Removal of Escherichia coli and Enterococcus faecalis after Hand Washing with Antimicrobial and Nonantimicrobial Soap and Persistence of These Bacteria in Rinsates.

PubMed

Pérez-Garza, J; García, S; Heredia, N

2017-10-01

Food handlers are important sources of contamination in the agricultural environment. This study was conducted (i) to evaluate the activity of antimicrobial soaps against Escherichia coli and Enterococcus faecalis using a hand washing model with soiled hands and (ii) to determine the survival and persistence of these bacteria in rinsates. Sterilized agricultural soil from tomato and pepper farms was inoculated with E. coli or E. faecalis at 10 3 or 10 6 CFU/g. Decontaminated hands were placed in contact with contaminated soil for 2 min and were then washed with soaps with or without antimicrobial compounds (citric extracts, chloroxylenol, triclosan, or chlorhexidine gluconate). As the control, hands were washed with sterile distilled water. The levels of bacteria remaining on the hands and recovered from the rinsates were determined using a membrane filtration method and selective media. Antimicrobial soaps removed levels of E. coli similar to those removed by distilled water and nonantimicrobial soap on hands contaminated with E. coli at 10 3 CFU/g. However, when hands were contaminated with E. coli at 10 6 CFU/g, more E. coli was removed with the chlorhexidine gluconate soap. When hands were contaminated with E. faecalis at 10 3 CFU/g, bacteria were removed more effectively with soaps containing chloroxylenol or chlorhexidine gluconate. When hands were contaminated with E. faecalis at 10 6 CFU/g, all of the antimicrobial soaps were more effective for removing the bacteria than were distilled water and nonantimicrobial soap. E. coli grew in all of the hand washing rinsates except that containing triclosan, whereas E. faecalis from the 10 6 CFU/g treatments grew in rinsates containing chlorhexidine gluconate and in the distilled water rinsates. Washing with antimicrobial soap was more effective for reducing bacteria on soiled hands than was washing with water or nonantimicrobial soap. However, persistence or growth of bacteria in these rinsates poses health risks.

Electrophoretic analysis of cyanide depletion by Pseudomonas alcaligenes.

PubMed

Zaugg, S E; Davidson, R A; Walker, J C; Walker, E B

1997-02-01

Bacterial-facilitated depletion of cyanide is under development for remediation of heap leach operations in the gold mining industry. Capillary electrophoresis was found to be a powerful tool for quantifying cyanide depletion. Changes in cyanide concentration in aqueous suspensions of Pseudomonas alcaligenes bacteria and cyanide at elevated pH were easily monitored by capillary electrophoresis. The resulting data can be used to study rates of cyanide depletion by this strain of bacteria. Concentrations of these bacteria at 10(5) cells/mL were found to reduce cyanide from 100 ppm to less than 8 ppm in four days. In addition, other ions of interest in cyanide metabolism, such as formate, can be simultaneously analyzed. Direct UV detection of cyanide at 192 nm further simplifies the analytical method for these ions.

Pathogenic features of heterotrophic plate count bacteria from drinking-water boreholes.

PubMed

Horn, Suranie; Pieters, Rialet; Bezuidenhout, Carlos

2016-12-01

Evidence suggests that heterotrophic plate count (HPC) bacteria may be hazardous to humans with weakened health. We investigated the pathogenic potential of HPC bacteria from untreated borehole water, consumed by humans, for: their haemolytic properties, the production of extracellular enzymes such as DNase, proteinase, lipase, lecithinase, hyaluronidase and chondroitinase, the effect simulated gastric fluid has on their survival, as well as the bacteria's antibiotic-susceptible profile. HuTu-80 cells acted as model for the human intestine and were exposed to the HPC isolates to determine their effects on the viability of the cells. Several HPC isolates were α- or β-haemolytic, produced two or more extracellular enzymes, survived the SGF treatment, and showed resistance against selected antibiotics. The isolates were also harmful to the human intestinal cells to varying degrees. A novel pathogen score was calculated for each isolate. Bacillus cereus had the highest pathogen index: the pathogenicity of the other bacteria declined as follows: Aeromonas taiwanensis > Aeromonas hydrophila > Bacillus thuringiensis > Alcaligenes faecalis > Pseudomonas sp. > Bacillus pumilus > Brevibacillus sp. > Bacillus subtilis > Bacillus sp. These results demonstrated that the prevailing standards for HPCs in drinking water may expose humans with compromised immune systems to undue risk.

Biosorption of heavy metal ions on Rhodobacter sphaeroides and Alcaligenes eutrophus H16

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seki, Hideshi; Suzuki, Akira; Mitsueda, Shinichiro

1998-01-15

A fundamental study of the application of bacteria to the recovery of toxic heavy metals from aqueous environments was carried out. The biosorption characteristics of cadmium and lead ions were determined with purple nonsulfur bacteria, Rhodobacter sphaeroides and hydrogen bacteria, Alcaligenes eutrophus H16 that were inactivated by steam sterilization. A simplified version of the metal binding model proposed by Plette et al. was used for the description of meal binding data. The results showed that the biosorption of bivalent metal ions to whole cell bodies of the bacteria was due to monodentate binding to two different types of acidic sites:more » carboxilic and phosphatic-type sites. The number of metal binding sites of A. eutrophus was 2.4-fold larger than that of R. sphaeroides.« less

Molecular weight-dependent degradation of D-lactate-containing polyesters by polyhydroxyalkanoate depolymerases from Variovorax sp. C34 and Alcaligenes faecalis T1.

PubMed

Sun, Jian; Matsumoto, Ken'ichiro; Tabata, Yuta; Kadoya, Ryosuke; Ooi, Toshihiko; Abe, Hideki; Taguchi, Seiichi

2015-11-01

Polyhydroxyalkanoate depolymerase derived from Variovorax sp. C34 (PhaZVs) was identified as the first enzyme that is capable of degrading isotactic P[67 mol% (R)-lactate(LA)-co-(R)-3-hydroxybutyrate(3HB)] [P(D-LA-co-D-3HB)]. This study aimed at analyzing the monomer sequence specificity of PhaZVs for hydrolyzing P(LA-co-3HB) in comparison with a P(3HB) depolymerase from Alcaligenes faecalis T1 (PhaZAf) that did not degrade the same copolymer. Degradation of P(LA-co-3HB) by action of PhaZVs generated dimers, 3HB-3HB, 3HB-LA, LA-3HB, and LA-LA, and the monomers, suggesting that PhaZVs cleaved the linkages between LA and 3HB units and between LA units. To provide a direct evidence for the hydrolysis of these sequences, the synthetic methyl trimers, 3HB-3HB-3HB, LA-LA-3HB, LA-3HB-LA, and 3HB-LA-LA, were treated with the PhaZs. Unexpectedly, not only PhaZVs but also PhaZAf hydrolyzed all of these substrates, namely PhaZAf also cleaved LA-LA linkage. Considering the fact that both PhaZs did not degrade P[(R)-LA] (PDLA) homopolymer, the cleavage capability of LA-LA linkage by PhaZs was supposed to depend on the length of the LA-clustering region in the polymer chain. To test this hypothesis, PDLA oligomers (6 to 40 mer) were subjected to the PhaZ assay, revealing that there was an inverse relationship between molecular weight of the substrates and their hydrolysis efficiency. Moreover, PhaZVs exhibited the degrading activity toward significantly longer PDLA oligomers compared to PhaZAf. Therefore, the cleaving capability of PhaZs used here toward the D-LA-based polymers containing the LA-clustering region was strongly associated with the substrate length, rather than the monomer sequence specificity of the enzyme.

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

Strain of alcaligenes latus bacteria used for the decomposition of polychlorinated biphenyls

DOEpatents

Dyadischev, Nikolai Romanovich; Zharikov, Gennady Alekseevich; Kapranov, Vladimir Vladimirovich

2001-09-11

Alcaligenes latus bacterial strain TXD-13 VKPM B 75-05 is capable of degrading polychlorinated biphenyls (PCBs). The strain may be employed to detoxicate environment media and PCB-containing industrial waste. To produce biomass, the strain is incubated on media which contain carbon sources, nitrogen sources and mineral salts. The strain is cultivated by a subsurface method up to a titer from 6.0.multidot.10.sup.8 to 2.0.times.10.sup.9 cells per cu cm. The produced biomass is used for degrading PCBs in concentrations from 10.sup.7 to 10.sup.8 cells per cu cm. The strain ensures from 35 to 50% reduction in PCB content in soil and water.

Alcaligenes xylosoxidans endophthalmitis following phacoemulsification and intraocular lens implantation.

PubMed

Robert, Pierre-Yves; Chainier, Delphine; Garnier, Fabien; Ploy, Marie-Cécile; Parneix, Pierre; Adenis, Jean-Paul; Martin, Christian

2008-01-01

Five consecutive cases of endophthalmitis that developed after cataract extraction by a single surgeon using the same operating room during one morning session are described. Following preoperative topical administration of ciprofloxacin, surgery consisted of phacoemulsification with peristaltic pump and fluid venting, polymethylmethacrylate intraocular lens implantation, and corneal suture. No complications occurred during surgery. All five patients developed endophthalmitis caused by infection with Alcaligenes xylosoxidans in less than 24 hours. Pulsed-field gel electrophoresis was used to prove similarity between strains. Bacterial inquiry on contamination of the operating room environment revealed massive colonization of phacoemulsifier irrigation channels by Pseudomonas fluorescens bacteria from an unestablished source. Four of the five patients ultimately recovered visual acuity better than 20/60.

Heterologous expression of glycopeptide resistance vanHAX gene clusters from soil bacteria in Enterococcus faecalis.

PubMed

Hasman, Henrik; Aarestrup, Frank M; Dalsgaard, Anders; Guardabassi, Luca

2006-04-01

The aim of the study was to determine whether glycopeptide resistance gene clusters from soil bacteria could be heterologously expressed in Enterococcus faecalis and adapt to the new host following exposure to vancomycin. The vanHAX clusters from Paenibacillus thiaminolyticus PT-2B1, Paenibacillus apiarius PA-B2B and Amycolatopsis coloradensis DSM 44225 were separately cloned in an appropriately constructed shuttle vector containing the two-component regulatory system (vanRS) of Tn1546. The complete vanA(PT) operon (vanRSHAXY) from P. thiaminolyticus PT-2B1 was cloned in the same shuttle vector lacking enterococcal vanRS. All plasmid constructs were electroporated into E. faecalis JH2-2 and the MICs of vancomycin and teicoplanin were determined for each recombinant strain before and following exposure to sublethal concentrations of vancomycin. The vanHAX clusters from P. thiaminolyticus and P. apiarius conferred high-level vancomycin resistance (MIC > or = 125 mg/L) in E. faecalis JH2-2. In contrast, cloning of the vanHAX cluster from A. coloradensis did not result in a significant increase of vancomycin resistance (MIC = 0.7 mg/L). Resistance to vancomycin was not observed after cloning the complete vanA(PT) operon from P. thiaminolyticus (MIC = 2 mg/L), but this recombinant rapidly adapted to high concentrations of vancomycin (MIC = 500 mg/L) following exposure to sub-lethal concentrations of this antibiotic. The results showed that vanA(PT) in P. thiaminolyticus is a possible ancestor of vanA-mediated glycopeptide resistance in enterococci. Experimental evidence supported the hypothesis that enterococci did not acquire glycopeptide resistance directly from glycopeptide-producing organisms such as A. coloradensis.

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

PubMed

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 (10(3) CFU/g)) and E. faecalis CECT7121 (10(4) 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.

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

Influence of irrigation regimens on the adherence of Enterococcus faecalis to root canal dentin.

PubMed

Kishen, Anil; Sum, Chee-Peng; Mathew, Shibi; Lim, Chwee-Teck

2008-07-01

Enterococcus faecalis is frequently associated with post-treatment endodontic infections. Because adherence of bacteria to a substrate is the earliest stage in biofilm formation, eliciting the factors that links adherence of this bacterium to dentin would help in understanding its association with treatment-failed root canals. This investigation aimed to study the effects of endodontic irrigants on the adherence of E. faecalis to dentin. The bacteria adherence assay was conducted by using fluorescence microscopy, and the adhesion force was measured by using atomic force microscopy. There were significant increases in adherence and adhesion force after irrigation of dentin with ethylenediaminetetraacetic acid (EDTA), whereas sodium hypochlorite (NaOCl) reduced it. With the use of chlorhexidine (CHX), the force of adhesion increased, but the adherence assay showed a reduction in the number of adhering bacteria. The irrigation regimen of EDTA, NaOCl, and CHX resulted in the least number of adhering E. faecalis cells. This study highlighted that chemicals that alter the physicochemical properties of dentin will influence the nature of adherence, adhesion force, and subsequent biofilm formation of E. faecalis to dentin.

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

Enterococcus faecalis Constitutes an Unusual Bacterial Model in Lysozyme Resistance▿

PubMed Central

Hébert, Laurent; Courtin, Pascal; Torelli, Riccardo; Sanguinetti, Maurizio; Chapot-Chartier, Marie-Pierre; Auffray, Yanick; Benachour, Abdellah

2007-01-01

Lysozyme is an important and widespread compound of the host constitutive defense system, and it is assumed that Enterococcus faecalis is one of the few bacteria that are almost completely lysozyme resistant. On the basis of the sequence analysis of the whole genome of E. faecalis V583 strain, we identified two genes that are potentially involved in lysozyme resistance, EF_0783 and EF_1843. Protein products of these two genes share significant homology with Staphylococcus aureus peptidoglycan O-acetyltransferase (OatA) and Streptococcus pneumoniae N-acetylglucosamine deacetylase (PgdA), respectively. In order to determine whether EF_0783 and EF_1843 are involved in lysozyme resistance, we constructed their corresponding mutants and a double mutant. The ΔEF_0783 mutant and ΔEF_0783 ΔEF_1843 double mutant were shown to be more sensitive to lysozyme than the parental E. faecalis JH2-2 strain and ΔEF_1843 mutant were. However, compared to other bacteria, such as Listeria monocytogenes or S. pneumoniae, the tolerance of ΔEF_0783 and ΔEF_0783 ΔEF_1843 mutants towards lysozyme remains very high. Peptidoglycan structure analysis showed that EF_0783 modifies the peptidoglycan by O acetylation of N-acetyl muramic acid, while the EF_1843 deletion has no obvious effect on peptidoglycan structure under the same conditions. Moreover, the EF_0783 and EF_1843 deletions seem to significantly affect the ability of E. faecalis to survive within murine macrophages. In all, while EF_0783 is currently involved in the lysozyme resistance of E. faecalis, peptidoglycan O acetylation and de-N-acetylation are not the main mechanisms conferring high levels of lysozyme resistance to E. faecalis. PMID:17785473

Characterization of an Indole-3-Acetamide Hydrolase from Alcaligenes faecalis subsp. parafaecalis and Its Application in Efficient Preparation of Both Enantiomers of Chiral Building Block 2,3-Dihydro-1,4-Benzodioxin-2-Carboxylic Acid.

PubMed

Mishra, Pradeep; Kaur, Suneet; Sharma, Amar Nath; Jolly, Ravinder S

2016-01-01

Both the enantiomers of 2,3-dihydro-1,4-benzodioxin-2-carboxylic acid are valuable chiral synthons for enantiospecific synthesis of therapeutic agents such as (S)-doxazosin mesylate, WB 4101, MKC 242, 2,3-dihydro-2-hydroxymethyl-1,4-benzodioxin, and N-[2,4-oxo-1,3-thiazolidin-3-yl]-2,3-dihydro-1,4-benzodioxin-2-carboxamide. Pharmaceutical applications require these enantiomers in optically pure form. However, currently available methods suffer from one drawback or other, such as low efficiency, uncommon and not so easily accessible chiral resolving agent and less than optimal enantiomeric purity. Our interest in finding a biocatalyst for efficient production of enantiomerically pure 2,3-dihydro-1,4-benzodioxin-2-carboxylic acid lead us to discover an amidase activity from Alcaligenes faecalis subsp. parafaecalis, which was able to kinetically resolve 2,3-dihydro-1,4-benzodioxin-2-carboxyamide with E value of >200. Thus, at about 50% conversion, (R)-2,3-dihydro-1,4-benzodioxin-2-carboxylic acid was produced in >99% e.e. The remaining amide had (S)-configuration and 99% e.e. The amide and acid were easily separated by aqueous (alkaline)-organic two phase extraction method. The same amidase was able to catalyse, albeit at much lower rate the hydrolysis of (S)-amide to (S)-acid without loss of e.e. The amidase activity was identified as indole-3-acetamide hydrolase (IaaH). IaaH is known to catalyse conversion of indole-3-acetamide (IAM) to indole-3-acetic acid (IAA), which is phytohormone of auxin class and is widespread among plants and bacteria that inhabit plant rhizosphere. IaaH exhibited high activity for 2,3-dihydro-1,4-benzodioxin-2-carboxamide, which was about 65% compared to its natural substrate, indole-3-acetamide. The natural substrate for IaaH indole-3-acetamide shared, at least in part a similar bicyclic structure with 2,3-dihydro-1,4-benzodioxin-2-carboxamide, which may account for high activity of enzyme towards this un-natural substrate. To the best of

Increased Enterococcus faecalis infection is associated with clinically active Crohn disease

PubMed Central

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

2016-01-01

Abstract 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

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.

Prevalence of tetracycline resistance genes among multi-drug resistant bacteria from selected water distribution systems in southwestern Nigeria.

PubMed

Adesoji, Ayodele T; Ogunjobi, Adeniyi A; Olatoye, Isaac O; Call, Douglas R; Douglas, Douglas R

2015-06-25

Antibiotic resistance genes [ARGs] in aquatic systems have drawn increasing attention they could be transferred horizontally to pathogenic bacteria. Water treatment plants (WTPs) are intended to provide quality and widely available water to the local populace they serve. However, WTPs in developing countries may not be dependable for clean water and they could serve as points of dissemination for antibiotic resistant bacteria. Only a few studies have investigated the occurrence of ARGs among these bacteria including tetracycline resistance genes in water distribution systems in Nigeria. Multi-drug resistant (MDR) bacteria, including resistance to tetracycline, were isolated from treated and untreated water distribution systems in southwest Nigeria. MDR bacteria were resistant to >3 classes of antibiotics based on break-point assays. Isolates were characterized using partial 16S rDNA sequencing and PCR assays for six tetracycline-resistance genes. Plasmid conjugation was evaluated using E. coli strain DH5α as the recipient strain. Out of the 105 bacteria, 85 (81 %) and 20 (19 %) were Gram- negative or Gram- positive, respectively. Twenty-nine isolates carried at least one of the targeted tetracycline resistance genes including strains of Aeromonas, Alcaligenes, Bacillus, Klebsiella, Leucobacter, Morganella, Proteus and a sequence matching a previously uncultured bacteria. Tet(A) was the most prevalent (16/29) followed by tet(E) (4/29) and tet30 (2/29). Tet(O) was not detected in any of the isolates. Tet(A) was mostly found with Alcaligenes strains (9/10) and a combination of more than one resistance gene was observed only amongst Alcaligenes strains [tet(A) + tet30 (2/10), tet(A) + tet(E) (3/10), tet(E) + tet(M) (1/10), tet(E) + tet30 (1/10)]. Tet(A) was transferred by conjugation for five Alcaligenes and two E. coli isolates. This study found a high prevalence of plasmid-encoded tet(A) among Alcaligenes isolates, raising the possibility that this

Presence of a new cytochrome b - like pigment with a peak at 567 nm in various aerobic bacteria.

PubMed

Jacobs, N J; O'Hara, J; Gray, C T

1983-09-01

Several physiological groups of bacteria were examined for the presence of a cytochrome b - like pigment which is demonstrable in dithionite-reduced minus substrate-reduced difference spectra. This pigment is characterized by an unusually high alpha band at 567 nm, a low concentration relative to conventional cytochromes, and an inability to be fully reduced by endogenous substrates or NADH. Previous studies with one denitrifying and nondenitrifying species of the genus Pseudomonas, in Paracoccus denitrificans, in Alcaligenes faecalis, in Azotobacter vinelandii, in Branhamella catarrhalis, and in Neisseria lactamicus. In all these organisms, the peak of the 567-nm pigment is accompanied by a peak of about equal height at approximately 559 nm, which exhibits similar properties to the 567-nm pigment. The 567-nm pigment was not demonstrable by this technique in Gluconobacter oxydans subspecies suboxydans, Bacillus subtilis, Bacillus licheniformis, Aeromonas hydrophilia, Escherichia coli, a Klebsiella species, Moraxella osloensis, Aquaspirillum itersonii, Micrococcus lysodeikticus, Micrococcus luteus, Agrobacterium tumefaciens, or Rhizobium meliloti.

Effect of gaseous ozone on Enterococcus faecalis biofilm-an in vitro study.

PubMed

Boch, Tanja; Tennert, Christian; Vach, Kirstin; Al-Ahmad, Ali; Hellwig, Elmar; Polydorou, Olga

2016-09-01

The aim of this study was to evaluate the antimicrobial effect of gaseous ozone compared to conventional methods against Enterococcus faecalis. One hundred twenty-five teeth were infected by E. faecalis and were incubated for 72 h to form biofilm. Teeth were distributed among five groups. In the first group, ozone was used; in the second group, teeth were rinsed with 20 % ethylenediaminetetraacetic acid (EDTA); in the third group, with 3 % sodium hypochlorite (NaOCl). Group 4 combined 20 % EDTA with ozone. NaOCl and ozone were combined in group 5. After treatment, the samples with paper points were taken, followed by dentin samples taken with K-file, and cultured for 24 h. Then bacterial colonies were counted. All treatments reduced significantly (p < 0.05) the bacteria. Paper points' samples showed 85.38 % reduction after ozone. The highest reduction was observed in NaOCl group (99.98 %). EDTA reduced bacteria by 80.64 %. Combination of NaOCl and ozone eradicated 99.95 % of the bacteria. Combination of EDTA and ozone reduced E. faecalis up to 91.33 %. The dentin chips showed the following: the highest CFU counts were observed in EDTA group, followed by ozone and NaOCl group. The lowest CFU counts were found in NaOCl-ozone group and EDTA-ozone group. Ozone reduced E. faecalis, even organised in a biofilm, however, lower than NaOCl. No treatment reduced totally the bacteria. Used as an adjuvant, ozone can increase the efficacy of conventional rinsing like EDTA and presents an alternative treatment when NaOCl cannot be used e.g. in teeth with a wide-open apical foramen.

Apical extrusion of Enterococcus faecalis using three different rotary instrumentation techniques: an in vitro study.

PubMed

Taneja, Sonali; Kumari, Manju; Barua, Madhumita; Dudeja, Chetna; Malik, Meeta

2015-01-01

To compare the apical extrusion of Enterococcus faecalis after instrumentation with three different Ni-Ti rotary instruments- An in vitro study. In vitro study Methods and Material: Forty freshly extracted mandibular premolars were mounted in bacteria collection apparatus and root canals were contaminated with a suspension of Enterococcus faecalis. The contaminated teeth were divided into 4 groups of 10 teeth each according to rotary system used for instrumentation: Group1: Hyflex files, Group 2: GTX files, Group 3: Protaper files and Group 4: control group (no instrumentation). Bacteria extruded after preparations were collected into vials and microbiological samples were incubated in BHI broth for 24 hrs. The colony forming units were determined for each sample. Statistical analysis was done using one way ANOVA followed by post hoc independent " t" test. GTX files extruded least amount of bacteria followed by Hyflex files. Maximum extrusion of E. faecalis was seen in rotary Protaper group. Least amount of extrusion was seen with GTX files followed by Hyflex files and then rotary Protaper system.

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

An Attenuated CRISPR-Cas System in Enterococcus faecalis Permits DNA Acquisition.

PubMed

Hullahalli, Karthik; Rodrigues, Marinelle; Nguyen, Uyen Thy; Palmer, Kelli

2018-05-01

Antibiotic-resistant bacteria are critical public health concerns. Among the prime causative factors for the spread of antibiotic resistance is horizontal gene transfer (HGT). A useful model organism for investigating the relationship between HGT and antibiotic resistance is the opportunistic pathogen Enterococcus faecalis , since the species possesses highly conjugative plasmids that readily disseminate antibiotic resistance genes and virulence factors in nature. Unlike many commensal E. faecalis strains, the genomes of multidrug-resistant (MDR) E. faecalis clinical isolates are enriched for mobile genetic elements (MGEs) and lack c lustered r egularly i nterspaced s hort p alindromic r epeats (CRISPR) and C RISPR- as sociated protein (Cas) genome defense systems. CRISPR-Cas systems cleave foreign DNA in a programmable, sequence-specific manner and are disadvantageous for MGE-derived genome expansion. An unexplored facet of CRISPR biology in E. faecalis is that MGEs that are targeted by native CRISPR-Cas systems can be maintained transiently. Here, we investigate the basis for this "CRISPR tolerance." We observe that E. faecalis can maintain self-targeting constructs that direct Cas9 to cleave the chromosome, but at a fitness cost. Interestingly, DNA repair genes were not upregulated during self-targeting, but integrated prophages were strongly induced. We determined that low cas9 expression contributes to this transient nonlethality and used this knowledge to develop a robust CRISPR-assisted genome-editing scheme. Our results suggest that E. faecalis has maximized the potential for DNA acquisition by attenuating its CRISPR machinery, thereby facilitating the acquisition of potentially beneficial MGEs that may otherwise be restricted by genome defense. IMPORTANCE CRISPR-Cas has provided a powerful toolkit to manipulate bacteria, resulting in improved genetic manipulations and novel antimicrobials. These powerful applications rely on the premise that CRISPR

Indole production provides limited benefit to Escherichia coli during co-culture with Enterococcus faecalis.

PubMed

Pringle, Shelly L; Palmer, Kelli L; McLean, Robert J C

2017-01-01

Escherichia coli lives in the gastrointestinal tract and elsewhere, where it coexists within a mixed population. Indole production enables E. coli to grow with other gram-negative bacteria as indole inhibits N-acyl-homoserine lactone (AHL) quorum regulation. We investigated whether E. coli indole production enhanced competition with gram-positive Enterococcus faecalis, wherein quorum signaling is mediated by small peptides. During planktonic co-culture with E. faecalis, the fitness and population density of E. coli tnaA mutants (unable to produce indole) equaled or surpassed that of E. coli wt. During biofilm growth, the fitness of both populations of E. coli stabilized around 100 %, whereas the fitness of E. faecalis declined over time to 85-90 %, suggesting that biofilm and planktonic populations have different competition strategies. Media supplementation with indole removed the competitive advantage of E. coli tnaA in planktonic populations but enhanced it in biofilm populations. E. coli wt and tnaA showed similar growth in Luria-Bertani (LB) broth. However, E. coli growth was inhibited in the presence of filter-sterilized spent LB from E. faecalis, with inhibition being enhanced by indole. Similarly, there was also an inhibition of E. faecalis growth by proteinaceous components (likely bacteriocins) from spent culture media from both E. coli strains. We conclude that E. coli indole production is not a universal competition strategy, but rather works against gram-negative, AHL-producing bacteria.

Enterococcus faecalis Gene Transfer under Natural Conditions in Municipal Sewage Water Treatment Plants†

PubMed Central

Marcinek, Herbert; Wirth, Reinhard; Muscholl-Silberhorn, Albrecht; Gauer, Matthias

1998-01-01

The ability of Enterococcus faecalis to transfer various genetic elements under natural conditions was tested in two municipal sewage water treatment plants. Experiments in activated sludge basins of the plants were performed in a microcosm which allowed us to work under sterile conditions; experiments in anoxic sludge digestors were performed in dialysis bags. We used the following naturally occurring genetic elements: pAD1 and pIP1017 (two so-called sex pheromone plasmids with restricted host ranges, which are transferred at high rates under laboratory conditions); pIP501 (a resistance plasmid possessing a broad host range for gram-positive bacteria, which is transferred at low rates under laboratory conditions); and Tn916 (a conjugative transposon which is transferred under laboratory conditions at low rates to gram-positive bacteria and at very low rates to gram-negative bacteria). The transfer rate between different strains of E. faecalis under natural conditions was, compared to that under laboratory conditions, at least 105-fold lower for the sex pheromone plasmids, at least 100-fold lower for pIP501, and at least 10-fold lower for Tn916. In no case was transfer from E. faecalis to another bacterial species detected. By determining the dependence of transfer rates for pIP1017 on bacterial concentration and extrapolating to actual concentrations in the sewage water treatment plant, we calculated that the maximum number of transfer events for the sex pheromone plasmids between different strains of E. faecalis in the municipal sewage water treatment plant of the city of Regensburg ranged from 105 to 108 events per 4 h, indicating that gene transfer should take place under natural conditions. PMID:9464401

Enterococcus faecalis infection causes inflammation, intracellular oxphos-independent ROS production, and DNA damage in human gastric cancer cells.

PubMed

Strickertsson, Jesper A B; Desler, Claus; Martin-Bertelsen, Tomas; Machado, Ana Manuel Dantas; Wadstrøm, Torkel; Winther, Ole; Rasmussen, Lene Juel; Friis-Hansen, Lennart

2013-01-01

Achlorhydria caused by e.g. atrophic gastritis allows for bacterial overgrowth, which induces chronic inflammation and damage to the mucosal cells of infected individuals driving gastric malignancies and cancer. Enterococcus faecalis (E. faecalis) can colonize achlohydric stomachs and we therefore wanted to study the impact of E. faecalis infection on inflammatory response, reactive oxygen species (ROS) formation, mitochondrial respiration, and mitochondrial genetic stability in gastric mucosal cells. To separate the changes induced by bacteria from those of the inflammatory cells we established an in vitro E. faecalis infection model system using the gastric carcinoma cell line MKN74. Total ROS and superoxide was measured by fluorescence microscopy. Cellular oxygen consumption was characterized non-invasively using XF24 microplate based respirometry. Gene expression was examined by microarray, and response pathways were identified by Gene Set Analysis (GSA). Selected gene transcripts were verified by quantitative real-time polymerase chain reaction (qRT-PCR). Mitochondrial mutations were determined by sequencing. Infection of MKN74 cells with E. faecalis induced intracellular ROS production through a pathway independent of oxidative phosphorylation (oxphos). Furthermore, E. faecalis infection induced mitochondrial DNA instability. Following infection, genes coding for inflammatory response proteins were transcriptionally up-regulated while DNA damage repair and cell cycle control genes were down-regulated. Cell growth slowed down when infected with viable E. faecalis and responded in a dose dependent manner to E. faecalis lysate. Infection by E. faecalis induced an oxphos-independent intracellular ROS response and damaged the mitochondrial genome in gastric cell culture. Finally the bacteria induced an NF-κB inflammatory response as well as impaired DNA damage response and cell cycle control gene expression. Array Express accession number E-MEXP-3496.

Efficacy of calcium oxide and calcium hydroxide nanoparticles on the elimination of Enterococcus faecalis in human root dentin.

PubMed

Louwakul, Phumisak; Saelo, Attapon; Khemaleelakul, Saengusa

2017-04-01

The objective of this study was to compare the antibacterial effect of calcium oxide nanoparticles (CONPs) and calcium hydroxide nanoparticles (CHNPs) against Enterococcus faecalis in a dentinal block model. E. faecalis strain JCM 7783 was introduced into dentinal tubules of semicylindrical dentin specimens by centrifugation and incubated for 1 week. Fifty microliters of CONPs or CHNPs was placed on the root canal side of the infected dentin specimens. The specimens were then incubated in aerobic condition at 37 °C and 100 % relative humidity for 1 week. The treated dentin specimens were subjected to fluorescent staining and confocal laser scanning microscopy (CLSM) to analyze the proportions of non-vital and vital bacterial cells inside the dentinal tubules. Scanning electron microscopy (SEM) was used to confirm the effect of the medicaments on the bacteria in the dentinal tubules. Calcium oxide (CO) and calcium hydroxide (CH) were used as controls. Based on the CLSM and SEM analyses, CHNPs were more efficient than CONPs in the elimination of the bacteria in the dentinal tubules. CONPs significantly killed more E. faecalis than CO and CH (P < .05). Neither CO nor CH was able to kill the bacteria. CHNPs were more effective than CONPs in the elimination of E. faecalis in dentinal tubules. CHNPs are effective nanoparticles in killing endodontic bacteria present in dentinal tubules. They have potential as an intracanal medicament, which may be beneficial in root canal therapy.

