma, Japan. To examine the drug susceptibility of Enterococcus faecalis and nosocomial infection caused by non-VRE strains, a retrospective surveillance was conducted in our university hospital. Molecular epidemiological analyses were performed on 1711 E. faecalis clinical isolates collected in our hospital over a 6-year period [1998-2003]. Of these isolates, 1241 (72.5%) were antibiotic resistant and 881 (51.5%) were resistant to two or more drugs. The incidence of multidrug resistant E. faecalis (MDR-Ef) isolates in the intensive care unit increased after enlargement and restructuring of the hospital. The major group of MDR-Ef strains consisted of 209 isolates (12.2%) resistant to the five drug combination tetracycline/erythromycin/kanamycin/streptomycin/gentamicin. Pulsed-field gel electrophoresis analysis of the major MDR-Ef isolates showed that nosocomial infections have been caused by MDR-Ef over a long period (more than 3 years). Multilocus sequence typing showed that these strains were mainly grouped into ST16 (CC58) or ST64 (CC8). Mating experiments suggested that the drug resistances were encoded on two conjugative transposons (integrative conjugative elements), one encoded tetracycline-resistance and the other erythromycin/kanamycin/streptomycin/gentamicin-resistance. To our knowledge, this is the first report of nosocomial infection caused by vancomycin-susceptible MDR-Ef strains over a long period in Japan.

  2. Histopathological changes induced in an animal model by potentially pathogenic Enterococcus faecalis strains recovered from ready-to-eat food outlets in Osun State, Nigeria

    PubMed Central

    Olawale, Adetunji Kola; David, Oluwole Moses; Oluyege, Adekemi Olubukunola; Osuntoyinbo, Richard Temitope; Laleye, Solomon Anjuwon; Famurewa, Oladiran

    2015-01-01

    Enterococci have been implicated as an emerging important cause of several diseases and multiple antibiotic resistance. However, there is little information about the prevalence of pathogenic and/or antibiotic-resistant Enterococcus faecalis in ready-to-eat foods in Nigeria. Here we report the pathogenic potential of three selected antibiotic-resistant E. faecalis strains isolated from food canteens and food outlets with different virulence determinant genes, including EFC 12 (with gel+, esp+, cylA+, and asa1+), EFT 148 (with gel+, ace+, and asa1+), and EFS 18 (with esp+ and cylA+) in an animal model. Enterococcemia, hematological parameters, and histopathological changes in organ tissues were examined in experimental animals. The results showed differences in enterococcemia and hematological parameters between the control group and experimental animal group. Enterococcemia was observed for 7 days, and the animal group infected with EFC 12 showed the highest growth rate, followed by EFT 148, with the lowest growth rate seen in the EFS 18-infected group. White blood cell count, packed cell volume, and platelets were significantly reduced (P<0.05) in the experimental animals compared with the controls. White blood cells decreased drastically during the study period in rats challenged with EFC 12 (from 7,800 to 6,120 per mm3) but levels remained higher in the control group (from 9,228 to 9,306 per mm3). Histopathological changes included areas of pronounced hemorrhage, necrosis, and distortion in liver tissues, which were more marked in rats infected with EFC 12, followed by EFT 148, then EFS 18. The results of this study suggest the presence of potentially pathogenic E. faecalis strains in food canteens and food outlets; hence, there is a need for strict adherence to good hygiene practices in the study area owing to the epidemiological significance of foods. PMID:26170700

  3. Depth-dependent inactivation of Escherichia coli and Enterococcus faecalis in soil after manure application and simulated rainfall

    Technology Transfer Automated Retrieval System (TEKTRAN)

    E.coli and Enterococcus serve as important water quality indicator organisms. Rainfall action on manured fields and pastures releases these organisms into soil with infiltrating water. They can then be released back to runoff during subsequent rainfall or irrigation events as soil solution interacts...

  4. Enterococcus faecalis from Food, Clinical Specimens, and Oral Sites: Prevalence of Virulence Factors in Association with Biofilm Formation

    PubMed Central

    Anderson, Annette C.; Jonas, Daniel; Huber, Ingrid; Karygianni, Lamprini; Wölber, Johan; Hellwig, Elmar; Arweiler, Nicole; Vach, Kirstin; Wittmer, Annette; Al-Ahmad, Ali

    2016-01-01

    Enterococci have gained significance as the cause of nosocomial infections; they occur as food contaminants and have also been linked to dental diseases. E. faecalis has a great potential to spread virulence as well as antibiotic resistance genes via horizontal gene transfer. The integration of food-borne enterococci into the oral biofilm in-vivo has been observed. Therefore, we investigated the virulence determinants and antibiotic resistance of 97 E. faecalis isolates from the oral cavity, food, and clinical specimens. In addition, phenotypic expression of gelatinase and cytolysin were tested, in-vitro biofilm formation was quantified and isolates were compared for strain relatedness via pulsed field gel electrophoresis (PFGE). Each isolate was found to possess two or more virulence genes, most frequently gelE, efaA, and asa1. Notably, plaque/saliva isolates possessed the highest abundance of virulence genes, the highest levels of phenotypic gelatinase and hemolysin activity and concurrently a high ability to form biofilm. The presence of asa1 was associated with biofilm formation. The biofilm formation capacity of clinical and plaque/saliva isolates was considerably higher than that of food isolates and they also showed similar antibiotic resistance patterns. These results indicate that the oral cavity can constitute a reservoir for virulent E. faecalis strains possessing antibiotic resistance traits and at the same time distinct biofilm formation capabilities facilitating exchange of genetic material. PMID:26793174

  5. Enterococcus faecalis from Food, Clinical Specimens, and Oral Sites: Prevalence of Virulence Factors in Association with Biofilm Formation.

    PubMed

    Anderson, Annette C; Jonas, Daniel; Huber, Ingrid; Karygianni, Lamprini; Wölber, Johan; Hellwig, Elmar; Arweiler, Nicole; Vach, Kirstin; Wittmer, Annette; Al-Ahmad, Ali

    2015-01-01

    Enterococci have gained significance as the cause of nosocomial infections; they occur as food contaminants and have also been linked to dental diseases. E. faecalis has a great potential to spread virulence as well as antibiotic resistance genes via horizontal gene transfer. The integration of food-borne enterococci into the oral biofilm in-vivo has been observed. Therefore, we investigated the virulence determinants and antibiotic resistance of 97 E. faecalis isolates from the oral cavity, food, and clinical specimens. In addition, phenotypic expression of gelatinase and cytolysin were tested, in-vitro biofilm formation was quantified and isolates were compared for strain relatedness via pulsed field gel electrophoresis (PFGE). Each isolate was found to possess two or more virulence genes, most frequently gelE, efaA, and asa1. Notably, plaque/saliva isolates possessed the highest abundance of virulence genes, the highest levels of phenotypic gelatinase and hemolysin activity and concurrently a high ability to form biofilm. The presence of asa1 was associated with biofilm formation. The biofilm formation capacity of clinical and plaque/saliva isolates was considerably higher than that of food isolates and they also showed similar antibiotic resistance patterns. These results indicate that the oral cavity can constitute a reservoir for virulent E. faecalis strains possessing antibiotic resistance traits and at the same time distinct biofilm formation capabilities facilitating exchange of genetic material.

  6. From (p)ppGpp to (pp)pGpp: Characterization of Regulatory Effects of pGpp Synthesized by the Small Alarmone Synthetase of Enterococcus faecalis

    PubMed Central

    Gaca, Anthony O.; Kudrin, Pavel; Colomer-Winter, Cristina; Beljantseva, Jelena; Liu, Kuanqing; Anderson, Brent; Wang, Jue D.; Rejman, Dominik; Potrykus, Katarzyna; Cashel, Michael

    2015-01-01

    ABSTRACT The bacterial stringent response (SR) is a conserved stress tolerance mechanism that orchestrates physiological alterations to enhance cell survival. This response is mediated by the intracellular accumulation of the alarmones pppGpp and ppGpp, collectively called (p)ppGpp. In Enterococcus faecalis, (p)ppGpp metabolism is carried out by the bifunctional synthetase/hydrolase E. faecalis Rel (RelEf) and the small alarmone synthetase (SAS) RelQEf. Although Rel is the main enzyme responsible for SR activation in Firmicutes, there is emerging evidence that SASs can make important contributions to bacterial homeostasis. Here, we showed that RelQEf synthesizes ppGpp more efficiently than pppGpp without the need for ribosomes, tRNA, or mRNA. In addition to (p)ppGpp synthesis from GDP and GTP, RelQEf also efficiently utilized GMP to form GMP 3′-diphosphate (pGpp). Based on this observation, we sought to determine if pGpp exerts regulatory effects on cellular processes affected by (p)ppGpp. We found that pGpp, like (p)ppGpp, strongly inhibits the activity of E. faecalis enzymes involved in GTP biosynthesis and, to a lesser extent, transcription of rrnB by Escherichia coli RNA polymerase. Activation of E. coli RelA synthetase activity was observed in the presence of both pGpp and ppGpp, while RelQEf was activated only by ppGpp. Furthermore, enzymatic activity of RelQEf is insensitive to relacin, a (p)ppGpp analog developed as an inhibitor of “long” RelA/SpoT homolog (RSH) enzymes. We conclude that pGpp can likely function as a bacterial alarmone with target-specific regulatory effects that are similar to what has been observed for (p)ppGpp. IMPORTANCE Accumulation of the nucleotide second messengers (p)ppGpp in bacteria is an important signal regulating genetic and physiological networks contributing to stress tolerance, antibiotic persistence, and virulence. Understanding the function and regulation of the enzymes involved in (p)ppGpp turnover is therefore

  7. Comparative Evaluation of Antimicrobial Activity of QMiX, 2.5% Sodium Hypochlorite, 2% Chlorhexidine, Guava Leaf Extract and Aloevera Extract Against Enterococcus faecalis and Candida albicans – An in-vitro Study

    PubMed Central

    Krishnamma, Shoba; Peedikayil, Faizal; Aman, Shibu; Tomy, Nithya; Mariodan, Jithin Pulickal

    2016-01-01

    Introduction Debridement and disinfection of the root canal system is a critical step in endodontic treatment. Most of the irrigants presently used in the endodontic treatment can have an impact on the microbes surviving in the biofilm but none of them are able to do all of the required tasks. Researches are going on its full swing in order to produce an endodontic irrigant having ideal properties. Aim To compare the antimicrobial efficacy of different irrigants like QMiX, guava leaf extract, aloevera extract, 2.5% sodium hypochlorite and 2% chlorhexidine gluconate against Enterococcus faecalis and Candida albicans. Materials and Methods The antimicrobial activity was determined using agar diffusion test. The solutions were divided into five groups: Group I- QMiX, Group II- Guava leaf extract and Group III-Aloevera extract, Group IV–2.5% Sodium hypochlorite and Group V-2% Chlorhexidine. The zones of inhibition of growth were recorded. Results Statistical analysis was performed using one way ANOVA with post-hoc Tukey’s HSD. Values obtained were statistically analyzed (p<0.05). QMiX showed maximum inhibitory effect against Enterococcus faecalis and Candida albicans followed by, 2% chlorhexidine, 2.5% sodium hypochlorite, guava leaf extract and aloevera extract. Results obtained were statistically significant. Conclusion Guava leaf extract showed significant inhibitory effects against Enterococcus faecalis and Candida albicans. QMiX demonstrated the best results among the tested solutions and can be considered as a potential alternative to existing root canal irrigants. PMID:27437354

  8. Using a Genome-Scale Metabolic Model of Enterococcus faecalis V583 To Assess Amino Acid Uptake and Its Impact on Central Metabolism

    PubMed Central

    Solheim, Margrete; van Grinsven, Koen W. A.; Olivier, Brett G.; Levering, Jennifer; Grosseholz, Ruth; Hugenholtz, Jeroen; Holo, Helge; Nes, Ingolf; Teusink, Bas; Kummer, Ursula

    2014-01-01

    Increasing antibiotic resistance in pathogenic bacteria necessitates the development of new medication strategies. Interfering with the metabolic network of the pathogen can provide novel drug targets but simultaneously requires a deeper and more detailed organism-specific understanding of the metabolism, which is often surprisingly sparse. In light of this, we reconstructed a genome-scale metabolic model of the pathogen Enterococcus faecalis V583. The manually curated metabolic network comprises 642 metabolites and 706 reactions. We experimentally determined metabolic profiles of E. faecalis grown in chemically defined medium in an anaerobic chemostat setup at different dilution rates and calculated the net uptake and product fluxes to constrain the model. We computed growth-associated energy and maintenance parameters and studied flux distributions through the metabolic network. Amino acid auxotrophies were identified experimentally for model validation and revealed seven essential amino acids. In addition, the important metabolic hub of glutamine/glutamate was altered by constructing a glutamine synthetase knockout mutant. The metabolic profile showed a slight shift in the fermentation pattern toward ethanol production and increased uptake rates of multiple amino acids, especially l-glutamine and l-glutamate. The model was used to understand the altered flux distributions in the mutant and provided an explanation for the experimentally observed redirection of the metabolic flux. We further highlighted the importance of gene-regulatory effects on the redirection of the metabolic fluxes upon perturbation. The genome-scale metabolic model presented here includes gene-protein-reaction associations, allowing a further use for biotechnological applications, for studying essential genes, proteins, or reactions, and the search for novel drug targets. PMID:25527553

  9. An assessment of antibacterial activity of three pulp capping materials on Enterococcus faecalis by a direct contact test: An in vitro study

    PubMed Central

    Koruyucu, Mine; Topcuoglu, Nursen; Tuna, E. Bahar; Ozel, Sevda; Gencay, Koray; Kulekci, Guven; Seymen, Figen

    2015-01-01

    Objective: The aim of this in vitro study was to evaluate antimicrobial activities of three different pulp capping materials; Biodentine, mineral trioxide aggregate (MTA) Angelus, and Dycal against Enterococcus faecalis and their durability with time. Materials and Methods: Direct contact test was used for the assessment. Three sets of sealers were mixed and placed on microtiter plate wells: One set was used within 20 min of recommended setting time while others were used after 24-h and 1-week. E. faecalis suspension was placed directly on the materials for 1 h and then transferred to another plate with fresh media. Nine wells of bacteria without the tested cements served as the positive control. One well of the tested cements without bacteria served as the negative control. Bacterial growth was evaluated by a temperature-controlled microplate spectrophotometer for 1-h intervals among 24 h. Data were analyzed using Kruskal–Wallis test. Results: All tested materials showed less bacterial density than the control group. MTA, Biodentine, and Dycal showed significantly higher bacterial density than the control group in freshly mixed samples (P < 0.05). And MTA showed significantly higher antibacterial activity than Dycal (P < 0.05). In 24 h, materials did not show any differences (P > 0.05). MTA and Biodentine samples showed significant differences than Dycal; MTA also showed higher antibacterial activity than control in 1-week samples (P < 0.05). Conclusion: While freshly mixed MTA showed the best antibacterial activity over time, Biodentine had shown similar antibacterial activity to MTA. PMID:26038657

  10. Development of solution phase hybridisation PCR-ELISA for the detection and quantification of Enterococcus faecalis and Pediococcus pentosaceus in Nurmi-type cultures.

    PubMed

    Waters, Sinéad M; Doyle, Sean; Murphy, Richard A; Power, Ronan F G

    2005-12-01

    Nurmi-type cultures (NTCs), derived from the fermentation of caecal contents of specifically pathogen-free (SPF) birds, have been used successfully to control salmonella colonisation in chicks. These cultures are undefined in nature and, consequently, it is difficult to obtain approval from regulatory agencies for their use as direct fed microbials (DFMs) for poultry. Progress towards the generation of effective defined probiotics requires further knowledge of the composition of these cultures. As such, species-specific, culture-independent quantification methodologies need to be developed to elucidate the concentration of specific bacterial constituents of NTCs. Quantification of specific bacterial species in such ill-defined complex cultures using conventional culturing methods is inaccurate due to low levels of sensitivity and reproducibility, in addition to slow turnaround times. Furthermore, these methods lack selectivity due to the nature of the accompanying microflora. This study describes the development of a rapid, sensitive, reliable, reproducible, and species-specific culture-independent, solution phase hybridisation PCR-ELISA procedure for the detection and quantification of Enterococcus faecalis and Pediococcus pentosaceus in NTCs. In this technique, biotin-labelled primers were designed to amplify a species-specific fragment of a marker gene of known copy number, in both species. Resulting amplicons were hybridised with a dinitrophenol (DNP)-labelled oligonucleotide probe in solution and were subsequently captured on a streptavidin-coated microtitre plate. The degree of binding was determined by the addition of IgG (anti-DNP)-horseradish peroxidase conjugate, which was subsequently visualised using a chromogenic substrate, tetramethylbenzidine. This novel quantitative method was capable of detecting E. faecalis and P. pentosaceus at levels as low as 5 CFU per PCR reaction. PMID:15949857

  11. Superbugs in the coming new decade; multidrug resistance and prospects for treatment of Staphylococcus aureus, Enterococcus spp. and Pseudomonas aeruginosa in 2010.