Antibiofilm efficacy of silver nanoparticles as a vehicle for calcium hydroxide medicament against Enterococcus faecalis.

PubMed

Afkhami, Farzaneh; Pourhashemi, Seyyed Jalal; Sadegh, Mona; Salehi, Yasaman; Fard, Mohammad Javad Kharrazi

2015-12-01

The aim of the present study was to investigate antibacterial characteristic and Enterococcus faecalis (E. faecalis) biofilm suppression effect of different vehicles of calcium hydroxide as intracanal medicaments in short and long-term. Fifty-four human single-root teeth were contaminated with E. faecalis bacteria. The teeth were randomly divided into three experimental (n=16) and one control group (n=6). Each group was then exposed to various intracanal medicaments, namely calcium hydroxide paste (group 1), calcium hydroxide with chlorhexidine (group 2), calcium hydroxide with silver nanoparticles suspension (AgNPs) (group 3), and saline as the control group (group 4). Cultures were made from each group after one week and one month, and the number of colonies was counted. Moreover, a sample of each group was examined under electron microscope. Kruskal-Wallis test served for inter-group comparisons, and Mann-Whitney test served for comparison between the two incubation periods. All the intracanal medicaments resulted in significant decrease in number of colonies compared to control group in both incubation periods. After one week, the mixture of calcium hydroxide and AgNPs was the most effective medicament against E. faecalis bacteria (p<.05). No significant difference in antibacterial effect of the medicaments existed after one month incubation period (p>.05). AgNPs was more effective on the E. faecalis biofilm than other tested vehicles in short-term medication. AgNPs seems to have a good potential to be used as an appropriate vehicle of calcium hydroxide in order to eliminate of E. faecalis biofilm from human dentine in short-term. Copyright © 2015 Elsevier Ltd. All rights reserved.

Photodynamic therapy with high-power LED mediated by erythrosine eliminates Enterococcus faecalis in planktonic forms.

PubMed

Borba, Alberto Sabin Moura; da Silva Pereira, Sângela Maria; Borba, Mellyna Cavalcante Mendes; Paschoal, Marco Aurélio Benini; de Jesus Tavarez, Rudys Rodolfo; de Castro Rizzi, Claudia; Ferreira, Meire Coelho; Maia Filho, Etevaldo Matos

2017-09-01

The failure of endodontic treatment is linked to the presence of microorganisms, particularly Enterococcus faecalis, in the root canals. This study evaluated the effectiveness of photodynamic therapy (PDT) using erythrosine irradiated by a high-power curing light on a planktonic suspension culture of E. faecalis. Bacterial suspensions of E. faecalis were adjusted and then mixed in a 1:1 proportion, in triplicate, in treatment groups by varying the length of irradiation time (120 and 240s) and the molarity of the erythrosine (5 and 10μM). In order to verify the post-treatment bactericidal effect, a count of the viable bacteria was performed (CFUmL -1 ) and transformed into Log10 CFU. The one-way ANOVA with Tukey post-hoc test was applied to check for differences between the groups. The bacteria were completely eradicated in the groups that used PDT with 5μM 240s, 10μM 120s and 10μM 240s (p≪0.001). The effect of the PDT 5μM 120s group was significant (p≪0.05) in comparison with the groups using only light or only erythrosine. Positive control (exposure to 2.5% NaClO for 120 and 240s) completely eradicated E. faecalis. The negative control (PBS) did not alter the quantities of E. faecalis CFU with 9.605 Log10 CFU at 120s and 9.621 Log10 CFU at 240s. PDT with erythrosine in a concentration of 10μM and high-power LED is capable of totally eliminating E. faecalis in planktonic suspension. Copyright © 2017 Elsevier B.V. All rights reserved.

Enterococcus faecalis Prophage Dynamics and Contributions to Pathogenic Traits

PubMed Central

Matos, Renata C.; Lapaque, Nicolas; Rigottier-Gois, Lionel; Debarbieux, Laurent; Meylheuc, Thierry; Gonzalez-Zorn, Bruno; Repoila, Francis; Lopes, Maria de Fatima; Serror, Pascale

2013-01-01

Polylysogeny is frequently considered to be the result of an adaptive evolutionary process in which prophages confer fitness and/or virulence factors, thus making them important for evolution of both bacterial populations and infectious diseases. The Enterococcus faecalis V583 isolate belongs to the high-risk clonal complex 2 that is particularly well adapted to the hospital environment. Its genome carries 7 prophage-like elements (V583-pp1 to -pp7), one of which is ubiquitous in the species. In this study, we investigated the activity of the V583 prophages and their contribution to E. faecalis biological traits. We systematically analyzed the ability of each prophage to excise from the bacterial chromosome, to replicate and to package its DNA. We also created a set of E. faecalis isogenic strains that lack from one to all six non-ubiquitous prophages by mimicking natural excision. Our work reveals that prophages of E. faecalis V583 excise from the bacterial chromosome in the presence of a fluoroquinolone, and are able to produce active phage progeny. Intricate interactions between V583 prophages were also unveiled: i) pp7, coined EfCIV583 for E. faecalis chromosomal island of V583, hijacks capsids from helper phage 1, leading to the formation of distinct virions, and ii) pp1, pp3 and pp5 inhibit excision of pp4 and pp6. The hijacking exerted by EfCIV583 on helper phage 1 capsids is the first example of molecular piracy in Gram positive bacteria other than staphylococci. Furthermore, prophages encoding platelet-binding-like proteins were found to be involved in adhesion to human platelets, considered as a first step towards the development of infective endocarditis. Our findings reveal not only a role of E. faecalis V583 prophages in pathogenicity, but also provide an explanation for the correlation between antibiotic usage and E. faecalis success as a nosocomial pathogen, as fluoriquinolone may provoke release of prophages and promote gene dissemination among

An Attenuated CRISPR-Cas System in Enterococcus faecalis Permits DNA Acquisition

PubMed Central

Hullahalli, Karthik; Rodrigues, Marinelle; Nguyen, Uyen Thy

2018-01-01

ABSTRACT Antibiotic-resistant bacteria are critical public health concerns. Among the prime causative factors for the spread of antibiotic resistance is horizontal gene transfer (HGT). A useful model organism for investigating the relationship between HGT and antibiotic resistance is the opportunistic pathogen Enterococcus faecalis, since the species possesses highly conjugative plasmids that readily disseminate antibiotic resistance genes and virulence factors in nature. Unlike many commensal E. faecalis strains, the genomes of multidrug-resistant (MDR) E. faecalis clinical isolates are enriched for mobile genetic elements (MGEs) and lack clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) genome defense systems. CRISPR-Cas systems cleave foreign DNA in a programmable, sequence-specific manner and are disadvantageous for MGE-derived genome expansion. An unexplored facet of CRISPR biology in E. faecalis is that MGEs that are targeted by native CRISPR-Cas systems can be maintained transiently. Here, we investigate the basis for this “CRISPR tolerance.” We observe that E. faecalis can maintain self-targeting constructs that direct Cas9 to cleave the chromosome, but at a fitness cost. Interestingly, DNA repair genes were not upregulated during self-targeting, but integrated prophages were strongly induced. We determined that low cas9 expression contributes to this transient nonlethality and used this knowledge to develop a robust CRISPR-assisted genome-editing scheme. Our results suggest that E. faecalis has maximized the potential for DNA acquisition by attenuating its CRISPR machinery, thereby facilitating the acquisition of potentially beneficial MGEs that may otherwise be restricted by genome defense. PMID:29717009

Antimicrobial activity of alexidine alone and associated with N-acetylcysteine against Enterococcus faecalis biofilm

PubMed Central

Silveira, Luiz Fernando Machado; Baca, Pilar; Arias-Moliz, María Teresa; Rodríguez-Archilla, Alberto; Ferrer-Luque, Carmen María

2013-01-01

The purpose of this study was to assess the efficacy of alexidine (ALX), alone and combined with N-acetylcysteine (NAC), in eradicating two Enterococcus faecalis strain biofilms. The biofilms of E. faecalis ATCC 29212 and the clinical isolate E. faecalis D1 were grown in the MBEC-high-throughput device for 24 h and were exposed to five twofold dilutions of ALX (2%–0.007 8%) alone and combined with 100 mg⋅mL−1 NAC, for 1 and 5 min. Eradication was defined as 100% kill of biofilm bacteria. The Student's t-test was used to compare the efficacy of the associations of the two irrigants. After 1-min contact time, ALX eradicated the biofilms at all concentrations except for 0.007 8% and 0.015 6%–0.007 8% with E. faecalis ATCC 29212 and E. faecalis D1, respectively. Similar results for eradication and concentration were obtained when it was combined with 100 mg⋅mL−1 NAC. After 5 min of contact time, ALX alone and combined with NAC eradicated all enterococci biofilms. ALX showed antimicrobial properties against the two E. faecalis strain biofilms tested at very low concentrations, and its combined use with NAC was not seen to enhance its activity. PMID:23970139

Chitosan-propolis nanoparticle formulation demonstrates anti-bacterial activity against Enterococcus faecalis biofilms

PubMed Central

Ong, Teik Hwa; Chitra, Ebenezer; Ramamurthy, Srinivasan; Siddalingam, Rajinikanth Paruvathanahalli; Yuen, Kah Hay; Ambu, Stephen Periathamby

2017-01-01

Propolis obtained from bee hives is a natural substance with antimicrobial properties. It is limited by its insolubility in aqueous solutions; hence ethanol and ethyl acetate extracts of Malaysian propolis were prepared. Both the extracts displayed antimicrobial and anti-biofilm properties against Enterococcus faecalis, a common bacterium associated with hospital-acquired infections. High performance liquid chromatography (HPLC) analysis of propolis revealed the presence of flavonoids like kaempferol and pinocembrin. This study investigated the role of propolis developed into nanoparticles with chitosan for its antimicrobial and anti-biofilm properties against E. faecalis. Bacteria that grow in a slimy layer of biofilm are resistant to penetration by antibacterial agents. The use of nanoparticles in medicine has received attention recently due to better bioavailability, enhanced penetrative capacity and improved efficacy. A chitosan-propolis nanoformulation was chosen based on ideal physicochemical properties such as particle size, zeta potential, polydispersity index, encapsulation efficiency and the rate of release of the active ingredients. This formulation inhibited E. faecalis biofilm formation and reduced the number of bacteria in the biofilm by ~90% at 200 μg/ml concentration. When tested on pre-formed biofilms, the formulation reduced bacterial number in the biofilm by ~40% and ~75% at 200 and 300 μg/ml, respectively. The formulation not only reduced bacterial numbers, but also physically disrupted the biofilm structure as observed by scanning electron microscopy. Treatment of biofilms with chitosan-propolis nanoparticles altered the expression of biofilm-associated genes in E. faecalis. The results of this study revealed that chitosan-propolis nanoformulation can be deemed as a potential anti-biofilm agent in resisting infections involving biofilm formation like chronic wounds and surgical site infections. PMID:28362873

Chitosan-propolis nanoparticle formulation demonstrates anti-bacterial activity against Enterococcus faecalis biofilms.

PubMed

Ong, Teik Hwa; Chitra, Ebenezer; Ramamurthy, Srinivasan; Siddalingam, Rajinikanth Paruvathanahalli; Yuen, Kah Hay; Ambu, Stephen Periathamby; Davamani, Fabian

2017-01-01

Propolis obtained from bee hives is a natural substance with antimicrobial properties. It is limited by its insolubility in aqueous solutions; hence ethanol and ethyl acetate extracts of Malaysian propolis were prepared. Both the extracts displayed antimicrobial and anti-biofilm properties against Enterococcus faecalis, a common bacterium associated with hospital-acquired infections. High performance liquid chromatography (HPLC) analysis of propolis revealed the presence of flavonoids like kaempferol and pinocembrin. This study investigated the role of propolis developed into nanoparticles with chitosan for its antimicrobial and anti-biofilm properties against E. faecalis. Bacteria that grow in a slimy layer of biofilm are resistant to penetration by antibacterial agents. The use of nanoparticles in medicine has received attention recently due to better bioavailability, enhanced penetrative capacity and improved efficacy. A chitosan-propolis nanoformulation was chosen based on ideal physicochemical properties such as particle size, zeta potential, polydispersity index, encapsulation efficiency and the rate of release of the active ingredients. This formulation inhibited E. faecalis biofilm formation and reduced the number of bacteria in the biofilm by ~90% at 200 μg/ml concentration. When tested on pre-formed biofilms, the formulation reduced bacterial number in the biofilm by ~40% and ~75% at 200 and 300 μg/ml, respectively. The formulation not only reduced bacterial numbers, but also physically disrupted the biofilm structure as observed by scanning electron microscopy. Treatment of biofilms with chitosan-propolis nanoparticles altered the expression of biofilm-associated genes in E. faecalis. The results of this study revealed that chitosan-propolis nanoformulation can be deemed as a potential anti-biofilm agent in resisting infections involving biofilm formation like chronic wounds and surgical site infections.

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. Copyright © 2016 Elsevier Inc. All rights reserved.

Antibacterial Potential of 2.5% Sodium Hypochlorite in Distinct Irrigation Protocols on Enterococcus faecalis Biofilm.

PubMed

Alves, Denise Ramos Silveira; Cunha, Rodrigo Sanches; da Silveira Bueno, Carlos Eduardo; de Alencar, Ana Helena Gonçalves; de Araújo Estrela, Cyntia Rodrigues; dos Santos, Tatiane Oliveira; Estrela, Carlos

2015-05-01

The aim of this study was to evaluate the effect of irrigation methods on antibacterial potential of 2.5% NaOCl on Enterococcus faecalis biofilm. Enterococcus faecalis biofilms were prepared during 60 days on 48 human root canals and randomized into control and experimental groups using positive and negative pressure irrigation. Bacterial growth was analyzed using turbidity of culture medium followed by UV spectrophotometry, and scanning electron microscopy (SEM) analyses were performed. Mean and standard deviations were used for evaluate the mean optical densities associated to the number of bacteria present culture, and Scheirer-Ray-Hare (an extension of the Kruskal-Wallis test) and Tamhane test to analyze the SEM images in the groups and thirds. Significance was set at 5%. Enterococcus faecalis was still present after root canal cleaning regardless of irrigation methods or bacterial identification methods. Positive and negative pressure irrigation protocols using 2.5% NaOCl show a similar capacity to reduce E. faecalis in infected root canals.

[Association of the pH change of vaginal environment in bacterial vaginosis with presence of Enterococcus faecalis in vagina].

PubMed

Jahić, Mahira; Nurkić, Mahmud; Fatusić, Zlatan

2006-01-01

Normal pH value of vagina from 3.8 to 4.2 has regulatory and protectors mechanisms of vaginal environment. The change in the pH value indicates to presence of disbalance in the ecosystem of vaginal environment. The value of pH above 4.0 is indicator of the decreased number of lactobacillus bacteria and the increased number of other microorganisms in the vaginal environment. This situation is present in the case of developing of bacterial vaginosis. One of the bacteria which is often isolated from vaginal swabs is Enterococcus faecalis. Aims of this study are to examine presence o f Enterococcus faecalis in vagina in healthy women and womenwith signs of bacterial vaginosis, the most often present signs in patients with bacterial vaginosis and isolated Enterococcus faecalis from vaginal swabs, and to determine whether the change of the pH value of vaginal environment could be indicator for bacterial vaginosis associated with Enterococcus faecalis. In this study there were included 90 patients. To all patients there were done: gynecological survey, determined pH of vaginal environment and color of vaginal secret, amino odor test, and taken vaginal swabs for microbiological examination. Enterococcus faecalis was found in the patients with pH 4.0 in 24.05 % cases, but in the patients with signs of bacterial vaginosis it was found in 52.78 %. Positive findings of Enterococcus faecalis was the most often associated with presence of all tree signs of bacterial vaginosis (pH>4.0, changed color of vaginal secret and positive amino odor test) it is in 60.78 6% cases. With two signs of bacterial vaginosis (pH>4.0, changed color of vaginal secret) Enterococcus faecalis was present in 60 % cases. The only presence of change in the pH>4.0 was associated with Enterococcus faecalis in 52.78 %. This study showed that pH change of vaginal environment was associated with Enterococcus faecalis in bacterial vaginosis in high percentage but it can not be used as the sure sign of presence

Purification and characteristics of a novel bacteriocin produced by Enterococcus faecalis L11 isolated from Chinese traditional fermented cucumber.

PubMed

Gao, Yurong; Li, Benling; Li, Dapeng; Zhang, Liyuan

2016-05-01

To purify and characterize a novel bacteriocin with broad inhibitory spectrum produced by an isolate of Enterococcus faecalis from Chinese fermented cucumber. E. faecalis L11 produced a bacteriocin with antimicrobial activity against both Escherichia coli and Staphylococcus aureus. The amino acid sequence of the purified bacteriocin, enterocin L11, was assayed by Edman degradation method. It differs from other class II bacteriocins and exhibited a broad antimicrobial activity against not only Gram-positive bacteria, including Bacillus subtilis, S. aureus, Listeria monocytogenes, Sarcina flava, Lactobacillus acidophilus, L. plantarum, L. delbrueckii subsp. delbrueckii, L. delbrueckii subsp. bulgaricus and Streptococcus thermophilus, but also some Gram-negative bacteria including Salmonella typhimurium, E. coli and Shigella flexneri. Enterocin L11 retained 91 % of its activity after holding at 121 °C for 30 min. It was also resistant to acids and alkalis. Enterocin L11 is a novel broad-spectrum Class II bacteriocin produced by E. faecalis L11, and may have potential as a food biopreservative.

Antimicrobial Susceptibility as a Diagnostic Aid in the Identification of Nonfermenting Gram-Negative Bacteria

PubMed Central

Gilardi, G. L.

1971-01-01

Antimicrobial susceptibility data regarding nonfermentative, gram-negative bacteria (Pseudomonas, Alcaligenes, Acinetobacter, Moraxella, Flavobacterium) are presented showing that the antibiograms of most species examined can be used as an important auxillary aid in their differentiation. PMID:5132093

Wheat Bran Enhances the Cytotoxicity of Immobilized Alcaligenes aquatilis F8 against Microcystis aeruginosa

PubMed Central

Sun, Pengfei; Lin, Hui; Wang, Guan; Zhang, Ximing; Zhang, Qichun; Zhao, Yuhua

2015-01-01

Algicidal bacteria offer a promising option for killing cyanobacteria. Therefore, a new Alcaligenes aquatilis strain F8 was isolated to control Microcystis aeruginosa in this study. The algicidal activity of strain F8 was dependent on the cell density of M. aeruginosa, and the maximal algicidal rate of the free bacterium reached 88.45% within 72 h. With a view to its application to the control of M. aeruginosa in the natural environment, strain F8 was immobilized in sodium alginate beads, but immobilization of the strain decreased its algicidal rate compared to that of the free bacterium. However, addition of wheat bran to the sodium alginate matrix used to immobilize strain F8 not only eliminated the adverse effects of immobilization on the bacteria but also resulted in an 8.83% higher algicidal rate of the immobilized than free bacteria. Exclusion and recovery methods were used to identify key ingredients of wheat bran and gain insight into the mechanism underlying the observed enhancement of algicidal activity. This analysis indicated that certain factors in wheat bran, including vitamins B1, B2, B9, and E were responsible for promoting bacterial growth and thereby improving the algicidal rate of immobilized strain F8. Our findings indicate that wheat bran is able to improve the algicidal efficiency of A. aquatilis strain F8 for killing M. aeruginosa and is a good source of not only carbon and nitrogen but also vitamins for bacteria. PMID:26295573

Antibacterial properties of silver nanoparticles as a root canal irrigant against Enterococcus faecalis biofilm and infected dentinal tubules.

PubMed

Rodrigues, C T; de Andrade, F B; de Vasconcelos, L R S M; Midena, R Z; Pereira, T C; Kuga, M C; Duarte, M A H; Bernardineli, N

2018-02-03

To evaluate the antimicrobial action of an irrigant containing silver nanoparticles in an aqueous vehicle (AgNp), sodium hypochlorite and chlorhexidine against Enterococcus faecalis biofilm and infected dentinal tubules. Bovine dentine blocks were used for E. faecalis biofilm development for 21 days and irrigated with 94 ppm AgNp solution, 2.5% NaOCl and 2% chlorhexidine for 5, 15 and 30 min. For infection of dentinal tubules with E. faecalis, dentine specimens from bovine incisors were submitted to a contamination protocol over 5 days, with eight centrifugation cycles on every alternate day, and irrigated with the same solutions and time intervals used for the biofilm. The specimens were stained with the Live/Dead technique and evaluated using a confocal laser scanning microscope (CLSM). The bioImage_L software was used for measurement of the total biovolume of biofilm in μm 3 and percentage of viable bacteria (green cells) in biofilm and in dentinal tubules found after the irrigation. Statistical analyses were performed using Kruskal-Wallis and Dunn's tests for quantification of viable cells in biofilm, the Friedman test for comparisons of viable bacteria in dentinal tubules in different areas of the root canal and the Mann-Whitney U-test to compare the action of the irrigants between the two methods (P < 0.05). The AgNp solution eliminated fewer bacteria, but was able to dissolve more biofilm compared with chlorhexidine (P < 0.05). NaOCl had the greatest antimicrobial activity and biofilm dissolution capacity. AgNp solution had less antimicrobial action in infected dentinal tubules compared with NaOCl (P < 0.05). The AgNp solution after 5 min was more effective in eliminating planktonic bacteria in dentinal tubules than in biofilm, but at 30 min fewer viable bacteria were observed in the biofilm compared with intratubular dentine (P < 0.05). AgNp irrigant was not as effective against E. faecalis compared to solutions commonly used in root canal

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

Biodegradation of 4-chlorophenol by adsorptive immobilized Alcaligenes sp. A 7-2 in soil.

PubMed

Balfanz, J; Rehm, H J

1991-08-01

Alcaligenes sp. A 7-2 immobilized on granular clay has been applied in a percolator to degrade 4-chlorophenol in sandy soil. Good adsorption rates on granular clay were achieved using cell suspensions with high titres and media at pH 8.0. The influence of various parameters such as aeration rate, pH, temperature, concentration of 4-chlorophenol and size of inoculum on the degradation rate were investigated. During fed-batch fermentations under optimal culture conditions, concentrations of 4-chlorophenol up to 160 mg.l-1 could be degraded. Semicontinuous culture experiments demonstrated that the degradation potential in soil could be well established and enhanced by the addition of immobilized bacteria. Continuous fermentation was performed with varying 4-chlorophenol concentrations in the feed and different input levels. The maximum degradation rate was 1.64 g.l-1.day-1.

[Development of an Enterococcus faecalis periapical biofilm model for in vitro morphological study].

PubMed

Cao, Ridan; Hou, Benxiang

2014-08-01

This study aims to develop and observe a model system of the periapical biofilm structure of Enterococcus faecalis (E. faecalis). A total of 24 intact human single-rooted premolars extracted for orthodontic reasons were collected and randomly divided into eight groups (n = 3). The specimens were subjected to ultraviolet disinfection, inoculated with E. faecalis (ATCC 29212) suspension adjusted to 1 x 10(8) CFU x mL(-1), and incubated at 37 degrees C for 1, 2, and 7 d. Specimen groups were prepared for scanning electron microscope to examine the biofilm formation. The specimens in the confocal laser scanning microscope (CLSM) groups were stained with propidium iodide (PI) and ConA-fluorescein isothiocyanate (ConA-FITC) to examine the biofilm formation. The images were randomized, and biofilm coverage (%) was assessed using Photoshop CS5. The biofilm coverage (%) on the cementum increased with increasing incubation period. The biofilm coverage of the 7 d group was significantly higher than those of the 1 and 2 d groups (P < 0.05). The values of the latter two groups were not significantly different (P > 0.05). Dense aggregations composed of E. faecalis and the amorphous matrix were observed on the root cementum surfaces of the specimens in the 7 d group. The bacteria were stained red by PI, and the matrix was stained green by ConA-FITC under CLSM observation. The biofilm coverage (%) on the samples in the 7 d group was 17.23% +/- 1.52%, showing multi-level space structure and water channels. E. faecalis forms bacterial biofilms on the root cementum surface in 7 d. The biofilms were composed of E. faecalis and the amorphous matrix.

Surveillance of multidrug resistant uropathogenic bacteria in hospitalized patients in Indian

PubMed Central

Mishra, Monali Priyadarsini; Debata, Nagen Kumar; Padhy, Rabindra Nath

2013-01-01

Objective To record surveillance, antibiotic resistance of uropathogens of hospitalized patients over a period of 18 months. Methods Urine samples from wards and cabins were used for isolating urinary tract infection (UTI)-causing bacteria that were cultured on suitable selective media and identified by biochemical tests; and their antibiograms were ascertained by Kirby-Bauer's disc diffusion method, in each 6-month interval of the study period, using 18 antibiotics of five different classes. Results From wards and cabins, 1 245 samples were collected, from which 996 strains of bacteria belonging to 11 species were isolated, during April 2011 to September 2012. Two Gram-positive, Staphylococcus aureus (S. aureus) and Enterococcus faecalis (E. faecalis), and nine Gram-negative bacteria, Acinetobacter baumannii, Citrobacter sp., Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Proteus vulgaris and Pseudomonas aeruginosa were isolated. Both S. aureus and E. faecalis were vancomycin resistant, and resistant-strains of all pathogens increased in each 6-month period of study. Particularly, all Gram-negatives were resistant to nitrofurantoin and co-trimoxazole, the most preferred antibiotics of empiric therapy for UTI. Conclusions Antibiograms of 11 UTI-causing bacteria recorded in this study indicated moderately higher numbers of strains resistant to each antibiotic studied, generating the fear of precipitating fervent episodes in public health particularly with bacteria, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and S. aureus. Moreover, vancomycin resistance in strains of S. aureus and E. faecalis is a matter of concern. PMID:23620859

Food isolate Listeria monocytogenes harboring tetM gene plasmid-mediated exchangeable to Enterococcus faecalis on the surface of processed cheese.

PubMed

Haubert, Louise; Cunha, Carlos Eduardo Pouey da; Lopes, Graciela Völz; Silva, Wladimir Padilha da

2018-05-01

The genetic basis of tetracycline resistance in a food isolate Listeria monocytogenes (Lm16) was evaluated. Resistance to tetracycline was associated with the presence of the tetM gene in plasmid DNA. The sequence of tetM showed 100% of similarity with the Enterococcus faecalis sequences found in the EMBL database, suggesting that Lm16 received this gene from E. faecalis. Various size bands were detected in the DNA plasmid analysis, the largest being approximately 54.38 kb. Transferability of the tetM gene was achieved in vitro by agar matings between Lm16 and E. faecalis JH2-2, proving the potential for the spread of tetM by horizontal gene transfer. Furthermore, the conjugation experiments were performed on the surface of processed cheese, confirming the transferability in a food matrix. PCR assays were used to confirm the identity of E. faecalis and to detect the tetM gene in transconjugant bacteria. Additionally, the minimal inhibitory concentration for tetracycline and rifampicin and plasmid profiling were performed. This is the first report of a food isolate L. monocytogenes carrying the tetM gene in plasmid DNA, and it highlights the potential risk of spreading antimicrobial resistance genes between different bacteria. Copyright © 2018 Elsevier Ltd. All rights reserved.

Functional Dissection of the CroRS Two-Component System Required for Resistance to Cell Wall Stressors in Enterococcus faecalis.

PubMed

Kellogg, Stephanie L; Kristich, Christopher J

2016-04-01

Bacteria use two-component signal transduction systems (TCSs) to sense and respond to environmental changes via a conserved phosphorelay between a sensor histidine kinase and its cognate response regulator. The opportunistic pathogen Enterococcus faecalis utilizes a TCS comprised of the histidine kinase CroS and the response regulator CroR to mediate resistance to cell wall stresses such as cephalosporin antibiotics, but the molecular details by which CroRS promotes cephalosporin resistance have not been elucidated. Here, we analyzed mutants of E. faecalis carrying substitutions in CroR and CroS to demonstrate that phosphorylated CroR drives resistance to cephalosporins, and that CroS exhibits kinase and phosphatase activities to control the level of CroR phosphorylation in vivo. Deletion of croS in various lineages of E. faecalis revealed a CroS-independent mechanism for CroR phosphorylation and led to the identification of a noncognate histidine kinase capable of influencing CroR (encoded by OG1RF_12162; here called cisS). Further analysis of this TCS network revealed that both systems respond to cell wall stress. TCSs allow bacteria to sense and respond to many different environmental conditions. The opportunistic pathogen Enterococcus faecalis utilizes the CroRS TCS to mediate resistance to cell wall stresses, including clinically relevant antibiotics such as cephalosporins and glycopeptides. In this study, we use genetic and biochemical means to investigate the relationship between CroRS signaling and cephalosporin resistance in E. faecalis cells. Through this, we uncovered a signaling network formed between the CroRS TCS and a previously uncharacterized TCS that also responds to cell wall stress. This study provides mechanistic insights into CroRS signaling and cephalosporin resistance in E. faecalis. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

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

Combined Antibacterial Effect of Sodium Hypochlorite and Root Canal Sealers against Enterococcus faecalis Biofilms in Dentin Canals.

PubMed

Du, Tianfeng; Wang, Zhejun; Shen, Ya; Ma, Jingzhi; Cao, Yingguang; Haapasalo, Markus

2015-08-01

The present study aimed to evaluate the antibacterial effect of the combined use of sodium hypochlorite (NaOCl) and root canal sealers on Enterococcus faecalis biofilms using a dentin infection model. Cells of E. faecalis were introduced into the dentinal tubules by centrifugation and incubated in brain-heart infusion for 3 weeks. The biofilms in dentin were first subjected to 5% NaOCl or sterile water for 10 minutes followed by an equal thickness of AH Plus (Dentsply International Inc, York, PA), Endosequence BC Sealer (Brasseler USA, Savannah, GA), or MTA Fillapex (Angelus Indústria de Produtos Odontológicos S/A, Londrina, Brazil) placed on the root canal wall of the dentin specimens for 7, 30, and 60 days. Gutta-percha and water were used in a similar manner as controls. The proportions of dead and live bacteria inside the dentinal tubules were assessed by confocal laser scanning microscopy and viability staining. The combined use of NaOCl and sealers (30 and 60 days) killed significantly more bacteria than NaOCl or sealers alone (P < .05). NaOCl + MTA Fillapex was the most effective antibacterial combination by killing 83% bacteria in dentin tubules in 60 days. Thirty and 60 days of exposure to the sealers resulted in significantly more dead bacteria in dentin biofilms than 7-day exposures (P < .05). The placement of root canal sealer after NaOCl treatment enhanced antibacterial effects against E. faecalis in the dentinal tubules. Little additional effect was obtained after 30 days of exposure to sealers. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

The Effect of Seed Soaking with Rhizobacteria Pseudomonas alcaligenes on the Growth of Swamp Cabbage (Ipomoea reptans Poir)

NASA Astrophysics Data System (ADS)

Widnyana, I. K.; Ngga, M.; Sapanca, P. L. Y.

2018-01-01

The research was conducted to determine the effect of seed soaking with suspense of P. alcaligenes isolate KtSl, TrN2, and TmAl to the growth of swamp cabbage. The research has been initially developed on tomatoes. In this research, Randomized Block Design was chosen as its model while the data analysis was performed by using SPSS v.17 for Windows. Three types of treatment were administered towards P. alcaligenes, namely isolating, soaking, and growing the medium. Some observed parameters were germination and growth. The results showed that seed soaking treatments with suspense P. alcaligenes fostered the germination 25% faster, enhanced the crop up to 24.4%, increased the number of leaves up until 23.15%, lengthen stems to 25%, lengthen the roots up to 46.90%, and increase the fresh weight of stems up until 67.07% and oven-dry weight of stem up to 84.21% compared to the control treatment. The best response of treatment for germination speed was soaking seeds with P. alcaligenes TrN2 for 20 minutes on both NB (Natrium Broth) and PDB (Potato Dextrose Broth) media.

Effects of Diode Laser, Gaseous Ozone, and Medical Dressings on Enterococcus faecalis Biofilms in the Root Canal Ex Vivo

PubMed Central

Vlassakidis, Alexander; Niepel, Mediha; Hoedke, Daniela; Schulze, Julia; Neumann, Konrad; Moter, Annette; Noetzel, Jörn

2017-01-01

The objective was to compare the antibacterial effects of adjunctive disinfection using diode laser and gaseous ozone compared to the medical dressings calcium hydroxide (Ca(OH)2) and chlorhexidine gel (CHX-Gel) on Enterococcus faecalis biofilms in human root canals ex vivo. Root canals of 180 human extracted teeth were infected by E. faecalis and divided into 3 main groups (G): G1, control; G2, instrumentation and irrigation using 0.9% NaCl; G3, instrumentation and irrigation using 1% NaOCl. In each main group, the following treatments were applied: gaseous ozone, diode laser, and medical dressings of Ca(OH)2 or CHX-Gel for 7 days (n = 15). Reduction of colony forming units (CFUs) inside the root canal of planktons and frequencies of adherent bacteria after treatment were calculated. Bacterial reduction was significantly affected by the irrigation protocol (p < 0.0005) and the disinfection method (p < 0.0005), and a significant interaction between both factors could be observed (p < 0.0005; ANOVA). In G3 (instrumentation using 1% NaOCl), no significant effect of disinfection methods could be demonstrated on planktonic bacteria (p = 0.062; ANOVA) and frequencies of adherent bacteria (p > 0.05; chi-square test). Instrumentation and irrigation using NaOCl combined with ozone or laser application resulted in comparable bacterial reduction on E. faecalis to the application of medical dressings. PMID:28567421

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

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

PubMed

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

2015-08-06

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.

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.

Molecular Determinants of the Thickened Matrix in a Dual-Species Pseudomonas aeruginosa and Enterococcus faecalis Biofilm

PubMed Central

Lee, Keehoon; Lee, Kang-Mu; Kim, Donggeun

2017-01-01

studies have reported that microbes in polymicrobial biofilms interact with each other and that the bacterial interactions result in elevated virulence, in terms of factors, such as infectivity and antibiotic resistance. Pseudomonas aeruginosa and Enterococcus faecalis are frequently isolated pathogens in chronic biofilm infections. Nevertheless, while both bacteria are known to be agents of numerous nosocomial infections and can cause serious diseases, interactions between the bacteria in biofilms have rarely been examined. In this investigation, we aimed to characterize P. aeruginosa and E. faecalis dual-species biofilms and to determine the molecular factors that cause synergistic effects, especially on the matrix thickening of the biofilm. We suspect that our findings will contribute to the development of more efficient methods for eradicating polymicrobial biofilm infections. PMID:28842537

Molecular Determinants of the Thickened Matrix in a Dual-Species Pseudomonas aeruginosa and Enterococcus faecalis Biofilm.

PubMed

Lee, Keehoon; Lee, Kang-Mu; Kim, Donggeun; Yoon, Sang Sun

2017-11-01

have reported that microbes in polymicrobial biofilms interact with each other and that the bacterial interactions result in elevated virulence, in terms of factors, such as infectivity and antibiotic resistance. Pseudomonas aeruginosa and Enterococcus faecalis are frequently isolated pathogens in chronic biofilm infections. Nevertheless, while both bacteria are known to be agents of numerous nosocomial infections and can cause serious diseases, interactions between the bacteria in biofilms have rarely been examined. In this investigation, we aimed to characterize P. aeruginosa and E. faecalis dual-species biofilms and to determine the molecular factors that cause synergistic effects, especially on the matrix thickening of the biofilm. We suspect that our findings will contribute to the development of more efficient methods for eradicating polymicrobial biofilm infections. Copyright © 2017 American Society for Microbiology.

Comparison of the antibacterial effect of sodium hypochlorite and aloe vera solutions as root canal irrigants in human extracted teeth contaminated with enterococcus faecalis.

PubMed

Sahebi, S; Khosravifar, N; Sedighshamsi, M; Motamedifar, M

2014-03-01

The main purpose of a root canal treatment is to eliminate the bacteria and their products from the pulp space. Sodium hypochlorite has excellent antibacterial properties, but also some negative features. The aim of the present study is to compare the antimicrobial effect of Aloe Vera solution with sodium hypochlorite on E.faecalis in the root canals of human extracted teeth. Sixty human extracted single rooted teeth were selected for this in vitro study. The teeth recruited in this study had no cracks, internal resorption, external resorption and calcification. Enterococcus faecalis was injected in the root canals of all teeth. The teeth were then divided into three groups randomly. Each group consisted of 20 teeth that were all rinsed with one of the following solutions: sodium hypochlorite 2.5%, Aloe vera and normal saline. Subsequent to rinsing, root canals of all teeth were sampled. The samples were cultured and growth of the bacteria was assessed after 48 hours. The number of colonies of the bacteria was then counted. The difference between the inhibitory effect of Aloe vera and normal saline on E.faecalis was not significant according to independent t-test (p= 0.966). The inhibitory effect of sodium hypochlorite on E.faecalis was much greater than that of Aloe vera and normal saline (p< 0.001). Aloe vera solution is not recommended as a root canal irrigator, but future studies are suggested to investigate the antibacterial effect of Aloe vera with longer duration of exposure and as an intra canal medicament.

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.

Targeting Enterococcus faecalis Biofilms with Phage Therapy

PubMed Central

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

2015-01-01

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. PMID:25662974

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.

Flow cytometry as a rapid test for detection of penicillin resistance directly in bacterial cells in Enterococcus faecalis and Staphylococcus aureus.

PubMed

Jarzembowski, T; Wiśniewska, K; Józwik, A; Bryl, E; Witkowski, J

2008-08-01

We studied the usefulness of flow cytometry for detection of penicillin resistance in E. faecalis and S. aureus by direct binding of commercially available fluorescent penicillin, Bocillin FL, to cells obtained from culture. There were significantly lower percentages of fluorescent cells and median and mean fluorescence values per particle in penicillin-resistant than in penicillin-sensitive strains of both species observed. The method allows rapid detection of penicillin resistance in S. aureus and E. faecalis. The results encourage further investigations on the detection of antibiotic resistance in bacteria using flow cytometry.

Antimicrobial effectiveness of intracanal medicaments on Enterococcus faecalis: chlorhexidine versus octenidine.

PubMed

de Lucena, J M V M; Decker, E M; Walter, C; Boeira, L S; Löst, C; Weiger, R

2013-01-01

To determine the viability of Enterococcus faecalis in infected human root dentine in vitro after exposure to root canal medicaments based on chlorhexidine and octenidine. Human root segments (n = 40) were infected with E. faecalis for 8 weeks. Root dentine samples (rd) collected at week 4 served as individual baseline values. At week 8, the root segments were randomly divided into four test groups (n = 10 each) for the placement of one of the following medicaments in the root canals: calcium hydroxide paste (CH), chlorhexidine gel (CHX-gel) (5.0%), chlorhexidine/gutta-percha points (CHX-GP) (active points(®) ; Roeko, Langenau, Germany) and octenidine gel (OCT-gel) (5.0%) followed by incubation for 4 weeks. The effect on E. faecalis viability was assessed by two fluorescent dyes (syto 9/propidium iodide) to determine the 'proportion of viable bacteria' (PVB%) and number of 'colony-forming units' (CFU). Mean values and 95% confidence intervals (CI) were calculated for PVB% and log CFU, and the difference between groups was established. Viable and dead bacterial cells were detected in all 'rd' samples at weeks 4 and 8. The treatment with CHX-gel, CHX-GP and OCT-gel resulted in significantly lower PVB% values with 15.4%, 3.5% and 0%, respectively. No growth (CFU) was recorded for these samples at week 12. When medicated by CH, the PVB% was increased without a corresponding change in CFUs. In contrast to calcium hydroxide, both CHX - and octenidine-based intracanal medicaments were effective in decreasing the viability of E. faecalis. OCT showed the most favourable results and may have potential as an endodontic medicament. © 2012 International Endodontic Journal.

Insight into the bacterial diversity of fermentation woad dye vats as revealed by PCR-DGGE and pyrosequencing.

PubMed

Milanović, Vesna; Osimani, Andrea; Taccari, Manuela; Garofalo, Cristiana; Butta, Alessandro; Clementi, Francesca; Aquilanti, Lucia

2017-07-01

The bacterial diversity in fermenting dye vats with woad (Isatis tinctoria L.) prepared and maintained in a functional state for approximately 12 months was examined using a combination of culture-dependent and -independent PCR-DGGE analyses and next-generation sequencing of 16S rRNA amplicons. An extremely complex ecosystem including taxa potentially contributing to both indigo reduction and formation, as well as indigo degradation was found. PCR-DGGE analyses revealed the presence of Paenibacillus lactis, Sporosarcina koreensis, Bacillus licheniformis, and Bacillus thermoamylovorans, while Bacillus thermolactis, Bacillus pumilus and Bacillus megaterium were also identified but with sequence identities lower than 97%. Dominant operational taxonomic units (OTUs) identified by pyrosequencing included Clostridium ultunense, Tissierella spp., Alcaligenes faecalis, Erysipelothrix spp., Enterococcus spp., Virgibacillus spp. and Virgibacillus panthothenicus, while sub-dominant OTUs included clostridia, alkaliphiles, halophiles, bacilli, moderately thermophilic bacteria, lactic acid bacteria, Enterobacteriaceae, aerobes, and even photosynthetic bacteria. Based on the current knowledge of indigo-reducing bacteria, it is considered that indigo-reducing bacteria constituted only a small fraction in the unique microcosm detected in the natural indigo dye vats.

Antimicrobial Activity of Photodynamic Therapy Against Enterococcus faecalis Before and After Reciprocating Instrumentation in Permanent Molars.

PubMed

Pinheiro, Sérgio Luiz; Azenha, Giuliana Rodrigues; Democh, Yasmin Marialva; Nunes, Daniela Camila; Provasi, Silvia; Fontanetti, Giovana Masiero; Duarte, Danilo Antônio; Fontana, Carlos Eduardo; da Silveira Bueno, Carlos Eduardo

2016-12-01

The present study sought to evaluate the antimicrobial activity against Enterococcus faecalis of photodynamic therapy applied before and after reciprocating instrumentation of permanent molars. Apical extrusion of debris can cause flare-ups due to introduction of bacteria into the periapical tissues. Eighteen mesial roots from permanent mandibular molars were selected. The crowns were removed to obtain a standard root length of 15 mm. The included mesial roots had an angulation of 10°-40° and canals with independent foramina. The orifice of each mesiolingual canal was sealed with light-curing resin, and the working length was established visually, 1 mm short of the apical foramen. The roots were rendered impermeable and sterilized, and the mesiobuccal canals were contaminated with a standard strain of E. faecalis for 21 days. Specimens were randomly divided into three groups (n = 6): G1, photodynamic therapy performed before instrumentation and irrigation with 0.9% NaCl (saline) solution; G2, photodynamic therapy performed after instrumentation and irrigation with 0.9% NaCl; and G3 (control), instrumentation and irrigation with 2.5% NaOCl (sodium hypochlorite) solution. Canals were shaped with a WaveOne primary file (25.08) and irrigated with 0.9% NaCl. E. faecalis samples were collected before and after each procedure, and the results were analyzed using descriptive statistics and the Kruskal-Wallis and Wilcoxon tests. Significant reductions in E. faecalis were observed when photodynamic therapy was performed before and after instrumentation of the root canal system (p < 0.05). Reciprocating instrumentation significantly reduced E. faecalis colonies in experimentally contaminated root canal systems (p < 0.05). Photodynamic therapy was effective in removing E. faecalis from the root canal system, whether performed before or after reciprocating instrumentation.

Subinhibitory concentrations of cell wall synthesis inhibitors promote biofilm formation of Enterococcus faecalis

NASA Astrophysics Data System (ADS)

Yu, Wen; Hallinen, Kelsey; Wood, Kevin

Enterococcus faecalis are commonly associated with hospital acquired infections, because they readily form biofilms on instruments and medical devices. Biofilms are inherently more resistant to killing by antibiotics compared to planktonic bacteria, in part because of their heterogeneous spatial structure. Surprisingly, however, subminimal inhibitory concentrations (sub-MICs) of some antibiotics can actually promote biofilm formation. Unfortunately, much is still unknown about how low drug doses affect the composition and spatial structure of the biofilm. In this work, we investigate the effects of sub-MICs of ampicillin on the formation of E. faecalis biofilms. First, we quantified biofilm mass using crystal violet staining in polystyrene microtiter plates. We found that total biofilm mass is increased over a narrow range of ampicillin concentrations before ultimately declining at higher concentrations. Second, we show that sub-MICs of ampicillin can increase mass of E. faecalis biofilms while simultaneously increasing extracellular DNA/RNA and changing total number of viable cells under confocal microscopy. Further, we use RNA-seq to identify genes differentially expressed under sub-MICs of ampicillin. Finally, we show a mathematical model to explain this phenomenon. This work was funded by The Hartwell Foundation Individual Biomedical Research Award and NSF CAREER 1553208 to KBW.

Comparative genomics of Enterococcus faecalis from healthy Norwegian infants

PubMed Central

Solheim, Margrete; Aakra, Ågot; Snipen, Lars G; Brede, Dag A; Nes, Ingolf F

2009-01-01

Background Enterococcus faecalis, traditionally considered a harmless commensal of the intestinal tract, is now ranked among the leading causes of nosocomial infections. In an attempt to gain insight into the genetic make-up of commensal E. faecalis, we have studied genomic variation in a collection of community-derived E. faecalis isolated from the feces of Norwegian infants. Results The E. faecalis isolates were first sequence typed by multilocus sequence typing (MLST) and characterized with respect to antibiotic resistance and properties associated with virulence. A subset of the isolates was compared to the vancomycin resistant strain E. faecalis V583 (V583) by whole genome microarray comparison (comparative genomic hybridization (CGH)). Several of the putative enterococcal virulence factors were found to be highly prevalent among the commensal baby isolates. The genomic variation as observed by CGH was less between isolates displaying the same MLST sequence type than between isolates belonging to different evolutionary lineages. Conclusion The variations in gene content observed among the investigated commensal E. faecalis is comparable to the genetic variation previously reported among strains of various origins thought to be representative of the major E. faecalis lineages. Previous MLST analysis of E. faecalis have identified so-called high-risk enterococcal clonal complexes (HiRECC), defined as genetically distinct subpopulations, epidemiologically associated with enterococcal infections. The observed correlation between CGH and MLST presented here, may offer a method for the identification of lineage-specific genes, and may therefore add clues on how to distinguish pathogenic from commensal E. faecalis. In this work, information on the core genome of E. faecalis is also substantially extended. PMID:19393078

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

PubMed

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

2016-04-01

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. 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. 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.

Investigation of the effect of rapid and slow external pH increases on Enterococcus faecalis biofilm grown on dentine.

PubMed

Stenhouse, M; Zilm, P; Ratnayake, J; Cathro, P

2018-01-11

Calcium hydroxide is a common endodontic medicament and has an antimicrobial effect by increasing the localized pH within the root canal. However, Enterococcus faecalis has shown some resistance to calcium hydroxide. A flow cell apparatus was used to grow an E. faecalis biofilm on dentine discs. Following 4 weeks growth in Todd Hewitt Broth, flow cells were exposed to either a rapid or slow increase to pH 11.5 or 12.5. Cellular viability was determined using serial plating and the number of colony-forming units was normalized against the cellular protein content. Scanning electron microscopy was carried out to qualitatively observe the effects of the different rates of pH increase. A significant difference in viability between the pH rapid and slow groups was not shown in this study. Compared with pH 11.5 solutions, pH 12.5 solutions were more effective at killing bacteria although some E. faecalis still survived. Enterococcus faecalis did not adapt and develop a greater resistance to high pH following a slow rise in pH compared with a rapid rise in pH. As expected, pH 12.5 was more effective in reducing bacterial numbers compared with pH 11.5 although E. faecalis was not completely eliminated. © 2018 Australian Dental Association.

Convergence of PASTA kinase and two-component signaling in response to cell wall stress in Enterococcus faecalis.

PubMed

Kellogg, Stephanie L; Kristich, Christopher J

2018-04-09

Two common signal transduction mechanisms used by bacteria to sense and respond to changing environments are two-component systems (TCSs) and eukaryotic-like Ser/Thr kinases and phosphatases (eSTK/Ps). Enterococcus faecalis is a Gram-positive bacterium and serious opportunistic pathogen that relies on both a TCS and an eSTK/P pathway for intrinsic resistance to cell wall-targeting antibiotics. The TCS consists of a histidine kinase (CroS) and response regulator (CroR) that become activated upon exposure of cells to cell wall-targeting antibiotics, leading to modulation of gene expression. The eSTK/P pathway consists of a transmembrane kinase (IreK) and its cognate phosphatase (IreP), which act antagonistically to mediate antibiotic resistance through an unknown mechanism. Because both CroS/R and IreK/P contribute to enterococcal resistance towards cell wall-targeting antibiotics, we hypothesized these signaling systems are intertwined. To test this hypothesis, we analyzed CroR phosphorylation and CroS/R-dependent gene expression to probe the influence of IreK and IreP on CroS/R signaling. In addition, we analyzed the phosphorylation state of CroS which revealed IreK-dependent phosphorylation of a Thr residue important for CroS function. Our results are consistent with a model in which IreK positively influences CroR-dependent gene expression through phosphorylation of CroS to promote antimicrobial resistance in E. faecalis Importance Two-component signaling systems (TCSs) and eukaryotic-like Ser/Thr kinases (eSTKs) are used by bacteria to sense and adapt to changing environments. Understanding how these pathways are regulated to promote bacterial survival is critical for a more complete understanding of bacterial stress responses and physiology. The opportunistic pathogen Enterococcus faecalis relies on both a TCS (CroS/R) and an eSTK (IreK) for intrinsic resistance to cell wall-targeting antibiotics. We probed the relationship between CroS/R and IreK, revealing

Calcium-silicate mesoporous nanoparticles loaded with chlorhexidine for both anti- Enterococcus faecalis and mineralization properties.

PubMed

Fan, Wei; Li, Yanyun; Sun, Qing; Ma, Tengjiao; Fan, Bing

2016-10-21

In infected periapical tissues, Enterococcus faecalis is one of the most common dominant bacteria. Chlorhexidine has been proved to show strong antibacterial ability against E. faecalis but is ineffective in promoting mineralization for tissues around root apex. Mesoporous calcium-silicate nanoparticles are newly synthesized biomaterials with excellent ability to promote mineralization and carry-release bioactive molecules in a controlled manner. In this study, mesoporous calcium-silicate nanoparticles were functionalized with chlorhexidine and their releasing profile, antibacterial ability, effect on cell proliferation and in vitro mineralization property were evaluated. The chlorhexidine was successfully incorporated into mesoporous calcium-silicate nanoparticles by a mixing-coupling method. The new material could release chlorhexidine as well as Ca 2+ and SiO 3 2- in a sustained manner with an alkaline pH value under different conditions. The antimicrobial ability against planktonic E. faecalis was dramatically improved after chlorhexidine incorporation. The nanoparticles with chlorhexidine showed no negative effect on cell proliferation with low concentrations. On dentin slices, the new synthesized material demonstrated a similar inhibitory effect on E. faecalis as the chlorhexidine. After being immersed in SBF for 9 days, numerous apatite crystals could be observed on surfaces of the material tablets. Mesoporous calcium-silicate nanoparticles loaded with chlorhexidine exhibited release of ions and chlorhexidine, low cytotoxicity, excellent antibacterial ability and in vitro mineralization. This material could be developed into a new effective intra-canal medication in dentistry or a new bone defect filling material for infected bone defects.

Evaluation of 4% Sodium Hypochlorite in eliminating Enterococcus faecalis from the Root Canal when Used with Three Irrigation Methods: An in vitro Study.

PubMed

Priyank, Harsh; Pandey, Vinisha; Bagul, Abhishek; Majety, Kishore Kumar; Verma, Parul; Choudhury, Basanta Kumar

2017-03-01

Endodontic treatment removes all pathogens, such as Enterococcus faecalis from pulp and root canals. The aim of this study is to assess the usefulness of sodium hypo-chlorite (NaOCl) in removing E. faecalis from the root canal used with three different irrigation methods. This study was conducted on freshly extracted maxillary incisors. After biomechanical preparation, root canals were injected with E. faecalis. Three groups were made which contained 30 teeth in each group; 2 mL of NaOCl solution was used for irrigation followed by agitation with K-files in group I; 2 mL of NaOCl solution was used for irrigation and ultrasonic agitation was done in group II. In group III, an alternate irrigation with NaOCl and 3% hydrogen peroxide was done. The fourth group (control) was irrigated with sterile saline solution. E. fae-calis bacteria were sampled to the root canals with paper points and were transferred to tubes that contained 5 mL of brain heart infusion broth. Tubes were incubated and the presence of broth turbidity was suggestive of bacteria remaining in the root canal. All three groups showed no statistically significant difference. However, difference existed between experimental groups and control groups. The author concluded that all three methods of application of NaOCl were effective in disinfecting the root canal than the saline solution. No single irrigant has 100% efficiency. Thus by this study, a best irrigating solution with maximum properties can be established.

Biosynthesis of selenium nanoparticles using Enterococcus faecalis and evaluation of their antibacterial activities.

PubMed

Shoeibi, Sara; Mashreghi, Mohammad

2017-01-01

Microorganisms are capable of synthesizing metal nanoparticles, and specifically Enterococcus faecalis bacteria were tested for its ability to synthesize selenium nanoparticles (Se-NPs) from sodium selenite. The biosynthesized Se-NPs were spherical in shape with the size range of 29-195nm. Also, the TEM microscopy showed the accumulation of nano-structures as extracellular deposits. The ability of the bacteria to tolerate high levels of toxic selenite was studied by changing with different concentrations of sodium selenite (0.19mM-2.97mM). Also, the effect of Se-NPs was studied on the growth profile of number of pathogenic Gram-positive and -negative bacteria. High concentrations of sodium selenite in the medium led to the production of small amounts of selenium nanostructures by bacteria. In addition, Se-NPs can be used as an anti-staphylococcal element to effectively prevent and treat S. aureus infections. Copyright © 2016 Elsevier GmbH. All rights reserved.

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

PubMed

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

2016-01-01

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). 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. 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). 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.

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.

Wide Distribution of Virulence Genes among Enterococcus faecium and Enterococcus faecalis Clinical Isolates

PubMed Central

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. PMID:25147855

The Starvation Resistance and Biofilm Formation of Enterococcus faecalis in Coexistence with Candida albicans, Streptococcus gordonii, Actinomyces viscosus, or Lactobacillus acidophilus.

PubMed

Gao, Yan; Jiang, Xiaoqiong; Lin, Dongjia; Chen, Yanhuo; Tong, Zhongchun

2016-08-01

Enterococcus faecalis is the most frequently detected species in root canal-treated teeth, and it is able to survive under starvation conditions. However, persistent periapical disease is often caused by multispecies. The aim of this study was to explore the survival of E. faecalis in starvation conditions and biofilm formation with the 4 common pathogenic species. A dual-species model of Candida albicans, Streptococcus gordonii, Actinomyces viscosus, or Lactobacillus acidophilus in combination with E. faecalis was established and allowed to grow in phosphate-buffered saline for the examination of starvation survival. Cefuroxime sodium and vancomycin at a concentration of 100 mg/L were added into brain-heart infusion plate agar to count the 2 bacteria separately in the dual species. Scanning electron microscopy was used to observe the dual species and multiple species on the root canal dentin of bovine teeth for 48 hours. A confocal laser scanning microscope was used to show the 4 groups of dual-species biofilms on substrates with glass bottoms for 48 hours. E. faecalis was more resistant to starvation in coexistence with C. albicans, S. gordonii, A. viscosus, or L. acidophilus, and S. gordonii was completely inhibited in coexistence with E. faecalis. The dual-species biofilm showed that E. faecalis formed thicker and denser biofilms on the root canal dentin and glass slides in coexistence with S. gordonii and A. viscosus than C. albicans and L. acidophilus. The multispecies community is conducive to the resistance to starvation of E. faecalis and biofilm formation in root canals. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Susceptibility of Enterococcus faecalis and Propionibacterium acnes to antimicrobial photodynamic therapy.

PubMed

de Annunzio, Sarah Raquel; de Freitas, Laura Marise; Blanco, Ana Lígia; da Costa, Mardoqueu Martins; Carmona-Vargas, Christian C; de Oliveira, Kleber Thiago; Fontana, Carla Raquel

2018-01-01

Bacterial resistance to available antibiotics nowadays is a global threat leading researchers around the world to study new treatment modalities for infections. Antimicrobial photodynamic therapy (aPDT) has been considered an effective and promising therapeutic alternative in this scenario. Briefly, this therapy is based on the activation of a non-toxic photosensitizing agent, known as photosensitizer (PS), by light at a specific wavelength generating cytotoxic singlet oxygen and free radicals. Virtually all studies related to aPDT involve a huge screening to identify ideal PS concentration and light dose combinations, a laborious and time-consuming process that is hardly disclosed in the literature. Herein, we describe an antimicrobial Photodynamic Therapy (aPDT) study against Enterococcus faecalis and Propionibacterium acnes employing methylene blue, chlorin-e6 or curcumin as PS. Similarities and discrepancies between the two bacterial species were pointed out in an attempt to speed up and facilitate futures studies against those clinical relevant strains. Susceptibility tests were performed by the broth microdilution method. Our results demonstrate that aPDT mediated by the three above-mentioned PS was effective in eliminating both gram-positive bacteria, although P. acnes showed remarkably higher susceptibility to aPDT when compared to E. faecalis. PS uptake assays revealed that P. acnes is 80 times more efficient than E. faecalis in internalizing all three PS molecules. Our results evidence that the cell wall structure is not a limiting feature when predicting bacterial susceptibility to aPDT treatment. Copyright © 2017. Published by Elsevier B.V.

Prevalence of Enterococcus faecalis and Porphyromonas gingivalis in infected root canals and their susceptibility to endodontic treatment procedures: a molecular study.

PubMed

Stojanović, Nikola; Krunić, Jelena; Popović, Branka; Stojičić, Sonja; Zivković, Slavoljub

2014-01-01

Because apical periodontitis is recognizably an infectious disease, elimination or reduction of intracanal bacteria is of utmost importance for optimum treatment outcome. The prevalence of Enterococcus faecalis and Porphyromonas gingivalis in infected root canals was studied Also, the effect of endodontic therapy by using intracanal medicaments, calcium hydroxide paste (CH) or gutta-percha points containing calcium hydroxide (CH-GP) or chlorhexidine (CHX-GP) on these microorganisms was assessed by polymerase chain reaction (PCR) assay. Fifty-one patients with chronic apical periodontitis were randomly allocated in one of the fol- lowing groups according to the intracanal medicament used: CH, CH-GP and CHX-GP group. Bacterial samples were taken upon access (S1), after chemomechanical instrumentation (S2) and after 15-day medication (S3). PCR assay was used to detect the presence of selected bacteria. E. faecalis was detected in 49% (25/51) and P. gingivalis in 17.6% (9/51) of the samples. Samples which showed no bacterial presence at S1 were excluded from further analysis. Overall analysis of all 29 samples revealed significant differences between S1 and S2 (p < 0.001), S2 and S3 (p < 0.05), and S1 and S3 (p < 0.001). When distinction was made between the intracanal medications, there was a significant difference in the number of PCR positive samples between S1 and 52, S1 and S3, but not between S2 and S3 samples. E. faecalis is more prevalent than P. gingivalis in primary endodontic infection. Intracanal medication in conduction with instrumentation and irrigation efficiently eliminates E. faecalis and P. gingivalis from infected root canals.

Study of the impact of cranberry extract on the virulence factors and biofilm formation by Enterococcus faecalis strains isolated from urinary tract infections.

PubMed

Wojnicz, Dorota; Tichaczek-Goska, Dorota; Korzekwa, Kamila; Kicia, Marta; Hendrich, Andrzej B

2016-12-01

Drinking of cranberry fruit juice and application of commercial preparations containing the cranberry extracts are recommended in the prevention and treatment of urinary tract infections (UTIs), especially in women with recurrent UTIs. Many studies focus on the activity of cranberries against uropathogenic Escherichia coli (E. coli) strains. However, the knowledge of the cranberry effect on Gram-positive Enterococcus faecalis (E. faecalis) is limited. Therefore, the aim of our study was to establish the activity of commercial concentrated cranberry extract on the growth, virulence factors and biofilm formation of E. faecalis strains isolated from urine. Minimal inhibitory concentrations (MICs) of cranberry extract were determined by the broth microdilution method. Disc diffusion method was used to determine antimicrobial susceptibility. The impact of cranberry extract on bacterial survival, hydrophobicity, synthesis of lipase, lecithinase, DNase, hemolysin, gelatinase and biofilm mass was determined. Results show that cranberry extract inhibits the growth, enzymatic activities of bacteria and limits biofilm formation. The antibacterial activities of the studied cranberry extract confirm that it could be successfully used in prevention of UTIs caused by E. faecalis.

A newly isolated probiotic Enterococcus faecalis strain from vagina microbiota enhances apoptosis of human cancer cells.

PubMed

Nami, Y; Abdullah, N; Haghshenas, B; Radiah, D; Rosli, R; Yari Khosroushahi, A

2014-08-01

This study aimed to describe probiotic properties and bio-therapeutic effects of newly isolated Enterococcus faecalis from the human vaginal tract. The Enterococcus faecalis strain was originally isolated from the vaginal microbiota of Iranian women and was molecularly identified using 16SrDNA gene sequencing. Some biochemical methodologies were preliminarily used to characterize the probiotic potential of Ent. faecalis, including antibiotic susceptibility, antimicrobial activity, as well as acid and bile resistance. The bio-therapeutic effects of this strain's secreted metabolites on four human cancer cell lines (AGS, HeLa, MCF-7 and HT-29) and one normal cell line (HUVEC) were evaluated by cytotoxicity assay and apoptosis scrutiny. The characterization results demonstrated into the isolated bacteria strain revealed probiotic properties, such as antibiotic susceptibility, antimicrobial activity and resistance under conditions similar to those in the gastrointestinal tract. Results of bio-therapeutic efficacy assessments illustrated acceptable apoptotic effects on four human cancer cell lines and negligible side effects on assayed normal cell line. Our findings revealed that the apoptotic effect of secreted metabolites mainly depended on proteins secreted by Ent. faecalis on different cancer cells. These proteins can induce the apoptosis of cancer cells. The metabolites produced by this vaginal Ent. faecalis strain can be used as alternative pharmaceutical compounds with promising therapeutic indices because they are not cytotoxic to normal mammalian cells. Accordingly, the physicochemical, structural and functional properties of the secreted anticancer substances should be further investigated before using them as anticancer therapeutics. This study aim to screen total bacterial secreted metabolites as a wealthy source to find the new active compounds to introduce as anticancer therapeutics in the future. © 2014 The Society for Applied Microbiology.