    PubMed

    Nordmann, Patrice; Naas, Thierry; Fortineau, Nicolas; Poirel, Laurent

    2007-10-01

    New resistance problems have emerged recently among hospital and community-acquired pathogens such as in Staphylococcus aureus, Enterococcus faecium and Pseudomonas aeruginosa. Hospital-acquired and now community-acquired methicillin-resistant S. aureus are emerging worldwide whereas vancomycin-resistant S. aureus remain extremely rare. Hospital-acquired outbreaks of vancomycin-resistant enterococci and multidrug resistant Pseudomonas aeruginosa infections are increasingly reported worldwide. Whereas novel molecules are being developed for treating Gram-positive infections, difficult to non possible-to-treat pandrug-resistant P. aeruginosa infections may become a therapeutic challenge soon.

  12. Effectiveness of N-acetyl cysteine, 2% chlorhexidine, and their combination as intracanal medicaments on Enterococcus faecalis biofilm

    PubMed Central

    Palaniswamy, Udayakumar; Lakkam, Surender Ram; Arya, Shikha; Aravelli, Swathi

    2016-01-01

    Aim: The purpose of this study was to evaluate the antibacterial efficacies of 2% chlorhexidine (CHX), N-acetyl cysteine (NAC) and assess their synergistic or antagonist action as intracanal medicament. Materials and Methods: Agar diffusion test was performed with 2% CHX, NAC, and their combination against E. faecalis planktonic cells. The diameters of the zones of bacterial inhibition were measured and recorded for each solution. The assay was further extended to 2 weeks old E. faecalis dentinal biofilm. Sixteen freshly extracted teeth were vertically sectioned into two halves resulting in a total of 32 samples. The samples were inoculated with bacterial suspension and incubated at 37°C for 2 weeks for biofilm formation. The samples were then divided into four experimental groups with 8 samples in each group. The samples were gently washed in saline and placed in culture wells containing the test solutions, i.e., 2% CHX, NAC, a combination of 2% CHX and NAC in 1:1 ratio, and a control group containing saline. The biofilm formed on the root canal surface were removed with a sterile scalpel and inoculated on blood agar plates to check for the formation of E. faecalis colonies. Statistical Analysis: For agar diffusion test, data were analyzed statistically using one-way analysis of variance and then by post-hoc Scheffe's test to compare the antimicrobial efficacy between the groups. Statistical analysis was not done for the cultures obtained from the biofilm as there was no growth in all the three test groups except the control group, i.e., saline. Results: In agar diffusion test, among the three groups tested, 2% CHX and NAC showed almost equal zones of inhibition whereas maximum inhibition was shown by a combination of NAC and 2% CHX suggesting a synergistic action. The results obtained were highly significant (P < 0.001) for the combination of medicament when compared to individual test group. In culture analysis, which was done for the biofilm, no growth was

  13. The annotated complete DNA sequence of Enterococcus faecalis bacteriophage φEf11 and its comparison with all available phage and predicted prophage genomes.

    PubMed

    Stevens, Roy H; Ektefaie, Mahmoud R; Fouts, Derrick E

    2011-04-01

    φEf11 is a temperate Siphoviridae bacteriophage isolated by induction from a lysogenic Enterococcus faecalis strain. The φEf11 DNA was completely sequenced and found to be 42,822 bp in length, with a G+C mol% of 34.4%. Genome analysis revealed 65 ORFs, accounting for 92.8% of the DNA content. All except for seven of the ORFs displayed sequence similarities to previously characterized proteins. The genes were arranged in functional modules, organized similar to that of several other phages of low GC Gram-positive bacteria; however, the number and arrangement of lysis-related genes were atypical of these bacteriophages. A 159 bp noncoding region between predicted cI and cro genes is highly similar to the functionally characterized early promoter region of lactococcal temperate phage TP901-1, and possessed a predicted stem-loop structure in between predicted P(L) and P(R) promoters, suggesting a novel mechanism of repression of these two bacteriophages from the λ paradigm. Comparison with all available phage and predicted prophage genomes revealed that the φEf11 genome displays unique features, suggesting that φEf11 may be a novel member of a larger family of temperate prophages that also includes lactococcal phages. Trees based on the blast score ratio grouped this family by tail fiber similarity, suggesting that these trees are useful for identifying phages with similar tail fibers. PMID:21204936

  14. Characterization of the Deoxynucleotide Triphosphate Triphosphohydrolase (dNTPase) Activity of the EF1143 Protein from Enterococcus faecalis and Crystal Structure of the Activator-Substrate Complex

    SciTech Connect

    Vorontsov, Ivan I.; Minasov, George; Kiryukhina, Olga; Brunzelle, Joseph S.; Shuvalova, Ludmilla; Anderson, Wayne F.

    2012-06-19

    The EF1143 protein from Enterococcus faecalis is a distant homolog of deoxynucleotide triphosphate triphosphohydrolases (dNTPases) from Escherichia coli and Thermus thermophilus. These dNTPases are important components in the regulation of the dNTP pool in bacteria. Biochemical assays of the EF1143 dNTPase activity demonstrated nonspecific hydrolysis of all canonical dNTPs in the presence of Mn{sup 2+}. In contrast, with Mg{sup 2+} hydrolysis required the presence of dGTP as an effector, activating the degradation of dATP and dCTP with dGTP also being consumed in the reaction with dATP. The crystal structure of EF1143 and dynamic light scattering measurements in solution revealed a tetrameric oligomer as the most probable biologically active unit. The tetramer contains four dGTP specific allosteric regulatory sites and four active sites. Examination of the active site with the dATP substrate suggests an in-line nucleophilic attack on the {alpha}-phosphate center as a possible mechanism of the hydrolysis and two highly conserved residues, His-129 and Glu-122, as an acid-base catalytic dyad. Structural differences between EF1143 apo and holo forms revealed mobility of the {alpha}3 helix that can regulate the size of the active site binding pocket and could be stabilized in the open conformation upon formation of the tetramer and dGTP effector binding.

  15. Cloning and molecular analysis of genes affecting expression of binding substance, the recipient-encoded receptor(s) mediating mating aggregate formation in Enterococcus faecalis.

    PubMed

    Bensing, B A; Dunny, G M

    1993-11-01

    Transfer of the conjugative plasmid pCF10 in Enterococcus faecalis strains involves production of a plasmid-encoded aggregation substance on the surface of donor cells in response to stimulation by a pheromone secreted by recipient cells. Aggregation substance then facilitates attachment to recipient cells via a chromosomally encoded receptor, termed binding substance (BS). A BS mutant, strain INY3000, generated by random Tn916 insertions, was previously found to carry copies of the transposon at four unique sites (K. M. Trotter and G. M. Dunny, Plasmid 24:57-67, 1990). In the present study, DNA flanking the Tn916 insertions was used to complement the BS mutation of INY3000 following Tn916 excision from cloned chromosomal fragments. Complementation results showed that three of the four regions mutated in INY3000 play some role in BS expression. Tn5 mutagenesis and DNA sequence analysis of the complementing fragment from one of these regions indicated the presence of three genes (ebsA, ebsB, and ebsC) that affect BS expression. The ebsA and ebsB genes encode peptides likely to function in cell wall metabolism, whereas ebsC may encode a product that suppresses the function or expression of EbsB.

  16. Cloning and molecular analysis of genes affecting expression of binding substance, the recipient-encoded receptor(s) mediating mating aggregate formation in Enterococcus faecalis.

    PubMed Central

    Bensing, B A; Dunny, G M

    1993-01-01

    Transfer of the conjugative plasmid pCF10 in Enterococcus faecalis strains involves production of a plasmid-encoded aggregation substance on the surface of donor cells in response to stimulation by a pheromone secreted by recipient cells. Aggregation substance then facilitates attachment to recipient cells via a chromosomally encoded receptor, termed binding substance (BS). A BS mutant, strain INY3000, generated by random Tn916 insertions, was previously found to carry copies of the transposon at four unique sites (K. M. Trotter and G. M. Dunny, Plasmid 24:57-67, 1990). In the present study, DNA flanking the Tn916 insertions was used to complement the BS mutation of INY3000 following Tn916 excision from cloned chromosomal fragments. Complementation results showed that three of the four regions mutated in INY3000 play some role in BS expression. Tn5 mutagenesis and DNA sequence analysis of the complementing fragment from one of these regions indicated the presence of three genes (ebsA, ebsB, and ebsC) that affect BS expression. The ebsA and ebsB genes encode peptides likely to function in cell wall metabolism, whereas ebsC may encode a product that suppresses the function or expression of EbsB. Images PMID:8226689

  17. Biotransformation of pinoresinol diglucoside to mammalian lignans by human intestinal microflora, and isolation of Enterococcus faecalis strain PDG-1 responsible for the transformation of (+)-pinoresinol to (+)-lariciresinol.

    PubMed

    Xie, Li-Hua; Akao, Teruaki; Hamasaki, Kenjiro; Deyama, Takeshi; Hattori, Masao

    2003-05-01

    By anaerobic incubation of pinoresinol diglucoside (1) from the bark of Eucommia ulmoides with a fecal suspension of humans, eleven metabolites were formed, and their structures were identified as (+)-pinoresinol (2), (+)-lariciresinol (3), 3'-demethyl-(+)-lariciresinol (4), (-)-secoisolariciresinol (5), (-)-3-(3", 4"-dihydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butane-1, 4-diol (6), 2-(3', 4'-dihydroxybenzyl)-3-(3", 4"-dihydroxybenzyl)butane-1, 4-diol (7), 3-(3"-hydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butane-1, 4-diol (8), 2-(3', 4'-dihydroxybenzyl)-3-(3"-hydroxybenzyl)butane-1, 4-diol (9), (-)-enterodiol (10), (-)-(2R, 3R)-3-(3", 4"-dihydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butyrolactone (11), (-)-(2R, 3R)-2-(3', 4'-dihydroxybenzyl)-3-(3", 4"-dihydroxybenzyl)butyrolactone (12), (-)-(2R, 3R)-3-(3"-hydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butyrolactone (13), 2-(3', 4'-dihydroxybenzyl)-3-(3"-hydroxybenzyl)butyrolactone (14), 2-(3'-hydroxybenzyl)-3-(3", 4"-dihydroxybenzyl)butyrolactone (15) and (-)-(2R, 3R)-enterolactone (16) by various spectroscopic means, including two dimensional (2D)-NMR, mass spectrometry and circular dichroism. A possible metabolic pathway was proposed on the basis of their structures and time course experiments monitored by thin-layer chromatography. Furthermore, a bacterial strain responsible for the transformation of (+)-pinoresinol to (+)-lariciresinol was isolated from a human fecal suspension and identified as Enterococcus faecalis strain PDG-1.

  18. DNA sequence heterogeneity in the three copies of the long 16S-23S rDNA spacer of Enterococcus faecalis isolates.

    PubMed

    Gürtler, V; Rao, Y; Pearson, S R; Bates, S M; Mayall, B C

    1999-07-01

    The possibility of intragenic heterogeneity between copies of the long intergenic (16S-23S rDNA) spacer region (LISR) was investigated by specific amplification of this region from 21 Enterococcus faecalis isolates. Three copies of the LISR (rrnA, B and C) were demonstrated by hybridization of the LISR to genomic DNA cleaved with I-Ceul and SmaI. When the LISR amplicon was digested with Tsp509I, two known nucleotide substitutions were detected, one 4 nt upstream from the 5' end of the tRNA(ala) gene (allele rrnB has the Tsp509I site and rrnA and C do not) and the other 22 nt downstream from the 3' end of the tRNA(ala) gene (rrnC has the Tsp509I site). Sequence differences at these sites were detected at the allelic level (alleles rrnA, B and C) and different combinations of these alleles were designated Tsp Types. Using densitometry to analyse bands from electrophoresis gels, the intra-isolate ratios of the separate alleles (rrnA:rrnB:rrnC) were determined in each Tsp Type: I (0:3:0), II (1:2:0), III (2:0:1), IV (3:0:0), V (2:1:0) and VI (1:1:1). Sequence variation between the three copies of the LISR was confirmed by the detection of at least five other intra-isolate nucleotide substitutions using heteroduplex analysis by conformation-sensitive gel electrophoresis (CSGE) that were not detected by Tsp509I cleavage. Perpendicular denaturing gradient gel electrophoresis was capable of resolving homoduplexes; six to seven out of a possible nine curves were obtained in some isolates. In the isolate where seven curves were obtained one or more further nucleotide substitutions, not detected by Tsp509I cleavage or CSGE, were detected. On the basis of LISR sequence heterogeneity, isolates were categorized into homogeneous (only one allele sequence present) and heterogeneous (two or three allele sequences present). The transition between homogeneous and heterogeneous LISRs may be useful in studying evolutionary mechanisms between E. faecalis isolates.

  19. Effects of pre-encapsulated and pro-encapsulated Enterococcus faecalis on growth performance, blood characteristics, and cecal microflora in broiler chickens.

    PubMed

    Zhang, L; Li, J; Yun, T T; Qi, W T; Liang, X X; Wang, Y W; Li, A K

    2015-11-01

    The effects of microencapsulation of Enterococcus faecalis on the growth performance, antioxidant activity, immune function, and cecal microbiota in broilers were investigated. Broilers (1-day-old) were assigned randomly as follows: 5 treatments, 5 replicate pens per treatment, and 20 broilers per pen. Treatments included (1) a basal diet (CON), (2) CON + Aureomycin (1 g/kg of diet) (ANT), (3) CON + free non-encapsulated probiotics (1 × 10(9) cfu/kg of diet) (FREE), (4) CON + pro-encapsulated probiotics (1 × 10(9) cfu/kg of diet) (PRO), and (5) CON + pre-encapsulated probiotics (1 × 10(9) cfu/kg of diet) (PRE). Feedings included starter (1 to 21 d) and grower (21 to 42 d) phases. In the starter phase, the ANT and the PRE groups had greater (P < 0.05) ADG than the CON groups, and the feed conversion ratio (FCR) for these 2 groups was decreased (P < 0.05). In the finisher phase, the PRE and PRO groups had greater (P < 0.05) ADG than the CON group and their FCR was decreased significantly (P < 0.05). During the entire feeding period, only the PRE group showed greater (P < 0.05) ADG and lower (P < 0.05) FCR. On day 21, only birds in the PRE group had greater (P < 0.05) total antioxidant capacity and number of Lactobacillus than the CON group. On day 42, The PRE group showed greater (P < 0.05) superoxide dismutase than the CON group. Serum IgA and IgM concentrations were increased (P < 0.05) in the PRE group. Serum IL-6 in the PRE group was greater (P < 0.05) than in the other groups with the exception of ANT. At the phylum level, Firmicutes was enriched (P < 0.05) and Proteobacteria was depleted (P < 0.05) only in the PRE group. At the genus level, only the PRE diets increased (P < 0.05) the number of both Lactobacillus and Enterococcus. The results indicate that pre-encapsulation assists the efficient functioning of probiotics in broilers. PMID:26371326

  20. Effects of pre-encapsulated and pro-encapsulated Enterococcus faecalis on growth performance, blood characteristics, and cecal microflora in broiler chickens.

    PubMed

    Zhang, L; Li, J; Yun, T T; Qi, W T; Liang, X X; Wang, Y W; Li, A K

    2015-11-01

    The effects of microencapsulation of Enterococcus faecalis on the growth performance, antioxidant activity, immune function, and cecal microbiota in broilers were investigated. Broilers (1-day-old) were assigned randomly as follows: 5 treatments, 5 replicate pens per treatment, and 20 broilers per pen. Treatments included (1) a basal diet (CON), (2) CON + Aureomycin (1 g/kg of diet) (ANT), (3) CON + free non-encapsulated probiotics (1 × 10(9) cfu/kg of diet) (FREE), (4) CON + pro-encapsulated probiotics (1 × 10(9) cfu/kg of diet) (PRO), and (5) CON + pre-encapsulated probiotics (1 × 10(9) cfu/kg of diet) (PRE). Feedings included starter (1 to 21 d) and grower (21 to 42 d) phases. In the starter phase, the ANT and the PRE groups had greater (P < 0.05) ADG than the CON groups, and the feed conversion ratio (FCR) for these 2 groups was decreased (P < 0.05). In the finisher phase, the PRE and PRO groups had greater (P < 0.05) ADG than the CON group and their FCR was decreased significantly (P < 0.05). During the entire feeding period, only the PRE group showed greater (P < 0.05) ADG and lower (P < 0.05) FCR. On day 21, only birds in the PRE group had greater (P < 0.05) total antioxidant capacity and number of Lactobacillus than the CON group. On day 42, The PRE group showed greater (P < 0.05) superoxide dismutase than the CON group. Serum IgA and IgM concentrations were increased (P < 0.05) in the PRE group. Serum IL-6 in the PRE group was greater (P < 0.05) than in the other groups with the exception of ANT. At the phylum level, Firmicutes was enriched (P < 0.05) and Proteobacteria was depleted (P < 0.05) only in the PRE group. At the genus level, only the PRE diets increased (P < 0.05) the number of both Lactobacillus and Enterococcus. The results indicate that pre-encapsulation assists the efficient functioning of probiotics in broilers.