Brief communication: treatment of Enterococcus faecalis endocarditis with ampicillin plus ceftriaxone.

PubMed

Gavaldà, Joan; Len, Oscar; Miró, José M; Muñoz, Patricia; Montejo, Miguel; Alarcón, Aristides; de la Torre-Cisneros, Julián; Peña, Carmen; Martínez-Lacasa, Xavier; Sarria, Cristina; Bou, Germán; Aguado, José M; Navas, Enrique; Romeu, Joan; Marco, Francesc; Torres, Carmen; Tornos, Pilar; Planes, Ana; Falcó, Vicenç; Almirante, Benito; Pahissa, Albert

2007-04-17

High-level aminoglycoside resistance (HLAR) that precludes bactericidal synergism with penicillins or glycopeptides and nephrotoxicity related to aminoglycoside treatment are major problems in treating Enterococcus faecalis endocarditis. To evaluate the efficacy and safety of ampicillin plus ceftriaxone for treating endocarditis due to E. faecalis with and without HLAR. Observational, open-label, nonrandomized, multicenter clinical trial. 13 centers in Spain. 21 patients with HLAR E. faecalis endocarditis and 22 patients with non-HLAR E. faecalis endocarditis. All were at risk for nephrotoxicity related to aminoglycoside use. 6-week course of intravenous ampicillin, 2 g every 4 hours, plus intravenous ceftriaxone, 2 g every 12 hours. Clinical and microbiological outcomes. The clinical cure rate at 3 months was 67.4% (29 of 43 patients) among all episodes. During treatment, 28.6% of patients with HLAR E. faecalis endocarditis and 18.2% of patients with non-HLAR E. faecalis endocarditis died of infection-related causes. The rate of clinical and microbiological cure in patients who completed the protocol was 100% in the HLAR E. faecalis endocarditis group. No episodes of breakthrough bacteremia occurred, although there were 2 relapses in the non-HLAR E. faecalis endocarditis group. Treatment was withdrawn in 1 case because of fever and skin rash. The study had a small sample and was observational. The combination of ampicillin and ceftriaxone is effective and safe for treating HLAR E. faecalis endocarditis and could be a reasonable alternative for patients with non-HLAR E. faecalis endocarditis who are at increased risk for nephrotoxicity.

Presence of the resistance genes vanC1 and pbp5 in phenotypically vancomycin and ampicillin susceptible Enterococcus faecalis.

PubMed

Schwaiger, Karin; Bauer, Johann; Hörmansdorfer, Stefan; Mölle, Gabriele; Preikschat, Petra; Kämpf, Peter; Bauer-Unkauf, Ilse; Bischoff, Meike; Hölzel, Christina

2012-08-01

Ampicillin and vancomycin are important antibiotics for the therapy of Enterococcus faecalis infections. The ampicillin resistance gene pbp5 is intrinsic in Enterococcus faecium. The vanC1 gene confers resistance to vancomycin and serves as a species marker for Enterococcus gallinarum. Both genes are chromosomally located. Resistance to ampicillin and vancomycin was determined in 484 E. faecalis of human and porcine origin by microdilution. Since E. faecalis are highly skilled to acquire resistance genes, all strains were investigated for the presence of pbp5 (and, in positive strains, for the penicillin-binding protein synthesis repressor gene psr) and vanC1 (and, in positive strains, for vanXYc and vanT) by using polymerase chain reaction (PCR). One porcine and one human isolate were phenotypically resistant to ampicillin; no strain was vancomycin resistant. Four E. faecalis (3/1 of porcine/human origin) carried pbp5 (MIC=1 mg/L), and four porcine strains were vanC1 positive (minimum inhibitory concentration [MIC]=1 mg/L). Real-time reverse transcriptase (RT)-PCR revealed that the genes were not expressed. The psr gene was absent in the four pbp5-positive strains; the vanXYc gene was absent in the four vanC1-positive strains. However, vanT of the vanC gene cluster was detected in two vanC1-positive strains. To our knowledge, this is the first report on the presence of pbp5, identical with the "E. faecium pbp5 gene," and of vanC1/vanT in E. faecalis. Even if resistance is not expressed in these strains, this study shows that E. faecalis have a strong ability to acquire resistance genes-and potentially to spread them to other bacteria. Therefore, close monitoring of this species should be continued.

Engineering solutions to improve the removal of fecal indicator bacteria by bioinfiltration systems during intermittent flow of stormwater.

PubMed

Mohanty, Sanjay K; Torkelson, Andrew A; Dodd, Hanna; Nelson, Kara L; Boehm, Alexandria B

2013-10-01

Bioinfiltration systems facilitate the infiltration of urban stormwater into soil and reduce high flow events and flooding. Stormwater carries a myriad of pollutants including fecal indicator bacteria (FIB). Significant knowledge gaps exist about the ability of bioinfiltration systems to remove and retain FIB. The present study investigates the ability of model, simplified bioinfiltration systems containing quartz sand and iron oxide-coated quartz sand (IOCS) to remove two FIB (Enterococcus faecalis and Escherichia coli) suspended in synthetic stormwater with and without natural organic matter (NOM) as well as the potential for accumulated FIB to be remobilized during intermittent flow. The experiments were conducted in two phases: (1) the saturated columns packed with either sand or IOCS were contaminated by injecting stormwater with bacteria followed by injection of sterile stormwater and (2) the contaminated columns were subjected to intermittent infiltration of sterile stormwater preceded by a pause during which columns were either kept saturated or drained by gravity. During intermittent flow, fewer bacteria were released from the saturated column compared to the column drained by gravity: 12% of attached E. coli and 3% of attached Ent. faecalis were mobilized from the drained sand column compared to 3% of attached E. coli and 2% attached Ent. faecalis mobilized from the saturated sand column. Dry and wet cycles introduce moving air-water interfaces that can scour bacteria from grain surfaces. During intermittent flows, less than 0.2% of attached bacteria were mobilized from IOCS, which bound both bacteria irreversibly in the absence of NOM. Addition of NOM, however, increased bacterial mobilization from IOCS: 50% of attached E. coli and 8% of attached Ent. faecalis were released from IOCS columns during draining and rewetting. Results indicate that using geomedia such as IOCS that promote irreversible attachment of bacteria, and maintaining saturated

Exploiting CRISPR-Cas to manipulate Enterococcus faecalis populations.

PubMed

Hullahalli, Karthik; Rodrigues, Marinelle; Palmer, Kelli L

2017-06-23

CRISPR-Cas provides a barrier to horizontal gene transfer in prokaryotes. It was previously observed that functional CRISPR-Cas systems are absent from multidrug-resistant (MDR) Enterococcus faecalis , which only possess an orphan CRISPR locus, termed CRISPR2, lacking cas genes. Here, we investigate how the interplay between CRISPR-Cas genome defense and antibiotic selection for mobile genetic elements shapes in vitro E. faecalis populations. We demonstrate that CRISPR2 can be reactivated for genome defense in MDR strains. Interestingly, we observe that E. faecalis transiently maintains CRISPR targets despite active CRISPR-Cas systems. Subsequently, if selection for the CRISPR target is present, toxic CRISPR spacers are lost over time, while in the absence of selection, CRISPR targets are lost over time. We find that forced maintenance of CRISPR targets induces a fitness cost that can be exploited to alter heterogeneous E. faecalis populations.

Reclassification of Alcaligenes latus strains IAM 12599T and IAM 12664 and Pseudomonas saccharophila as Azohydromonas lata gen. nov., comb. nov., Azohydromonas australica sp. nov. and Pelomonas saccharophila gen. nov., comb. nov., respectively.

PubMed

Xie, Cheng-Hui; Yokota, Akira

2005-11-01

The aim of this study was to clarify the taxonomic position of the nitrogen-fixing and hydrogen-oxidizing bacteria Alcaligenes latus strains IAM 12599T, IAM 12664 and IAM 12665 and Pseudomonas saccharophila IAM 14368T. It was found that the type strain of Alcaligenes latus, IAM 12599T, showed 99 x 9 and 96 x 1 % 16S rRNA gene sequence similarity to strains IAM 12665 and IAM 12664, respectively. A comparison using DNA-DNA hybridization suggested that strains IAM 12599T and IAM 12665 belong to a single species (89 x 7 %) and that strain IAM 12664 (35 x 1 %) forms a separate species. The phenotypic characteristics also support the conclusion that these bacteria should be identified as two species of a new genus: Azohydromonas lata gen. nov., comb. nov. (type strain IAM 12599T=DSM 1122T=LMG 3321T=ATCC 29712T; reference strain IAM 12665=DSM 1123=LMG 3325=ATCC 29714) and Azohydromonas australica sp. nov. (type strain IAM 12664T=DSM 1124T=LMG 3324T=ATCC 29713T). Pseudomonas saccharophila IAM 14368T was found to be closely related to the phototrophic bacterium Roseateles depolymerans, with 96 x 8 % 16S rRNA gene sequence similarity, but the two bacteria are quite different with respect to their metabolism and some significant phenotypic characteristics, suggesting that they cannot be included in a single genus. Further studies on their nifH gene sequences, G+C content of the DNA and cellular fatty acid composition confirm that Pseudomonas saccharophila should be reclassified: the name Pelomonas saccharophila gen. nov., comb. nov. is proposed, with the type strain IAM 14368T (=LMG 2256T=ATCC 15946T).

Bacteriocinogenic potential and virulence traits of Enterococcus faecium and E. faecalis isolated from human milk

PubMed Central

Khalkhali, Soodabeh; Mojgani, Naheed

2017-01-01

Background and Objectives: Human milk is a continuous supply of Lactic Acid bacteria (LAB), including enterococci with probiotic potentials. The aim of this study was to analyze two Enterococcus species, isolated from human milk for their probiotic potential, bacteriocin producing ability and virulence traits. Materials and Methods: Enterococcus faecium TA0033 and E. faecalis TA102 were tested for acid and bile tolerance, survival in simulated gastric and intestinal conditions. The antibacterial spectrum of the isolates was tested by agar well diffusion assay. The antagonistic agent was characterized by physico-chemical methods. The enterocin structural genes, virulence determinants, vancomycin resistance and biogenic amine genes, such as hdc1, hdc2, tdc, ldc and odc were also determined. Results: The tested isolates survived acidic conditions, high bile salt (1%), simulated gastric and intestinal conditions. The culture supernatant fluids of the two isolates inhibited the growth of Escherichia coli, Listeria monocytogenes, Salmonella typhi, Staphylococcus aureus, Shigella dysenteriae and Streptococcus agalactiae. The antagonistic activity was lost in the presence of proteolytic enzymes but tolerated the action of catalase, lysozyme and lipase. In contrast to enterocin TA102, enterocin TA0033 possessed bactericidal mode of action. Bacteriocin structural genes, entA and entB were present in the genome of the two isolates, while E. faecalis TA102 additionally harboured entP and bac31 genes. The phenotypic and genotypic virulence assessment studies indicated hyaluronidase (hyl) production and vancomycin resistance in E. faecalis TA102 while, none of the isolates harboured the biogenic amine genes. Conclusion: The presence of virulence genes in E. faecalis TA102 calls for careful monitoring of Enterococcus isolates for their safety parameters. PMID:29238458

Novel de novo synthesized phosphate carrier compound ABA-PEG20k-Pi20 suppresses collagenase production in Enterococcus faecalis and prevents colonic anastomotic leak in an experimental model.

PubMed

Wiegerinck, M; Hyoju, S K; Mao, J; Zaborin, A; Adriaansens, C; Salzman, E; Hyman, N H; Zaborina, O; van Goor, H; Alverdy, J C

2018-04-16

Previous work has demonstrated that anastomotic leak can be caused by collagenolytic bacteria such as Enterococcus faecalis via an effect on wound collagen. In humans, E. faecalis is the organism cultured most commonly from a leaking anastomosis, and is not routinely eliminated by standard oral or intravenous antibiotics. Novel strategies are needed to contain the virulence of this pathogen when present on anastomotic tissues. Polyphosphorylated polymer ABA-PEG20k-Pi20 was tested in mice for its ability to prevent anastomotic leak caused by collagenolytic E. faecalis. The study design included a distal colonic resection and anastomosis followed by introduction of E. faecalis to anastomotic tissues via enema. Mice were assigned randomly to receive either ABA-PEG20-Pi20 or its unphosphorylated precursor ABA-PEG20k in their drinking water. The development of anastomotic leak was determined after the animals had been killed. Overnight incubation of two different E. faecalis collagenolytic strains with 2 mmol/l of ABA-PEG20k-Pi20 led to near complete inhibition of collagenase production (from 21 000 to 1000 and from 68 000 to 5000 units; P < 0·001; 6 samples per group) without suppressing bacterial growth. In mice drinking 1 per cent ABA-PEG20k-Pi20, the phosphate concentration in the distal colonic mucosa increased twofold and leak rates decreased from eight of 15 to three of 15 animals (P < 0·001). In mice drinking ABA-PEG20k-Pi20, the percentage of collagenolytic colonies among E. faecalis populations present at anastomotic tissue sites was decreased by 6-4800-fold (P = 0·008; 5 animals). These data indicate that oral intake of ABA-PEG20k-Pi20 may be an effective agent to contain the virulence of E. faecalis and may prevent anastomotic leak caused by this organism. Clinical relevance Progress in understanding the pathogenesis of anastomotic leak continues to point to intestinal bacteria as key causative agents. The presence of pathogens such as Enterococcus

Impact of Manganese, Copper and Zinc Ions on the Transcriptome of the Nosocomial Pathogen Enterococcus faecalis V583

PubMed Central

Coelho Abrantes, Marta; Lopes, Maria de Fátima; Kok, Jan

2011-01-01

Mechanisms that enable Enterococcus to cope with different environmental stresses and their contribution to the switch from commensalism to pathogenicity of this organism are still poorly understood. Maintenance of intracellular homeostasis of metal ions is crucial for survival of these bacteria. In particular Zn2+, Mn2+ and Cu2+ are very important metal ions as they are co-factors of many enzymes, are involved in oxidative stress defense and have a role in the immune system of the host. Their concentrations inside the human body vary hugely, which makes it imperative for Enterococcus to fine-tune metal ion homeostasis in order to survive inside the host and colonize it. Little is known about metal regulation in Enterococcus faecalis. Here we present the first genome-wide description of gene expression of E. faecalis V583 growing in the presence of high concentrations of zinc, manganese or copper ions. The DNA microarray experiments revealed that mostly transporters are involved in the responses of E. faecalis to prolonged exposure to high metal concentrations although genes involved in cellular processes, in energy and amino acid metabolisms and genes related to the cell envelope also seem to play important roles. PMID:22053193

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.

Characterization and application of enterocin RM6, a bacteriocin from Enterococcus faecalis.

PubMed

Huang, En; Zhang, Liwen; Chung, Yoon-Kyung; Zheng, Zuoxing; Yousef, Ahmed E

2013-01-01

Use of bacteriocins in food preservation has received great attention in recent years. The goal of this study is to characterize enterocin RM6 from Enterococcus faecalis OSY-RM6 and investigate its efficacy against Listeria monocytogenes in cottage cheese. Enterocin RM6 was purified from E. faecalis culture supernatant using ion exchange column, multiple C18-silica cartridges, followed by reverse-phase high-performance liquid chromatography. The molecular weight of enterocin RM6 is 7145.0823 as determined by mass spectrometry (MS). Tandem mass spectrometry (MS/MS) analysis revealed that enterocin RM6 is a 70-residue cyclic peptide with a head-to-tail linkage between methionine and tryptophan residues. The peptide sequence of enterocin RM6 was further confirmed by sequencing the structural gene of the peptide. Enterocin RM6 is active against Gram-positive bacteria, including L. monocytogenes, Bacillus cereus, and methicillin-resistant Staphylococcus aureus (MRSA). Enterocin RM6 (final concentration in cottage cheese, 80 AU/mL) caused a 4-log reduction in population of L. monocytogenes inoculated in cottage cheese within 30 min of treatment. Therefore, enterocin RM6 has potential applications as a potent antimicrobial peptide against foodborne pathogens in food.

Sewage sludge and liquid pig manure as possible sources of antibiotic resistant bacteria.

PubMed

Hölzel, Christina S; Schwaiger, Karin; Harms, Katrin; Küchenhoff, Helmut; Kunz, Anne; Meyer, Karsten; Müller, Christa; Bauer, Johann

2010-05-01

Within the last decades, the environmental spread of antibiotic resistant bacteria has become a topic of concern. In this study, liquid pig manure (n=305) and sewage sludge (n=111) - used as agricultural fertilizers between 2002 and 2005 - were investigated for the presence of Escherichia coli, Enterococcus faecalis and Enterococcus faecium. Bacteria were tested for their resistance against 40 chemotherapeutics including several "reserve drugs". E. coli (n=613) from pig manure were at a significantly higher degree resistant to streptomycin, doxycycline, spectinomycin, cotrimoxazole, and chloramphenicol than E. coli (n=116) from sewage sludge. Enterococci (Ent. faecalis, n=387, and Ent. faecium, n=183) from pig manure were significantly more often resistant to high levels of doxycycline, rifampicin, erythromycin, and streptomycin than Ent. faecalis (n=44) and Ent. faecium (n=125) from sewage sludge. Significant differences in enterococcal resistance were also seen for tylosin, chloramphenicol, gentamicin high level, fosfomycin, clindamicin, enrofloxacin, moxifloxacin, nitrofurantoin, and quinupristin/dalfopristin. By contrast, aminopenicillins were more effective in enterococci from pig manure, and mean MIC-values of piperacillin+tazobactam and third generation cefalosporines were significantly lower in E. coli from pig manure than in E. coli from sewage sludge. 13.4% (E. coli) to 25.3% (Ent. faecium) of pig manure isolates were high-level multiresistant to substances from more than three different classes of antimicrobial agents. In sewage sludge, high-level-multiresistance reached from 0% (Ent. faecalis) to 16% (Ent. faecium). High rates of (multi-) resistant bacteria in pig manure emphasize the need for a prudent - cautious - use of antibiotics in farm animals. Copyright 2010 Elsevier Inc. All rights reserved.

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.

Do different strains of E. faecalis have the same behavior towards intracanal medications in in vitro research?

PubMed

Zancan, Rafaela Fernandes; Canali, Lyz Cristina Furquim; Tartari, Talita; Andrade, Flaviana Bombarda de; Vivan, Rodrigo Ricci; Duarte, Marco Antonio Hungaro

2018-05-24

The aim of this study was to evaluate the antimicrobial action of different endodontic pastes against Enterococcus faecalis ATCC 29212, isolated from the urinary tract, and compare the action with E. faecalis ATCC 4083, isolated from the root canal. For this purpose, dentin blocks were infected for 21 days with both bacteria at different time-intervals to ensure there would be no cross contamination. After this period, blocks were immersed in the test medications for 7 days, according to the following groups: CH/S, CH/P, CH/CMCP, CH/CHX, CH/DAP and TAP. Images of the samples were captured with a confocal microscope and the percentage of live cells was computed by means of the Bioimage program. The ATCC 29212 strain was shown to be more resistant to CH/SS, Calen, CH/DAP, and TAP than the ATCC 4083 strain. The antimicrobial action of the medications against each strain were divergent concerning the order of susceptibility. The authors concluded that the strains behaved in a different manner: in general, those extracted from the urinary tract were more resistant to the tested medications. Therefore, when E. faecalis must be used for in vitro research in endodontics, we suggest the use of ATCC 4083 strain to obtain results that are closer to the clinical reality.

Uptake of photosensitizers by bacteria is influenced by the presence of cations

NASA Astrophysics Data System (ADS)

Kishen, A.; George, S.

2007-05-01

This investigation studies the influence of cations on photosensitizer uptake by Enterococcus faecalis (gram positive) and Actinobacillus actinomycetemcomitans (gram negative). Methods- The uptake of Methylene blue (MB) and Indocyanine Green (ICG), by bacteria were studied under the influence of divalent cations (CaCl II & MgCl II) and EDTA. Further, E. faecalis cells subjected to trypsinisation and calcium channel blocker (verapamil) were also analysed for MB and ICG uptake inorder to understand the mechanism of photosensitizer uptake. Results- Uptake of ICG was enhanced in the presence of divalent cations in E. faecalis and A. actinomycetemcomitans. Treating cells with EDTA had no significant effect on the photosensitizer uptake, although the highest concentration tested showed an enhancement of uptake. In contrast to ICG, MB showed a decreased uptake by bacterial cells on subjecting them to divalent cations and EDTA. Calcium channel blocker had no significant inhibitory effect on photosensitizers uptake. However, trypsin treatment resulted in significant reduction of ICG uptake. The result suggested that ICG uptake by bacteria is mediated through specific transporter protein while MB is associated with the outer surface structures of bacterial cells.

Enterococcus faecalis urinary-tract infections: Do they have a zoonotic origin?

PubMed

Abat, Cédric; Huart, Michael; Garcia, Vincent; Dubourg, Grégory; Raoult, Didier

2016-10-01

Major human pathogens are frequently isolated from meat-producing animals, particularly poultry. Among them is Enterococcus faecalis, which is known to be one of the main cause of human urinary-tract infections worldwide. Early in 2015, we detected several, consecutive abnormal increases in the weekly number of human E. faecalis infections in various medical settings in the Provence-Alpes-Côte d'Azur region of France, especially including community-acquired urinary-tract infections. Speculating that this region-wide epidemiological event may have originated from animal-based food, we initiated this work to provide an overview of the epidemiology of E. faecalis, with a particular focus on the possible link between E. faecalis clones isolated from food-producing animals and those responsible for human urinary-tract infections. At that time, only one study had clearly identified strong epidemiological links between E. faecalis clones isolated from food-producing animals and human E. faecalis urinary-tract infections. This observation, coupled with our region-wide epidemiological experience, leads us to strongly believe that E. faecalis is a real zoonotic pathogen with potentially highly significant impact on human health. This is of particular concern because of its ability to acquire antibiotic-resistance genes and to infect animals and humans. Various strategies must be urgently implemented to address this public health threat, in particular through the development and implementation of large integrated automated surveillance systems based on animal and human health data to enable us to detect E. faecalis epidemiological events. Copyright © 2016 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

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.

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

Biodegradation Capability of Some Bacteria Isolates to Use Lubricant Oil in Vitro

NASA Astrophysics Data System (ADS)

Ahda, Y.; Azhar, M.; Fitri, L.; Afnida, A.; Adha, G. S.; Alifa, W. N.; Handayani, D.; Putri, D. H.; Irdawati, I.; Chatri, M.

2018-04-01

Our previous study identified three species of bacteria, i.e. Alcaligenes sp., Bacillus spl, and Bacillus sp2 isolated from using lubricant oil-contaminated soil in a Padang’s workshop. However, its ability to degrade hydrocarbon were not known yet. In this extension study, we explore a wider area to find more hydrocarbonoclastic bacteria and examined its capability to degrade hydrocarbon in vitro. Seventeen isolates were characterized its capability using NA + used lubricant oil + tween + neutral red medium. Isolates A1, B2, D1 and D4 shows the high degradation index, whereas isolates A2, A3, A5, D2, B1, B3 and isolates A4, B4, D3 have medium and low degradation index, respectively. These potential hydrocarbonoclastic bacteria need in situ characterization to know their actual activities for bioremediation.

Characterization and Application of Enterocin RM6, a Bacteriocin from Enterococcus faecalis

PubMed Central

Chung, Yoon-Kyung; Yousef, Ahmed E.

2013-01-01

Use of bacteriocins in food preservation has received great attention in recent years. The goal of this study is to characterize enterocin RM6 from Enterococcus faecalis OSY-RM6 and investigate its efficacy against Listeria monocytogenes in cottage cheese. Enterocin RM6 was purified from E. faecalis culture supernatant using ion exchange column, multiple C18-silica cartridges, followed by reverse-phase high-performance liquid chromatography. The molecular weight of enterocin RM6 is 7145.0823 as determined by mass spectrometry (MS). Tandem mass spectrometry (MS/MS) analysis revealed that enterocin RM6 is a 70-residue cyclic peptide with a head-to-tail linkage between methionine and tryptophan residues. The peptide sequence of enterocin RM6 was further confirmed by sequencing the structural gene of the peptide. Enterocin RM6 is active against Gram-positive bacteria, including L. monocytogenes, Bacillus cereus, and methicillin-resistant Staphylococcus aureus (MRSA). Enterocin RM6 (final concentration in cottage cheese, 80 AU/mL) caused a 4-log reduction in population of L. monocytogenes inoculated in cottage cheese within 30 min of treatment. Therefore, enterocin RM6 has potential applications as a potent antimicrobial peptide against foodborne pathogens in food. PMID:23844357

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

Buwchitin: a ruminal peptide with antimicrobial potential against Enterococcus faecalis

NASA Astrophysics Data System (ADS)

Oyama, Linda B.; Crochet, Jean-Adrien; Edwards, Joan E.; Girdwood, Susan E.; Cookson, Alan R.; Fernandez-Fuentes, Narcis; Hilpert, Kai; Golyshin, Peter N.; Golyshina, Olga V.; Privé, Florence; Hess, Matthias; Mantovani, Hilario C.; Creevey, Christopher J.; Huws, Sharon A.

2017-07-01

Antimicrobial peptides (AMPs) are gaining popularity as alternatives for treatment of bacterial infections and recent advances in omics technologies provide new platforms for AMP discovery. We sought to determine the antibacterial activity of a novel antimicrobial peptide, buwchitin, against Enterococcus faecalis. Buwchitin was identified from a rumen bacterial metagenome library, cloned, expressed and purified. The antimicrobial activity of the recombinant peptide was assessed using a broth microdilution susceptibility assay to determine the peptide's killing kinetics against selected bacterial strains. The killing mechanism of buwchitin was investigated further by monitoring its ability to cause membrane depolarization (diSC3(5) method) and morphological changes in E. faecalis cells. Transmission electron micrographs of buwchitin treated E. faecalis cells showed intact outer membranes with blebbing, but no major damaging effects and cell morphology changes. Buwchitin had negligible cytotoxicity against defibrinated sheep erythrocytes. Although no significant membrane leakage and depolarization was observed, buwchitin at minimum inhibitory concentration (MIC) was bacteriostatic against E. faecalis cells and inhibited growth in vitro by 70% when compared to untreated cells. These findings suggest that buwchitin, a rumen derived peptide, has potential for antimicrobial activity against E. faecalis.

Partial Diversity Generates Effector Immunity Specificity of the Bac41-Like Bacteriocins of Enterococcus faecalis Clinical Strains.

PubMed

Kurushima, Jun; Ike, Yasuyoshi; Tomita, Haruyoshi

2016-09-01

Bacteriocin 41 (Bac41) is the plasmid-encoded bacteriocin produced by the opportunistic pathogen Enterococcus faecalis Its genetic determinant consists of bacL1 (effector), bacL2 (regulator), bacA (effector), and bacI (immunity). The secreted effectors BacL1 and BacA coordinate to induce the lytic cell death of E. faecalis Meanwhile, the immunity factor BacI provides self-resistance to the Bac41 producer, E. faecalis, against the action of BacL1 and BacA. In this study, we demonstrated that more than half of the 327 clinical strains of E. faecalis screened had functional Bac41 genes. Analysis of the genetic structure of the Bac41 genes in the DNA sequences of the E. faecalis strains revealed that the Bac41-like genes consist of a relatively conserved region and a variable region located downstream from bacA Based on similarities in the variable region, the Bac41-like genes could be classified into type I, type IIa, and type IIb. Interestingly, the distinct Bac41 types had specific immunity factors for self-resistance, BacI1 or BacI2, and did not show cross-immunity to the other type of effector. We also demonstrated experimentally that the specificity of the immunity was determined by the combination of the C-terminal region of BacA and the presence of the unique BacI1 or BacI2 factor. These observations suggested that Bac41-like bacteriocin genes are extensively disseminated among E. faecalis strains in the clinical environment and can be grouped into at least three types. It was also indicated that the partial diversity results in specificity of self-resistance which may offer these strains a competitive advantage. Bacteriocins are antibacterial effectors produced by bacteria. In general, a bacteriocin-coding gene is accompanied by a cognate immunity gene that confers self-resistance on the bacteriocin-producing bacterium itself. We demonstrated that one of the bacteriocins, Bac41, is disseminated among E. faecalis clinical strains and the Bac41 subtypes with

Bioaugmented sulfur-oxidizing denitrification system with Alcaligenes defragrans B21 for high nitrate containing wastewater treatment.

PubMed

Flores, Angel; Nisola, Grace M; Cho, Eulsaeng; Gwon, Eun-Mi; Kim, Hern; Lee, Changhee; Park, Shinjung; Chung, Wook-Jin

2007-05-01

The performance of enriched sludge augmented with the B21 strain of Alcaligenes defragrans was compared with that of enriched sludge, as well as with pure Alcaligenes defragrans B21, in the context of a sulfur-oxidizing denitrification (SOD) process. In synthetic wastewater treatment containing 100-1,000 mg NO3-N/L, the single strain-seeded system exhibited superior performance, featuring higher efficiency and a shorter startup period, provided nitrate loading rate was less than 0.2 kg NO3-N/m(3) per day. At nitrate loading rate of more than 0.5 kg NO3-N/m(3) per day, the bioaugmented sludge system showed higher resistance to shock loading than two other systems. However, no advantage of the bioaugmented system over the enriched sludge system without B21 strain was observed in overall efficiency of denitrification. Both the bioaugmented sludge and enriched sludge systems obtained stable denitrification performance of more than 80% at nitrate loading rate of up to 2 kg NO3-N/m(3) per day.

Evaluation of instrumentation systems in reducing E. faecalis from root canals: TF adaptive and ProTaper next versus single file systems.

PubMed

Karataş, Ertuğrul; Gültekin, Esra; Arslan, Hakan; Kirici, Damla Özsu; Alsancak, Meltem; Topçu, Meltem Çolak

2015-03-01

To compare the effect of the TF Adaptive, ProTaper Next, OneShape, WaveOne, Reciproc, (SAF) on the reduction of E. faecalis in experimentally infected root canals. 70 human mandibular incisor teeth with straight roots and single root canals were selected for this experiment and the root canals of the selected teeth were infected with E. faecalis. After contamination, all the root canals were randomly divided into 7 groups: control, ProTaper Next, TF Adaptive, SAF, WaveOne, Reciproc, and OneShape. After the irrigation procedures, samples were taken from root canals with paper points and incubated in blood agar plates. The colonies grown on the blood agar were counted and interpreted as colony forming units per milliliter. Analysis of results showed that all instrumentation systems were more effective in reducing the number of bacteria than the control (P<.001). The ProTaper Next, TF Adaptive, WaveOne, Reciproc, and OneShape systems were significantly more effective than the SAF system in reducing E. faecalis within the root canals (P<.001). All instrumentation systems used in this study provided a significant reduction in bacterial populations.

Genes Important for Catalase Activity in Enterococcus faecalis

PubMed Central

Baureder, Michael; Hederstedt, Lars

2012-01-01

Little in general is known about how heme proteins are assembled from their constituents in cells. The Gram-positive bacterium Enterococcus faecalis cannot synthesize heme and does not depend on it for growth. However, when supplied with heme in the growth medium the cells can synthesize two heme proteins; catalase (KatA) and cytochrome bd (CydAB). To identify novel factors important for catalase biogenesis libraries of E. faecalis gene insertion mutants were generated using two different types of transposons. The libraries of mutants were screened for clones deficient in catalase activity using a colony zymogram staining procedure. Analysis of obtained clones identified, in addition to katA (encoding the catalase enzyme protein), nine genes distributed over five different chromosomal loci. No factors with a dedicated essential role in catalase biogenesis or heme trafficking were revealed, but the results indicate the RNA degradosome (srmB, rnjA), an ABC-type oligopeptide transporter (oppBC), a two-component signal transducer (etaR), and NADH peroxidase (npr) as being important for expression of catalase activity in E. faecalis. It is demonstrated that catalase biogenesis in E. faecalis is independent of the CydABCD proteins and that a conserved proline residue in the N-terminal region of KatA is important for catalase assembly. PMID:22590595

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

Differential effector responses by circulating/blood and tissue/peritoneal neutrophils following burn combined with Enterococcus faecalis infection.

PubMed

Fazal, Nadeem; Shelip, Alla; Siddiqui, Erum; Ali, Ashraf; Azim, Anser C; Al-Ghoul, Walid M

2012-03-01

Recently we found that superimposition of Enterococcus faecalis infection on burn injury caused an eruption of host mortality not seen with either individual challenge. We hypothesized that the Enterococcus bacteria, and/or factors related to these organisms, aggravate burn-induced modulations in host defense by neutrophils. Our study focuses on alterations in neutrophils' oxidative, proteolytic, and adhesive functions and transendothelial migration of neutrophils in burn rats inoculated with E. faecalis. Rats were subjected to burn (30% total body surface area) and then intra-abdominally inoculated with E. faecalis (10(4)CFU kg(-1) b.w). Polymorphonuclear neutrophils (PMNs) were harvested from circulating/blood and tissue/peritoneal cavity at day-2 post injury. Extracellular release of O(-)(2) anion production was determined by luminometry, and intracellular production of reactive oxygen species was measured by digital imaging technique. Fluoroscan analysis and confocal microscopy determined intracellular elastase production. The expression of adhesion molecule CD11b/CD18 was performed by flow cytometry. Calcein AM-labeled PMNs were co-cultured with TNF-α-stimulated rat lung microvascular endothelial cells, and their ability to adhere was assessed by fluorometry and digital imaging and finally, chemotaxis was measured by neutrophil transmigration assays. The results showed differential effector responses by circulatory and/or tissue PMNs. Tissue/peritoneal PMNs produced more O(-)(2), less intracellular elastase, and increased expression of CD11b/CD18 accompanied with increased adhesivity of MIP-2-stimulated PMNs to endothelial cells as compared to circulatory/blood PMNs. This differential effect was more pronounced following burn plus E. faecalis infection, indicating that the combined injury changed neutrophil functions. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Staphylococcus aureus undergoes major transcriptional reorganization during growth with Enterococcus faecalis in milk.

PubMed

Viçosa, Gabriela Nogueira; Botta, Cristian; Ferrocino, Ilario; Bertolino, Marta; Ventura, Marco; Nero, Luís Augusto; Cocolin, Luca

2018-08-01

Previous studies have demonstrated the antagonistic potential of lactic acid bacteria (LAB) present in raw milk microbiota over Staphylococcus aureus, albeit the molecular mechanisms underlying this inhibitory effect are not fully understood. In this study, we compared the behavior of S. aureus ATCC 29213 alone and in the presence of a cheese-isolated LAB strain, Enterococcus faecalis 41FL1 in skimmed milk at 30 °C for 24 h using phenotypical and molecular approaches. Phenotypic analysis showed the absence of classical staphylococcal enterotoxins in co-culture with a 1.2-log decrease in S. aureus final population compared to single culture. Transcriptional activity of several exotoxins and global regulators, including agr, was negatively impacted in co-culture, contrasting with the accumulation of transcripts coding for surface proteins. After 24 h, the number of transcripts coding for several metabolite responsive elements, as well as enzymes involved in glycolysis and acetoin metabolism was increased in co-culture. The present study discusses the complexity of the transcriptomic mechanisms possibly leading to S. aureus attenuated virulence in the presence of E. faecalis and provides insights into this interspecies interaction in a simulated food context. Copyright © 2018 Elsevier Ltd. All rights reserved.

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

Agrochemicals indirectly increase survival of E. coli O157:H7 and indicator bacteria by reducing ecosystem services.

PubMed

Staley, Zachery R; Rohr, Jason R; Senkbeil, Jacob K; Harwood, Valerie J

Storm water and agricultural runoff frequently contain agrochemicals, fecal indicator bacteria (FIB), and zoonotic pathogens. Entry of such contaminants into aquatic ecosystems may affect ecology and human health. This study tested the hypothesis that the herbicide atrazine and the fungicide chlorothalonil indirectly affect the survival of FIB (Escherichia coli and Enterococcus faecalis) and a pathogen (E. coli O157:H7) by altering densities of protozoan predators or by altering competition from autochthonous bacteria. Streptomycin-resistant E. coli, En. faecalis, and E. coli O157:H7 were added to microcosms composed of Florida river water containing natural protozoan and bacterial populations. FIB, pathogen, and protozoan densities were monitored over six days. Known metabolic inhibitors, cycloheximide and streptomycin, were used to inhibit autochthonous protozoa or bacteria, respectively. The inhibitors made it possible to isolate the effects of predation or competition on survival of allochthonous bacteria, and each treatment increased the survival of FIB and pathogens. Chlorothalonil's effect was similar to that of cycloheximide, significantly reducing protozoan densities and elevating densities of FIB and pathogens relative to the control. Atrazine treatment did not affect protozoan densities, but, through an effect on competition, resulted in significantly greater densities of En. faecalis and E. coli O157:H7. Hence, by reducing predaceous protozoa and bacterial competitors that facilitate purifying water bodies of FIBs and human pathogens, chlorothalonil and atrazine indirectly diminished an ecosystem service of fresh water.

Enterococcus faecalis phosphomevalonate kinase

PubMed Central

Doun, Stephanie S.; Burgner, John W.; Briggs, Scott D.; Rodwell, Victor W.

2005-01-01

The six enzymes of the mevalonate pathway of isopentenyl diphosphate biosynthesis represent potential for addressing a pressing human health concern, the development of antibiotics against resistant strains of the Gram-positive streptococci. We previously characterized the first four of the mevalonate pathway enzymes of Enterococcus faecalis, and here characterize the fifth, phosphomevalonate kinase (E.C. 2.7.4.2). E. faecalis genomic DNA and the polymerase chain reaction were used to clone DNA thought to encode phosphomevalonate kinase into pET28b(+). Double-stranded DNA sequencing verified the sequence of the recombinant gene. The encoded N-terminal hexahistidine-tagged protein was expressed in Escherichia coli with induction by isopropylthiogalactoside and purified by Ni++ affinity chromatography, yield 20 mg protein per liter. Analysis of the purified protein by MALDI-TOF mass spectrometry established it as E. faecalis phosphomevalonate kinase. Analytical ultracentrifugation revealed that the kinase exists in solution primarily as a dimer. Assay for phosphomevalonate kinase activity used pyruvate kinase and lactate dehydrogenase to couple the formation of ADP to the oxidation of NADH. Optimal activity occurred at pH 8.0 and at 37°C. The activation energy was ~5.6 kcal/mol. Activity with Mn++, the preferred cation, was optimal at about 4 mM. Relative rates using different phosphoryl donors were 100 (ATP), 3.6 (GTP), 1.6 (TTP), and 0.4 (CTP). Km values were 0.17 mM for ATP and 0.19 mM for (R,S)-5-phosphomevalonate. The specific activity of the purified enzyme was 3.9 μmol substrate converted per minute per milligram protein. Applications to an immobilized enzyme bioreactor and to drug screening and design are discussed. PMID:15802646

Magnesium and iron nanoparticles production using microorganisms and various salts

NASA Astrophysics Data System (ADS)

Kaul, R. K.; Kumar, P.; Burman, U.; Joshi, P.; Agrawal, A.; Raliya, R.; Tarafdar, J. C.

2012-09-01

Response of five fungi and two bacteria to different salts of magnesium and iron for production of nanoparticles was studied. Pochonia chlamydosporium, and Aspergillus fumigatus were exposed to three salts of magnesium while Curvularia lunata, Chaetomium globosum, A. fumigatus, A. wentii and the bacteria Alcaligenes faecalis and Bacillus coagulans were exposed to two salts of iron for nanoparticle production. The results revealed that P. chlamydosporium induces development of extracellular nanoparticles in MgCl2 solution while A. fumigatus produces also intracellular nanoparticles when exposed to MgSO4 solution. C. globosum was found as the most effective in producing nanoparticles when exposed to Fe2O3 solution. The FTIR analysis of the nanoparticles obtained from Fe2O3 solution showed the peaks similar to iron (Fe). In general, the species of the tested microbes were selective to different chemicals in their response for synthesis of nanoparticles. Further studies on their characterization and improving the efficiency of promising species of fungi need to be undertaken before tapping their potential as nanonutrients for plants.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen Wei; Huang Jun; Wang Xingquan

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 amore » significant role in the sterilization process.« less

Qualitative comparison of sonic or laser energisation of 4% sodium hypochlorite on an Enterococcus faecalis biofilm grown in vitro.

PubMed

Seet, Aaron N; Zilm, Peter S; Gully, Neville J; Cathro, Peter R

2012-12-01

The effectiveness of sonic activation, laser activation and syringe irrigation of 4% sodium hypochlorite in removing an Enterococcus faecalis biofilm was compared. Biofilms were grown in extracted human single rooted teeth using a flow cell apparatus. After 4 weeks' growth, teeth were subjected to each treatment using 4% sodium hypochlorite and radicular dentinal surfaces of the root canals were analysed by scanning electron microscopy. Results showed that sonic activation and syringe irrigation with sodium hypochlorite showed reduced numbers of bacterial cells on the radicular dentine but were not effective in eliminating E. faecalis in the dentinal tubules. Laser activation of sodium hypochlorite resulted in clean dentine walls and undetectable levels of bacteria within dentinal tubules. Qualitatively, sonic or laser activation of 4% NaOCl resulted in greater bacterial reduction compared with syringe irrigation, with laser activation producing the greatest overall reduction. © 2012 The Authors. Australian Endodontic Journal © 2012 Australian Society of Endodontology.

Root Canal Irrigation: Chemical Agents and Plant Extracts Against Enterococcus faecalis

PubMed Central

Borzini, Letizia; Condò, Roberta; De Dominicis, Paolo; Casaglia, Adriano; Cerroni, Loredana

2016-01-01

Background: There are various microorganisms related to intra and extra-radicular infections and many of these are involved in persistent infections. Bacterial elimination from the root canal is achieved by means of the mechanical action of instruments and irrigation as well as the antibacterial effects of the irrigating solutions. Enterococcus faecalis can frequently be isolated from root canals in cases of failed root canal treatments. Antimicrobial agents have often been developed and optimized for their activity against endodontic bacteria. An ideal root canal irrigant should be biocompatible, because of its close contact with the periodontal tissues during endodontic treatment. Sodium hypoclorite (NaOCl) is one of the most widely recommended and used endodontic irrigants but it is highly toxic to periapical tissues. Objectives: To analyze the literature on the chemotherapeutic agent and plant extracts studied as root canal irrigants. In particularly, the study is focused on their effect on Enterococcus faecalis. Method: Literature search was performed electronically in PubMed (PubMed Central, MEDLINE) for articles published in English from 1982 to April 2015. The searched keywords were “endodontic irrigants” and “Enterococcus faecalis” and “essential oil” and “plant extracts”. Results: Many of the studied chemotherapeutic agents and plant extracts have shown promising results in vitro. Conclusion: Some of the considered phytotherapic substances, could be a potential alternative to NaOCl for the biomechanical treatment of the endodontic space. PMID:28217184

Comprehensive analysis of phospholipids and glycolipids in the opportunistic pathogen Enterococcus faecalis

PubMed Central

Gao, Iris H.; Nair, Zeus J.; Kumar, Jaspal K.; Gao, Liang; Kline, Kimberly A.; Wenk, Markus R.

2017-01-01

Enterococcus faecalis is a Gram-positive, opportunistic, pathogenic bacterium that causes a significant number of antibiotic-resistant infections in hospitalized patients. The development of antibiotic resistance in hospital-associated pathogens is a formidable public health threat. In E. faecalis and other Gram-positive pathogens, correlations exist between lipid composition and antibiotic resistance. Resistance to the last-resort antibiotic daptomycin is accompanied by a decrease in phosphatidylglycerol (PG) levels, whereas multiple peptide resistance factor (MprF) converts anionic PG into cationic lysyl-PG via a trans-esterification reaction, providing resistance to cationic antimicrobial peptides. Unlike previous studies that relied on thin layer chromatography and spectrophotometry, we have performed liquid chromatography-tandem mass spectrometry (LC-MS/MS) directly on lipids extracted from E. faecalis, and quantified the phospholipids through multiple reaction monitoring (MRM). In the daptomycin-sensitive E. faecalis strain OG1RF, we have identified 17 PGs, 8 lysyl-PGs (LPGs), 23 cardiolipins (CL), 3 glycerophospho-diglucosyl-diacylglycerols (GPDGDAG), 5 diglucosyl-diacylglycerols (DGDAG), 3 diacylglycerols (DAGs), and 4 triacylglycerols (TAGs). We have quantified PG and shown that PG levels vary during growth of E. faecalis in vitro. We also show that two daptomycin-resistant (DapR) strains of E. faecalis have substantially lower levels of PG and LPG levels. Since LPG levels in these strains are lower, daptomycin resistance is likely due to the reduction in PG. This lipidome map is the first comprehensive analysis of membrane phospholipids and glycolipids in the important human pathogen E. faecalis, for which antimicrobial resistance and altered lipid homeostasis have been intimately linked. PMID:28423018

Differential Microbial Diversity in Drosophila melanogaster: Are Fruit Flies Potential Vectors of Opportunistic Pathogens?

PubMed Central

Maldonado-Morales, Génesis; Bayman, Paul

2017-01-01

Drosophila melanogaster has become a model system to study interactions between innate immunity and microbial pathogens, yet many aspects regarding its microbial community and interactions with pathogens remain unclear. In this study wild D. melanogaster were collected from tropical fruits in Puerto Rico to test how the microbiota is distributed and to compare the culturable diversity of fungi and bacteria. Additionally, we investigated whether flies are potential vectors of human and plant pathogens. Eighteen species of fungi and twelve species of bacteria were isolated from wild flies. The most abundant microorganisms identified were the yeast Candida inconspicua and the bacterium Klebsiella sp. The yeast Issatchenkia hanoiensis was significantly more common internally than externally in flies. Species richness was higher in fungi than in bacteria, but diversity was lower in fungi than in bacteria. The microbial composition of flies was similar internally and externally. We identified a variety of opportunistic human and plant pathogens in flies such as Alcaligenes faecalis, Aspergillus flavus, A. fumigatus, A. niger, Fusarium equiseti/oxysporum, Geotrichum candidum, Klebsiella oxytoca, Microbacterium oxydans, and Stenotrophomonas maltophilia. Despite its utility as a model system, D. melanogaster can be a vector of microorganisms that represent a potential risk to plant and public health. PMID:29234354

Enterococcus faecalis lipoteichoic acid regulates macrophages autophagy via PI3K/Akt/mTOR pathway.

PubMed

Lin, Dongjia; Gao, Yan; Zhao, Luodan; Chen, Yanhuo; An, Shaofeng; Peng, Zhixiang

2018-04-15

Enterococcus faecalis (E. faecalis) infection is considered an important etiological factor for the development of persistent apical periodontitis (PAP), but the exact mechanisms of autophagy between E. faecalis and immune cells remain unknown. In this study, we elucidated how E. faecalis lipoteichoic acid (LTA) is associated with macrophages autophagy. We found that E. faecalis LTA apparently activated macrophage autophagy with significant increase of autophagosomes and autophagy relative protein. Meanwhile, we noticed significantly decreasing expression of p-Akt and p-mTOR. However, these effect were absent in macrophages knockdown of Beclin1. In summary, these findings suggested E. faecalis LTA may increased macrophages autophagy via inhibiting PI3K/Akt/mTOR pathway and this process was Beclin1 dependent. Copyright © 2018 Elsevier Inc. All rights reserved.

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

Outbreak of mastitis in sheep caused by multi-drug resistant Enterococcus faecalis in Sardinia, Italy.

PubMed

Sanciu, G; Marogna, G; Paglietti, B; Cappuccinelli, P; Leori, G; Rappelli, P

2013-03-01

An outbreak of infective mastitis due to Enterococcus faecalis occurred in an intensive sheep farm in north Sardinia (Italy). E. faecalis, which is only rarely isolated from sheep milk, was unexpectedly found in 22·3% of positive samples at microbiological examination. Forty-five out of the 48 E. faecalis isolates showed the same multi-drug resistance pattern (cloxacillin, streptomycin, kanamycin, clindamycin, oxytetracycline). E. faecalis isolates were analysed by pulsed-field gel electrophoresis, and all 45 multi-drug resistant strains showed an indistinguishable macrorestiction profile, indicating their clonal origin. To our knowledge, this is the first report of an outbreak of mastitis in sheep caused by E. faecalis.

Chemical resistance of the gram-negative bacteria to different sanitizers in a water purification system

PubMed Central

Mazzola, Priscila G; Martins, Alzira MS; Penna, Thereza CV

2006-01-01

Background Purified water for pharmaceutical purposes must be free of microbial contamination and pyrogens. Even with the additional sanitary and disinfecting treatments applied to the system (sequential operational stages), Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas alcaligenes, Pseudomonas picketti, Flavobacterium aureum, Acinetobacter lowffi and Pseudomonas diminuta were isolated and identified from a thirteen-stage purification system. To evaluate the efficacy of the chemical agents used in the disinfecting process along with those used to adjust chemical characteristics of the system, over the identified bacteria, the kinetic parameter of killing time (D-value) necessary to inactivate 90% of the initial bioburden (decimal reduction time) was experimentally determined. Methods Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas alcaligenes, Pseudomonas picketti, Flavobacterium aureum, Acinetobacter lowffi and Pseudomonas diminuta were called in house (wild) bacteria. Pseudomonas diminuta ATCC 11568, Pseudomonas alcaligenes INCQS , Pseudomonas aeruginosa ATCC 15442, Pseudomonas fluorescens ATCC 3178, Pseudomonas picketti ATCC 5031, Bacillus subtilis ATCC 937 and Escherichia coli ATCC 25922 were used as 'standard' bacteria to evaluate resistance at 25°C against either 0.5% citric acid, 0.5% hydrochloric acid, 70% ethanol, 0.5% sodium bisulfite, 0.4% sodium hydroxide, 0.5% sodium hypochlorite, or a mixture of 2.2% hydrogen peroxide (H2O2) and 0.45% peracetic acid. Results The efficacy of the sanitizers varied with concentration and contact time to reduce decimal logarithmic (log10) population (n cycles). To kill 90% of the initial population (or one log10 cycle), the necessary time (D-value) was for P. aeruginosa into: (i) 0.5% citric acid, D = 3.8 min; (ii) 0.5% hydrochloric acid, D = 6.9 min; (iii) 70% ethanol, D = 9.7 min; (iv) 0.5% sodium bisulfite, D = 5.3 min; (v) 0.4% sodium hydroxide, D = 14.2 min; (vi) 0.5% sodium hypochlorite

SEQUENCE SIMILARITIES IN THE GENES ENCODING POLY- CHLORINATED BIPHENYL DEGRADATION BY PSEUDOMONAS STRAIN LB400 AND ALCALIGENES EUTROPHUS H850

EPA Science Inventory

DNA-DNA hybridization was used to compare the Pseudomonas strain LB400 genes for polychlorinated biphenyl (PCB) degradation with those from seven other PCB-degrading strains. Significant hybridization was detected to the genome of Alcaligenes eutrophus H850, a strain similar to L...

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

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ragumani, S.; Sauder, J; Burley, 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.more » 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.« less

2,4-Dichlorophenoxyacetic acid-degrading bacteria contain mosaics of catabolic genes.

PubMed Central

Fulthorpe, R R; McGowan, C; Maltseva, O V; Holben, W E; Tiedje, J M

1995-01-01

DNA from 32 2,4-dichlorophenoxyacetic acid (2,4-D)-degrading bacteria from diverse locations was probed with the first three genes of the well-known 2,4-D degradation pathway found in Alcaligenes eutrophus JMP134(pJP4). The majority of strains did not show high levels of homology to the first three genes of the 2,4-D degradation pathway, tfdA, -B, and -C. Most strains showed combinations of tfdA-, B-, and C-like elements that exhibited various degrees of homology to the gene probes. Strains having the same genomic fingerprints (as determined by repetitive extragenic palindromic PCR) exhibited the same hybridization pattern regardless of the geographic origin of the strain, with the exception of a strain isolated from Puerto Rico. This strain had the same genomic fingerprint as that of numerous other strains in the collection but differed in its hybridization against the tfdA gene probe. Members of the beta subdivision of the Proteobacteria class, specifically Alcaligenes, Burkholderia, and Rhodoferax species, carried DNA fragments with 60% or more sequence similarity to tfdA of pJP4, and most carried fragments showing at least 60% homology to tfdB. However, many strains did not hybridize with tfdC, although they exhibited chlorocatechol dioxygenase activity. Members of the alpha subdivision of the Proteobacteria class, mostly of the genus Sphingomonas, did not hybridize to either tfdA or tfdC, but some hybridized at low stringency to tfdB. The data suggest that extensive interspecies transfer of a variety of homologous degradative genes has been involved in the evolution of 2,4-D-degrading bacteria. PMID:7574638

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

Lack of direct effects of agrochemicals on zoonotic pathogens and fecal indicator bacteria.

PubMed

Staley, Zachery R; Senkbeil, Jacob K; Rohr, Jason R; Harwood, Valerie J

2012-11-01

Agrochemicals, fecal indicator bacteria (FIB), and pathogens frequently contaminate water simultaneously. No significant direct effects of fertilizer, atrazine, malathion, and chlorothalonil on the survival of Escherichia coli, Enterococcus faecalis, Salmonella enterica, human polyomaviruses, and adenovirus were detected, supporting the assertion that previously observed effects of agrochemicals on FIB were indirect.

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

Partial purification and characterization of the mode of action of enterocin S37: a bacteriocin produced by Enterococcus faecalis S37 isolated from poultry feces.

PubMed

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 80(o)C 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, alpha-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 K(ATP) channels. This study contributed to gain more insights into the mode of action of enterocins.

Antimicrobial-resistant bacteria in wild game in Slovenia

NASA Astrophysics Data System (ADS)

Križman, M.; Kirbiš, A.; Jamnikar-Ciglenečki, U.

2017-09-01

Wildlife is usually not exposed to clinically-used antimicrobial agents but can acquire antimicrobial resistance throughout contact with humans, domesticated animals and environments. Samples of faeces from intestines (80 in total) were collected from roe deer (52), wild boars (11), chamois (10) red deer (6) and moufflon (1). After culture on ChromID extended spectrum β-lactamase (ESBL) plates to select for growth of ESBL-producing bacteria, 25 samples produced bacterial colonies for further study. Six species of bacteria were identified from the 25 samples: Stenotrophomonas maltophilia, Serratia fonticola, Stenotrophomonas nitritireducens, Enterococcus faecium, Enterococcus faecalis and Escherichia coli. Two ESBL enzymes were amplified from group TEM and three from group CTX-M-1. Undercooked game meat and salami can be a source of resistant bacteria when animals are not eviscerated properly.

Fate of pathogenic bacteria in microcosms mimicking human body sites.

PubMed

Castellani, Francesco; Ghidini, Valentina; Tafi, Maria Carla; Boaretti, Marzia; Lleo, Maria M

2013-07-01

During the infectious process, pathogens may reach anatomical sites where they are exposed to substances interfering with their growth. These substances can include molecules produced by the host, and his resident microbial population, as well as exogenous antibacterial drugs. Suboptimal concentrations of inhibitory molecules and stress conditions found in vivo (high or low temperatures, lack of oxygen, extreme pH) might induce in bacteria the activation of survival mechanisms blocking their division capability but allowing them to stay alive. These "dormant" bacteria can be reactivated in particular circumstances and would be able to express their virulence traits. In this study, it was evaluated the effect of some environmental conditions, such as optimal and suboptimal temperatures, direct light and antibiotic sub-inhibitory concentrations doses of antibiotic, on the human pathogens Escherichia coli and Enterococcus faecalis when incubated in fluids accumulated in the body of patients with different pathologies. It is shown that inoculation in a number of accumulated body fluids and the presence of gentamicin, reliable conditions encountered during pathological states, induce stress-responding strategies enabling bacteria to persist in microcosms mimicking the human body. Significant differences were detected in Gram-negative and Gram-positive species with E. faecalis surviving, as starved or viable but non-culturable forms, in any microcosm and condition tested and E. coli activating a viable but non-culturable state only in some clinical samples. The persistence of bacteria under these conditions, being non-culturable, might explain some recurrent infections without isolation of the causative agent after application of the standard microbiological methods.

Response to alkaline stress by root canal bacteria in biofilms.

PubMed

Chávez de Paz, L E; Bergenholtz, G; Dahlén, G; Svensäter, G

2007-05-01

To determine whether bacteria isolated from infected root canals survive alkaline shifts better in biofilms than in planktonic cultures. Clinical isolates of Enterococcus faecalis, Lactobacillus paracasei, Olsenella uli, Streptococcus anginosus, S. gordonii, S. oralis and Fusobacterium nucleatum in biofilm and planktonic cultures were stressed at pH 10.5 for 4 h, and cell viability determined using the fluorescent staining LIVE/DEAD BacLight bacterial viability kit. In addition, proteins released into extracellular culture fluids were identified by Western blotting. Enterococcus faecalis, L. paracasei, O. uli and S. gordonii survived in high numbers in both planktonic cultures and in biofilms after alkaline challenge. S. anginosus, S. oralis and F. nucleatum showed increased viability in biofilms compared with planktonic cultures. Alkaline exposure caused all planktonic cultures to aggregate into clusters and resulted in a greater extrusion of cellular proteins compared with cells in biofilms. Increased levels of DnaK, HPr and fructose-1,6-bisphosphate aldolase were observed in culture fluids, especially amongst streptococci. In general, bacteria isolated from infected roots canals resisted alkaline stress better in biofilms than in planktonic cultures, however, planktonic cells appeared to use aggregation and the extracellular transport of specific proteins as survival mechanisms.

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.

Autolytic defective mutant of Streptococcus faecalis.

PubMed Central

Cornett, J B; Redman, B E; Shockman, G D

1978-01-01

Properties of a variant of Streptococcus faecalis ATCC 9790 with defective cellular autolysis are described. The mutant strain was selected as a survivor from a mutagenized cell population simultaneously challenged with two antibiotics which inhibit cell wall biosynthesis, penicillin G and cycloserine. Compared to the parental strain, the mutant strain exhibited: (i) a thermosensitive pattern of cellular autolysis; (ii) an autolytic enzyme activity that had only a slightly increased thermolability when tested in solution in the absence of wall substrate; and (iii) an isolated autolysin that had hydrolytic activity on isolated S. faecalis wall substrate indistinguishable from that of the parental strain, but that was inactive when tested on walls of Micrococcus lysodeikticus as a substrate. These data indicate an alteration in the substrate specificity of the autolytic enzyme of the mutant which appears to result from the synthesis of an altered form of autolytic enzyme. PMID:415045

Lack of Direct Effects of Agrochemicals on Zoonotic Pathogens and Fecal Indicator Bacteria

PubMed Central

Staley, Zachery R.; Senkbeil, Jacob K.; Rohr, Jason R.

2012-01-01

Agrochemicals, fecal indicator bacteria (FIB), and pathogens frequently contaminate water simultaneously. No significant direct effects of fertilizer, atrazine, malathion, and chlorothalonil on the survival of Escherichia coli, Enterococcus faecalis, Salmonella enterica, human polyomaviruses, and adenovirus were detected, supporting the assertion that previously observed effects of agrochemicals on FIB were indirect. PMID:22961900

Oxidation/Biodegradation of Solid Propellants Used in Legacy Chemical Rounds

DTIC Science & Technology

2007-08-01

Bioreactor Sample Source Sample Number Similarity Index Genus Species ICB M28-1 Sample 1A 0.771 Kluyvera cryocrescenes 0.704 Enterobacter cloacae...0.678 Photorhabdus luminencent 0.676 Entrobacter aerogenes Sample 1B 0.901 Alcaligenes faecalis Sample 2 0.894 Pseudomonas stutzeri 0.807 Pseudomonas...et. al. 13 has also described the role of Enterobacter cloacae NADH in the degradation of nitro aromatic compounds. Paracoccus denitrificans, commonly

Displacement of Enterococcus faecalis from hydrophobic and hydrophilic substrata by Lactobacillus and Streptococcus spp. as studied in a parallel plate flow chamber.

PubMed Central

Millsap, K; Reid, G; van der Mei, H C; Busscher, H J

1994-01-01

The displacement of Enterococcus faecalis 1131 from hydrophobic and hydrophilic substrata by isolates of Lactobacillus casei 36 and Streptococcus hyointestinalis KM1 was studied in a parallel plate flow chamber. The experiments were conducted with either 10 mM potassium phosphate buffer or human urine as the suspending fluid, and adhesion and displacement were measured by real-time in situ image analysis. The results showed that E. faecalis 1131 was displaced by lactobacilli (31%) and streptococci (74%) from fluorinated ethylene propylene in buffer and that displacement by lactobacilli was even more effective on a glass substratum in urine (54%). The passage of an air-liquid interface significantly impacted on adhesion, especially when the surface had been challenged with lactobacilli (up to 100% displacement) or streptococci (up to 94% displacement). These results showed that the parallel plate flow system with real-time in situ image analysis was effective for studying bacterial adhesion and that uropathogenic enterococci can be displaced by indigenous bacteria. Images PMID:8031082

An agmatine-inducible system for the expression of recombinant proteins in Enterococcus faecalis.

PubMed

Linares, Daniel M; Perez, Marta; Ladero, Victor; Del Rio, Beatriz; Redruello, Begoña; Martin, M Cruz; Fernandez, María; Alvarez, Miguel A

2014-12-04

Scientific interest in Enterococcus faecalis has increased greatly over recent decades. Some strains are involved in food fermentation and offer health benefits, whereas others are vancomycin-resistant and cause infections that are difficult to treat. The limited availability of vectors able to express cloned genes efficiently in E. faecalis has hindered biotechnological studies on the bacterium's regulatory and pathogenicity-related genes. The agmatine deiminase (AGDI) pathway of E. faecalis, involved in the conversion of agmatine into putrescine, is driven by a response inducer gene aguR. This study describes that the exposure to the induction factor (agmatine) results in the transcription of genes under the control of the aguB promoter, including the aguBDAC operon. A novel E. faecalis expression vector, named pAGEnt, combining the aguR inducer gene and the aguB promoter followed by a cloning site and a stop codon was constructed. pAGEnt was designed for the overexpression and purification of a protein fused to a 10-amino-acid His-tag at the C-terminus. The use of GFP as a reporter of gene expression in E. faecalis revealed that under induction with 60 mM agmatine, fluorescence reached 40 arbitrary units compared to 0 in uninduced cells. pAGEnt vector can be used for the overexpression of recombinant proteins under the induction of agmatine in E. faecalis, with a close correlation between agmatine concentration and fluorescence when GFP was used as reporter.

Extracellular Gelatinase of Enterococcus faecalis Destroys a Defense System in Insect Hemolymph and Human Serum▿

PubMed Central

Park, Shin Yong; Kim, Kyoung Mi; Lee, Joon Ha; Seo, Sook Jae; Lee, In Hee

2007-01-01

We isolated Enterococcus faecalis from the body fluids of dead larvae of the greater wax moth, Galleria mellonella. Extracellular gelatinase (GelE) and serine protease (SprE), both of which are considered putative virulence factors of E. faecalis, were purified from the culture supernatant of E. faecalis. In an attempt to elucidate their virulence mechanisms, purified GelE and SprE were injected into hemolymph of G. mellonella and evaluated with regard to their effects on the immune system of insect hemolymph. As a result, it was determined that E. faecalis GelE degraded an inducible antimicrobial peptide (Gm cecropin) which is known to perform a critical role in host defense during the early phase of microbial infection. The results obtained from the G. mellonella-E. faecalis infection model compelled us to assess the virulence activity of GelE against the complement system in human serum. E. faecalis GelE hydrolyzed C3a and also mediated the degradation of the alpha chain of C3b, thereby inhibiting opsonization and the formation of the membrane attack complex resultant from the activation of the complement cascade triggered by C3 activation. In contrast, E. faecalis SprE exhibited no virulence effect against the immune system of insect hemolymph or human serum tested in this study. PMID:17261598

Antibacterial activity of essential oils from Australian native plants.