  1. Bactericidal effect of Er:YAG laser combined with sodium hypochlorite irrigation against Enterococcus faecalis deep inside dentinal tubules in experimentally infected root canals.

    PubMed

    Cheng, X; Chen, B; Qiu, J; He, W; Lv, H; Qu, T; Yu, Q; Tian, Y

    2016-02-01

    This study evaluated the bactericidal effect of Er:YAG laser radiation combined with sodium hypochlorite (NaOCl) irrigation in the treatment of Enterococcus faecalis deep inside dentinal tubules. The Er:YAG laser was activated, respectively, at 0.3, 0.5 and 1.0 W for either 20 or 30 s; 52.5 g l(-1) NaOCl and normal saline were used for the control groups. Root canals before and after treatments were examined using scanning electron microscopy (SEM). Bacterial reductions both on the root canal walls and at 100, 200, 300, 400 and 500 μm inside the dentinal tubules were analysed using a one-way analysis of variance. SEM results showed that the Er:YAG laser combined with NaOCl disinfected the dentinal tubules from 200 to over 500 μm depth as irradiation power and time increased. This combination killed significantly more bacteria than both the negative control group at each level tested and the positive control group at 300, 400 and 500 μm inside the dentinal tubules. It reached 100% in all experimental groups, both on the root canal walls and at 100 and 200 μm inside the dentinal tubules. However, at 300, 400 and 500 μm inside the dentinal tubules, only the groups treated with 0.5 and 1.0 W for 30s exhibited no bacterial growth. Of the two groups in which no bacteria were detected at all tested depths, Er:YAG laser irradiation at 0.5 W for 30 s combined with NaOCl irrigation was preferable because of the lower emission power and shorter irradiation time, and may serve as a new option for effective root canal disinfection. PMID:26645354

  2. Anti-breast cancer effects of live, heat-killed and cytoplasmic fractions of Enterococcus faecalis and Staphylococcus hominis isolated from human breast milk.

    PubMed

    Hassan, Zubaida; Mustafa, Shuhaimi; Rahim, Raha Abdul; Isa, Nurulfiza Mat

    2016-03-01

    Development of tumour that is resistant to chemotherapeutics and synthetic drugs, coupled with their life-threatening side effects and the adverse effects of surgery and hormone therapies, led to increased research on probiotics' anticancer potentials. The current study investigated the potential of live, heat-killed cells (HKC) and the cytoplasmic fractions (CF) of Enterococcus faecalis and Staphylococcus hominis as anti-breast cancer agents. MCF-7 cell line was treated with 25, 50, 100 and 200 μg/mL each of live, HKC and CF of the bacteria; and cytotoxicity was evaluated for 24, 48 and 72 h using MTT assay. The morphological features of the treated cells were examined by fluorescence microscopy. The stage of cell cycle arrest and apoptosis were quantified by flow cytometry. The bacterial effect on non-malignant breast epithelial cell line, MCF-10A, was assessed using MTT assay for 24, 48 and 72 h. All the three forms of the bacteria caused a significant decrease in MCF-7 (up to 33.29%) cell proliferation in concentration- and time-dependent manner. Morphological features of apoptosis like cell death, cell shrinkage and membrane blebbing were observed. Flow cytometry analyses suggested that about 34.60% of treated MCF-7 was undergoing apoptosis. A strong anti-proliferative activity was efficiently induced through sub-G1 accumulation (up to 83.17%) in treated MCF-7 and decreased number in the G0/G1 phase (74.39%). MCF-10A cells treated with both bacteria showed no significant difference with the untreated (>90% viability). These bacteria can be used as good alternative nutraceutical with promising therapeutic indexes for breast cancer because of their non-cytotoxic effects to normal cells.

  3. Characterization of the chitinolytic machinery of Enterococcus faecalis V583 and high-resolution structure of its oxidative CBM33 enzyme.

    PubMed

    Vaaje-Kolstad, Gustav; Bøhle, Liv Anette; Gåseidnes, Sigrid; Dalhus, Bjørn; Bjørås, Magnar; Mathiesen, Geir; Eijsink, Vincent G H

    2012-02-17

    Little information exists for the ability of enterococci to utilize chitin as a carbon source. We show that Enterococcus faecalis V583 can grow on chitin, and we describe two proteins, a family 18 chitinase (ef0361; EfChi18A) and a family 33 CBM (carbohydrate binding module) (ef0362; EfCBM33A) that catalyze chitin conversion in vitro. Various types of enzyme activity assays showed that EfChi18A has functional properties characteristic of an endochitinase. EfCBM33A belongs to a recently discovered family of enzymes that cleave glycosidic bonds via an oxidative mechanism and that act synergistically with classical hydrolytic enzymes (i.e., chitinases). The structure and function of this protein were probed in detail. An ultra-high-resolution crystal structure of EfCBM33A revealed details of a conserved binding surface that is optimized to interact with chitin and contains the catalytic center. Chromatography and mass spectrometry analyses of product formation showed that EfCBM33A cleaves chitin via the oxidative mechanism previously described for CBP21 from Serratia marcescens. Metal-depletion studies showed that EfCBM33A is a copper enzyme. In the presence of an external electron donor, EfCBM33A boosted the activity of EfChi18A, and combining the two enzymes led to rapid and complete conversion of β-chitin to chitobiose. This study provides insight into the structure and function of the CBM33 family of enzymes, which, together with their fungal counterpart called GH61, currently receive considerable attention in the biomass processing field. PMID:22210154

  4. Enterococcus faecalis PrgJ, a VirB4-Like ATPase, Mediates pCF10 Conjugative Transfer through Substrate Binding

    PubMed Central

    Li, Feng; Alvarez-Martinez, Cristina; Chen, Yuqing; Choi, Kyoung-Jae; Yeo, Hye-Jeong

    2012-01-01

    The Enterococcus faecalis prg and pcf genes of plasmid pCF10 encode a type IV secretion system (T4SS) required for conjugative transfer. PrgJ is a member of the VirB4 family of ATPases that are universally associated with T4SSs. Here, we report that purified PrgJ dimers displayed ATP binding and hydrolysis activities. A PrgJ nucleoside triphosphate (NTP) binding site mutation (K471E) slightly diminished ATP binding but abolished ATP hydrolysis in vitro and blocked pCF10 transfer in vivo. As shown with affinity pulldown assays, PrgJ and the K471E mutant protein interacted with the substrate receptor PcfC and with relaxase PcfG and accessory factor PcfF, which together form the relaxosome at the oriT sequence to initiate plasmid processing. The purified PrgJ and K471E proteins also bound single- and double-stranded DNA substrates without sequence specificity in vitro, and both PrgJ derivatives bound pCF10 in vivo by a mechanism dependent on an intact oriT sequence and cosynthesis of PcfC, PcfF, and PcfG, as shown by a formaldehyde-cross-linking assay. Our findings support a model in which the PcfC receptor coordinates with the PrgJ ATPase to drive early steps of pCF10 processing/transfer: (i) PcfC first binds the pCF10 relaxosome through contacts with PcfF, PcfG, and DNA; (ii) PcfC delivers the plasmid substrate to PrgJ; and (iii) PrgJ catalyzes substrate transfer to the membrane translocase. Substrate engagement with a VirB4-like subunit has not been previously described; consequently, our studies point to a novel function for these signature T4SS ATPases in mediating early steps of type IV secretion. PMID:22636769

  5. Characterization of Antimicrobial Substances Produced by Enterococcus faecalis MRR 10-3, Isolated from the Uropygial Gland of the Hoopoe (Upupa epops)

    PubMed Central

    Martín-Platero, Antonio M.; Valdivia, Eva; Ruíz-Rodríguez, Magdalena; Soler, Juan J.; Martín-Vivaldi, Manuel; Maqueda, Mercedes; Martínez-Bueno, Manuel

    2006-01-01

    The uropygial gland (preen gland) is a holocrine secretory gland situated at the base of the tail in birds which produces a hydrophobic fatty secretion. In certain birds, such as the hoopoe, Upupa epops, the composition of this secretion is influenced by both seasonal and sexual factors, becoming darker and more malodorous in females and in their nestlings during the nesting phase. The secretion is spread throughout the plumage when the bird preens itself, leaving its feathers flexible and waterproof. It is also thought to play a role in defending the bird against predators and parasites. We have isolated from the uropygial secretion of a nestling a bacterium that grows in monospecific culture which we have identified unambiguously by phenotypic and genotypic means as Enterococcus faecalis. The strain in question produces antibacterial substances that are active against all gram-positive bacteria assayed and also against some gram-negative strains. Its peptide nature identifies it as a bacteriocin within the group known as enterocins. Two peptides were purified to homogeneity (MR10A and MR10B), and matrix-assisted laser desorption ionization-time of flight (mass spectrometry) analysis showed masses of 5201.58 and 5207.7 Da, respectively. Amino acid sequencing of both peptides revealed high similarity with enterocin L50A and L50B (L. M. Cintas, P. Casaus, H. Holo, P. E. Hernández, I. F. Nes, and L. S. Håvarstein, J. Bacteriol. 180:1988-1994, 1998). PCR amplification of total DNA from strain MRR10-3 with primers for the L50A/B structural genes and sequencing of the amplified fragment revealed almost identical sequences, except for a single conservative change in residue 38 (Glu→Asp) in MR10A and two changes in residues 9 (Thr→Ala) and 15 (Leu→Phe) in MR10B. This is the first time that the production of bacteriocins by a bacterium isolated from the uropygial gland has been described. The production of these broad-spectrum antibacterial substances by an

  6. Changes in the treatment of Enterococcus faecalis infective endocarditis in Spain in the last 15 years: from ampicillin plus gentamicin to ampicillin plus ceftriaxone.

    PubMed

    Pericas, J M; Cervera, C; del Rio, A; Moreno, A; Garcia de la Maria, C; Almela, M; Falces, C; Ninot, S; Castañeda, X; Armero, Y; Soy, D; Gatell, J M; Marco, F; Mestres, C A; Miro, J M

    2014-12-01

    The aim of this study was to assess changes in antibiotic resistance, epidemiology and outcome among patients with Enterococcus faecalis infective endocarditis (EFIE) and to compare the efficacy and safety of the combination of ampicillin and gentamicin (A+G) with that of ampicillin plus ceftriaxone (A+C). The study was a retrospective analysis of a prospective cohort of EFIE patients treated in our centre from 1997 to 2011. Thirty patients were initially treated with A+G (ampicillin 2 g/4 h and gentamicin 3 mg/kg/day) and 39 with A+C (ampicillin 2 g/4 h and ceftriaxone 2 g/12 h) for 4-6 weeks. Increased rates of high-level aminoglycoside resistance (HLAR; gentamicin MIC ≥512 mg/L, streptomycin MIC ≥1024 mg/L or both) were observed in recent years (24% in 1997-2006 and 49% in 2007-2011; p 0.03). The use of A+C increased over time: 1997-2001, 4/18 (22%); 2002-2006, 5/16 (31%); 2007-2011, 30/35 (86%) (p <0.001). Renal failure developed in 65% of the A+G group and in 34% of the A+C group (p 0.014). Thirteen patients (43%) in the A+G group had to discontinue treatment, whereas only one patient (3%) treated with A+C had to discontinue treatment (p <0.001). Only development of heart failure and previous chronic renal failure were independently associated with 1-year mortality, while the individual antibiotic regimen (A+C vs. A+G) did not affect outcome (OR, 0.7; 95% CI, 0.2-2.2; p 0.549). Our study shows that the prevalence of HLAR EFIE has increased significantly in recent years and that alternative treatment with A+C is safer than A+G, with similar clinical outcomes, although the sample size is too small to draw firm conclusions. Randomized controlled studies are needed to confirm these results.

  7. L-lactate production from biodiesel-derived crude glycerol by metabolically engineered Enterococcus faecalis: cytotoxic evaluation of biodiesel waste and development of a glycerol-inducible gene expression system.

    PubMed

    Doi, Yuki

    2015-03-01

    Biodiesel waste is a by-product of the biodiesel production process that contains a large amount of crude glycerol. To reuse the crude glycerol, a novel bioconversion process using Enterococcus faecalis was developed through physiological studies. The E. faecalis strain W11 could use biodiesel waste as a carbon source, although cell growth was significantly inhibited by the oil component in the biodiesel waste, which decreased the cellular NADH/NAD(+) ratio and then induced oxidative stress to cells. When W11 was cultured with glycerol, the maximum culture density (optical density at 600 nm [OD600]) under anaerobic conditions was decreased 8-fold by the oil component compared with that under aerobic conditions. Furthermore, W11 cultured with dihydroxyacetone (DHA) could show slight or no growth in the presence of the oil component with or without oxygen. These results indicated that the DHA kinase reaction in the glycerol metabolic pathway was sensitive to the oil component as an oxidant. The lactate dehydrogenase (Ldh) activity of W11 during anaerobic glycerol metabolism was 4.1-fold lower than that during aerobic glycerol metabolism, which was one of the causes of low l-lactate productivity. The E. faecalis pflB gene disruptant (Δpfl mutant) expressing the ldhL1LP gene produced 300 mM l-lactate from glycerol/crude glycerol with a yield of >99% within 48 h and reached a maximum productivity of 18 mM h(-1) (1.6 g liter(-1) h(-1)). Thus, our study demonstrates that metabolically engineered E. faecalis can convert crude glycerol to l-lactate at high conversion efficiency and provides critical information on the recycling process for biodiesel waste.

  8. A genomic virulence reference map of Enterococcus faecalis reveals an important contribution of phage03-like elements in nosocomial genetic lineages to pathogenicity in a Caenorhabditis elegans infection model.

    PubMed

    La Rosa, Sabina Leanti; Snipen, Lars-Gustav; Murray, Barbara E; Willems, Rob J L; Gilmore, Michael S; Diep, Dzung B; Nes, Ingolf F; Brede, Dag Anders

    2015-05-01

    In the present study, the commensal and pathogenic host-microbe interaction of Enterococcus faecalis was explored using a Caenorhabditis elegans model system. The virulence of 28 E. faecalis isolates representing 24 multilocus sequence types (MLSTs), including human commensal and clinical isolates as well as isolates from animals and of insect origin, was investigated using C. elegans strain glp-4 (bn2ts); sek-1 (km4). This revealed that 6 E. faecalis isolates behaved in a commensal manner with no nematocidal effect, while the remaining strains showed a time to 50% lethality ranging from 47 to 120 h. Principal component analysis showed that the difference in nematocidal activity explained 94% of the variance in the data. Assessment of known virulence traits revealed that gelatinase and cytolysin production accounted for 40.8% and 36.5% of the observed pathogenicity, respectively. However, coproduction of gelatinase and cytolysin did not increase virulence additively, accounting for 50.6% of the pathogenicity and therefore indicating a significant (26.7%) saturation effect. We employed a comparative genomic analysis approach using the 28 isolates comprising a collection of 82,356 annotated coding sequences (CDS) to identify 2,325 patterns of presence or absence among the investigated strains. Univariate statistical analysis of variance (ANOVA) established that individual patterns positively correlated (n = 61) with virulence. The patterns were investigated to identify potential new virulence traits, among which we found five patterns consisting of the phage03-like gene clusters. Strains harboring phage03 showed, on average, 17% higher killing of C. elegans (P = 4.4e(-6)). The phage03 gene cluster was also present in gelatinase-and-cytolysin-negative strain E. faecalis JH2-2. Deletion of this phage element from the JH2-2 clinical strain rendered the mutant apathogenic in C. elegans, and a similar mutant of the nosocomial V583 isolate showed significantly attenuated

  9. Characterization of the Pheromone Response of the Enterococcus faecalis Conjugative Plasmid pCF10: Complete Sequence and Comparative Analysis of the Transcriptional and Phenotypic Responses of pCF10-Containing Cells to Pheromone Induction†

    PubMed Central

    Hirt, Helmut; Manias, Dawn A.; Bryan, Edward M.; Klein, Joanna R.; Marklund, Jesper K.; Staddon, Jack H.; Paustian, Michael L.; Kapur, Vivek; Dunny, Gary M.