PubMed

Wilkinson, Jenny M; Cavanagh, Heather M A

2005-07-01

To date, of the Australian essential oils, only tea tree (Melaleuca alternifolia) and Eucalyptus spp. have undergone extensive investigation. In this study a range of Australian essential oils, including those from Anethole anisata, Callistris glaucophyllia, Melaleuca spp. and Thyptomine calycina, were assayed for in vitro antibacterial activity. M. alternifolia was also included for comparison purposes. Activity was determined using standard disc diffusion assays with each oil assayed at 100%, 10% and 1% against five bacteria (Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Pseudomonas aeruginosa and Alcaligenes faecalis) and the yeast, Candida albicans. All bacteria, with the exception of Ps. aeruginosa, were susceptible to one or more of the essential oils at 100%, with only Eremophilia mitchelli inhibiting the growth of any bacteria at 1% (inhibition of Sal. typhimurium). Where multiple samples of a single oil variety were tested variability in activity profiles were noted. This suggests that different methods of preparation of essential oils, together with variability in plant chemical profiles has an impact on whether or not the essential oil is of use as an antimicrobial agent. These results show that essential oils from Australian plants may be valuable antimicrobial agents for use alone or incorporated into cosmetics, cleaning agents and pharmaceutical products.

Extracellular dextran and DNA affect the formation of Enterococcus faecalis biofilms and their susceptibility to 2% chlorhexidine.

PubMed

Li, Weilan; Liu, Hongyan; Xu, Qiong

2012-07-01

Enterococcus faecalis is frequently recovered from root-filled teeth with refractory apical periodontitis. The ability of E. faecalis to form a matrix-encased biofilm contributes to its pathogenicity; however, the role of extracellular dextran and DNA in biofilm formation and its effect on the susceptibility of the biofilm to chlorhexidine remains poorly understood. E. faecalis biofilms were incubated on dentin blocks. The effect of a dextran-degrading enzyme (dextranase) and DNase I on the adhesion of E. faecalis to dentin was measured using the colony-forming unit (CFU) counting method. CFU assays and confocal laser scanning microscopy were used to investigate the influence of dextranase and DNase I on the antimicrobial activity of 2% chlorhexidine. The CFU count assays indicated that the formation of biofilms by E. faecalis was reduced in cells treated with dextranase or DNase I compared with that in untreated cells (P < .05). In addition, we found that treating E. faecalis biofilms with dextranase or DNase I effectively sensitized the biofilms to 2% chlorhexidine (P < .05). Both dextranase and DNase I decrease the adhesion of E. faecalis to dentin and sensitized E. faecalis biofilms to 2% chlorhexidine. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Evidence for significantly enhancing reduction of Azo dyes in Escherichia coli by expressed cytoplasmic Azoreductase (AzoA) of Enterococcus faecalis.

PubMed

Feng, J; Heinze, T M; Xu, H; Cerniglia, C E; Chen, H

2010-05-01

Although cytoplasmic azoreductases have been purified and characterized from various bacteria, little evidence demonstrating that these azoreductases are directly involved in azo dye reduction in vivo is known. In order to evaluate the contribution of the enzyme to azo dye reduction in vivo, experiments were conducted to determine the effect of a recombinant cytoplasmic azoreductase (AzoA) from Enterococcus faecalis expressed in Escherichia coli on the rate of metabolism of Methyl Red, Ponceau BS and Orange II. The intact cells that contained IPTG induced AzoA had a higher rate of dye reduction with increases of 2 (Methyl Red), 4 (Ponceau BS) and 2.6 (Orange II)-fold compared to noninduced cells, respectively. Metabolites of Methyl Red isolated from induced cultures were identified as N,N-dimethyl-p-phenylenediamine and 2-aminobenzoic acid through liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) analyses. In conclusion, our data demonstrate that AzoA from Ent. faecalis is capable of increasing the reduction of azo dyes in intact E. coli cells and that cytoplasmic azoreductase is involved in bacterial dye degradation in vivo.

New intracanal formulations containing doxycycline or chlorhexidine against Enterococcus faecalis.

PubMed

Silva, Ana Rita Marques da; Pinto, Shelon Cristina Souza; Santos, Elizabete Brasil dos; Santos, Fábio André dos; Farago, Paulo Vitor; Gomes, João Carlos; Pina-Vaz, Irene; Carvalho, Manuel Fontes

2014-01-01

The present study aims to evaluate the antimicrobial effect of two new intracanal preparations against E. faecalis. Thirty single-rooted human canine teeth were used. The crowns were removed and the roots were instrumented using a conventional technique. Three groups of ten teeth each were infected with 108 CFU/ ml of E. faecalis for 21 days. The root canals were flled with new intracanal medications containing 3% doxycycline hydrochloride (DX) or 2% chlorhexidine digluconate (CHX). Ten teeth received no medication (NM)-negative control. Microbial samples were obtained 21 days after contamination: 14 days under the effect of the intracanal medications and 7 days after replacing the medications by BHI broth. The samples were homogenized, diluted, seeded on BHI agar and incubated for 48h/36°C. The number of colony forming units (CFU/ml) was obtained and analyzed statistically. All intracanal dressings significantly reduced the number of bacterial cells in the root canal after 14 days with medication. After the period with 7 days with BHI broth, the CFU counts of E. faecalis remained at low values. However, the NM group showed a significant increase of CFU in this period to similar values of the initial contamination. 3% doxycycline hydrochloride gel and 2% CHX gel were effective to eliminate E. faecalis from the root canal system.

Comparative evaluation of antimicrobial activity of hydroalcoholic extract of Aloe vera, garlic, and 5% sodium hypochlorite as root canal irrigants against Enterococcus faecalis: An in vitro study.

PubMed

Karkare, Swati Ramesh; Ahire, Nivedita Pramod; Khedkar, Smita Uday

2015-01-01

Enterococcus faecalis are the most resistant and predominant microorganisms recovered from root canals of teeth where previous treatment has failed. Over the past decade, interest in drugs derived from medicinal plants has markedly increased. In dentistry, phytomedicines has been used as an anti-inflammatory, antibiotic, analgesic, sedative, and also as an endodontic irrigant. In endodontics, because of the cytotoxic reactions of most of the commercial intracanal medicaments and their inability to eliminate bacteria completely from dentinal tubules, the trend is shifting toward use of biologic medication extracted from natural plants. To compare the antimicrobial efficacy of newer irrigating agents which would probably be as effective or more and at the same time less irritating to the tissues than sodium hypochlorite (NaOCl). The objective of this study was to compare the antimicrobial activity of saturated and diluted (1:1) hydroalcoholic extract of Aloe vera, garlic, and 5% NaOCl against E. faecalis using the commonly used agar diffusion method. Saturated hydroalcoholic extract of A. vera showed the highest zone of inhibition against E. faecalis. NaOCl, which is considered as gold standard, also showed higher zones of inhibition.

Growth inhibition of foodborne pathogens and food spoilage organisms by select raw honeys.

PubMed

Mundo, Melissa A; Padilla-Zakour, Olga I; Worobo, Randy W

2004-12-01

Twenty-seven honey samples from different floral sources and geographical locations were evaluated for their ability to inhibit the growth of seven food spoilage organisms (Alcaligenes faecalis, Aspergillus niger, Bacillus stearothermophilus, Geotrichum candidum, Lactobacillus acidophilus, Penicillium expansum, Pseudomonas fluorescens) and five foodborne pathogens (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica Ser. Typhimurium, and Staphylococcus aureus) using an overlay inhibition assay. They were also tested for specific activity against S. aureus 9144 and B. stearothermophilus using the equivalent percent phenol test--a well diffusion assay corresponding to a dilute phenol standard curve. Honey inhibited bacterial growth due to high sugar concentration (reduced water activity), hydrogen peroxide generation, and proteinaceous compounds present in the honey. Some antibacterial activity was due to other unidentified components. The ability of honey to inhibit the growth of microorganisms varies widely, and could not be attributed to a specific floral source or demographic region produced in this study. Antibacterially active samples in this study included Montana buckwheat, tarweed, manuka, melaleuca, and saw palmetto. Furthermore, the bacteria were not uniformly affected by honey. Varying sensitivities to the antimicrobial properties were observed with four strains of S. aureus thus emphasizing the variability in the antibacterial effect of honey samples. Mold growth was not inhibited by any of the honeys tested. B. stearothermophilus, a heat-resistant spoilage bacteria, was shown to be highly sensitive to honey in both the overlay and well diffusion assays; other sensitive bacteria included A. faecalis and L. acidophilus. Non-peroxide antibacterial activity was observed in both assays; the highest instance was observed in the specific activity assay against B. stearothermophilus. Further research could indicate whether

Investigating the mobilome in clinically important lineages of Enterococcus faecium and Enterococcus faecalis.

PubMed

Mikalsen, Theresa; Pedersen, Torunn; Willems, Rob; Coque, Teresa M; Werner, Guido; Sadowy, Ewa; van Schaik, Willem; Jensen, Lars Bogø; Sundsfjord, Arnfinn; Hegstad, Kristin

2015-04-10

The success of Enterococcus faecium and E. faecalis evolving as multi-resistant nosocomial pathogens is associated with their ability to acquire and share adaptive traits, including antimicrobial resistance genes encoded by mobile genetic elements (MGEs). Here, we investigate this mobilome in successful hospital associated genetic lineages, E. faecium sequence type (ST)17 (n=10) and ST78 (n=10), E. faecalis ST6 (n=10) and ST40 (n=10) by DNA microarray analyses. The hybridization patterns of 272 representative targets including plasmid backbones (n=85), transposable elements (n=85), resistance determinants (n=67), prophages (n=29) and clustered regularly interspaced short palindromic repeats (CRISPR)-cas sequences (n=6) separated the strains according to species, and for E. faecalis also according to STs. RCR-, Rep_3-, RepA_N- and Inc18-family plasmids were highly prevalent and with the exception of Rep_3, evenly distributed between the species. There was a considerable difference in the replicon profile, with rep 17/pRUM , rep 2/pRE25 , rep 14/EFNP1 and rep 20/pLG1 dominating in E. faecium and rep 9/pCF10 , rep 2/pRE25 and rep 7 in E. faecalis strains. We observed an overall high correlation between the presence and absence of genes coding for resistance towards antibiotics, metals, biocides and their corresponding MGEs as well as their phenotypic antimicrobial susceptibility pattern. Although most IS families were represented in both E. faecalis and E. faecium, specific IS elements within these families were distributed in only one species. The prevalence of IS256-, IS3-, ISL3-, IS200/IS605-, IS110-, IS982- and IS4-transposases was significantly higher in E. faecium than E. faecalis, and that of IS110-, IS982- and IS1182-transposases in E. faecalis ST6 compared to ST40. Notably, the transposases of IS981, ISEfm1 and IS1678 that have only been reported in few enterococcal isolates were well represented in the E. faecium strains. E. faecalis ST40 strains harboured

Prevalence and Genetic Diversity of Enterococcus faecalis Isolates from Mineral Water and Spring Water in China.

PubMed

Wei, Lei; Wu, Qingping; Zhang, Jumei; Guo, Weipeng; Chen, Moutong; Xue, Liang; Wang, Juan; Ma, Lianying

2017-01-01

Enterococcus faecalis is an important opportunistic pathogen which is frequently detected in mineral water and spring water for human consumption and causes human urinary tract infections, endocarditis and neonatal sepsis. The aim of this study was to determine the prevalence, virulence genes, antimicrobial resistance and genetic diversity of E. faecalis from mineral water and spring water in China. Of 314 water samples collected from January 2013 to January 2014, 48 samples (15.3%) were contaminated E. faecalis . The highest contamination rate occurred in activated carbon filtered water of spring water (34.5%), followed by source water of spring water (32.3%) and source water of mineral water (6.4%). The virulence gene test of 58 E. faecalis isolates showed that the detection rates of asa1 , ace , cylA , gelE and hyl were 79.3, 39.7, 0, 100, 0%, respectively. All 58 E. faecalis isolates were not resistant to 12 kinds of antibiotics (penicillin, ampicillin, linezolid, quinupristin/dalfopristin, vancomycin, gentamicin, streptomycin, ciprofloxacin, levofloxacin, norfloxacin, nitrofurantoin, and tetracycline). Enterobacterial repetitive intergenic consensus-PCR classified 58 isolates and three reference strains into nine clusters with a similarity of 75%. This study is the first to investigate the prevalence of E. faecalis in mineral water and spring water in China. The results of this study suggested that spring water could be potential vehicles for transmission of E. faecalis .

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.

Quantitative and Qualitative Analysis of Bacteria in Er(III) Solution by Thin-Film Magnetopheresis

PubMed Central

Zborowski, Maciej; Tada, Yoko; Malchesky, Paul S.; Hall, Geraldine S.

1993-01-01

Magnetic deposition, quantitation, and identification of bacteria reacting with the paramagnetic trivalent lanthanide ion, Er3+, was evaluated. The magnetic deposition method was dubbed thin-film magnetopheresis. The optimization of the magnetic deposition protocol was accomplished with Escherichia coli as a model organism in 150 mM NaCl and 5 mM ErCl3 solution. Three gram-positive bacteria, Staphylococcus epidermidis, Staphylococcus saprophyticus, and Enterococcus faecalis, and four gram-negative bacteria, E. coli, Pseudomonas aeruginosa, Proteus mirabilis, and Klebsiella pneumoniae, were subsequently investigated. Quantitative analysis consisted of the microscopic cell count and a scattered-light scanning of the magnetically deposited material aided by the computer data acquisition system. Qualitative analysis consisted of Gram stain differentiation and fluorescein isothiocyanate staining in combination with selected antisera against specific types of bacteria on the solid substrate. The magnetic deposition protocol allowed quantitative detection of E. coli down to the concentration of 105 CFU ml-1, significant in clinical diagnosis applications such as urinary tract infections. Er3+ did not interfere with the typical appearance of the Gram-stained bacteria nor with the antigen recognition by the antibody in the immunohistological evaluations. Indirect antiserum-fluorescein isothiocyanate labelling correctly revealed the presence of E. faecalis and P. aeruginosa in the magnetically deposited material obtained from the mixture of these two bacterial species. On average, the reaction of gram-positive organisms was significantly stronger to the magnetic field in the presence of Er3+ than the reaction of gram-negative organisms. The thin-film magnetophoresis offers promise as a rapid method for quantitative and qualitative analysis of bacteria in solutions such as urine or environmental water. Images PMID:16348916

Interactions between Pteris vittata L. genotypes and a polycyclic aromatic hydrocarbon (PAH)-degrading bacterium (Alcaligenes sp.) in arsenic uptake and PAH-dissipation.

PubMed

Sun, Lu; Zhu, Ganghui; Liao, Xiaoyong; Yan, Xiulan

2017-11-01

The effects of two Pteris vittata L. accessions and a polycyclic aromatic hydrocarbon (PAH)-degrading bacterium (Alcaligenes sp.) on arsenic (As) uptake and phenanthrene dissipation were studied. The Alcaligenes sp. survived in the rhizosphere and improved soil As bioavailability with co-exposure. However, bacterial inoculation altered Pteris vittata L. stress tolerance, and substantially affected the As distribution in the rhizosphere of the two P. vittata accessions. Bacterial inoculation was beneficial to protect the Guangxi accession against the toxic effects, and significantly increased plant As and phenanthrene removal ratios by 27.8% and 2.89%, respectively. In contrast, As removal was reduced by 29.8% in the Hunan accession, when compared with corresponding non-inoculated treatments. We conclude that plant genotype selection is critically important for successful microorganism-assisted phytoremediation of soil co-contaminated with As and PAHs, and appropriate genotype selection may enhance remediation efficiency. Copyright © 2017 Elsevier Ltd. All rights reserved.

Conjugal transfer of aac(6')Ie-aph(2″)Ia gene from native species and mechanism of regulation and cross resistance in Enterococcus faecalis MCC3063 by real time-PCR.

PubMed

Jaimee, G; Halami, P M

2017-09-01

High level aminoglycoside resistance (HLAR) in the lactic acid bacteria (LAB) derived from food animals is detrimental. The aim of this study was to investigate the localization and conjugal transfer of aminoglycoside resistance genes, aac(6')Ie-aph(2″)Ia and aph(3')IIIa in different Enterococcus species. The cross resistance patterns in Enterococcus faecalis MCC3063 to clinically important aminoglycosides by real time PCR were also studied. Southern hybridization experiments revealed the presence of aac(6')Ie-aph(2 ″ )Ia and aph(3')IIIa genes conferring HLAR in high molecular weight plasmids except in Lactobacillus plantarum. The plasmid encoded bifunctional aac(6')Ie-aph(2″)Ia gene was transferable from Enterococcus avium (n = 2), E. cecorum (n = 1), E. faecalis (n = 1) and Pediococcus lolii (n = 1) species into the recipient strain; E. faecalis JH2-2 by filter mating experiments thus indicating the possible risks of gene transfer into pathogenic strains. Molecular analysis of cross resistance patterns in native isolate of E. faecalis MCC3063 carrying aac(6')Ie-aph(2″)Ia and aph(3')IIIa gene was displayed by quantification of the mRNA levels in this study. For this, the culture was induced with increasing concentrations of gentamicin, kanamycin and streptomycin (2048, 4096, 8192, 16384 μg/mL) individually. The increasing concentrations of gentamicin and kanamycin induced the expression of the aac(6')Ie-aph(2″)Ia and aph(3')IIIa resistance genes, respectively. Interestingly, it was observed that induction with streptomycin triggered a significant fold increase in the expression of the aph(3')IIIa gene which otherwise was not known to modify the aminoglycoside. This is noteworthy as streptomycin was found to confer cross resistance to structurally unrelated kanamycin. Also, expression of the aph(3')IIIa gene when induced with streptomycin, revealed that bacteria harbouring this gene will be able to overcome streptomycin bactericidal action at

CARRIAGE OF MULTIDRUG RESISTANT ENTEROCOCCUS FAECIUM AND ENTEROCOCCUS FAECALIS AMONG APPARENTLY HEALTHY HUMANS.

PubMed

Adesida, Solayide A; Ezenta, Cynthia C; Adagbada, Ajoke O; Aladesokan, Amudat A; Coker, Akitoye O

2017-01-01

Enterococci are indigenous flora of the gastro-intestinal tracts of animals and humans. Recently, interest in two major species, E. faecium and E. faecalis , has heightened because of their ability to cause serious infections and their intrinsic resistance to antimicrobials. This study was aimed at determining the prevalence of E . faecium and E . faecalis in human faecal samples and evaluating the susceptibility of the isolates to antibiotics. One hundred faecal samples were collected from apparently healthy individuals and analysed using conventionalbacteriological methods. The susceptibility profile of the isolates to nine antibiotics were determined using disk diffusion method. Seventy-three (73) Enterococcus were phenotypically identified and 65 of the isolates were differentiated into 36 (55.4%) E. faecium and 29 (44.6%) E. faecalis . Eight (8) isolates could not be identified by the conventional biochemical methods employed. No dual colonization by the E. faecalis and E. faecium was observed and isolation rate was not dependent on sex of the participants. All the isolates were resistant to ceftriaxone, cefuroxime and ceftizoxime. Enterococcus faecium exhibited resistance toerythromycin (88.9%), gentamicin (77.8%), amoxicillin-clavulanate (63.9%), ofloxacin (44.4%), teicoplanin (19.4%) and vancomycin (16.7%). Enterococcus faecalis showed the least resistance to vancomycin (13.8%) and teicoplanin (27.7%). Remarkable multiple antibiotic resistances to the classes of antibiotic tested were observed among the two species. The high carriage rate of antibiotic resistant E. faecium and E. faecalis in this study provides information on the local antibiotic patterns of our enterococci isolates thereby suggesting that they could present as important reservoir and vehicle for dissemination of resistant genes in our community.

Regulation of the Activity of Lactate Dehydrogenases from Four Lactic Acid Bacteria*

PubMed Central

Feldman-Salit, Anna; Hering, Silvio; Messiha, Hanan L.; Veith, Nadine; Cojocaru, Vlad; Sieg, Antje; Westerhoff, Hans V.; Kreikemeyer, Bernd; Wade, Rebecca C.; Fiedler, Tomas

2013-01-01

Despite high similarity in sequence and catalytic properties, the l-lactate dehydrogenases (LDHs) in lactic acid bacteria (LAB) display differences in their regulation that may arise from their adaptation to different habitats. We combined experimental and computational approaches to investigate the effects of fructose 1,6-bisphosphate (FBP), phosphate (Pi), and ionic strength (NaCl concentration) on six LDHs from four LABs studied at pH 6 and pH 7. We found that 1) the extent of activation by FBP (Kact) differs. Lactobacillus plantarum LDH is not regulated by FBP, but the other LDHs are activated with increasing sensitivity in the following order: Enterococcus faecalis LDH2 ≤ Lactococcus lactis LDH2 < E. faecalis LDH1 < L. lactis LDH1 ≤ Streptococcus pyogenes LDH. This trend reflects the electrostatic properties in the allosteric binding site of the LDH enzymes. 2) For L. plantarum, S. pyogenes, and E. faecalis, the effects of Pi are distinguishable from the effect of changing ionic strength by adding NaCl. 3) Addition of Pi inhibits E. faecalis LDH2, whereas in the absence of FBP, Pi is an activator of S. pyogenes LDH, E. faecalis LDH1, and L. lactis LDH1 and LDH2 at pH 6. These effects can be interpreted by considering the computed binding affinities of Pi to the catalytic and allosteric binding sites of the enzymes modeled in protonation states corresponding to pH 6 and pH 7. Overall, the results show a subtle interplay among the effects of Pi, FBP, and pH that results in different regulatory effects on the LDHs of different LABs. PMID:23720742

Laser scanning confocal microscopy and laser tweezers based experiments to understand dentine-bacteria interactions

NASA Astrophysics Data System (ADS)

Peng, Sum Chee; Mohanty, Samarendra; Gupta, P. K.; Kishen, Anil

2007-02-01

Failure of endodontic treatment is commonly due to Enterococcal infection. In this study influence of chemical treatments of type-I collagen membrane by chemical agents commonly used in endodontic treatment on Enterococcus faecalis cell adherence was evaluated. In order to determine the change in number of adhering bacteria after chemical treatment, confocal laser scanning microscopy was used. For this, overnight culture of E faecalis in All Culture broth was applied to chemically treated type-I collagen membrane. It was found that Ca(OH) II treated groups had statistically significant (p value=0.05) increase in population of bacteria adherence. The change in adhesion force between bacteria and collagen was determined by using optical tweezers (1064 nm). For this experiment, Type-I collagen membrane was soaked for 5 mins in a media that contained 50% all culture media and 50% saturated Ca(OH) II . The membrane was spread on the coverslip, on which diluted bacterial suspension was added. The force of laser tweezers on the bacteria was estimated at different trap power levels using viscous drag method and trapping stiffness was calculated using Equipartition theorem method. Presence of Ca(OH) II was found to increase the cell-substrate adherence force from 0.38pN to >2.1pN. Together, these experiments show that it was highly probable that the increase in adherence to collagen was due to a stronger adhesion in the presence of Ca (OH) II.

Isolation and identification of Enterococcus faecalis from necrotic root canals using multiplex PCR.

PubMed

Mahmoudpour, Ali; Rahimi, Saeed; Sina, Mahmood; Soroush, Mohammad H; Shahi, Shahriar; Shahisa, Shahriar; Asl-Aminabadi, Naser

2007-09-01

This study was designed to survey the incidence of Enterococcus faecalis infection in symptomatic and asymptomatic root canals of necrotic teeth using PCR and to isolate the bacterium for further screening. Sixty patients categorized according to their clinical symptoms were used for sampling by insertion of paper points into the root canals and absorbing all the fluids present within them. The samples were incubated in 1.0 ml 2xYT (containing 16 g bacto tryptone, 10 g yeast extract and 5.0 g NaCl per liter) for 24 h at 37 degrees C without aeration prior to multiplex PCR analysis. To assist the isolation of E. faecalis, sub-samples were further grown in the same medium supplemented with 6.5% NaCl and back-inoculated into bile esculin. Using multiple cultivation-dependent and PCR analyses, 6 cases (10%) of E. faecalis were identified. Four isolates were obtained from asymptomatic cases of chronic apical periodontitis, and the other two were associated with phoenix abscess and acute apical abscess, respectively. No E. faecalis infection was found in 5 patients with acute apical periodontitis or in 9 with chronic suppurative periodontitis. Our results indicate that there is no significant difference in the incidence of E. faecalis between symptomatic and asymptomatic necrotic dental root canals (P > 0.05).

In vitro antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis in root canals

PubMed Central

VALERA, Marcia Carneiro; MAEKAWA, Lilian Eiko; de OLIVEIRA, Luciane Dias; JORGE, Antonio Olavo Cardoso; SHYGEI, Érika; CARVALHO, Cláudio Antonio Talge

2013-01-01

Objective: The aim of this study was to evaluate the antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis inoculated in root canals. Material and Methods: Seventy-two human tooth roots were contaminated with C. albicans and E. faecalis for 21 days. The groups were divided according to the auxiliary chemical substance into: G1) 2.5% sodium hypochlorite (NaOCl), G2) 2% chlorhexidine gel (CHX), G3) castor oil, G4) glycolic Aloe vera extract, G5) glycolic ginger extract, and G6) sterile saline (control). The samples of the root canal were collected at different intervals: confirmation collection, at 21 days after contamination; 1st collection, after instrumentation; and 2nd collection, seven days after instrumentation. Microbiological samples were grown in culture medium and incubated at 37º C for 48 hours. Results: The results were submitted to the Kruskal-Wallis and Dunn (5%) statistical tests. NaOCl and CHX completely eliminated the microorganisms of the root canals. Castor oil and ginger significantly reduced the number of CFU of the tested bacteria. Reduction of CFU/mL at the 1st and 2nd collections for groups G1, G2, G3 and G4 was greater in comparison to groups G5 and G6. Conclusion: It was concluded that 2.5% sodium hypochlorite and 2% chlorhexidine gel were more effective in eliminating C. albicans and E. faecalis, followed by the castor oil and glycolic ginger extract. The Aloe vera extract showed no antimicrobial activity. PMID:23739849

In vitro antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis in root canals.

PubMed

Valera, Marcia Carneiro; Maekawa, Lilian Eiko; de Oliveira, Luciane Dias; Jorge, Antonio Olavo Cardoso; Shygei, Érika; Carvalho, Cláudio Antonio Talge

2013-01-01

The aim of this study was to evaluate the antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis inoculated in root canals. Seventy-two human tooth roots were contaminated with C. albicans and E. faecalis for 21 days. The groups were divided according to the auxiliary chemical substance into: G1) 2.5% sodium hypochlorite (NaOCl), G2) 2% chlorhexidine gel (CHX), G3) castor oil, G4) glycolic Aloe vera extract, G5) glycolic ginger extract, and G6) sterile saline (control). The samples of the root canal were collected at different intervals: confirmation collection, at 21 days after contamination; 1st collection, after instrumentation; and 2nd collection, seven days after instrumentation. Microbiological samples were grown in culture medium and incubated at 37°C for 48 hours. The results were submitted to the Kruskal-Wallis and Dunn (5%) statistical tests. NaOCl and CHX completely eliminated the microorganisms of the root canals. Castor oil and ginger significantly reduced the number of CFU of the tested bacteria. Reduction of CFU/mL at the 1st and 2nd collections for groups G1, G2, G3 and G4 was greater in comparison to groups G5 and G6. It was concluded that 2.5% sodium hypochlorite and 2% chlorhexidine gel were more effective in eliminating C. albicans and E. faecalis, followed by the castor oil and glycolic ginger extract. The Aloe vera extract showed no antimicrobial activity.

Detection of endocarditis bacteria in tonsillar mucosa of Afghan population.

PubMed

Ruggiero, F; Carbone, D; Mugavero, R; Palmieri, A; Lauritano, D; Baggi, L; Nardone, M; Carinci, F; Martinelli, M

2018-01-01

Endocarditis is a cardiovascular disease caused by the inflammation of the inner tissues of the heart, the endocardium, usually of the valves. Bacteraemia is essential in the development of endocarditis, and there are some findings that the main pathogens of endocarditis are viridans group streptococci: Streptococcus oralis, Streptococcus sanguinis, and Enterococcus faecalis. There is strong evidence that endocarditis bacteria are present in the tonsillar microbiota, so that tonsillar infection is associated with an increased risk of endocarditis. The aim of this manuscript is to investigate the presence of the main pathogens of endocarditis in tonsillar microbiota of an Afghan population group. A sample of 80 tonsil swabs were analyzed by quantitative real time PCR to detect endocarditis pathogens and an estimation of the total bacterial load. The median bacterial load in PCR reaction was 1.4x106 (interquartile range 4,7x105 - 2,9x106). Three species, S. Oralis, S. Sanguinis, and E. Faecalis were found in large amounts in all specimens. On the other hand, S. Mitis was never detected. The S. Aureus was found in 3 samples with a prevalence of 0.04 (C.I. 0.01-0.10). The S. Mutans was found in 33 samples with a prevalence of 0.41 (C.I. 0.31-0.52). Endocarditis bacteria has been found into the tonsillar microbiota, so there is sufficient evidence to justify that the oral cavity is a reservoir of endocarditis bacteria that can have a significant impact on the cardiovascular function.

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.

Evaluation of Enterococcus faecalis adhesion, penetration, and method to prevent the penetration of Enterococcus faecalis into root cementum: Confocal laser scanning microscope and scanning electron microscope analysis.

PubMed

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

2016-01-01

To ascertain the role of Enterococcus faecalis in persistent infection and a possible method to prevent the penetration of E. faecalis into root cementum. One hundred and twenty human single-rooted extracted teeth divided into five groups. Group I (control): intact teeth, Group II: no apical treatment done, Group III divided into two subgroups. In Groups IIIa and IIIb, root apex treated with lactic acid of acidic and neutral pH, respectively. Group IV: apical root cementum exposed to lactic acid and roughened to mimic the apical resorption. Group V: apical treatment done same as Group IV and root-end filling done using mineral trioxide aggregate (MTA). Apical one-third of all samples immersed in E. faecalis broth for 8 weeks followed by bone morphogenetic protein and obturation and again immersed into broth for 8 weeks. Teeth split into two halves and observed under confocal laser scanning microscope and scanning electron microscope, organism identified by culture and polymerase chain reaction techniques. Adhesion and penetration was observed in Group IIIa and Group IV. Only adhesion in Group II and IIIB and no adhesion and penetration in Group I and V. Adhesion and penetration of E. faecalis into root cementum providing a long-term nidus for subsequent infection are the possible reason for persistent infection and root-end filling with MTA prevents the adhesion and penetration.

Effect of Er:YAG laser-activated irrigation solution on Enterococcus Faecalis biofilm in an ex-vivo root canal model.