    2005-01-01

    The sex pheromone plasmids in Enterococcus faecalis are one of the most efficient conjugative plasmid transfer systems known in bacteria. Plasmid transfer rates can reach or exceed 10−1 transconjugants per donor in vivo and under laboratory conditions. We report the completion of the DNA sequence of plasmid pCF10 and the analysis of the transcription profile of plasmid genes, relative to conjugative transfer ability following pheromone induction. These experiments employed a mini-microarray containing all 57 open reading frames of pCF10 and a set of selected chromosomal genes. A clear peak of transcription activity was observed 30 to 60 min after pheromone addition, with transcription subsiding 2 h after pheromone induction. The transcript activity correlated with the ability of donor cells to transfer pCF10 to recipient cells. Remarkably, aggregation substance (Asc10, encoded by the prgB gene) was present on the cell surface for a long period of time after pheromone-induced transcription of prgB and plasmid transfer ability had ceased. This observation could have relevance for the virulence of E. faecalis. PMID:15659682

  10. Humoral Immunity to Commensal Oral Bacteria in Human Infants: Salivary Secretory Immunoglobulin A Antibodies Reactive with Streptococcus mitis biovar 1, Streptococcus oralis, Streptococcus mutans, and Enterococcus faecalis during the First Two Years of Life

    PubMed Central

    Cole, Michael F.; Bryan, Stacey; Evans, Mishell K.; Pearce, Cheryl L.; Sheridan, Michael J.; Sura, Patricia A.; Wientzen, Raoul L.; Bowden, George H. W.

    1999-01-01

    Secretory immunoglobulin A (SIgA) antibodies reactive with the pioneer oral streptococci Streptococcus mitis biovar 1 and Streptococcus oralis, the late oral colonizer Streptococcus mutans, and the pioneer enteric bacterium Enterococcus faecalis in saliva samples from 10 human infants from birth to age 2 years were analyzed. Low levels of salivary SIgA1 and SIgA2 antibodies reactive with whole cells of all four species were detected within the first month after birth, even though S. mutans and E. faecalis were not recovered from the mouths of the infants during the study period. Although there was a fivefold increase in the concentration of SIgA between birth and age 2 years, there were no differences between the concentrations of SIgA1 and SIgA2 antibodies reactive with the four species over this time period. When the concentrations of SIgA1 and SIgA2 antibodies reactive with all four species were normalized to the concentrations of SIgA1 and SIgA2 in saliva, SIgA1 and SIgA2 antibodies reactive with these bacteria showed a significant decrease from birth to 2 years of age. Adsorption of each infant’s saliva with cells of one species produced a dramatic reduction of antibodies recognizing the other three species. Sequential adsorption of saliva samples removed all SIgA antibody to the bacteria, indicating that the SIgA antibodies were directed to antigens shared by all four species. The induction by the host of a limited immune response to common antigens that are likely not involved in adherence may be among the mechanisms that commensal streptococci employ to persist in the oral cavity. PMID:10085031

  11. Killing of VRE Enterococcus faecalis by commensal strains: Evidence for evolution and accumulation of mobile elements in the absence of competition.

    PubMed

    Gaca, Anthony O; Gilmore, Michael S

    2016-01-01

    Enterococci are members of the gastrointestinal tract of humans and most animals that, over the past 3 decades, have emerged as leading causes of multidrug resistant hospital acquired infection (HAI). In addition to their general hardiness, many traits have entered enterococcal lineages through horizontal gene transfer, which has led to the evolution of pathogenic hospital-associated lineages uniquely adapted for survival and proliferation in the antibiotic perturbed ecology of the gastrointestinal tract. We recently observed that the accretion of mobile genetic elements in the prototype vancomycin resistant E. faecalis, clinical isolate V583, renders it unable to co-exist with native enterococci in healthy human fecal flora. In this addendum, we discuss how these findings inform our understanding of how multidrug resistant enterococci evolve, and the implications for the development of treatments that limit colonization and spread of highly antibiotic refractory microbes of this type. PMID:26939857

  12. Effect of NaCl-induced osmotic stress on intracellular concentrations of glycine betaine and potassium in Escherichia coli, Enterococcus faecalis, and staphylococci.

    PubMed

    Kunin, C M; Rudy, J

    1991-09-01

    Staphylococci are more salt tolerant than are enterococci or Escherichia coli. They have a more rigid cell wall and higher internal turgor pressure. The mechanisms of NaCl-induced osmotic tolerance among these bacteria were examined by determining the generation of osmoprotective activity of cellular extracts and intracellular concentrations of glycine betaine and potassium (K+) in response to graded amounts of NaCl. Staphylococci as well as E. coli were shown to require choline or glycine betaine to achieve maximal salt tolerance. In response to 0.9 mol/L NaCl, E. coli exhibited a marked increase in osmoprotective activity, a 168-fold rise in glycine betaine, and a 2.3-fold rise in K+. Enterococcus faealis exhibited a small increase in osmoprotective activity, a 9.3-fold increase in glycine betaine, and a twofold increase in K+. In contrast, strains of Staphylococcus aureus, S. epidermidis, and S. saprophyticus were found to have considerably greater osmoprotective activity, glycine betaine, and K+ than other organisms, even in the absence of external osmotic stress. Glycine betaine rose in some strains, but K+ remained virtually unchanged as the concentration of NaCl was increased. The high concentrations of glycine betaine and K+ in staphylococci, even in the absence of osmotic stress, may explain in part their remarkable salt tolerance and high turgor pressure.

  13. Effect of NaCl-induced osmotic stress on intracellular concentrations of glycine betaine and potassium in Escherichia coli, Enterococcus faecalis, and staphylococci.

    PubMed

    Kunin, C M; Rudy, J

    1991-09-01

    Staphylococci are more salt tolerant than are enterococci or Escherichia coli. They have a more rigid cell wall and higher internal turgor pressure. The mechanisms of NaCl-induced osmotic tolerance among these bacteria were examined by determining the generation of osmoprotective activity of cellular extracts and intracellular concentrations of glycine betaine and potassium (K+) in response to graded amounts of NaCl. Staphylococci as well as E. coli were shown to require choline or glycine betaine to achieve maximal salt tolerance. In response to 0.9 mol/L NaCl, E. coli exhibited a marked increase in osmoprotective activity, a 168-fold rise in glycine betaine, and a 2.3-fold rise in K+. Enterococcus faealis exhibited a small increase in osmoprotective activity, a 9.3-fold increase in glycine betaine, and a twofold increase in K+. In contrast, strains of Staphylococcus aureus, S. epidermidis, and S. saprophyticus were found to have considerably greater osmoprotective activity, glycine betaine, and K+ than other organisms, even in the absence of external osmotic stress. Glycine betaine rose in some strains, but K+ remained virtually unchanged as the concentration of NaCl was increased. The high concentrations of glycine betaine and K+ in staphylococci, even in the absence of osmotic stress, may explain in part their remarkable salt tolerance and high turgor pressure. PMID:1919294

  14. The All-Alpha Domains of Coupling Proteins from the Agrobacterium tumefaciens VirB/VirD4 and Enterococcus faecalis pCF10-Encoded Type IV Secretion Systems Confer Specificity to Binding of Cognate DNA Substrates

    PubMed Central

    Whitaker, Neal; Chen, Yuqing; Jakubowski, Simon J.; Sarkar, Mayukh K.; Li, Feng

    2015-01-01

    ABSTRACT Bacterial type IV coupling proteins (T4CPs) bind and mediate the delivery of DNA substrates through associated type IV secretion systems (T4SSs). T4CPs consist of a transmembrane domain, a conserved nucleotide-binding domain (NBD), and a sequence-variable helical bundle called the all-alpha domain (AAD). In the T4CP structural prototype, plasmid R388-encoded TrwB, the NBD assembles as a homohexamer resembling RecA and DNA ring helicases, and the AAD, which sits at the channel entrance of the homohexamer, is structurally similar to N-terminal domain 1 of recombinase XerD. Here, we defined the contributions of AADs from the Agrobacterium tumefaciens VirD4 and Enterococcus faecalis PcfC T4CPs to DNA substrate binding. AAD deletions abolished DNA transfer, whereas production of the AAD in otherwise wild-type donor strains diminished the transfer of cognate but not heterologous substrates. Reciprocal swaps of AADs between PcfC and VirD4 abolished the transfer of cognate DNA substrates, although strikingly, the VirD4-AADPcfC chimera (VirD4 with the PcfC AAD) supported the transfer of a mobilizable plasmid. Purified AADs from both T4CPs bound DNA substrates without sequence preference but specifically bound cognate processing proteins required for cleavage at origin-of-transfer sequences. The soluble domains of VirD4 and PcfC lacking their AADs neither exerted negative dominance in vivo nor specifically bound cognate processing proteins in vitro. Our findings support a model in which the T4CP AADs contribute to DNA substrate selection through binding of associated processing proteins. Furthermore, MOBQ plasmids have evolved a docking mechanism that bypasses the AAD substrate discrimination checkpoint, which might account for their capacity to promiscuously transfer through many different T4SSs. IMPORTANCE For conjugative transfer of mobile DNA elements, members of the VirD4/TraG/TrwB receptor superfamily bind cognate DNA substrates through mechanisms that are

  15. Rapid detection of Enterococcus spp. direct from blood culture bottles using Enterococcus QuickFISH method: a multicenter investigation.

    PubMed

    Deck, Melissa K; Anderson, Erica S; Buckner, Rebecca J; Colasante, Georgia; Davis, Thomas E; Coull, James M; Crystal, Benjamin; Latta, Phyllis Della; Fuchs, Martin; Fuller, Deanna; Harris, Will; Hazen, Kevin; Klimas, Lisa L; Lindao, Daniel; Meltzer, Michelle C; Morgan, Margie; Shepard, Janeen; Stevens, Sharon; Wu, Fann; Fiandaca, Mark J

    2014-04-01

    The performance of a diagnostic method for detection and identification of Enterococcus spp. directly from positive blood culture was evaluated in a clinical study. The method, Enterococcus QuickFISH BC, is a second-generation peptide nucleic acid (PNA) fluorescence in situ hybridization (FISH) test, which uses a simplified, faster assay procedure. The test uses fluorescently labeled PNA probes targeting 16S rRNA to differentiate Enterococcus faecalis from other Enterococcus spp. by the color of the cellular fluorescence. Three hundred fifty-six routine blood culture samples were tested; only 2 discordant results were recorded. The sensitivities for detection of Enterococcus faecalis and non-faecalis Enterococcus were 100% (106/106) and 97.0% (65/67), respectively, and the combined specificity of the assay was 100%. The combined positive and negative predictive values of the assay were 100% (171/171) and 98.9% (185/187), respectively.

  16. Disclosure of the quackery: testing of the bactericidal action of products based on the "hydronic" technology ("informed glass") on ATCC strains of Enterococcus faecalis, Salmonella enteritidis and Candida albicans.

    PubMed

    Aleksandar, Racz; Josip, Cipriš; Olivera, Petrak

    2011-01-01

    To disclose a quackery called "revitalisation of tired water by hydronic technology", scientific experiments have been conducted with drinking water kept in "ordinary, everyday-use" drinking glasses and so-called 'informed' glasses, a patent-protected product supposed to have an effect on the "structure, vitality and memory of water". Drinking "informed" water is claimed to have a wide range of positive revitalising health effects (blue informed glass), to facilitate weight loss (red informed glass) and to have a stress-relieving action (green informed glass). Allegedly, by the use of the "orgon methodology", information is coded into the glass, which action is additionally enforced by the addition of the "magic life" symbol - a specially designed energy condenser which, together with the selected information, is permanently introduced into the liquid contained in the glass. Since the manufacturer claimed the products to have a broad bactericidal action, regardless of the external conditions and completely independent from additional factor that would lead to the activation of the system, the efficacy of the informed drinking glass was tested using standardised, microbiological tests. Respecting the principle of a single-blind test for each of 5 samples of each type of the informed glass, growth reduction factor (RF) (difference log cfu/ml - colony per unit/ml of control glass and log cfu/ml of each informed glass) was determined after 0,2,4,6 and 8 h in spring water experimentally contaminated with standardised ATCC strains of two types of bacteria and one yeast. The results showed a statistically significant bactericidal action of the blue informed glass with all strains-Enterococcus faecalis (RF 0.62/0.76), Salmonella enteritidis (RF 0.87/0.97), and Candida albicans (RF 0.5/0.60) - as opposed to the red and green glasses where this effect was negligible (RF < 0.1). However, when the tests were repeated in complete darkness, none of the three informed glasses

  17. Molecular and genetic analysis of a region of plasmid pCF10 containing positive control genes and structural genes encoding surface proteins involved in pheromone-inducible conjugation in Enterococcus faecalis.

    PubMed Central

    Kao, S M; Olmsted, S B; Viksnins, A S; Gallo, J C; Dunny, G M

    1991-01-01

    Exposure of Enterococcus faecalis cells carrying the tetracycline resistance plasmid pCF10 to the heptapeptide pheromone cCF10 results in an increase in conjugal transfer frequency by as much as 10(6)-fold. Pheromone-induced donor cells also express at least two plasmid-encoded surface proteins, the 130-kDa Sec 10 protein, which is involved in surface exclusion, and the 150-kDa Asc10 protein, which has been associated with the formation of mating aggregates. Previous subcloning and transposon mutagenesis studies indicated that the adjacent EcoRI c (7.5 kb) and e (4.5 kb) fragments of pCF10 encode the structural genes for these proteins and that the EcoRI c fragment also encodes at least two regulatory genes involved in activation of the expression of the genes encoding Asc10 and Sec10. In this paper, the results of physical and genetic analysis of this region of pCF10, along with the complete DNA sequences of the EcoRI c and e fragments, are reported. The results of the genetic studies indicate the location of the structural genes for the surface proteins and reveal important features of their transcription. In addition, we provide evidence here and in the accompanying paper (S. B. Olmsted, S.-M. Kao, L. J. van Putte, J. C. Gallo, and G. M. Dunny, J. Bacteriol. 173:7665-7672, 1991) for a role of Asc10 in mating aggregate formation. The data also reveal a complex positive control system that acts at distances of at least 3 to 6 kb to activate expression of Asc10. DNA sequence analysis presented here reveals the positions of a number of specific genes, termed prg (pheromone-responsive genes) in this region of pCF10. The genes mapped include prgA (encoding Sec10) and prgB (encoding Asc10), as well as four putative regulatory genes, prgX, -R, -S, and -T. Although the predicted amino acid sequences of Sec10 and Asc10 have some structural features in common with a number of surface proteins of gram-positive cocci, and the Asc10 sequence is highly similar to that of a

  18. Intestinal Translocation of Clinical Isolates of Vancomycin-Resistant Enterococcus faecalis and ESBL-Producing Escherichia coli in a Rat Model of Bacterial Colonization and Liver Ischemia/Reperfusion Injury

    PubMed Central

    van der Heijden, Karin M.; van der Heijden, Inneke M.; Galvao, Flavio H.; Lopes, Camila G.; Costa, Silvia F.; Abdala, Edson; D’Albuquerque, Luiz A.; Levin, Anna S.

    2014-01-01

    The objectives of this study were to develop a rat model of gastrointestinal colonization with vancomycin-resistant Enterococcus faecalis (VRE) and extended-spectrum beta-lactamase (ESBL)-producing E. coli and to evaluate intestinal translocation to blood and tissues after total and partial hepatic ischemia. Methods - We developed a model of rat colonization with VRE and ESBL-E coli. Then we studied four groups of colonized rats: Group I (with hepatic pedicle occlusion causing complete liver ischemia and intestinal stasis); Group II (with partial liver ischemia without intestinal stasis); Group III (surgical manipulation without hepatic ischemia or intestinal stasis); Group IV (anesthetized without surgical manipulation). After sacrifice, portal and systemic blood, large intestine, small intestine, spleen, liver, lungs, and cervical and mesenteric lymph nodes were cultured. Endotoxin concentrations in portal and systemic blood were determined. Results – The best inocula were: VRE: 2.4×1010 cfu and ESBL-E. coli: 1.12×1010 cfu. The best results occurred 24 hours after inoculation and antibiotic doses of 750 µg/mL of water for vancomycin and 2.1 mg/mL for ceftriaxone. There was a significantly higher proportion of positive cultures for ESBL-E. coli in the lungs in Groups I, II and III when compared with Group IV (67%; 60%; 75% and 13%, respectively; p:0.04). VRE growth was more frequent in mesenteric lymph nodes for Groups I (67%) and III (38%) than for Groups II (13%) and IV (none) (p:0.002). LPS was significantly higher in systemic blood of Group I (9.761±13.804 EU/mL−p:0.01). No differences for endotoxin occurred in portal blood. Conclusion –We developed a model of rats colonized with resistant bacteria useful to study intestinal translocation. Translocation occurred in surgical procedures with and without hepatic ischemia-reperfusion and probably occurred via the bloodstream. Translocation was probably lymphatic in the ischemia-reperfusion groups

  19. Investigation of the effectiveness of disinfectants against planktonic and biofilm forms of P. aeruginosa and E. faecalis cells using a compilation of cultivation, microscopic and flow cytometric techniques.