PubMed

Sahar-Helft, Sharonit; Stabholtz, Adam; Moshonov, Joshua; Gutkin, Vitaly; Redenski, Idan; Steinberg, Doron

2013-07-01

Abstract Objective: The purpose of this study was to evaluate mineral content and surface morphology of root canals coated with Enterococcus faecalis biofilm after treatment with several endodontic irrigation solutions, with and without Er:YAG laser-activated irrigation (LAI). LAI has been introduced as a powerful method for root canal irrigation resulting in smear-layer removal from the root canal wall. Distal and palatal roots from 60 freshly extracted human molars were used in this study. The coronal of each tooth was removed. Roots were split longitudinally and placed in an ultrasonic bath to remove the smear layer, creating conditions for the formation of E. faecalis biofilm. After incubation, the two halves were reassembled in impression material to simulate clinical conditions. Specimens were divided into two main groups: roots rinsed with irrigation solutions and roots subjected to laser irradiation combined with irrigation solutions. Solutions tested were 2% chlorhexidine and 17% ethylenediaminetetraacetic acid (EDTA) and saline. Surface morphology: 17% EDTA irrigant solution combined with Er:YAG laser showed the best results for removing bacteria from the root canal walls. Chemical analysis: all samples treated with combined laser irradiation and irrigation solution had low surface levels of Ca compared with samples treated with irrigation alone. The Ca/P ratio was highest in the laser-EDTA group. Overall, mineral changes caused by laser with irrigation solutions were minimal, and statistically nonsignificant. In vitro irrigation solutions, combined with Er:YAG laser irradiation, were effective in removing E. faecalis biofilm from root canal walls. Irrigation solutions without laser irradiation were less effective, leaving a layer of biofilm on the dentin surface.

Mechanical reduction of the intracanal Enterococcus faecalis population by Hyflex CM, K3XF, ProTaper Next, and two manual instrument systems: an in vitro comparative study.

PubMed

Tewari, Rajendra K; Ali, Sajid; Mishra, Surendra K; Kumar, Ashok; Andrabi, Syed Mukhtar-Un-Nisar; Zoya, Asma; Alam, Sharique

2016-05-01

In the present study, the effectiveness of three rotary and two manual nickel titanium instrument systems on mechanical reduction of the intracanal Enterococcus faecalis population was evaluated. Mandibular premolars with straight roots were selected. Teeth were decoronated and instrumented until 20 K file and irrigated with physiological saline. After sterilization by ethylene oxide gas, root canals were inoculated with Enterococcus faecalis. The specimens were randomly divided into five groups for canal instrumentation: Manual Nitiflex and Hero Shaper nickel titanium files, and rotary Hyflex CM, ProTaper Next, and K3XF nickel titanium files. Intracanal bacterial sampling was done before and after instrumentation. After serial dilution, samples were plated onto the Mitis Salivarius agar. The c.f.u. grown were counted, and log10 transformation was calculated. All instrumentation systems significantly reduced the intracanal bacterial population after root canal preparation. ProTaper Next was found to be significantly more effective than Hyflex CM and manual Nitiflex and Hero Shaper. However, ProTaper Next showed no significant difference with K3XF. Canal instrumentation by all the file systems significantly reduced the intracanal Enterococcus faecalis counts. ProTaper Next was found to be most effective in reducing the number of bacteria than other rotary or hand instruments. © 2014 Wiley Publishing Asia Pty Ltd.

Evidence for Significantly Enhancing Reduction of Azo Dyes in Escherichia coli by Expressed Cytoplasmic Azoreductase (AzoA) of Enterococcus faecalis

PubMed Central

Feng, Jinhui; Heinze, Thomas M.; Xu, Haiyan; Cerniglia, Carl E.; Chen, Huizhong

2018-01-01

Although cytoplasmic azoreductases have been purified and characterized from various bacteria, little evidence demonstrating that these azoreductases are directly involved in azo dye reduction in vivo is known. In order to evaluate the contribution of the enzyme to azo dye reduction in vivo, experiments were conducted to determine the effect of a recombinant cytoplasmic azoreductase (AzoA) from Enterococcus faecalis expressed in Escherichia coli on the rate of metabolism of Methyl Red, Ponceau BS and Orange II. The intact cells that contained IPTG induced AzoA had a higher rate of dye reduction with increases of 2 (Methyl Red), 4 (Ponceau BS) and 2.6 (Orange II)-fold compared to noninduced cells, respectively. Metabolites of Methyl Red isolated from induced cultures were identified as N,N-dimethyl-p-phenylenediamine and 2-aminobenzoic acid through liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) analyses. In conclusion, our data demonstrate that AzoA from Ent. faecalis is capable of increasing the reduction of azo dyes in intact E. coli cells and that cytoplasmic azoreductase is involved in bacterial dye degradation in vivo. PMID:19663804

[Study on the antibacterial activity of four kinds of nano-hydroxyapatite composites against Enterococcus faecalis].

PubMed

Liu, Yi; Zhou, Rongjing; Wu, Hongkun

2015-06-01

This study aims to compare and determine a kind of nano-hydroxyapatite composite material with good antibacterial efficacy on Enterococcusfaecalis (E. faecalis) in vitro. We investigated the antimicrobial activity of four kinds of nano-hydroxyapatite composites, namely, silver/hydroxyapatite composite nanoparticles (Ag/nHA), yttrium/hydroxyapatite composite nanoparticles (Yi/nHA), cerium/hydroxyapatite composite nanoparticles (Ce/nHA), and hydroxyapatite nanoparticles (nHA), against E. faecalis in vitro using the agar diffusion and broth dilution method by measuring the growth inhibition zone and the minimum inhibitory concentration (MIC), respectively. The agar diffusion test results showed that Ag/nHA displayed an obvious growth inhibition zone, whereas Yi/nHA, Ce/nHA, and nHA showed no influence on E. faecalis. The MIC value of Ag/nHA was 1.0 g.L-1, and the three other materials had no effect on E.faecalis even at the high concentration of 32.0 g.L-1. Ag/nHA display a potential antimicrobial efficacy to planktonic E.faecalis. Whereas, the three other kinds of nano-hydroxyapatite composites (Yi/nHA, Ce/nHA, nHA) show no influence.

The transcriptome of the nosocomial pathogen Enterococcus faecalis V583 reveals adaptive responses to growth in blood.

PubMed

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

2009-11-04

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. 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. 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.

Clinical and molecular epidemiology of hospital Enterococcus faecalis isolates in eastern France.

PubMed

Mulin, Blandine; Bailly, Pascale; Thouverez, Michelle; Cailleaux, Vincent; Cornette, Christian; Dupont, Marie-Jeanne; Talon, Daniel

1999-03-01

OBJECTIVE: To report on the occurrence of Enterococcus faecalis hospital isolates obtained during 1 year in hospitals in the Franche-Comté region of France. METHODS: Clinical isolates of E. faecalis of different antibiotic susceptibility phenotypes from hospitalized patients were characterized by pulsed-field gel electrophoresis. Patients with positive cultures were investigated by three case-control studies to identify risk factors for colonization/infection. RESULTS: The crude incidence of colonization/infection was 2.37%, and 4-day and 7-day colonization rates after admission were 10.0% and 6.36%, respectively. The rates of high-level resistance to kanamycin (HLKR) and to gentamicin (HLGR) were 47.1% and 7.1%, respectively. No isolate was resistant to glycopeptides or produced beta-lactamase. The 209 hospital isolates obtained during the study yielded 98 major DNA patterns, of which two were major epidemic patterns including HLKR isolates. No single factor was significantly associated with colonization/infection by HLKR isolates. The length of hospitalization before isolation was associated with colonization by HLGR isolates. CONCLUSIONS: The isolation frequency of E. faecalis strains with acquired resistance to aminoglycoside antibiotics, and the wide dissemination of resistant strains with characteristics that allow them to persist and spread, argue for further large prospective surveys of clinical isolates of E. faecalis in hospitals.

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

Selective grazing by protists upon enteric bacteria in an aquatic system.

PubMed

Domínguez, María S; Escalante, Alicia H; Folabella, Alicia M; Zamora, Angela S

2012-01-01

It is well known that protozoan grazing can be an important agent of mortality for suspended bacteria, both in marine and freshwater environments. Considering that the presence of fecal contamination is a frequent phenomenon in tríese environments, and that Escherichia coli and the genus Enterococcus are indicators of microbiological water quality, the effect of protozoan grazing on E. coli and Enterococcus faecalis in Los Padres Lagoon waters (Buenos Aires, Argentina, 37° 56'30" S, 57° 44'30" W) was herein analyzed. Microcosm assays were carried out, simulating lacustrine conditions, confronting suspensions of autochthonous bacterivorous protozoans with suspensions of autochthonous and collection strains of E. coli and E. faecalis, combined and individually. Daily counts were made for evaluating bacterial survival and the number of ciliates. The results obtained indicate that there is a preferential sequence for bacterial removal in the water, where E. faecalis is more grazing-resistant than E. coli. Moreover, it was noted that the origin of bacterial strains influenced their sensitivity for grazing, at least in the short term (e.g. the collection strains were less affected). We conclude that protozoan grazing can modify the relative abundance of fecal indicator microorganisms, thus altering the results of water quality studies.

Effect of carbon sources on the aggregation of photo fermentative bacteria induced by L-cysteine for enhancing hydrogen production.

PubMed

Xie, Guo-Jun; Liu, Bing-Feng; Ding, Jie; Wang, Qilin; Ma, Chao; Zhou, Xu; Ren, Nan-Qi

2016-12-01

Poor flocculation of photo fermentative bacteria resulting in continuous biomass washout from photobioreactor is a critical challenge to achieve rapid and stable hydrogen production. In this work, the aggregation of Rhodopseudomonas faecalis RLD-53 was successfully developed in a photobioreactor and the effects of different carbon sources on hydrogen production and aggregation ability were investigated. Extracellular polymeric substances (EPS) production by R. faecalis RLD-53 cultivated using different carbon sources were stimulated by addition of L-cysteine. The absolute ζ potentials of R. faecalis RLD-53 were considerably decreased with addition of L-cysteine, and aggregation barriers based on DLVO dropped to 15-43 % of that in control groups. Thus, R. faecalis RLD-53 flocculated effectively, and aggregation abilities of strain RLD-53 cultivated with acetate, propionate, lactate and malate reached 29.35, 32.34, 26.07 and 24.86 %, respectively. In the continuous test, hydrogen-producing activity was also promoted and reached 2.45 mol H 2 /mol lactate, 3.87 mol H 2 /mol propionate and 5.10 mol H 2 /mol malate, respectively. Therefore, the aggregation of R. faecalis RLD-53 induced by L-cysteine is independent on the substrate types, which ensures the wide application of this technology to enhance hydrogen recovery from wastewater dominated by different organic substrates.

Brazilian artisanal ripened cheeses as sources of proteolytic lactic acid bacteria capable of reducing cow milk allergy.

PubMed

Biscola, V; Choiset, Y; Rabesona, H; Chobert, J-M; Haertlé, T; Franco, B D G M

2018-04-13

The objective was to obtain lactic acid bacteria (LAB) capable of hydrolysing immunoreactive proteins in milk, to optimize the hydrolysis, to determine the proteolysis kinetics and to test the safety of the best hydrolytic strain. Brazilian cheese was used as source of LAB capable of hydrolysing main milk allergens. Proteolytic isolates were submitted to RAPD-PCR for the characterization of clonal diversity. Optimized hydrolysis was strain and protein fraction dependent. 16S rDNA sequencing identified three proteolytic strains: Enterococcus faecalis VB43, that hydrolysed α S1 -, α S2 - and β-caseins, α-lactalbumin and β-lactoglobulin (partial hydrolysis), and Pediococcus acidilactici VB90 and Weissella viridescens VB111, that caused partial hydrolysis of α S1 - and α S2 -caseins. Enterococcus faecalis VB43 tested negative for virulence genes asa1, agg, efaA, hyl, esp, cylL L and cylL S but positive for genes ace and gelE. Ethylenediamine tetra-acetic acid inhibited the proteolysis, indicating that the main proteases of E. faecalis VB43 are metalloproteases. Brazilian artisanal cheese is a good source of LAB capable of hydrolysing allergenic proteins in milk. One isolate (E. faecalis VB43) presented outstanding activity against these proteins and lacked most of the tested virulence genes. Enterococcus faecalis VB43 presents good potential for the manufacture of hypoallergenic dairy products. © 2018 The Society for Applied Microbiology.

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

Recovery of metallo-tolerant and antibiotic resistant psychrophilic bacteria from Siachen glacier, Pakistan.

PubMed

Rafiq, Muhammad; Hayat, Muhammad; Anesio, Alexandre M; Jamil, Syed Umair Ullah; Hassan, Noor; Shah, Aamer Ali; Hasan, Fariha

2017-01-01

Cultureable bacterial diversity of previously unexplored Siachen glacier, Pakistan, was studied. Out of 50 isolates 33 (66%) were Gram negative and 17 (34%) Gram positive. About half of the isolates were pigment producers and were able to grow at 4-37°C. 16S rRNA gene sequences revealed Gram negative bacteria dominated by Proteobacteria (especially γ-proteobacteria and β-proteobacteria) and Flavobacteria. The genus Pseudomonas (51.51%, 17) was dominant among γ- proteobacteria. β-proteobacteria constituted 4 (12.12%) Alcaligenes and 4 (12.12%) Janthinobacterium strains. Among Gram positive bacteria, phylum Actinobacteria, Rhodococcus (23.52%, 4) and Arthrobacter (23.52%, 4) were the dominating genra. Other bacteria belonged to Phylum Firmicutes with representative genus Carnobacterium (11.76%, 2) and 4 isolates represented 4 genera Bacillus, Lysinibacillus, Staphylococcus and Planomicrobium. Most of the Gram negative bacteria were moderate halophiles, while most of the Gram positives were extreme halophiles and were able to grow up to 6.12 M of NaCl. More than 2/3 of the isolates showed antimicrobial activity against multidrug resistant S. aureus, E. coli, Klebsiella pneumonia, Enterococcus faecium, Candida albicans, Aspergillus flavus and Aspergillus fumigatus and ATCC strains. Gram positive bacteria (94.11%) were more resistant to heavy metals as compared to Gram negative (78.79%) and showed maximum tolerance against iron and least tolerance against mercury.

Recovery of metallo-tolerant and antibiotic resistant psychrophilic bacteria from Siachen glacier, Pakistan

PubMed Central

Rafiq, Muhammad; Hayat, Muhammad; Anesio, Alexandre M.; Jamil, Syed Umair Ullah; Hassan, Noor; Shah, Aamer Ali

2017-01-01

Cultureable bacterial diversity of previously unexplored Siachen glacier, Pakistan, was studied. Out of 50 isolates 33 (66%) were Gram negative and 17 (34%) Gram positive. About half of the isolates were pigment producers and were able to grow at 4–37°C. 16S rRNA gene sequences revealed Gram negative bacteria dominated by Proteobacteria (especially γ-proteobacteria and β-proteobacteria) and Flavobacteria. The genus Pseudomonas (51.51%, 17) was dominant among γ- proteobacteria. β-proteobacteria constituted 4 (12.12%) Alcaligenes and 4 (12.12%) Janthinobacterium strains. Among Gram positive bacteria, phylum Actinobacteria, Rhodococcus (23.52%, 4) and Arthrobacter (23.52%, 4) were the dominating genra. Other bacteria belonged to Phylum Firmicutes with representative genus Carnobacterium (11.76%, 2) and 4 isolates represented 4 genera Bacillus, Lysinibacillus, Staphylococcus and Planomicrobium. Most of the Gram negative bacteria were moderate halophiles, while most of the Gram positives were extreme halophiles and were able to grow up to 6.12 M of NaCl. More than 2/3 of the isolates showed antimicrobial activity against multidrug resistant S. aureus, E. coli, Klebsiella pneumonia, Enterococcus faecium, Candida albicans, Aspergillus flavus and Aspergillus fumigatus and ATCC strains. Gram positive bacteria (94.11%) were more resistant to heavy metals as compared to Gram negative (78.79%) and showed maximum tolerance against iron and least tolerance against mercury. PMID:28746396

Enterococcus faecalis and Candida albicans in the dental root canal and periapical infections.

PubMed

Kovac, J; Kovac, D; Slobodnikova, L; Kotulova, D

2013-01-01

The aim of the present study was to examine the prevalence of Enterococcus faecalis and Candida albicans in endodontic infections. Samples for microbiological examination were collected from 32 patients with deep dental caries, infected dental root canal, or periapical infection. Cultivation of the dental samples yielded four strains of Enterococcus faecalis (12.5 %), and three strains of Candida albicans (9.4 %). All Enterococcus faecalis isolates were susceptible to ampicillin, one isolate was resistant to tetracycline, two to erythromycin and azithromycin (additional 2 had intermediate susceptibility), and one strain had intermediate susceptibility to ciprofloxacin and moxifloxacin. We conclude that Enterococcus faecalis and Candida albicans can participate in the dental root canal and periapical infections, and the use of effective irrigant solutions and intracanal medicaments active against these microbes is important in order to prevent endodontic therapy failures. Unexpected was the isolation of C. albicans from a nine-year-old child with periodontitis apicalis. This finding must draw attention to the possibility that even at such a young age, this microorganism could be a potential etiological agent in endodontic infections (Tab. 2, Ref. 34). Text in PDF www.elis.sk.

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

Bacteria in the apical root canals of teeth with apical periodontitis.

PubMed

Lee, Li-Wan; Lee, Ya-Ling; Hsiao, Sheng-Huang; Lin, Hung-Pin

2017-06-01

Bacteria in the tooth root canal may cause apical periodontitis. This study examined the bacterial species present in the apical root canal of teeth with apical periodontitis. Antibiotic sensitivity tests were performed to evaluate whether these identified bacterial species were susceptible to specific kinds of antibiotics. Selective media plating and biochemical tests were used first to detect the bacterial species in samples taken from the apical portion of root canals of 62 teeth with apical periodontitis. The isolated bacterial species were further confirmed by matrix-assisted laser desorption ionization-time of flight mass spectrometry. We found concomitant presence of two (32 teeth) or three species (18 teeth) of bacteria in 50 (80.6%) out of 62 tested teeth. However, only 34 bacterial species were identified. Of a total of 118 bacterial isolates (83 anaerobes and 35 aerobes), Prophyromonas endodontalis was detected in 10; Bacteroides, Dialister invisus or Fusobacterium nucleatum in 9; Treponema denticola or Enterococcus faecalis in 8; Peptostreptococcus or Olsenella uli in 6; and Veillonella in 5 teeth. The other 25 bacterial species were detected in fewer than five teeth. Approximately 80-95% of bacterial isolates of anaerobes were sensitive to ampicillin/sulbactam (Unasyn), amoxicillin/clavulanate (Augmentin), cefoxitin, and clindamycin. For E. faecalis, 85-90% of bacterial isolates were sensitive to gentamicin and linezolid. Root canal infections are usually caused by a mixture of two or three species of bacteria. Specific kinds of antibiotic can be selected to control these bacterial infections after antibiotic sensitivity testing. Copyright © 2016. Published by Elsevier B.V.

How long can culturable bacteria and total DNA persist in environmental waters? The role of sunlight and solid particles.

PubMed

Gutiérrez-Cacciabue, Dolores; Cid, Alicia G; Rajal, Verónica B

2016-01-01

In this work, sunlight inactivation of two indicator bacteria in freshwater, with and without solid particles, was studied and the persistence of culturable cells and total DNA was compared. Environmental water was used to prepare two matrices, with and without solid particles, which were spiked with Escherichia coli and Enterococcus faecalis. These matrices were used to prepare microcosm bags that were placed in two containers: one exposed to sunlight and the other in the dark. During one month, samples were removed from each container and detection was done by membrane filter technique and real-time PCR. Kinetic parameters were calculated to assess sunlight effect. Indicator bacteria without solid particles exposed to sunlight suffered an immediate decay (<4h) compared with the ones which were shielded from them. In addition, the survival of both bacteria with solid particles varied depending on the situation analyzed (T99 from 3 up to 60days), being always culturable E. coli more persistent than E. faecalis. On the other side, E. faecalis DNA persisted much longer than culturable cells (T99>40h in the dark with particles). In this case active cells were more prone to sunlight than total DNA and the protective effect of solid particles was also observed. Results highlight that the effects caused by the parameters which describe the behavior of culturable microorganisms and total DNA in water are different and must be included in simulation models but without forgetting that these parameters will also depend on bacterial properties, sensitizers, composition, type, and uses of the aquatic environment under assessment. Copyright © 2015 Elsevier B.V. All rights reserved.

Predominant bacteria in an activated sludge reactor for the degradation of cutting fluids

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baker, C.A.; Claus, G.W.; Taylor, P.A.

1983-01-01

For the first time, an activated sludge reactor, established for the degradation of cutting fluids, was examined for predominant bacteria. In addition, both total and viable numbers of bacteria in the reactor were determined so that the percentage of each predominant type in the total reactor population could be determined. Three samples were studied, and a total of 15 genera were detected. In each sample, the genus Pseudomonas and the genus Microcyclus were present in high numbers. Three other genera, Acinetobacter, Alcaligenes, and Corynebacterium, were also found in every sample but in lower numbers. In one sample, numerous appendage bacteriamore » were present, and one of these, the genus Seliberia, was the most predominant organism in that sample. However, in the other two samples no appendage bacteria were detected. Six genera were found in this reactor which have not been previously reported in either cutting fluids in use or in other activated sludge systems. These genera were Aeromonas, Hyphomonas, Listeria, Microcyclus, Moraxella, and Spirosoma. None of the predominant bacterial belonged to groups of strict pathogens. 22 references, 6 figures, 3 tables.« less

Envelope Structures of Gram-Positive Bacteria

PubMed Central

Rajagopal, Mithila; Walker, Suzanne

2016-01-01

Gram-positive organisms, including the pathogens Staphylococcus aureus, Streptococcus pneumoniae and Enterococcus faecalis, have dynamic cell envelopes that mediate interactions with the environment and serve as the first line of defense against toxic molecules. Major components of the cell envelope include peptidoglycan, which is a well-established target for antibiotics, teichoic acids, capsular polysaccharides, surface proteins, and phospholipids. These components can undergo modification to promote pathogenesis, decrease susceptibility to antibiotics and host immune defenses, and enhance survival in hostile environments. This chapter will cover the structure, biosynthesis and important functions of major cell envelope components in Gram-positive bacteria. Possible targets for new antimicrobials will be noted. PMID:26919863

Incidence of virulence determinants in clinical Enterococcus faecalis and Enterococcus faecium isolates collected in Bulgaria.

PubMed

Strateva, Tanya; Atanasova, Daniela; Savov, Encho; Petrova, Guergana; Mitov, Ivan

2016-01-01

To evaluate the prevalence of some virulence genes among 510 clinical Enterococcus spp. isolates and to assess the association of those genes with the species, infection site, and patient group (inpatients/outpatients). Adhesins genes (aggregation substances agg and asa1 of Enterococcus faecalis and Enterococcus faecium, respectively), enterococcal surface protein (esp), endocarditis-specific antigen A (efaA), collagen-binding proteins (ace/acm)); invasins (hyaluronidase (hyl) and gelatinase (gelE)); cytotoxines (activation of cytolysin (cylA) in E. faecalis); and modulators of the host immunity and inflammation (enhanced expression pheromone (eep) in E. faecalis) were detected by polymerase chain reaction. The overall prevalence was: esp - 44.3%, agg/asa1 - 38.4%, ace/acm - 64.3%, efaA - 85.9%, eep - 69.4%, gelE - 64.3%, hyl - 25.1%, and cylA - 47.1%. E. faecalis isolates had significantly higher frequency of adhesin genes (esp and agg/asa1) and gelatinase in comparison to E. faecium. Multiple virulence genes in E. faecalis were significantly more prevalent than in E. faecium isolates. Domination of E. faecium with or without only one gene compared to the isolates of E. faecalis were found. Enterococcus spp. isolates obtained from outpatients compared to inpatients isolates had significantly higher frequency of agg/asa1, eep, gelE and cylA. Some adhesins genes (esp, agg/asa1 and efaA) had higher prevalence among the non-invasive Enterococcus spp. isolates compared to those causing invasive bacteremia, while ace/acm revealed higher dissemination in isolates causing invasive infections compared to non-invasive isolates. Most E. faecalis attaches to abiotic surfaces in hospital environment, which correlates with higher prevalence of gene encoding for virulence factors involved in biofilm formation, such as enterococcal surface protein, aggregation substance, and gelatinase. The intestinal tract is an important reservoir for opportunistic enterococcal pathogens and

Integration of non-oral bacteria into in vitro oral biofilms.

PubMed

Thurnheer, Thomas; Belibasakis, Georgios N

2015-01-01

Biofilms are polymicrobial communities that grow on surfaces in nature. Oral bacteria can spontaneously form biofilms on the surface of teeth, which may compromise the health of the teeth, or their surrounding (periodontal) tissues. While the oral bacteria exhibit high tropism for their specialized ecological niche, it is not clear if bacteria that are not part of the normal oral microbiota can efficiently colonize and grow within oral biofilms. By using an in vitro "supragingival" biofilm model of 6 oral species, this study aimed to investigate if 3 individual bacterial species that are not part of the normal oral microbiota (Eschericia coli, Staphylococcus aureus, Enterococcus faecails) and one not previously tested oral species (Aggregatibacter actinomycetemcomitans) can be incorporated into this established supragingival biofilm model. Staphylococcus aureus and A. actinomycetemcomitans were able to grow efficiently in the biofilm, without disrupting the growth of the remaining species. They localized in sparse small aggregates within the biofilm mass. Enterococcus faecalis and E. coli were both able to populate the biofilm at high numbers, and suppressed the growth of A. oris and S. mutants. Enterococcus faecalis was arranged in a chain-like conformation, whereas E. coli was densely and evenly spread throughout the biofilm mass. In conclusion, it is possible for selected species that are not part of the normal oral microbiota to be introduced into an oral biofilm, under the given experimental micro-environmental conditions. Moreover, the equilibrated incorporation of A. actinomycetemcomitans and S. aureus in this oral biofilm model could be a useful tool in the study of aggressive periodontitis and peri-implantitis, in which these organisms are involved, respectively.

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. Copyright © 2012 Elsevier GmbH. All rights reserved.

The effects of sodium hypochlorite and chlorhexidine irrigants on the antibacterial activities of alkaline media against Enterococcus faecalis.

PubMed

Ma, Jinglei; Tong, Zhongchun; Ling, Junqi; Liu, Hongyan; Wei, Xi

2015-07-01

Sodium hypochlorite (NaOCl), chlorhexidine (CHX) and calcium hydroxide are common intracanal medicaments. The present study aimed to evaluate the effects of NaOCl and CHX on the antibacterial activities of alkaline media against Enterococcus faecalis. The survival rates of planktonic and biofilm E. faecalis were evaluated by plate counts after 1 min of pretreatment with NaOCl and CHX, and time-kill assays were then used to assess subsequent pH alkaline challenges. Dead and living cells in the E. faecalis biofilm were assessed with SYTO 9 and PI staining in combination with confocal laser scanning microscopy following exposure to NaOCl or CHX and subsequent alkaline challenges by common root canal irrigation and dressing procedures. One minute of pretreatment with 2% CHX, 0.2% CHX, or 5.25% NaOCl in combination with a subsequent alkaline challenge significantly decreased planktonic E. faecalis survival rates, but pretreatment with 1% NaOCl did not. The E. faecalis biofilm survival rates were reduced in the subsequent alkaline challenge following CHX pretreatment but gradually increased following NaOCl pretreatment. Similarly, CLSM analysis revealed that the greatest proportions of dead E. faecalis cells in the biofilms were presented in the CHX and alkaline treatment group. CHX might be more effective in improving the antibacterial activities of alkaline root canal medicaments against E. faecalis than NaOCl during routine root canal therapy procedures. Copyright © 2015 Elsevier Ltd. All rights reserved.

Sudan azo dyes and Para Red degradation by prevalent bacteria of the human gastrointestinal tract☆

PubMed Central

Xu, Haiyan; Heinze, Thomas M.; Paine, Donald D.; Cerniglia, Carl E.; Chen, Huizhong

2018-01-01

Sudan azo dyes have genotoxic effects and ingestion of food products contaminated with Sudan I, II, III, IV, and Para Red could lead to exposure in the human gastrointestinal tract. In this study, we examined thirty-five prevalent species of human intestinal bacteria to evaluate their capacity to degrade Sudan dyes and Para Red. Among these tested bacterial strains, 23, 13, 33, 30, and 29 out of 35 species tested were able to reduce Sudan I, II, III, IV, and Para Red, respectively, to some extent. Bifidobacterium infantis, Clostridium indolis, Enterococcus faecalis, Lactobacillus rhamnosus, and Ruminococcus obeum were able to reduce completely all four tested Sudan dyes and Para Red. Escherichia coli and Peptostreptococcus magnus were the only two strains that were not able to reduce any of the tested Sudan dyes and Para Red to any significant extent. Metabolites of the reduction of the tested Sudan dyes and Para Red by E. faecalis were isolated and identified by HPLC and LC/ESI-MS analyses and compared with authentic standards. Thus it appears that the ability to reduce Sudan dyes and Para Red except Sudan II is common among bacteria in the human colon. PMID:19580882

Separation and characterization of effective demulsifying substances from surface of Alcaligenes sp. S-XJ-1 and its application in water-in-kerosene emulsion.

PubMed

Huang, Xiangfeng; Peng, Kaiming; Feng, Yi; Liu, Jia; Lu, Lijun

2013-07-01

The main goal of this work was to analyze the effect of surface substances on demulsifying capability of the demulsifying strain Alcaligenes sp. S-XJ-1. The demulsifying substances were successfully separated from the cell surface with dichloromethane-alkali treatment, and exhibited 67.5% of the demulsification ratio for water-in-kerosene emulsions at a dosage of 356mg/L. FT-IR, TLC and ESI-MS analysis confirmed the presence of a carbohydrate-protein-lipid complex in the demulsifying substances with the major molecular ions from mass-to-charge ratio (m/z) 165 to 814. After the substances separated, the cell morphology changed from aggregated to dispersed, and the concentration of cell surface functional groups decreased. Cell surface hydrophobicity and the ability of cell adhesion to hydrophobic surface of the treated cells was also reduced compared with original cell. It was proved that the demulsifying substances had a significant effect on cell surface properties and accordingly with demulsifying capability of Alcaligenes sp. S-XJ-1. Copyright © 2013 Elsevier Ltd. All rights reserved.

[Spectrum and drug sensitivity of pathogenic bacteria in children with nephrotic syndrome complicated by urinary tract infection: an analysis of 97 cases].

PubMed

Song, Shao-Na; Zhang, Bi-Li; Wang, Wen-Hong; Zhang, Xuan

2012-09-01

To investigate the spectrum and drug sensitivity of pathogenic bacteria in children with nephrotic syndrome (NS) complicated by urinary tract infection (UTI). A retrospective analysis was performed on the spectrum and drug sensitivity of pathogenic bacteria in 97 children with NS complicated by UTI, who hospitalized from January to December, 2011. The incidence of UTI in children with NS was 36.5%. It was significantly more common in children with recurrent NS than in those with primary NS (44.0% vs 31.9%; P<0.05). These cases mainly presented with asymptomatic bacteriuria. Enterococcus was the most common pathogenic bacteria (50.5%), including Enterococcus faecium (29.4%) and Enterococcus faecalis (21.1%), followed by Gram-negative bacteria, such as Escherichia coli (15.6%) and Klebsiella pneumoniae (14.7%). Enterococcus was highly sensitive to nitrofurantoin, vacomycin and linezolid, but was highly resistant to tetracycline and moxifloxacin. More multi-resistant strains were detected in Enterococcus faecium than in Enterococcus faecalis (72% vs 17%; P<0.05). Escherichia coli and Klebsiella pneumoniae were highly sensitive to amikacin, imipenem and piperacillin/tazobactam. Of the Gram-negative bacteria, 25% produced extended spectrum β-lactamases (ESBLs). ESBLs-producing bacteria had 100% sensitivity to imipenem, amikacin and piperacillin/tazobactam but were highly resistant to ampicillin, cefazolin and ceftriaxone. Children with recurrent NS are more susceptible to UTI than those with primary NS. Enterococcus is becoming major pathogenic bacteria for UTI in children with NS and has relatively high drug resistance, and most strains of Enterococcus faecium are multi-resistant.