    PubMed

    Juzwa, Wojciech; Myszka, Kamila; Białas, Wojciech; Dobrucka, Renata; Konieczny, Piotr; Czaczyk, Katarzyna

    2015-01-01

    This study evaluated the effectiveness of selected disinfectants against bacterial cells within a biofilm using flow cytometry, the conventional total viable count test and scanning electron microscopy (SEM). A flow cytometric procedure based on measurement of the cellular redox potential (CRP) was demonstrated to have potential for the rapid evaluation of activity against biofilm and planktonic forms of microbes. Quaternary ammonium compound-based disinfectant (QACB) demonstrated a higher level of anti-microbial activity than a performic acid preparation (PAP), with mean CRP values against P. aeruginosa cells of 2 and 1.33 relative fluorescence units (RFU) vs 63.33 and 61.33 RFU for 8 and 24 h cultures respectively. Flow cytometric evaluation of the anti-biofilm activity demonstrated a higher efficacy of QACB compared to PAP for P. aeruginosa cells of 1 and 0.66 RFU vs 18.33 and 22.66 RFU for 8 and 24 h cultures respectively. SEM images of treated P. aeruginosa cells demonstrated disinfectant-specific effects on cell morphology.

  20. Antimicrobial Resistance of Escherichia coli, Enterococci, Pseudomonas aeruginosa, and Staphylococcus aureus from Raw Fish and Seafood Imported into Switzerland.

    PubMed

    Boss, Renate; Overesch, Gudrun; Baumgartner, Andreas

    2016-07-01

    A total of 44 samples of salmon, pangasius (shark catfish), shrimps, and oysters were tested for the presence of Escherichia coli, enterococci, Pseudomonas aeruginosa, and Staphylococcus aureus, which are indicator organisms commonly used in programs to monitor antibiotic resistance. The isolated bacterial strains, confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectroscopy, were tested against a panel of 29 antimicrobial agents to obtain MICs. Across the four sample types, Enterococcus faecalis (59%) was most common, followed by E. coli (55%), P. aeruginosa (27%), and S. aureus (9%). All bacterial species were resistant to some antibiotics. The highest rates of resistance were in E. faecalis to tetracycline (16%), in E. coli to ciprofloxacin (22%), and in S. aureus to penicillin (56%). Antibiotic resistance was found among all sample types, but salmon and oysters were less burdened than were shrimps and pangasius. Multidrug-resistant (MDR) strains were exclusively found in shrimps and pangasius: 17% of pangasius samples (MDR E. coli and S. aureus) and 64% of shrimps (MDR E. coli, E. faecalis, and S. aureus). Two of these MDR E. coli isolates from shrimps (one from an organic sample) were resistant to seven antimicrobial agents. Based on these findings, E. coli in pangasius, shrimps, and oysters, E. faecalis in pangasius, shrimps, and salmon, and P. aeruginosa in pangasius and shrimps are potential candidates for programs monitoring antimicrobial resistance. Enrichment methods for the detection of MDR bacteria of special public health concern, such as methicillin-resistant S. aureus and E. coli producing extended-spectrum β-lactamases and carbapenemases, should be implemented.

  1. Antimicrobial Resistance of Escherichia coli, Enterococci, Pseudomonas aeruginosa, and Staphylococcus aureus from Raw Fish and Seafood Imported into Switzerland.

    PubMed

    Boss, Renate; Overesch, Gudrun; Baumgartner, Andreas

    2016-07-01

    A total of 44 samples of salmon, pangasius (shark catfish), shrimps, and oysters were tested for the presence of Escherichia coli, enterococci, Pseudomonas aeruginosa, and Staphylococcus aureus, which are indicator organisms commonly used in programs to monitor antibiotic resistance. The isolated bacterial strains, confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectroscopy, were tested against a panel of 29 antimicrobial agents to obtain MICs. Across the four sample types, Enterococcus faecalis (59%) was most common, followed by E. coli (55%), P. aeruginosa (27%), and S. aureus (9%). All bacterial species were resistant to some antibiotics. The highest rates of resistance were in E. faecalis to tetracycline (16%), in E. coli to ciprofloxacin (22%), and in S. aureus to penicillin (56%). Antibiotic resistance was found among all sample types, but salmon and oysters were less burdened than were shrimps and pangasius. Multidrug-resistant (MDR) strains were exclusively found in shrimps and pangasius: 17% of pangasius samples (MDR E. coli and S. aureus) and 64% of shrimps (MDR E. coli, E. faecalis, and S. aureus). Two of these MDR E. coli isolates from shrimps (one from an organic sample) were resistant to seven antimicrobial agents. Based on these findings, E. coli in pangasius, shrimps, and oysters, E. faecalis in pangasius, shrimps, and salmon, and P. aeruginosa in pangasius and shrimps are potential candidates for programs monitoring antimicrobial resistance. Enrichment methods for the detection of MDR bacteria of special public health concern, such as methicillin-resistant S. aureus and E. coli producing extended-spectrum β-lactamases and carbapenemases, should be implemented. PMID:27357045

  2. Detection of vancomycin-resistant Enterococcus faecalis ST6-vanB2 and E. faecium ST915-vanA in faecal samples of wild Rattus rattus in Spain.

    PubMed

    Lozano, Carmen; González-Barrio, David; García, Jesús T; Ceballos, Sara; Olea, Pedro P; Ruiz-Fons, Francisco; Torres, Carmen

    2015-05-15

    The detection of vancomycin-resistant-enterococci (VRE) among wild animals represents a worrisome public health concern. The objectives of the study were to determine the possible presence of VRE in faecal samples of wild small mammals in Spain, to characterize the vancomycin resistance mechanisms and genetic lineages of recovered isolates and to know the diversity of enterococcal species in these animals. A total of 155 faecal samples from small mammals were inoculated in Slanetz-Bartley agar supplemented or not with vancomycin (Van-SB/SB plates). The antimicrobial susceptibility profile to 12 antimicrobials and the presence of 20 antimicrobial resistance genes was analyzed. The structure of Tn1546 and the presence of gelE, cylA, asa, esp and hyl genes was studied. Multilocus-sequence-typing (MLST) technique was also performed. VRE isolates were recovered in Van-SB plates in 11 samples. Two samples contained vanB2-positive E. faecalis isolates of lineage ST6, which showed a multiresistance phenotype and harboured the virulence genes gelE and asa. One sample contained a vancomycin-resistant E. faecium isolate of the new lineage ST915, with the vanA gene included into Tn1546 (truncated with IS1542 and IS1216 elements). The vanB2 and vanA isolates were obtained from Rattus rattus. The remaining eight VRE-positive samples contained species with intrinsic vancomycin-resistance mechanisms: E. casseliflavus (n=5) and E. gallinarum (n=3). One hundred and forty-seven vancomycin-susceptible-enterococcal isolates were obtained in SB plates, and E. faecalis and E. faecium were the most frequent detected species. This is the first report of vanB2-containing enterococci in wild animals.

  3. An antibacterial and antiviral peptide produced by Enterococcus mundtii ST4V isolated from soya beans.

    PubMed

    Todorov, Svetoslav D; Wachsman, Mónica B; Knoetze, Hendriëtte; Meincken, Martina; Dicks, Leon M T

    2005-06-01

    Enterococcus mundtii ST4V, isolated from soya beans, produces a 3950Da antibacterial peptide active against Gram-positive and Gram-negative bacteria, including Enterococcus faecalis, Streptococcus spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae and Staphylococcus aureus. The peptide also inactivated the herpes simplex viruses HSV-1 (strain F) and HSV-2 (strain G), a polio virus (PV3, strain Sabin) and a measles virus (strain MV/BRAZIL/001/91, an attenuated strain of MV). MV, HSV-1 and HSV-2 were 95.5%-99.9% inactivated by peptide ST4V at 400 microg/ml. Monkey kidney Vero cells were not inactivated, even at four times the level peptide ST4V displayed antiviral activity, indicating that the effect was not due to cytotoxicity. Complete inactivation or significant reduction in antimicrobial activity was observed after treatment of peptide ST4V with Proteinase K, pronase, pepsin and trypsin. No change in antimicrobial activity was recorded after treatment with alpha-amylase, suggesting that peptide ST4V was not glycosylated. This is the first description of an antibacterial and antiviral peptide with such broad-spectrum of activity, produced by a lactic acid bacterium.

  4. An antibacterial and antiviral peptide produced by Enterococcus mundtii ST4V isolated from soya beans.

    PubMed

    Todorov, Svetoslav D; Wachsman, Mónica B; Knoetze, Hendriëtte; Meincken, Martina; Dicks, Leon M T

    2005-06-01

    Enterococcus mundtii ST4V, isolated from soya beans, produces a 3950Da antibacterial peptide active against Gram-positive and Gram-negative bacteria, including Enterococcus faecalis, Streptococcus spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae and Staphylococcus aureus. The peptide also inactivated the herpes simplex viruses HSV-1 (strain F) and HSV-2 (strain G), a polio virus (PV3, strain Sabin) and a measles virus (strain MV/BRAZIL/001/91, an attenuated strain of MV). MV, HSV-1 and HSV-2 were 95.5%-99.9% inactivated by peptide ST4V at 400 microg/ml. Monkey kidney Vero cells were not inactivated, even at four times the level peptide ST4V displayed antiviral activity, indicating that the effect was not due to cytotoxicity. Complete inactivation or significant reduction in antimicrobial activity was observed after treatment of peptide ST4V with Proteinase K, pronase, pepsin and trypsin. No change in antimicrobial activity was recorded after treatment with alpha-amylase, suggesting that peptide ST4V was not glycosylated. This is the first description of an antibacterial and antiviral peptide with such broad-spectrum of activity, produced by a lactic acid bacterium. PMID:15869868

  5. Prevalence of enterococcus species and their virulence genes in fresh water prior to and after storm events.

    PubMed

    Sidhu, J P S; Skelly, E; Hodgers, L; Ahmed, W; Li, Y; Toze, S

    2014-01-01

    Enterococcus spp. isolates (n = 286) collected from six surface water bodies in subtropical Brisbane, Australia, prior to and after storm events, were identified to species level and tested for the presence of seven clinically important virulence genes (VGs). Enterococcus faecalis (48%), Enterococcus faecium (14%), Enterococcus mundtii (13%), and Enterococcus casseliflavus (13%) were frequently detected at all sites. The frequency of E. faecium occurrence increased from 6% in the dry period to 18% after the wet period. The endocarditis antigen (efaA), gelatinase (gelE), collagen-binding protein (ace), and aggregation substance (asa1) were detected in 61%, 43%, 43%, and 23% of Enterococcus isolates, respectively. The chances of occurrence of ace, gelE, efaA, and asa1 genes in E. faecalis were found to be much higher compared to the other Enterococcus spp. The observed odds ratio of occurrence of ace and gelE genes in E. faecalis was much higher at 7.96 and 6.40 times, respectively. The hyl gene was 3.84 times more likely to be detected in E. casseliflavus. The presence of multiple VGs in most of the E. faecalis isolates underscores the importance of E. faecalis as a reservoir of VGs in the fresh water aquatic environment. Consequently, if contaminated surface water is to be used for production of potable and nonpotable water some degree of treatment depending upon intended use such as detention in basins prior to use or chlorination is required.

  6. Antibacterial, anti-inflammatory and probiotic potential of Enterococcus hirae isolated from the rumen of Bos primigenius.

    PubMed

    Arokiyaraj, Selvaraj; Hairul Islam, Villianur Ibrahim; Bharanidharan, R; Raveendar, Sebastian; Lee, Jinwook; Kim, Do Hyung; Oh, Young Kyoon; Kim, Eun-Kyung; Kim, Kyoung Hoon

    2014-07-01

    In the present study bacterial strains were isolated from the rumen fluids of Bos primigenius and investigated their in vitro probiotic properties with potent antibacterial activity and anti-inflammatory effects. 9 g positive bacterial isolates were obtained and three isolates could able to tolerate gastric conditions, high bile salt concentrations and exhibited significant bactericidal effect against the enteric pathogens Vibrio cholera, Enterococcus faecalis, Enterobacter aerogens, Pseudomonas aeruginosa, Escherichia coli and Salmonella typhi. Moreover it showed above 70% cell surface hydrophobicity, significant low-invasion ability and potential adherence capacity in Caco-2 cells when compared with the control. The proinflammatory cytokines (TNF-α) was greatly reduced in rumen bacteria treatment and ARBS-1 modulate the immune response by activating the IL-4 secretion in parallel to TNF-α suppression. The 16s rRNA gene sequence of the active isolates were identified as Enterococcus hirae (ARBS-1), Pediococcus acidilactici (ARBS-4) and Bacillus licheniformis (ARBS-7). This study revealed the probiotic bactericidal properties of E. hirae obtained from the rumen of B. primigenius with potential antibacterial and anti-inflammatory effects. Future studies with the strains may yield some novel probiotic product for livestock's.

  7. Identification of vancomycin-susceptible major clones of clinical Enterococcus from Algeria.

    PubMed

    Bourafa, Nadjette; Abat, Cédric; Loucif, Lotfi; Olaitan, Abiola Olumuyiwa; Bentorki, Ahmed Aimen; Boutefnouchet, Nafissa; Rolain, Jean-Marc

    2016-09-01

    The main objectives of this study were to characterize clinical strains of Enterococcus spp. isolated from Algerian inpatients and outpatients, to investigate their susceptibility to antibiotics and to analyse their phylogenetic relatedness. A total of 85 non-duplicate Enterococcus spp. isolates collected between 2010 and 2013 from various clinical samples, including urine, vaginal swab, pus, blood and semen, from Algerian inpatients (n=62) and outpatients (n=23) were identified using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/MS). Antibiotic susceptibility testing was performed using disk diffusion and Etest methods. Clonal relatedness was analysed using multilocus sequence typing (MLST). Enterococcus faecalis was the most predominant species (75.3%), followed by Enterococcus faecium (21.2%), Enterococcus gallinarum (2.4%) and Enterococcus casseliflavus (1.2%). High-level resistance to aminoglycosides was significantly more prevalent in hospitalized patients than in outpatients. None of the E. faecalis and E. faecium isolates were resistant to vancomycin. High genetic diversity was observed among the E. faecalis isolates, with the identification of a new clonal complex (CC256), as well as the detection of E. faecalis ST6 and E. faecium lineages ST17, ST18 and ST78 associated with hospital isolates. This is the first report of E. faecalis ST6 and E. faecium ST17 and ST18 in Algeria. Although acquired vancomycin resistance was not observed among the enterococcal strains, there is a continued need to monitor the level of antibiotic resistance among enterococci as well as the evolution of the E. faecalis/E. faecium ratio.

  8. Identification of vancomycin-susceptible major clones of clinical Enterococcus from Algeria.

    PubMed

    Bourafa, Nadjette; Abat, Cédric; Loucif, Lotfi; Olaitan, Abiola Olumuyiwa; Bentorki, Ahmed Aimen; Boutefnouchet, Nafissa; Rolain, Jean-Marc

    2016-09-01

    The main objectives of this study were to characterize clinical strains of Enterococcus spp. isolated from Algerian inpatients and outpatients, to investigate their susceptibility to antibiotics and to analyse their phylogenetic relatedness. A total of 85 non-duplicate Enterococcus spp. isolates collected between 2010 and 2013 from various clinical samples, including urine, vaginal swab, pus, blood and semen, from Algerian inpatients (n=62) and outpatients (n=23) were identified using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/MS). Antibiotic susceptibility testing was performed using disk diffusion and Etest methods. Clonal relatedness was analysed using multilocus sequence typing (MLST). Enterococcus faecalis was the most predominant species (75.3%), followed by Enterococcus faecium (21.2%), Enterococcus gallinarum (2.4%) and Enterococcus casseliflavus (1.2%). High-level resistance to aminoglycosides was significantly more prevalent in hospitalized patients than in outpatients. None of the E. faecalis and E. faecium isolates were resistant to vancomycin. High genetic diversity was observed among the E. faecalis isolates, with the identification of a new clonal complex (CC256), as well as the detection of E. faecalis ST6 and E. faecium lineages ST17, ST18 and ST78 associated with hospital isolates. This is the first report of E. faecalis ST6 and E. faecium ST17 and ST18 in Algeria. Although acquired vancomycin resistance was not observed among the enterococcal strains, there is a continued need to monitor the level of antibiotic resistance among enterococci as well as the evolution of the E. faecalis/E. faecium ratio. PMID:27530845

  9. Silver Nanoparticles: Biosynthesis Using an ATCC Reference Strain of Pseudomonas aeruginosa and Activity as Broad Spectrum Clinical Antibacterial Agents

    PubMed Central

    Quinteros, Melisa A.; Aiassa Martínez, Ivana M.; Dalmasso, Pablo R.; Páez, Paulina L.

    2016-01-01

    Currently, the biosynthesis of silver-based nanomaterials attracts enormous attention owing to the documented antimicrobial properties of these ones. This study reports the extracellular biosynthesis of silver nanoparticles (Ag-NPs) using a Pseudomonas aeruginosa strain from a reference culture collection. A greenish culture supernatant of P. aeruginosa incubated at 37°C with a silver nitrate solution for 24 h changed to a yellowish brown color, indicating the formation of Ag-NPs, which was confirmed by UV-vis spectroscopy, transmission electron microscopy, and X-ray diffraction. TEM analysis showed spherical and pseudospherical nanoparticles with a distributed size mainly between 25 and 45 nm, and the XRD pattern revealed the crystalline nature of Ag-NPs. Also it provides an evaluation of the antimicrobial activity of the biosynthesized Ag-NPs against human pathogenic and opportunistic microorganisms, namely, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Proteus mirabilis, Acinetobacter baumannii, Escherichia coli, P. aeruginosa, and Klebsiella pneumonia. Ag-NPs were found to be bioactive at picomolar concentration levels showing bactericidal effects against both Gram-positive and Gram-negative bacterial strains. This work demonstrates the first helpful use of biosynthesized Ag-NPs as broad spectrum bactericidal agents for clinical strains of pathogenic multidrug-resistant bacteria such as methicillin-resistant S. aureus, A. baumannii, and E. coli. In addition, these Ag-NPs showed negligible cytotoxic effect in human neutrophils suggesting low toxicity to the host. PMID:27340405

  10. Eight-year Surveillance of Antimicrobial Resistance among Enterococcus Spp. Isolated in the First Bethune Hospital

    NASA Astrophysics Data System (ADS)

    Xu, Jiancheng; Wang, Liqiang; Wang, Kai; Zhou, Qi

    This study was to investigate the antimicrobial resistance of Enterococcus spp. isolated in 8 consecutive years in the First Bethune Hospital. Disk diffusion test was used to study the antimicrobial resistance. The data were analyzed by WHONET 5 software according to Clinical and Laboratory Standards Institute (CLSI). Most of 1446 strains of Enterococcus spp. were collected from urine 640 (44.3%), sputum 315 (21.8%), secretions and pus 265 (18.3%) during the past 8 years. The rates of high-level aminoglycoside resistance in Enterococcus faecalis and Enterococcus faecium were 57.4%∼75.9% and 69.0%∼93.8% during the past 8 years, respectively. No Enterococcus spp. was resistant to vancomycin. The antimicrobial resistance of Enterococcus spp. had increased in recent 8 years. The change of the antimicrobial resistance should be investigated in order to direct rational drug usage in the clinic and prevent bacterial strain of drug resistance from being transmitted.

  11. Diversity, distribution and antibiotic resistance of Enterococcus spp. recovered from tomatoes, leaves, water and soil on U.S. Mid-Atlantic farms.

    PubMed

    Micallef, Shirley A; Goldstein, Rachel E Rosenberg; George, Ashish; Ewing, Laura; Tall, Ben D; Boyer, Marc S; Joseph, Sam W; Sapkota, Amy R

    2013-12-01

    Antibiotic-resistant enterococci are important opportunistic pathogens and have been recovered from retail tomatoes. However, it is unclear where and how tomatoes are contaminated along the farm-to-fork continuum. Specifically, the degree of pre-harvest contamination with enterococci is unknown. We evaluated the prevalence, diversity and antimicrobial susceptibilities of enterococci collected from tomato farms in the Mid-Atlantic United States. Tomatoes, leaves, groundwater, pond water, irrigation ditch water, and soil were sampled and tested for enterococci using standard methods. Antimicrobial susceptibility testing was performed using the Sensititre microbroth dilution system. Enterococcus faecalis isolates were characterized using amplified fragment length polymorphism to assess dispersal potential. Enterococci (n = 307) occurred in all habitats and colonization of tomatoes was common. Seven species were identified: Enterococcus casseliflavus, E. faecalis, Enterococcus gallinarum, Enterococcus faecium, Enterococcus avis, Enterococcus hirae and Enterococcus raffinosus. E. casseliflavus predominated in soil and on tomatoes and leaves, and E. faecalis predominated in pond water. On plants, distance from the ground influenced presence of enterococci. E. faecalis from samples within a farm were more closely related than those from samples between farms. Resistance to rifampicin, quinupristin/dalfopristin, ciprofloxacin and levofloxacin was prevalent. Consumption of raw tomatoes as a potential exposure risk for antibiotic-resistant Enterococcus spp. deserves further attention.

  12. Review of virulence factors of enterococcus: an emerging nosocomial pathogen.

    PubMed

    Giridhara Upadhyaya, P M; Ravikumar, K L; Umapathy, B L

    2009-01-01

    Enterococcus, considered a normal commensal of intestinal tract, is fast emerging as a pathogen causing serious and life threatening hospital borne infections. This is attributed to acquisition of multi drug resistance and virulence factors of the organisms. The sequencing of Enterococcus faecalis has given a lot of insight into its genetic makeup. The E. faecalis strain V583, which has been sequenced, contains a total of 3182 open reading frames (ORFs) with 1760 of these showing similarity to known proteins and 221 of unknown functions. Strikingly unique to this genome is the fact that over 25% of the genome is made up of mobile and exogenously acquired DNA which includes a number of conjugative and composite transposons, a pathogenicity island, integrated plasmid genes and phage regions, and a high number of insertion sequence (IS) elements. This review addresses the genomic arrangement and the study of virulence factors that have occurred since the sequencing of the genome. PMID:19736397

  13. USE OF TAQMAN TO ENUMERATE ENTEROCOCCUS FAECALIS IN WATER

    EPA Science Inventory

    The Polymerase Chain Reaction (PCR) has become a useful tool in the detection of microorganisms. However, conventional PCR is somewhat time-consuming considering that additional steps (e.g., gel electrophoresis and gene sequencing) are required to confirm the presence of the tar...

  14. Enterococcus species composition determined by capillary electrophoresis of the groESL gene spacer region DNA.

    PubMed

    Yasuda, M; Paar, J; Doolittle, M; Brochi, J; Pancorbo, O C; Tang, R J; Stoner, R E; Shiaris, M P

    2010-07-01

    Marine recreational beaches are monitored for fecal contamination by Enterococcus spp. (ENT) counts. Although different ENT species in the environment tend to thrive in and originate from distinct hosts, the current monitoring method does not differentiate among species. Time-consuming isolation-based species identification precludes routine analysis of environmental ENT communities. Therefore, an isolation-independent DNA fingerprinting method was developed to characterize environmental ENT communities using DNA length polymorphism of the spacer region between the groES and groEL genes common to most ENT species. Capillary electrophoresis resulted in distinct peak sizes of PCR products that carried polymorphic groESL spacers (300-335 bp in length) among 8 different ENT species (Enterococcus avium, Enterococcus gallinarum, Enterococcus casseliflavus, Enterococcus mundtii, Enterococcus hirae, Enterococcus faecium, Enterococcus durans, and Enterococcus faecalis). Distortions in true species ratios observed in electropherograms were caused by PCR biases arising in a mixed ENT community DNA template. E. faecalis was overestimated and E. avium and E. faecium were underestimated compared to the original species ratios in the mixed community. The PCR product bias was constant between species, so good approximation of the species ratio in ENT communities is possible. In environmental samples, a high percentage of E. faecalis (96%) together with high total ENT counts were observed in samples collected from a sewer line and from several sites in a storm drain system where sewage leaks were suspected. In contrast, samples with <400 CFU 100 ml-1 ENT were either dominated by E. mundtii or had 4 or more ENT species. The latter ENT community profiles are considered to be signatures of enterococci rarely associated with animals with low or of non-fecal origin.

  15. Comparative evaluation of antimicrobial efficacy of three herbal irrigants in reducing intracanal E. faecalis populations: An in vitro study

    PubMed Central

    Wadhwa, Jitesh; Duhan, Jigyasa

    2016-01-01

    Background The present study aimed to evaluate the intracanal bacterial reduction promoted by chemomechanical preparation using three different herbal extracts named Ocimum sanctum (OS), Cinnamomum zeylanicum (CZ), Syzygium aromaticum (SA) against Enterococcus faecalis. Material and Methods Root canals from extracted teeth were contaminated with Enterococcus faecalis ATCC 29212 for 7 days and then randomly distributed into 3 experimental groups of 10 teeth each: which includes conventional irrigation with OS, CZ and SA. The control groups included 5 teeth each consisting of NaOCl (positive control) and distilled water (negative control). Samples taken before and after chemomechanical procedures were cultured, and the colony-forming units (CFUs) were counted. Bacterial identification was performed using Polymerase chain reaction technique. The statistical analyses were performed with various tests. Results Reduction in the intracanal bacterial populations was highly significant for all the experimental groups. CZ and SA showed 80 to 85% intracanal bacterial reduction while O. Sanctum revealed only 70 to 75 % reduction. NaOCl showed 96 to 100 % bacterial reduction on the other hand distilled water showed very minimal bacterial reduction i.e 10 to 16%. Conclusions Cinnamomum zeylanicum, Syzygium aromaticum and Ocimum sanctum showed intracanal bacterial reduction against Enterococcus faecalis. The 3 experimental groups were less effective in terms of intracanal bacterial reduction as compare to NaOCl but more effective than distilled water. Key words:Antimicrobial activity, Cinnamomum zeylanicum, Enterococcus faecalis, Ocimum sanctum, Syzygium aromaticum, herbal extracts. PMID:27398170

  16. Enterococcus infection biology: lessons from invertebrate host models.

    PubMed

    Yuen, Grace J; Ausubel, Frederick M

    2014-03-01

    The enterococci are commensals of the gastrointestinal tract of many metazoans, from insects to humans. While they normally do not cause disease in the intestine, they can become pathogenic when they infect sites outside of the gut. Recently, the enterococci have become important nosocomial pathogens, with the majority of human enterococcal infections caused by two species, Enterococcus faecalis and Enterococcus faecium. Studies using invertebrate infection models have revealed insights into the biology of enterococcal infections, as well as general principles underlying host innate immune defense. This review highlights recent findings on Enterococcus infection biology from two invertebrate infection models, the greater wax moth Galleria mellonella and the free-living bacteriovorous nematode Caenorhabditis elegans. PMID:24585051

  17. Microbiologic Evaluation of Matricaria and Chlorhexidine against E. faecalis and C. albicans

    PubMed Central

    Rahman, Hena; Chandra, Anil

    2015-01-01

    Objective: To evaluate the antimicrobial activity of different concentrations of Matricaria chamomilla and Chlorhexidine gel against Candida albicans and Enterococcus faecalis. Materials and Methods: The agar diffusion test was used to evaluate the antimicrobial activity of 15%, 25% Matricaria chamomilla in aq. base and 2% chlorhexidine gel against C. albicans (ATCC 24433) and E. faecalis (ATCC 24212) strains. Vancomycin was used as the positive control for E. faecalis and fluconazole for C. albicans . The agar plates were incubated at 37°C for 48 h after which the zone of inhibition were measured separately for each material. Data thus obtained were statistically analyzed using the Wilcoxon rank–order test. Results: 2% chlorhexidine showed maximum inhibitory zone for C. albicans (33.26 mm) and E. faecalis (24.54 mm). 25% Matricaria showed zones of 24.16 mm and 20.62 mm for C. albicans and E. faecalis, respectively. 15% Matricaria did not show any antimicrobial activity (0 mm). Conclusion: The results of the current in vitro study suggest that 25% Matricaria can be used as an antimicrobial agent, but it is less effective than 2% chlorhexidine gluconate gel against C. albicans and E. faecalis. Matricaria at a lesser concentration of 15% aq. base is ineffective against both the microorganisms. PMID:26097333

  18. AN EFFICIENT IMMUNOMAGNETIC CAPTURE SYSTEM FOR ENTEROCOCCUS FAECALIS AND ENTEROCOCCUS FAECIUM

    EPA Science Inventory

    Enterococci detection is one of the two approved procedures by the US Environmental Protection Agency (EPA) used for the assessment of the microbiological quality of recreational waters. The action levels established by the EPA for enterococci are 35 pr 100 ml in marine recreati...

  19. Enterococcus populations in artisanal Manchego cheese: biodiversity, technological and safety aspects.

    PubMed

    Nieto-Arribas, Pedro; Seseña, Susana; Poveda, Justa M; Chicón, Rosa; Cabezas, Lourdes; Palop, Llanos

    2011-08-01

    Enterococci represent a considerable proportion of the microbiota in Manchego cheeses. In this study, a total of 132 enterococci isolated from good quality Manchego cheeses from two dairies at different ripening times were genotypically characterized and identified using molecular techniques. Representative isolates from the clusters obtained after genotyping were assayed for some enzymatic activities considered to have a potential role in cheese ripening, and for 2,3-butanedione and acetoin production, evaluation of odor intensity and appearance in milk and safety evaluation. Enterococcus faecalis was the predominant specie, accounting for 81.8% of the total isolates, while Enterococcus faecium, Enterococcus hirae and Enterococcus avium were present in low proportions. The number of genotypes involved at each ripening time varied both between dairies and with the ripening times; genotype E. faecalis Q1 being present in almost all the samples from both dairies. Eight isolates showed a higher proteolytic activity and 3 isolates produced high quantities of acetoin-diacetyl, for which reason they are interesting from a technological standpoint. A low antibiotic resistance was found and almost all the strains were susceptible to clinically important antibiotics. On the contrary, only four isolates (E. faecalis C4W1 and N0W5, and E. faecium N32W1 and C16W2) did not harbor some of the virulence genes assayed. PMID:21569931

  20. Identification and antimicrobial resistance of Enterococcus spp. isolated from the river and coastal waters in northern Iran.

    PubMed

    Alipour, Majid; Hajiesmaili, Reza; Talebjannat, Maryam; Yahyapour, Yousef

    2014-01-01

    As fecal streptococci commonly inhabit the intestinal tract of humans and warm blooded animals, and daily detection of all pathogenic bacteria in coastal water is not practical, thus these bacteria are used to detect the fecal contamination of water. The present study examined the presence and the antibiotic resistance patterns of Enterococcus spp. isolated from the Babolrud River in Babol and coastal waters in Babolsar. Seventy samples of water were collected in various regions of the Babolrud and coastal waters. Isolated bacteria were identified to the species level using standard biochemical tests and PCR technique. In total, 70 Enterococcus spp. were isolated from the Babolrud River and coastal waters of Babolsar. Enterococcus faecalis (68.6%) and Enterococcus faecium (20%) were the most prevalent species. Resistance to chloramphenicol, ciprofloxacin, and tetracyclin was prevalent. The presence of resistant Enterococcus spp. in coastal waters may transmit resistant genes to other bacteria; therefore, swimming in such environments is not suitable.

  1. Australian Group on Antimicrobial Resistance Enterococcus Surveillance Programme annual report, 2010.

    PubMed

    Coombs, Geoffrey W; Pearson, Julie C; Christiansen, Keryn; Gottlieb, Thomas; Bell, Jan M; George, Narelle; Turnidge, John D

    2013-09-01

    In 2010, 15 institutions around Australia conducted a period prevalence study of key resistances in isolates of Enterococcus species associated with a range of clinical disease amongst in- and outpatients. Each institution collected up to 100 consecutive isolates and tested these for susceptibility to commonly used antimicrobials using standardised methods. Vancomycin-resistant Enterococcus faecium and Enterococcus faecalis were characterised by pulsed-field gel electrophoresis. Multilocus sequence typing was performed on representative pulsotypes of E. faecium. Susceptibility results were compared with similar surveys conducted in 1995, 1999, 2003, 2005, 2007 and 2009. In the 2010 survey, E. faecalis (1,201 isolates) and E. faecium (170 isolates) made up 98.9% of the 1,386 isolates tested. Ampicillin resistance was very common (85.3%) in E. faecium and absent in E. faecalis. Non-susceptibility to vancomycin was 36.5% in E. faecium (similar to the 35.2% in 2009 but up from 15.4% in the 2007 survey) and 0.5% in E. faecalis. There were significant differences in the proportion of vancomycin-resistant E. faecium between the states ranging from 0% in Western Australia to 54.4% in South Australia. The vanB gene was detected in 62 E. faecium and 3 E. faecalis isolates. The vanA gene was detected in 1 E. faecium isolate. All vancomycin-resistant E. faecium belonged to clonal complex 17. The most common sequence type (ST) was ST203, which was found in all regions that had reports of vancomycin resistant enterococci. ST341 was detected only in New South Wales/Australian Capital Territory and ST414 only in South Australia and Victoria. High-level resistance to gentamicin was 34.1% in E. faecalis and 66.1% in E. faecium. A subset of isolates was tested against high-level streptomycin, linezolid and quinupristin/dalfopristin. High-level streptomycin resistance was found in 8.2% of E. faecalis isolates and 43.8% of E. faecium isolates. Linezolid non

  2. Prevalence and antibiotic resistance of Enterococcus strains isolated from poultry.

    PubMed

    Stępień-Pyśniak, Dagmara; Marek, Agnieszka; Banach, Tomasz; Adaszek, Łukasz; Pyzik, Ewelina; Wilczyński, Jarosław; Winiarczyk, Stanisław

    2016-06-01

    The aim of this study was to evaluate the frequency of occurrence of bacteria of the genus Enterococcus in poultry, to identify them by means of matrixassisted laser desorption/ionisation time-of-flight mass spectrometry (MALDITOF MS), and to analyse the antimicrobial susceptibility of the isolated strains to the drugs most frequently used in poultry. The material for the bacteriological tests was obtained mainly from the heart (97%) of the birds investigated. Of a total of 2,970 samples tested, 911 (30.7%) tested positive for Enterococcus spp. Enterococci were detected in broilers (88.1%), laying hens (5.3%), turkeys (3.9%), breeding hens (2.2%), and geese (0.4%). The most commonly identified species were Enterococcus (E.) faecalis (74.7%), E. faecium (10.1%), E. gallinarum (5.5%), E. hirae (4.6%), and E. cecorum (4.1%). The most frequent resistance properties were resistance to sulphamethoxazole/trimethoprim (88%), tylosin (71.4%), enrofloxacin (69.4%), doxycycline (67.3%), and lincomycin/spectinomycin (56.1%). Only one vancomycin-resistant Enterococcus, E. cecorum from a broiler, was found. PMID:27342087

  3. Prevalence and antibiotic resistance of Enterococcus strains isolated from poultry.

    PubMed

    Stępień-Pyśniak, Dagmara; Marek, Agnieszka; Banach, Tomasz; Adaszek, Łukasz; Pyzik, Ewelina; Wilczyński, Jarosław; Winiarczyk, Stanisław

    2016-06-01

    The aim of this study was to evaluate the frequency of occurrence of bacteria of the genus Enterococcus in poultry, to identify them by means of matrixassisted laser desorption/ionisation time-of-flight mass spectrometry (MALDITOF MS), and to analyse the antimicrobial susceptibility of the isolated strains to the drugs most frequently used in poultry. The material for the bacteriological tests was obtained mainly from the heart (97%) of the birds investigated. Of a total of 2,970 samples tested, 911 (30.7%) tested positive for Enterococcus spp. Enterococci were detected in broilers (88.1%), laying hens (5.3%), turkeys (3.9%), breeding hens (2.2%), and geese (0.4%). The most commonly identified species were Enterococcus (E.) faecalis (74.7%), E. faecium (10.1%), E. gallinarum (5.5%), E. hirae (4.6%), and E. cecorum (4.1%). The most frequent resistance properties were resistance to sulphamethoxazole/trimethoprim (88%), tylosin (71.4%), enrofloxacin (69.4%), doxycycline (67.3%), and lincomycin/spectinomycin (56.1%). Only one vancomycin-resistant Enterococcus, E. cecorum from a broiler, was found.

  4. Experimental study of the impact of antimicrobial treatments on Campylobacter, Enterococcus and PCR-capillary electrophoresis single-strand conformation polymorphism profiles of the gut microbiota of chickens.

    PubMed

    Mourand, Gwenaëlle; Jouy, Eric; Bougeard, Stéphanie; Dheilly, Alexandra; Kérouanton, Annaëlle; Zeitouni, Salman; Kempf, Isabelle

    2014-11-01

    An experiment was conducted to compare the impact of antimicrobial treatments on the susceptibility of Campylobacter, Enterococcus faecium and Enterococcus faecalis, and on the diversity of broiler microbiota. Specific-pathogen-free chickens were first orally inoculated with strains of Campylobacter and Enterococcus faecium. Birds were then orally treated with recommended doses of oxytetracycline, sulfadimethoxine/trimethoprim, amoxicillin or enrofloxacin. Faecal samples were collected before, during and after antimicrobial treatment. The susceptibility of Campylobacter, Enterococcus faecium and Enterococcus faecalis strains isolated on supplemented or non-supplemented media was studied and PCR-capillary electrophoresis single-strand conformation polymorphism (CE-SSCP) profiles of the gut microbiota were analysed. Enrofloxacin-resistant Campylobacter were selected in the enrofloxacin-treated group and showed the Thr86Ile mutation in the gyrA gene. Acquisition of the tetO gene in Campylobacter coli isolates was significantly more frequent in birds given oxytetracycline. No impact of amoxicillin treatment on the susceptibility of Campylobacter could be detected. Ampicillin- and sulfadimethoxine/trimethoprim-resistant Enterococcus faecium were selected in amoxicillin-treated broilers, but no selection of the inoculated vancomycin-resistant Enterococcus faecium could be detected, although it was also resistant to tetracycline and sulfadimethoxine/trimethoprim. PCR-CE-SSCP revealed significant variations in a few peaks in treated birds as compared with non-treated chickens. In conclusion, antimicrobial treatments perturbed chicken gut microbiota, and certain antimicrobial treatments selected or co-selected resistant strains of Campylobacter and Enterococcus.

  5. Transferable multiresistance plasmids carrying cfr in Enterococcus spp. from swine and farm environment.

    PubMed

    Liu, Yang; Wang, Yang; Schwarz, Stefan; Li, Yun; Shen, Zhangqi; Zhang, Qijing; Wu, Congming; Shen, Jianzhong

    2013-01-01

    Seventy-seven porcine Enterococcus isolates with florfenicol MICs of ≥16 μg of were/ml screened for the presence of the multiresistance gene cfr, its location on plasmids, and its genetic environment. Three isolates-Enterococcus thailandicus 3-38 (from a porcine rectal swab collected at a pig farm), Enterococcus thailandicus W3, and Enterococcus faecalis W9-2 (the latter two from sewage at a different farm), carried the cfr gene. The SmaI pulsed-field gel electrophoresis patterns of the three isolates differed distinctly. In addition, E. faecalis W9-2 was assigned to a new multilocus sequence type ST469. Mating experiments and Southern blot analysis indicated that cfr is located on conjugative plasmids pW3 (∼75 kb) from E. thailandicus W3, p3-38 (∼72 kb) from E. thailandicus 3-38, and pW9-2 (∼55 kb) from E. faecalis W9-2; these plasmids differed in their sizes, additional resistance genes, and the analysis of the segments encompassing the cfr gene. Sequence analysis revealed that all plasmids harbored a 4,447-bp central region, in which cfr was bracketed by two copies of the novel insertion sequence ISEnfa4 located in the same orientation. The sequences flanking the central regions of these plasmids, including the partial tra gene regions and a ω-ε-ζ toxin-antitoxin module, exhibited >95% nucleotide sequence identity to the conjugative plasmid pAMβ1 from E. faecalis. Conjugative plasmids carrying cfr appear to play an important role in the dissemination and maintenance of the multiresistance gene cfr among enterococcal isolates and possibly other species of Gram-positive bacteria.

  6. Transferable multiresistance plasmids carrying cfr in Enterococcus spp. from swine and farm environment.

    PubMed

    Liu, Yang; Wang, Yang; Schwarz, Stefan; Li, Yun; Shen, Zhangqi; Zhang, Qijing; Wu, Congming; Shen, Jianzhong

    2013-01-01

    Seventy-seven porcine Enterococcus isolates with florfenicol MICs of ≥16 μg of were/ml screened for the presence of the multiresistance gene cfr, its location on plasmids, and its genetic environment. Three isolates-Enterococcus thailandicus 3-38 (from a porcine rectal swab collected at a pig farm), Enterococcus thailandicus W3, and Enterococcus faecalis W9-2 (the latter two from sewage at a different farm), carried the cfr gene. The SmaI pulsed-field gel electrophoresis patterns of the three isolates differed distinctly. In addition, E. faecalis W9-2 was assigned to a new multilocus sequence type ST469. Mating experiments and Southern blot analysis indicated that cfr is located on conjugative plasmids pW3 (∼75 kb) from E. thailandicus W3, p3-38 (∼72 kb) from E. thailandicus 3-38, and pW9-2 (∼55 kb) from E. faecalis W9-2; these plasmids differed in their sizes, additional resistance genes, and the analysis of the segments encompassing the cfr gene. Sequence analysis revealed that all plasmids harbored a 4,447-bp central region, in which cfr was bracketed by two copies of the novel insertion sequence ISEnfa4 located in the same orientation. The sequences flanking the central regions of these plasmids, including the partial tra gene regions and a ω-ε-ζ toxin-antitoxin module, exhibited >95% nucleotide sequence identity to the conjugative plasmid pAMβ1 from E. faecalis. Conjugative plasmids carrying cfr appear to play an important role in the dissemination and maintenance of the multiresistance gene cfr among enterococcal isolates and possibly other species of Gram-positive bacteria. PMID:23070165

  7. Molecular Screening of Enterococcus Virulence Determinants and Potential for Genetic Exchange between Food and Medical Isolates

    PubMed Central

    Eaton, Tracy J.; Gasson, Michael J.

    2001-01-01

    Enterococci are used as starter and probiotic cultures in foods, and they occur as natural food contaminants. The genus Enterococcus is of increased significance as a cause of nosocomial infections, and this trend is exacerbated by the development of antibiotic resistance. In this study, we investigated the incidence of known virulence determinants in starter, food, and medical strains of Enterococcus faecalis, E. faecium, and E. durans. PCR and gene probe strategies were used to screen enterococcal isolates from both food and medical sources. Different and distinct patterns of incidence of virulence determinants were found for the E. faecalis and E. faecium strains. Medical E. faecalis strains had more virulence determinants than did food strains, which, in turn, had more than did starter strains. All of the E. faecalis strains tested possessed multiple determinants (between 6 and 11). E. faecium strains were generally free of virulence determinants, with notable exceptions. Significantly, esp and gelE determinants were identified in E. faecium medical strains. These virulence determinants have not previously been identified in E. faecium strains and may result from regional differences or the evolution of pathogenic E. faecium. Phenotypic testing revealed the existence of apparently silent gelE and cyl genes. In E. faecalis, the trend in these silent genes mirrors that of the expressed determinants. The potential for starter strains to acquire virulence determinants by natural conjugation mechanisms was investigated. Transconjugation in which starter strains acquired additional virulence determinants from medical strains was demonstrated. In addition, multiple pheromone-encoding genes were identified in both food and starter strains, indicating their potential to acquire other sex pheromone plasmids. These results suggest that the use of Enterococcus spp. in foods requires careful safety evaluation. PMID:11282615

  8. Draft Genome Sequences of Pseudomonas aeruginosa Isolates from Wounded Military Personnel.

    PubMed

    Arivett, Brock A; Ream, Dave C; Fiester, Steven E; Kidane, Destaalem; Actis, Luis A

    2016-08-11

    Pseudomonas aeruginosa, a Gram-negative bacterium that causes severe hospital-acquired infections, is grouped as an ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogen because of its extensive drug resistance phenotypes and effects on human health worldwide. Five multidrug resistant P. aeruginosa strains isolated from wounded military personnel were sequenced and annotated in this work.

  9. Draft Genome Sequences of Pseudomonas aeruginosa Isolates from Wounded Military Personnel

    PubMed Central

    Arivett, Brock A.; Ream, Dave C.; Fiester, Steven E.; Kidane, Destaalem

    2016-01-01

    Pseudomonas aeruginosa, a Gram-negative bacterium that causes severe hospital-acquired infections, is grouped as an ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogen because of its extensive drug resistance phenotypes and effects on human health worldwide. Five multidrug resistant P. aeruginosa strains isolated from wounded military personnel were sequenced and annotated in this work. PMID:27516516

  10. Draft Genome Sequences of Pseudomonas aeruginosa Isolates from Wounded Military Personnel.

    PubMed

    Arivett, Brock A; Ream, Dave C; Fiester, Steven E; Kidane, Destaalem; Actis, Luis A

    2016-01-01

    Pseudomonas aeruginosa, a Gram-negative bacterium that causes severe hospital-acquired infections, is grouped as an ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogen because of its extensive drug resistance phenotypes and effects on human health worldwide. Five multidrug resistant P. aeruginosa strains isolated from wounded military personnel were sequenced and annotated in this work. PMID:27516516

  11. Prevalence and Antimicrobial Resistance of Enterococcus Species: A Hospital-Based Study in China

    PubMed Central

    Jia, Wei; Li, Gang; Wang, Wen

    2014-01-01

    Objective: to investigate the prevalence and antimicrobial resistance of Enterococcus species isolated from a university hospital, and explore the mechanisms underlying the antimicrobial resistance, so as to provide clinical evidence for the inappropriate clinical use of antimicrobial agents and the control and prevention of enterococcal infections. Methods: a total of 1,157 enterococcal strains isolated from various clinical specimens from January 2010 to December 2012 in the General Hospital of Ningxia Medical University were identified to species level with a VITEK-2 COMPACT fully automated microbiological system, and the antimicrobial susceptibility of Enterococcus species was determined using the Kirby-Bauer disc diffusion method. The multiple-drug resistant enterococcal isolates were screened from the clinical isolates of Enterococcus species from the burns department. The minimal inhibitory concentration (MIC) of Enterococcus species to the three fluoroquinolones, including ciprofloxacin, gatifloxacin and levofloxacin was determined with the agar dilution method, and the changes in the MIC of Enterococcus species to the three fluoroquinolones following reserpine treatment were evaluated. The β-lactam, aminoglycoside, tetracycline, macrolide, glycopeptide resistance genes and the efflux pump emeA genes were detected in the enterococcal isolates using a polymerase chain reaction (PCR) assay. Results: the 1,157 clinical isolates of Enterococcus species included 679 E. faecium isolates (58.7%), 382 E. faecalis isolates (33%), 26 E. casseliflavus isolates (2.2%), 24 E. avium isolates (2.1%), and 46 isolates of other Enterococcus species (4%). The prevalence of antimicrobial resistance varied significantly between E. faecium and E. faecalis, and ≤1.1% of these two Enterococcus species were found to be resistant to vancomycin, teicoplanin or linezolid. In addition, the Enterococcus species isolated from different departments of the hospital exhibited various

  12. Comparative Evaluation of Antimicrobial Efficacy of MTAD, 3% NaOCI and Propolis Against E Faecalis

    PubMed Central

    Dhanu; Chandra, Prakash; Anandakrishna, Latha; Dhananjaya

    2010-01-01

    Aim The present study sought to compare the antimicrobial efficacy of 3% NaOCl, Biopure MTAD (Tulsa Dentsply, Tulsa, OK) and Brazilian ethanolic extract of propolis (EEP) against Enterococcus faecalis (E. faecalis). Methodology The study utilized 55 extracted human permanent teeth with single root canal. The samples were decoronated, instrumented and sterilized. The teeth were infected with E faecalis for 48 hours. The teeth were divided randomly into 3 groups according to the irrigants used and kept in contact with the respective irrigant for 5 minutes. All the samples were incubated in brain heart infusion (BHI) broth for 96 hours. Disinfection of the samples was determined based on presence or absence of turbidity in the BHI broth 96 hours later. Statistical analysis was done using Chi-square test. Results All the samples treated with MTAD showed complete absence of turbidity, while all the 15 teeth treated with propolis showed presence of turbidity, 8 out of 15 teeth treated with NaOCl showed presence of turbidity. Statistical analysis of the data using chi-square test showed significant difference between the groups (P < 0.05). Conclusion The study concluded that MTAD was more effective than 3% NaOCl and propolis against E. faecalis.

  13. Study of invasion and colonization of E. faecalis in microtubes by a novel device.

    PubMed

    Sun, Xiaoqiang; Wang, Shujing; Yang, Yue; Luo, Chunxiong; Hou, Benxiang

    2016-10-01

    Enterococcus faecalis (E. faecalis) is a species that has frequently been isolated from root canal of patients suffering from persistent periodontitis. To a great degree, the resistance of E. faecalis to irrigating solutions and intracanal medicaments is due to its invasion into the dentinal tubules. In this study, we developed a device to observe the dynamic process of the bacterial invasion into microtubes. According to the diameter of the dentinal tubules and other microstructures in the root canals, we designed four different size microtubes with different lengths in this device. As expected, E. faecalis is able to steadily grow in this device and penetrate into the microtubes, and a continuous observation is achieved. We found that the depth and speed of bacterial penetration, the extent of colonization and the arrangement of the bacteria in the microtubes are strongly influenced by the size of the microtube. The length of the microtube also influences the speed and depth of the bacterial invasion. Bacteria in microtubes with a similar diameter to the real dentinal tubules showed a discontinuous distribution, which is consistent with the final bacterial distribution in the native dentinal tubules. Considering the device's advantages such as its ability to provide real-time observations, its ability to be modified as necessary, and its standardized operation, it has great potential to be widely used as a platform for the observation of the interaction of different bacteria during an invasion course and to test the efficacy of new antibacterial agents in dentistry. PMID:27540728

  14. Identification of Multiple Bacteriocins in Enterococcus spp. Using an Enterococcus-Specific Bacteriocin PCR Array

    PubMed Central

    Henning, Chris; Gautam, Dhiraj; Muriana, Peter

    2015-01-01

    Twenty-two bacteriocin-producing Enterococcus isolates obtained from food and animal sources, and demonstrating activity against Listeria monocytogenes, were screened for bacteriocin-related genes using a bacteriocin PCR array based on known enterococcal bacteriocin gene sequences in the NCBI GenBank database. The 22 bacteriocin-positive (Bac+) enterococci included En. durans (1), En. faecalis (4), En. faecium (12), En. hirae (3), and En. thailandicus (2). Enterocin A (entA), enterocins mr10A and mr10B (mr10AB), and bacteriocin T8 (bacA) were the most commonly found structural genes in order of decreasing prevalence. Forty-five bacteriocin genes were identified within the 22 Bac+ isolates, each containing at least one of the screened structural genes. Of the 22 Bac+ isolates, 15 possessed two bacteriocin genes, seven isolates contained three different bacteriocins, and three isolates contained as many as four different bacteriocin genes. These results may explain the high degree of bactericidal activity observed with various Bac+ Enterococcus spp. Antimicrobial activity against wild-type L. monocytogenes and a bacteriocin-resistant variant demonstrated bacteriocins having different modes-of-action. Mixtures of bacteriocins, especially those with different modes-of-action and having activity against foodborne pathogens, such as L. monocytogenes, may play a promising role in the preservation of food.

  15. Prevalence and antibiotic resistance of Enterococcus spp. isolated from retail cheese, ready-to-eat salads, ham, and raw meat.

    PubMed

    Pesavento, G; Calonico, C; Ducci, B; Magnanini, A; Lo Nostro, A

    2014-08-01

    Food specimens were analyzed in order to research Enterococcus spp.: 636 samples of raw meat (227 beef, 238 poultry, and 171 pork), 278 samples of cheese (110 fresh soft cheese and 168 mozzarella cheese), 214 samples of ready-to-eat salads, and 187 samples of ham. 312 strains of Enterococcus spp samples were isolated, then identified and submitted to susceptibility tests against 11 antimicrobial agents. The predominant species were Enterococcus faecalis in raw meat and Enterococcus faecium in retail products. Low percentages of microorganisms were resistant to vancomycin (3.53%), teicoplanin (2.24%), linezolid (0.32%), and amoxicillin in combination with clavulanic acid (0.32%). A high percentage of resistance was noted in E. faecalis at high level gentamicin (21.9%) and tetracycline (60.6%). In general, strains of E. faecalis were more resistant than E. faecium. Enterococci should be considered not only potential pathogens, but also a reservoir of genes encoding antibiotic resistance which can be transferred to other microorganisms. Continuous monitoring of their incidence and emerging resistance is important in order to identify foods which potentially represent a real risk to the population, and to ensure effective treatment of human enterococcal infections. PMID:24750807

  16. Prevalence and antibiotic resistance of Enterococcus spp. isolated from retail cheese, ready-to-eat salads, ham, and raw meat.

    PubMed

    Pesavento, G; Calonico, C; Ducci, B; Magnanini, A; Lo Nostro, A

    2014-08-01

    Food specimens were analyzed in order to research Enterococcus spp.: 636 samples of raw meat (227 beef, 238 poultry, and 171 pork), 278 samples of cheese (110 fresh soft cheese and 168 mozzarella cheese), 214 samples of ready-to-eat salads, and 187 samples of ham. 312 strains of Enterococcus spp samples were isolated, then identified and submitted to susceptibility tests against 11 antimicrobial agents. The predominant species were Enterococcus faecalis in raw meat and Enterococcus faecium in retail products. Low percentages of microorganisms were resistant to vancomycin (3.53%), teicoplanin (2.24%), linezolid (0.32%), and amoxicillin in combination with clavulanic acid (0.32%). A high percentage of resistance was noted in E. faecalis at high level gentamicin (21.9%) and tetracycline (60.6%). In general, strains of E. faecalis were more resistant than E. faecium. Enterococci should be considered not only potential pathogens, but also a reservoir of genes encoding antibiotic resistance which can be transferred to other microorganisms. Continuous monitoring of their incidence and emerging resistance is important in order to identify foods which potentially represent a real risk to the population, and to ensure effective treatment of human enterococcal infections.

  17. Performance of Vitek 2 for antimicrobial susceptibility testing of Staphylococcus spp. and Enterococcus spp.

    PubMed

    Bobenchik, April M; Hindler, Janet A; Giltner, Carmen L; Saeki, Sandra; Humphries, Romney M

    2014-02-01

    Vitek 2 (bioMérieux, Inc., Durham, NC) is a widely used commercial antimicrobial susceptibility testing system. We compared MIC results obtained by Vitek 2 to those obtained by the Clinical and Laboratory Standards Institute (CLSI) broth microdilution (BMD) reference method for 134 staphylococcal and 84 enterococcal clinical isolates. Nineteen agents were evaluated, including all those available on Vitek 2 for testing staphylococci and enterococci. The resistance phenotypes tested included methicillin-resistant Staphylococcus aureus (MRSA) (n = 58), S. aureus with inducible clindamycin resistance (ICR) (n = 30), trimethoprim-sulfamethoxazole-resistant MRSA (n = 10), vancomycin-resistant Enterococcus (n = 37), high-level gentamicin-resistant Enterococcus (n = 15), linezolid-resistant Enterococcus (n = 5), and daptomycin-nonsusceptible Enterococcus faecalis (n = 6). For the staphylococci, there was 98.9% categorical agreement (CA). There was one very major error (VME) for gentamicin in a Staphylococcus hominis isolate, six VMEs for inducible clindamycin in S. aureus isolates, and two major errors (ME) for daptomycin in an S. aureus and a Staphylococcus epidermidis isolate. For enterococci, there was 97.3% CA. Two VMEs were observed for daptomycin in isolates of E. faecalis and 2 ME, 1 for high-level gentamicin resistance and 1 for nitrofurantoin, in E. faecium isolates. Overall, there was 98.3% CA and 99% essential agreement for the testing of staphylococci and enterococci by the Vitek 2. With the exception of detecting ICR in S. aureus, Vitek 2 performed reliably for antimicrobial susceptibility testing of staphylococci and enterococci.

  18. The life and times of the Enterococcus.

    PubMed Central

    Murray, B E

    1990-01-01

    Enterococci are important human pathogens that are increasingly resistant to antimicrobial agents. These organisms were previously considered part of the genus Streptococcus but have recently been reclassified into their own genus, called Enterococcus. To date, 12 species pathogenic for humans have been described, including the most common human isolates, Enterococcus faecalis and E. faecium. Enterococci cause between 5 and 15% of cases of endocarditis, which is best treated by the combination of a cell wall-active agent (such as penicillin or vancomycin, neither of which alone is usually bactericidal) and an aminoglycoside to which the organism is not highly resistant; this characteristically results in a synergistic bactericidal effect. High-level resistance (MIC, greater than or equal to 2,000 micrograms/ml) to the aminoglycoside eliminates the expected bactericidal effect, and such resistance has now been described for all aminoglycosides. Enterococci can also cause urinary tract infections; intraabdominal, pelvic, and wound infections; superinfections (particularly in patients receiving expanded-spectrum cephalosporins); and bacteremias (often together with other organisms). They are now the third most common organism seen in nosocomial infections. For most of these infections, single-drug therapy, most often with penicillin, ampicillin, or vancomycin, is adequate. Enterococci have a large number of both inherent and acquired resistance traits, including resistance to cephalosporins, clindamycin, tetracycline, and penicillinase-resistant penicillins such as oxacillin, among others. The most recent resistance traits reported are penicillinase resistance (apparently acquired from staphylococci) and vancomycin resistance, both of which can be transferred to other enterococci. It appears likely that we will soon be faced with increasing numbers of enterococci for which there is no adequate therapy. PMID:2404568

  19. Enterococcus rivorum sp. nov., from water of pristine brooks.

    PubMed

    Niemi, R Maarit; Ollinkangas, Tuula; Paulin, Lars; Svec, Pavel; Vandamme, Peter; Karkman, Antti; Kosina, Marcel; Lindström, Kristina

    2012-09-01

    A significant number of Enterococcus strains from pristine waters of two brooks in Finland formed a distinct cluster on the basis of whole-cell protein fingerprinting by one-dimensional SDS-PAGE. The strains shared the following characteristics. Cells were ovoid, Gram-positive-staining and non-spore-forming, appearing singly or in pairs or chains. They were facultatively anaerobic and catalase-negative. Growth in broth containing 6.5 % NaCl or at 45 °C was weak or absent. Production of D antigen was variable. The strains tolerated 60 °C for 30 min, 40 % bile and tellurite, hydrolysed aesculin strongly and gelatin weakly, produced no acid from hippurate and did not reduce it, grew weakly at 10 °C, showed a strong reaction for the Voges-Proskauer test and produced acid from methyl α-d-glucoside, mannitol, sorbitol and sucrose, with weak or no production of acid from methyl α-d-mannoside, l-arabinose, gluconate and l-xylose. Several of the strains were selected for identification on the basis of sequencing of almost the whole 16S rRNA gene and partial atpA and pheS genes and of (GTG)(5)-PCR fingerprints. Partial atpA and pheS gene sequencing was also performed for those type strains of Enterococcus species without available sequences in the database. The pristine brook isolates formed a novel species, for which the name Enterococcus rivorum sp. nov. (type strain S299(T) = HAMBI 3055(T) = LMG 25899(T) = CCM 7986(T)) is proposed. On the basis of 16S rRNA gene sequence similarity, E. rivorum sp. nov. is related to the Enterococcus faecalis genogoup. It is distinguished from described Enterococcus species on the basis of 16S rRNA, atpA and pheS gene sequences and whole-cell protein and (GTG)(5)-PCR fingerprints. It is most closely related to E. faecalis, but DNA-DNA hybridization confirms it to represent a novel species.

  20. Application of bacteriocinogenic Enterococcus mundtii CRL35 and Enterococcus faecium ST88Ch in the control of Listeria monocytogenes in fresh Minas cheese.

    PubMed

    Vera Pingitore, Esteban; Todorov, Svetoslav Dimitrov; Sesma, Fernando; Franco, Bernadette Dora Gombossy de Melo

    2012-10-01

    Several strains of Enterococcus spp. are capable of producing bacteriocins with antimicrobial activity against important bacterial pathogens in dairy products. In this study, the bacteriocins produced by two Enterococcus strains (Enterococcus mundtii CRL35 and Enterococcus faecium ST88Ch), isolated from cheeses, were characterized and tested for their capability to control growth of Listeria monocytogenes 426 in experimentally contaminated fresh Minas cheese during refrigerated storage. Both strains were active against a variety of pathogenic and non-pathogenic microorganisms and bacteriocin absorption to various L. monocytogenes, Enterococcus faecalis ATCC 19443 and Lactobacillus sakei ATCC 15521 varied according to the strain and the testing conditions (pH, temperature, presence of salts and surfactants). Growth of L. monocytogenes 426 was inhibited in cheeses containing E. mundtii CRL35 up to 12 days at 8 °C, evidencing a bacteriostatic effect. E. faecium ST88Ch was less effective, as the bacteriostatic affect occurred only after 6 days at 8 °C. In cheeses containing nisin (12.5 mg/kg), less than one log reduction was observed. This research underlines the potential application of E. mundtii CRL35 in the control of L. monocytogenes in Minas cheese.

  1. Speciation and frequency of virulence genes of Enterococcus spp. isolated from rainwater tank samples in Southeast Queensland, Australia.

    PubMed

    Ahmed, W; Sidhu, J P S; Toze, S

    2012-06-19

    In this study, 212 Enterococcus isolates from 23 rainwater tank samples in Southeast Queensland (SEQ), Australia were identified to the species level. The isolates were also tested for the presence of 6 virulence genes associated with Enterococcus related infections. Among the 23 rainwater tank samples, 20 (90%), 10 (44%), 7 (30%), 5 (22%), 4 (17%), 2 (9%), and 1 (4%) samples yielded E. faecalis, E. mundtii, E. casseliflavus, E. faecium, E. hirae, E. avium, and E. durans, respectively. Among the 6 virulence genes tested, gelE and efaA were most prevalent, detected in 19 (83%) and 18 (78%) of 23 rainwater tank samples, respectively. Virulence gene ace was also detected in 14 (61%) rainwater tank samples followed by AS, esp (E. faecalis variant), and cylA genes which were detected in 3 (13%), 2 (9%), and 1 (4%) samples, respectively. In all, 120 (57%) Enterococcus isolates from 20 rainwater tank samples harbored virulence genes. Among these tank water samples, Enterococcus spp. from 5 (25%) samples harbored a single virulence gene and 15 (75%) samples were harboring two or more virulence genes. The significance of these strains in terms of health implications remains to be assessed. The potential sources of these strains need to be identified for the improved management of captured rainwater quality. Finally, it is recommended that Enterococcus spp. should be used as an additional fecal indicator bacterium in conjunction with E. coli for the microbiological assessment of rainwater tanks.

  2. Macrolide, glycopeptide resistance and virulence genes in Enterococcus species isolates from dairy cattle.

    PubMed

    Iweriebor, Benson C; Obi, Larry C; Okoh, Anthony I

    2016-07-01

    The genus Enterococcus is known to possess the capacity to acquire and disseminate antimicrobial resistant determinants alongside the ability to produce various virulence genes that enables it to establish infections. We assessed the prevalence and antibiogram profiles of Enterococcus spp. in faecal samples of dairy cattle. Faecal swab samples were collected from 400 dairy cattle from two commercial cattle farms in two rural communities in the Eastern Cape, South Africa. Confirmation of enterococci isolates was carried out by PCR targeting of the tuf gene. Species delineation was by species-specific primers targeting the superoxide dismutase (sod A) gene in a multiplex PCR assay. Isolates were screened for the presence of the following virulence genes (ace, gel E, esp, efa A, cyl A and hyl E) and antimicrobial resistance determinants to erythromycin, vancomycin and streptomycin were evaluated molecularly. A total of 340 isolates were confirmed as belonging to the genus Enterococcus . Species distribution among the isolates consisted of Enterococcus faecium (52.94 %) and Enterococcus durans (23.53 %) in preponderance compared to the three other species, namely Enterococcus faecalis (8.8 %), Enterococcus hirae (8.6 %) and Enterococcus casseliflavus (5.9 %). All were resistant to vancomycin, while 99 % showed resistance to aminoglycoside and 94 % to macrolide. Three virulence genes (ace, gel E and esp) were detected in almost all the confirmed isolates. The resistance determinants van B (19.7 %), van C1 (25 %), van C2/3 (26.3 %) erm B (40.29 %) and str A (50.88 %) were detected among the isolates. A high prevalence of multidrug-resistant enterococci isolates was detected in this study and the genetic repertoire to survive in the presence of antimicrobial agents was present in these organisms.

  3. Macrolide, glycopeptide resistance and virulence genes in Enterococcus species isolates from dairy cattle.

    PubMed

    Iweriebor, Benson C; Obi, Larry C; Okoh, Anthony I

    2016-07-01

    The genus Enterococcus is known to possess the capacity to acquire and disseminate antimicrobial resistant determinants alongside the ability to produce various virulence genes that enables it to establish infections.