Rhizosphere colonization and arsenic translocation in sunflower (Helianthus annuus L.) by arsenate reducing Alcaligenes sp. strain Dhal-L.

PubMed

Cavalca, Lucia; Corsini, Anna; Bachate, Sachin Prabhakar; Andreoni, Vincenza

2013-10-01

In the present study, six arsenic-resistant strains previously isolated were tested for their plant growth promoting characteristics and heavy metal resistance, in order to choose one model strain as an inoculum for sunflower plants in pot experiments. The aim was to investigate the effect of arsenic-resistant strain on sunflower growth and on arsenic uptake from arsenic contaminated soil. Based on plant growth promoting characteristics and heavy metal resistance, Alcaligenes sp. strain Dhal-L was chosen as an inoculum. Beside the ability to reduce arsenate to arsenite via an Ars operon, the strain exhibited 1-amino-cyclopropane-1-carboxylic acid deaminase activity and it was also able to produce siderophore and indole acetic acid. Pot experiments were conducted with an agricultural soil contaminated with arsenic (214 mg kg⁻¹). A real time PCR method was set up based on the quantification of ACR3(2) type of arsenite efflux pump carried by Alcaligenes sp. strain Dhal-L, in order to monitor presence and colonisation of the strain in the bulk and rhizospheric soil. As a result of strain inoculation, arsenic uptake by plants was increased by 53 %, whereas ACR3(2) gene copy number in rhizospheric soil was 100 times higher in inoculated than in control pots, indicating the colonisation of strain. The results indicated that the presence of arsenate reducing strains in the rhizosphere of sunflower influences arsenic mobilization and promotes arsenic uptake by plant.

Rapid Identification of Vancomycin Resistant Enterococcus Faecalis Clinical Isolates using a Sugar Fermentation Method

PubMed Central

Raeisi, Javad; Saifi, Mahnaz; Pourshafie, Mohammad Reza; Habibi, Mehri; Mohajerani, Hamid Reza; Akbari, Neda

2017-01-01

Introduction Vancomycin Resistant Enterococci (VRE) can be found all over the world. Thus, rapid detection of the isolates could be of high importance in the treatment or prevention of the associated disease. Aim To measure the turanose fermentation in Enterococcus faecalis clinical isolates for rapid differentiation of VRE and Vancomycin-Susceptible E. faecalis (VSE) isolates. Materials and Methods Forty E. faecalis samples were isolated from 200 clinical samples in Tehran Medical Center, Iran, from October 2012 to December 2012. These isolates were detected according to the standard microbial and biochemical tests. Detection of VRE isolates was originally performed by disk diffusion using 1 μg vancomycin disk, followed by Polymerase Chain Reaction (PCR) amplification of the vanA gene. Finally, the turanose consumption in 1%, 0.7% and 0.5% dilutions was detected by a phenotypic method. Results Among the 40 E. faecalis isolates, 20 vancomycin-susceptible and 20 vancomycin-resistant E. faecalis were isolated according to the disk diffusion and PCR of the vanA gene. There was a considerable difference between VRE and VSE isolates in 0.7% dilution of turanose. However, there was no significant difference between VRE and VSE in 1% and 0.5% dilutions of turanose. Conclusion Since detection of VRE isolates is of high importance, especially in nosocomial infections, phenotypic methods may be highly useful for this purpose. In conclusion, our data indicate that VRE isolated from clinical samples could be distinguished from VSE isolates by turanose fermentation at dilution 0.7%. PMID:28511382

Survival and activity of Streptococcus faecalis and Escherichia coli in petroleum-contaminated tropical marine waters

DOE Office of Scientific and Technical Information (OSTI.GOV)

Santo Domingo, J.W.; Fuentes, F.A.; Hazen, T.C.

1987-12-31

The in situ survival and activity of Streptococcus faecalis and Escherichia coli were studied using membrane diffusion chambers in tropical marine waters receiving oil refinery effluents. Protein synthesis, DNA synthesis, respiration or fermentation, INT reduced per cell, and ATP per cell were used to measure physiological activity. Cell densities decreased significantly over time at both sites for both S. faecalis and E. coli; however, no significant differences in survival pattern were observed between S. faecalis and E.coli. Differences in protein synthesis between the two were only observed at a study site which was not heavily oiled. Although fecal streptococci havemore » been suggested as a better indicator of fecal contamination than fecal coliforms in marine waters, in this study both E. coli and S. faecalis survived and remained physiologically active for extended periods of time. These results suggest that the fecal streptococci group is not a better indicator of fecal contamination in tropical marine waters than the fecal coliform group, especially when that environment is high in long-chained hydrocarbons.« less

Myrcia ovata Cambessedes essential oils: A proposal for a novel natural antimicrobial against foodborne bacteria.

PubMed

de Jesus, Isabela Cristina; Santos Frazão, Gladslene Góes; Blank, Arie Fitzgerald; de Aquino Santana, Luciana Cristina Lins

2016-10-01

This paper reports the innovative antibacterial activity of essential oils (EOs) from nine Myrcia ovata Cambessedes plants against eight foodborne bacteria. Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Enterococcus faecalis and Pseudomonas aeruginosa were the most susceptible bacteria to EOs. In particular, the P. aeruginosa, which is usually resistant to antimicrobials agents, was extremely sensitive to some EOs. The gram-positive and gram-negative bacteria were inhibited and eliminated with minimum EOs concentrations ranging from 0.78 to 25 μL/mL. The Serratia marcensces and Escherichia coli were less susceptible to EOs alone. Consequently, some EOs combinations were investigated by checkerboard method against these bacteria and a synergistic effect was obtained. Myrcia ovata Cambessedes EOs showed high inhibitory and bactericidal effects against foodborne bacteria might be an interesting alternative for future applications as natural antimicrobials in food systems. Copyright © 2016 Elsevier Ltd. All rights reserved.

Nonspecific Bacterial Flora Isolated from the Body Surface and Inside Ixodes ricinus Ticks.

PubMed

Okła, Hubert; Sosnowska, Malwina; Jasik, Krzysztof P; Słodki, Jan; Wojtyczka, Robert D

2012-09-28

Ixodes ricinus and other representatives of the order Ixodida are vectors of typical pathogens: Borrelia burgdorferi sensu lato, Anaplasma phagocytophilium, Babesia spp., a tick-borne encephalitis virus, and other microorganisms which are important from a medical and veterinary point of view. The presented study focuses on the verification of nonspecific bacterial flora of I. ricinus. We analyzed ticks collected in a forest region in Silesia, an industrial district in Poland. Methods of classical microbiology and biochemical assays (API 20 NE test, API Staph test and MICRONAUT System) were used for isolation and identification of microorganisms living on the body surface of I. ricinus and inside ticks. The results show the presence of various bacteria on the surface and inside ticks' bodies. During the study, we isolated Acinetobacter lwoffi, Pseudomonas fluorescens, Aeromonas hydrophila, Achromobacter denitrificans, Alcaligenes faecalis, Stenotrophomonas maltophilia, Pseudomonas oryzihabitans, Micrococcus spp., Kocuria varians, Staphylococcus lentus, Kocuria kristinae, Streptococcus pneumoniae, Rhizobium radiobacter, Staphylococcus xylosus. Majority of the isolated species are non-pathogenic environmental microorganisms, but some of the isolated bacterial strains could cause severe infections.

Biotransformation of arsenite and bacterial aox activity in drinking water produced from surface water of floating houses: Arsenic contamination in Cambodia.

PubMed

Chang, Jin-Soo

2015-11-01

The potential arsenite bioteansformation activity of arsenic was investigated by examining bacterial arsenic arsenite-oxidizing gene such as aoxS, aoxR, aoxA, aoxB, aoxC, and aoxD in high arsenic-contaminated drinking water produced from the surface water of floating houses. There is a biogeochemical cycle of activity involving arsenite oxidase aox system and the ars (arsenic resistance system) gene operon and aoxR leader gene activity in Alcaligenes faecalis SRR-11 and aoxS leader gene activity in Achromobacter xylosoxidans TSL-66. Batch experiments showed that SRR-11 and TSL-66 completely oxidized 1 mM of As (III) to As (V) within 35-40 h. The leaders of aoxS and aoxR are important for gene activity, and their effects in arsenic bioremediation and mobility in natural water has a significant ecological role because it allows arsenite oxidase in bacteria to control the biogeochemical cycle of arsenic-contaminated drinking water produced from surface water of floating houses. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enterococcus faecalis Responds to Individual Exogenous Fatty Acids Independently of Their Degree of Saturation or Chain Length

PubMed Central

2017-01-01

ABSTRACT Enterococcus faecalis is a commensal of the human gastrointestinal tract that can persist in the external environment and is a leading cause of hospital-acquired infections. Given its diverse habitats, the organism has developed numerous strategies to survive a multitude of environmental conditions. Previous studies have demonstrated that E. faecalis will incorporate fatty acids from bile and serum into its membrane, resulting in an induced tolerance to membrane-damaging agents. To discern whether all fatty acids induce membrane stress protection, we examined how E. faecalis responded to individually supplied fatty acids. E. faecalis readily incorporated fatty acids 14 to 18 carbons in length into its membrane but poorly incorporated fatty acids shorter or longer than this length. Supplementation with saturated fatty acids tended to increase generation time and lead to altered cellular morphology in most cases. Further, exogenously supplied saturated fatty acids did not induce tolerance to the membrane-damaging antibiotic daptomycin. Supplementation with unsaturated fatty acids produced variable growth effects, with some impacting generation time and morphology. Exogenously supplied unsaturated fatty acids that are normally produced by E. faecalis and those that are found in bile or serum could restore growth in the presence of a fatty acid biosynthetic inhibitor. However, only the eukaryote-derived fatty acids oleic acid and linoleic acid provided protection from daptomycin. Thus, exogenous fatty acids do not lead to a common physiological effect on E. faecalis. The organism responds uniquely to each, and only host-derived fatty acids induce membrane protection. IMPORTANCE Enterococcus faecalis is a commonly acquired hospital infectious agent with resistance to many antibiotics, including those that target its cellular membrane. We previously demonstrated that E. faecalis will incorporate fatty acids found in human fluids, like serum, into its cellular

Characterization of Dihydrofolate Reductase Genes from Trimethoprim-Susceptible and Trimethoprim-Resistant Strains of Enterococcus faecalis

PubMed Central

Coque, Teresa M.; Singh, Kavindra V.; Weinstock, George M.; Murray, Barbara E.

1999-01-01

Enterococci are usually susceptible in vitro to trimethoprim; however, high-level resistance (HLR) (MICs, >1,024 μg/ml) has been reported. We studied Enterococcus faecalis DEL, for which the trimethoprim MIC was >1,024 μg/ml. No transfer of resistance was achieved by broth or filter matings. Two different genes that conferred trimethoprim resistance when they were cloned in Escherichia coli (MICs, 128 and >1,024 μg/ml) were studied. One gene that coded for a polypeptide of 165 amino acids (MIC, 128 μg/ml for E. coli) was identical to dfr homologs that we cloned from a trimethoprim-susceptible E. faecalis strain, and it is presumed to be the intrinsic E. faecalis dfr gene (which causes resistance in E. coli when cloned in multiple copies); this gene was designated dfrE. The nucleotide sequence 5′ to this dfr gene showed similarity to thymidylate synthetase genes, suggesting that the dfr and thy genes from E. faecalis are located in tandem. The E. faecalis gene that conferred HLR to trimethoprim in E. coli, designated dfrF, codes for a predicted polypeptide of 165 amino acids with 38 to 64% similarity with other dihydrofolate reductases from gram-positive and gram-negative organisms. The nucleotide sequence 5′ to dfrF did not show similarity to the thy sequences. A DNA probe for dfrF hybridized under high-stringency conditions only to colony lysates of enterococci for which the trimethoprim MIC was >1,024 μg/ml; there was no hybridization to plasmid DNA from the strain of origin. To confirm that this gene causes trimethoprim resistance in enterococci, we cloned it into the integrative vector pAT113 and electroporated it into RH110 (E. faecalis OG1RF::Tn916ΔEm) (trimethoprim MIC, 0.5 μg/ml), which resulted in RH110 derivatives for which the trimethoprim MIC was >1,024 μg/ml. These results indicate that dfrF is an acquired but probably chromosomally located gene which is responsible for in vitro HLR to trimethoprim in E. faecalis. PMID:9869579

Enterocin 96, a Novel Class II Bacteriocin Produced by Enterococcus faecalis WHE 96, Isolated from Munster Cheese▿

PubMed Central

Izquierdo, Esther; Wagner, Camille; Marchioni, Eric; Aoude-Werner, Dalal; Ennahar, Saïd

2009-01-01

Enterococcus faecalis WHE 96, a strain isolated from soft cheese based on its anti-Listeria activity, produced a 5,494-Da bacteriocin that was purified to homogeneity by ultrafiltration and cation-exchange and reversed-phase chromatographies. The amino acid sequence of this bacteriocin, named enterocin 96, was determined by Edman degradation, and its structural gene was sequenced, revealing a double-glycine leader peptide. After a comparison with other bacteriocins, it was shown that enterocin 96 was a new class II bacteriocin that showed very little similarity with known structures. Enterocin 96 was indeed a new bacteriocin belonging to class II bacteriocins. The activity spectrum of enterocin 96 covered a wide range of bacteria, with strong activity against most gram-positive strains but very little or no activity against gram-negative strains. PMID:19411428

Enterocin 96, a novel class II bacteriocin produced by Enterococcus faecalis WHE 96, isolated from Munster cheese.

PubMed

Izquierdo, Esther; Wagner, Camille; Marchioni, Eric; Aoude-Werner, Dalal; Ennahar, Saïd

2009-07-01

Enterococcus faecalis WHE 96, a strain isolated from soft cheese based on its anti-Listeria activity, produced a 5,494-Da bacteriocin that was purified to homogeneity by ultrafiltration and cation-exchange and reversed-phase chromatographies. The amino acid sequence of this bacteriocin, named enterocin 96, was determined by Edman degradation, and its structural gene was sequenced, revealing a double-glycine leader peptide. After a comparison with other bacteriocins, it was shown that enterocin 96 was a new class II bacteriocin that showed very little similarity with known structures. Enterocin 96 was indeed a new bacteriocin belonging to class II bacteriocins. The activity spectrum of enterocin 96 covered a wide range of bacteria, with strong activity against most gram-positive strains but very little or no activity against gram-negative strains.

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

Apical Extrusion of Intracanal Bacteria with Single File and Multifile Rotary Instrumentation Systems.

PubMed

Saberi, Eshaghali; Zahedani, Shahram Shahraki; Ebrahimipour, Sediqe

2017-01-01

Instrumentation techniques may cause extrusion of microorganisms and their products into the periapical region resulting inflammation and treatment failure. The aim of this ex vivo study was comparing the apical bacterial extrusion in canals prepared with single file versus multiple file rotary systems. Ninety-two human single-rooted mandibular first premolars were used. Endodontic access cavities were prepared, and root canals were contaminated with an Enterococcus faecalis ( E. faecalis ) suspension. The samples were incubated at 37°C for 30 days; the contaminated teeth were divided into four groups of 20 specimens each (1: Reciproc, 2: Mtwo, 3: Neoniti A1, 4: Safesider). Six teeth were not infected and each were prepared with one of the above instruments were considered as negative and six teeth which had been previously infected, were used as positive control groups. Extruded bacteria from the apical foramen during instrumentation were collected into vials containing 0.9% NaCl. The microbial samples were taken from the vials and incubated in brain heart agar medium for 24 h. The resulting bacterial titer, in colony-forming units per mL, was determined. The data entered into SPSS 18 software and were analyzed by Kruskal-Wallis and Mann-Whitney U-tests at 0.05 significance level. Mtwo multifile system showed significantly less bacterial extrusion than Safesider ( P = 0.015) and Neoniti A1 ( P = 0.042) but did not show significant difference with Reciproc system ( P = 0.25). All instrumentation systems extruded bacteria beyond the apical foramen. However, this study showed that Mtwo multifile rotary system extruded fewer bacteria.

Soymilk fermentation by Enterococcus faecalis VB43 leads to reduction in the immunoreactivity of allergenic proteins β-conglycinin (7S) and glycinin (11S).

PubMed

Biscola, V; de Olmos, A Rodriguez; Choiset, Y; Rabesona, H; Garro, M S; Mozzi, F; Chobert, J-M; Drouet, M; Haertlé, T; Franco, B D G M

2017-08-24

Food allergies represent a serious problem affecting human health and soy proteins rank among the most allergenic proteins from food origin. The proteolytic enzymes produced by lactic acid bacteria (LAB) can hydrolyse the major allergens present in soybean, reducing their immunoreactivity. Many studies have reported the ability of LAB to ferment soy-based products; while the majority of them focus on the improvement of the sensory characteristics and functionality of soy proteins, a lack of information about the role of lactic fermentation in the reduction of immunoreactivity of these proteins exists. The aim of the present study was to evaluate the capability of the proteolytic strain Enterococcus faecalis VB43 to hydrolyse the main allergenic proteins present in soymilk and to determine the immunoreactivity of the obtained hydrolysates. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) results of fermented soymilk demonstrated complete hydrolysis of the β-subunit from β-conglycinin and the acidic polypeptide from glycinin. Reversed phase high performance liquid chromatography (RP-HPLC) analysis of the peptides released after hydrolysis revealed the appearance of new peptides and the disappearance of non-hydrolysed proteins, indicating extensive hydrolysis of the substrate. Results from competitive enzyme-linked immunosorbent assay (ELISA) tests clearly indicated a reduction in the immunoreactivity (more than one logarithmic unit) in the fermented sample as compared to the non-fermented control. Our results suggest that the soymilk fermented by E. faecalis VB43 may induce lower allergic responses in sensitive individuals. The strain E. faecalis VB43 may be considered as an excellent candidate to efficiently reduce the immunoreactivity of soymilk proteins.

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). © 2014 Australian Society of Endodontology.

Extracellular polysaccharides produced by cooling water tower biofilm bacteria and their possible degradation.

PubMed

Ceyhan, Nur; Ozdemir, Guven

2008-01-01

The extracellular polymers (EPS) of biofilm bacteria that can cause heat and mass transfer problems in cooling water towers in the petrochemical industry were investigated. In addition, these microorganisms were screened for their ability to grow and degrade their own EPS and the EPS of other species. Twelve bacteria producing the most EPS were isolated from cooling water towers and characterized biochemically by classic and commercial systems. These were species of Pseudomonas, Burkholderia, Aeromonas, Pasteurella, Pantoea, Alcaligenes and Sphingomonas. EPS of these species were obtained by propan-2-ol precipitation and centrifugation from bacterial cultures in media enriched with glucose, sucrose or galactose. EPS yields were of 1.68-4.95 g l(-1). These EPS materials were characterized for total sugar and protein contents. Their total sugar content ranged from 24 to 56% (g sugar g(-1) EPS), and their total protein content ranged from 10 to 28% (g protein g(-1) EPS). The monosaccharide compositions of EPS were determined by HPLC. Generally, these compositions were enriched in galactose and glucose, with lesser amounts of mannose, rhamnose, fructose and arabinose. All bacteria were investigated in terms of EPS degradation. Eight of the bacteria were able to utilize EPS from Burkholderia cepacia, seven of the bacteria were able to utilize EPS from Pseudomonas sp. and Sphingomonas paucimobilis. The greatest viscosity reduction of B. cepacia was obtained with Pseudomonas sp. The results show that the bacteria in this study are able to degrade EPS from biofilms in cooling towers.

Restructuring of Enterococcus faecalis biofilm architecture in response to antibiotic-induced stress

DOE PAGES

Dale, Jennifer L.; Nilson, Jennifer L.; Barnes, Aaron M. T.; ...

2017-06-30

Bacterial biofilms are intrinsically resistant to antimicrobial treatment, which contributes to microbial persistence in clinical infections. Enterococcus faecalis is an opportunistic pathogen that readily forms biofilms and is the most prevalent enterococcal species identified in healthcare-associated infections. Since intrinsic resistance to multiple antibiotics is common for enterococci, and antibiotic resistance is elevated in biofilm populations, it is imperative to understand the mechanisms involved. Previously, we identified two glycosyltransferase genes whose disruption resulted in impaired nascent biofilm formation in the presence of antibiotic concentrations subinhibitory for parent growth and biofilm formation. The glycosyltransferases are involved in synthesis of the cell-wall-associated rhamnopolysaccharidemore » Epa. Here we examined the effect of epa mutations on the temporal development of E. faecalis biofilms, and on the effects of antibiotics on pre-formed biofilms using scanning electron microscopy. We show that ΔepaOX mutant cells arrange into complex multidimensional biofilms independent of antibiotic exposure, while parent cells form biofilms that are monolayers in the absence of antibiotics. Remarkably, upon exposure to antibiotics parent biofilm cells restructure into complex three-dimensional biofilms resembling those of the ΔepaOX mutant without antibiotics. All biofilms exhibiting complex cellular architectures were less structurally stable than monolayer biofilms, with the biofilm cells exhibiting increased detachment. Our results indicate that E. faecalis biofilms restructure in response to cellular stress whether induced by antibiotics in the case of parent cells, or by deficiencies in Epa composition for the ΔepaOX strain. The data demonstrate a link between cellular architecture and antibiotic resistance of E. faecalis biofilms.« less

Restructuring of Enterococcus faecalis biofilm architecture in response to antibiotic-induced stress

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dale, Jennifer L.; Nilson, Jennifer L.; Barnes, Aaron M. T.

Bacterial biofilms are intrinsically resistant to antimicrobial treatment, which contributes to microbial persistence in clinical infections. Enterococcus faecalis is an opportunistic pathogen that readily forms biofilms and is the most prevalent enterococcal species identified in healthcare-associated infections. Since intrinsic resistance to multiple antibiotics is common for enterococci, and antibiotic resistance is elevated in biofilm populations, it is imperative to understand the mechanisms involved. Previously, we identified two glycosyltransferase genes whose disruption resulted in impaired nascent biofilm formation in the presence of antibiotic concentrations subinhibitory for parent growth and biofilm formation. The glycosyltransferases are involved in synthesis of the cell-wall-associated rhamnopolysaccharidemore » Epa. Here we examined the effect of epa mutations on the temporal development of E. faecalis biofilms, and on the effects of antibiotics on pre-formed biofilms using scanning electron microscopy. We show that ΔepaOX mutant cells arrange into complex multidimensional biofilms independent of antibiotic exposure, while parent cells form biofilms that are monolayers in the absence of antibiotics. Remarkably, upon exposure to antibiotics parent biofilm cells restructure into complex three-dimensional biofilms resembling those of the ΔepaOX mutant without antibiotics. All biofilms exhibiting complex cellular architectures were less structurally stable than monolayer biofilms, with the biofilm cells exhibiting increased detachment. Our results indicate that E. faecalis biofilms restructure in response to cellular stress whether induced by antibiotics in the case of parent cells, or by deficiencies in Epa composition for the ΔepaOX strain. The data demonstrate a link between cellular architecture and antibiotic resistance of E. faecalis biofilms.« less

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.

Photodynamic UVA-riboflavin bacterial elimination in antibiotic-resistant bacteria.

PubMed

Makdoumi, Karim; Bäckman, Anders

2016-09-01

To evaluate the bactericidal effect of clinical ultraviolet A (UVA) settings used in photoactivated chromophore for infectious keratitis (PACK)-collagen cross-linking (CXL) in antibiotic-resistant and non-resistant bacterial strains. Well-characterized bacterial strains from clinical isolates, without and with antibiotic resistance, were studied in a pairwise comparison. The evaluated pathogens were Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, and Enterococcus faecalis. Bacteria were dispersed in PBS and diluted to a concentration of approximately 4 × 10 5 /ml. Riboflavin was added to a concentration of 0.01%. By spreading the solution on a microscope slide, a fluid film layer, with a thickness of around 400 mm, was formed and UVA exposure followed. Eight separate exposures were made for each strain (n = 8). The degree of elimination in resistant and non-resistant pathogens was compared. The bactericidal efficacy of exposure differed between the tested microorganisms, and the mean elimination ranged between 60 and 92%, being most extensive in both of the evaluated Pseudomonas strains and least in the E. faecalis strains. Similar reductions were seen in antibiotic-resistant and non-resistant strains, with the exception of S. aureus, in which the resistant strain metchicillin-resistant Staphylococcus aureus (MRSA) was eradicated in a greater extent than the non-resistant strain (P = 0.030). UVA-riboflavin settings used in PACK-CXL are effective in reducing both antibiotic-resistant and non-resistant bacteria. Antibiotic resistance does not appear to be protective against the photooxidative exposure. © 2016 Royal Australian and New Zealand College of Ophthalmologists.

Isolation and identification of symbiotic bacteria from the skin, mouth, and rectum of wild and captive tree shrews.

PubMed

Li, Gui; Lai, Ren; Duan, Gang; Lyu, Long-Bao; Zhang, Zhi-Ye; Liu, Huang; Xiang, Xun

2014-11-18

Endosymbionts influence many aspects of their hosts' health conditions, including physiology, development, immunity, metabolism, etc. Tree shrews (Tupaia belangeri chinensis) have attracted increasing attention in modeling human diseases and therapeutic responses due to their close relationship with primates. To clarify the situation of symbiotic bacteria from their body surface, oral cavity, and anus, 12 wild and 12 the third generation of captive tree shrews were examined. Based on morphological and cultural characteristics, physiological and biochemical tests, as well as the 16S rDNA full sequence analysis, 12 bacteria strains were isolated and identified from the wild tree shrews: body surface: Bacillus subtilis (detection rate 42%), Pseudomonas aeruginosa (25%), Staphlococcus aureus (33%), S. Epidermidis (75%), Micrococcus luteus (25%), Kurthia gibsonii (17%); oral cavity: Neisseria mucosa (58%), Streptococcus pneumonia (17%); anus: Enterococcus faecalis (17%), Lactococus lactis (33%), Escherichia coli (92%), Salmonella typhosa (17%); whereas, four were indentified from the third generation captive tree shrews: body surface: S. epidermidis (75%); oral cavity: N.mucosa (67%); anus: L. lactis (33%), E. coli (100%). These results indicate that S. epidermidis, N. mucosa, L. lactis and E. coli were major bacteria in tree shrews, whereas, S. aureus, M. luteus, K. gibsonii, E. faecalis and S. typhosa were species-specific flora. This study facilitates the future use of tree shrews as a standard experimental animal and improves our understanding of the relationship between endosymbionts and their hosts.

Temperature effect on bacterial azo bond reduction kinetics: an Arrhenius plot analysis.

PubMed

Angelova, Blaga; Avramova, Tatyana; Stefanova, Lilyana; Mutafov, Sava

2008-06-01

Studied was the effect of temperature in the range 12-46 degrees C on the rate of bacterial decolorization of the mono-azo dye Acid Orange 7 by Alcaligenes faecalis 6132 and Rhodococcus erythropolis 24. With both strains the raise of temperature led to a corresponding raise of decolorization rate better manifested by R. erythropolis. The analysis of the Arrhenius plot revealed a break near the middle of the temperature range. The regression analysis showed practically complete identity of the observed break point temperatures (T (BP)): 20.7 degrees C for Alc. faecalis and 20.8 degrees C for R. erythropolis. The values of the activation energy of the decolorization reaction (E (a)) were found to depend on both the organism and the temperature range. In the range below T (BP) the estimated values of E (a) were 138 +/- 7 kJ mol(-1) for Alc. faecalis and 160 +/- 8 kJ mol(-1) for R. erythropolis. In the range above T (BP) they were 54.2 +/- 1.8 kJ mol(-1) for Alc. faecalis and 37.6 +/- 4.1 kJ mol(-1) for R. erythropolis. Discussed are the possible reasons for the observed abrupt change of the activation energy.

Replacement of Tyrosine 181 by Phenylalanine in Gentisate 1,2-Dioxygenase I from Pseudomonas alcaligenes NCIMB 9867 Enhances Catalytic Activities

PubMed Central

Tan, Chew Ling; Yeo, Chew Chieng; Khoo, Hoon Eng; Poh, Chit Laa

2005-01-01

xlnE, encoding gentisate 1,2-dioxygenase (EC 1.13.11.4), from Pseudomonas alcaligenes (P25X) was mutagenized by site-directed mutagenesis. The mutant enzyme, Y181F, demonstrated 4-, 3-, 6-, and 16-fold increases in relative activity towards gentisate and 3-fluoro-, 4-methyl-, and 3-methylgentisate, respectively. The specific mutation conferred a 13-fold higher catalytic efficiency (kcat/Km) on Y181F towards 3-methylgentisate than that of the wild-type enzyme. PMID:16237038

[Study of marine actinomycetes isolated from the central coast of Peru and their antibacterial activity against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecalis].

PubMed

León, Jorge; Aponte, Juan José; Rojas, Rosario; Cuadra, D'Lourdes; Ayala, Nathaly; Tomás, Gloria; Guerrero, Marco

2011-06-01

To determine the antimicrobial potential of marine actinomycetes against drug-resistant pathogens represented by strains of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE). Strains of actinomycetes (29) isolated from marine sediment were evaluated by their characteristics in two culture media and by testing their inhibitory capacity by in vitro antagonism against multi-drug resistant (MDR) pathogenic bacteria for MRSA and VRE. Organic extracts of 3 selected actinomicetes were processed to determine the minimum inhibitory concentration (MIC) of the active compound. Most isolated actinomycetes belong to a homogeneous group of write-gray actinomycetes with a good growth in Marine Agar. The inhibitory rates of the isolates were above 85% for both pathogens with inhibition zones greater than 69 and 78 mm in diameter for MRSA and VRE respectively. Dichloromethane extracts of 3 isolates (I-400A, B1-T61, M10-77) showed strong inhibitory activity of both pathogens, M10-77 being the highest actinomycete strain with antibiotic activity against methicillin-resistant S. aureus ATCC 43300 and vancomycin-resistant E. faecalis ATCC 51299 with a minimum inhibitory concentrations (MIC) of 7.9 and 31.7 μg/ml respectively. Phylogenetic analysis of M10-77 strain showed 99% similarity with the marine species Streptomyces erythrogriseus. Marine sediments of the central coast of Peru, are a source of actinomycetes strains showing high capacity to produce bioactive compounds able to inhibit pathogens classified as multi-drug-resistant such as methicillin-resistant S. aureus and vancomycin-resistant E. faecalis.

Detection and identification of intestinal pathogenic bacteria by hybridization to oligonucleotide microarrays

PubMed Central

Jin, Lian-Qun; Li, Jun-Wen; Wang, Sheng-Qi; Chao, Fu-Huan; Wang, Xin-Wei; Yuan, Zheng-Quan

2005-01-01

AIM: To detect the common intestinal pathogenic bacteria quickly and accurately. METHODS: A rapid (<3 h) experimental procedure was set up based upon the gene chip technology. Target genes were amplified and hybridized by oligonucleotide microarrays. RESULTS: One hundred and seventy strains of bacteria in pure culture belonging to 11 genera were successfully discriminated under comparatively same conditions, and a series of specific hybridization maps corresponding to each kind of bacteria were obtained. When this method was applied to 26 divided cultures, 25 (96.2%) were identified. CONCLUSION: Salmonella sp., Escherichia coli, Shigella sp., Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus, Proteus sp., Bacillus cereus, Vibrio cholerae, Enterococcus faecalis, Yersinia enterocolitica, and Campylobacter jejuni can be detected and identified by our microarrays. The accuracy, range, and discrimination power of this assay can be continually improved by adding further oligonucleotides to the arrays without any significant increase of complexity or cost. PMID:16437687

The Effect of Diode Laser on Planktonic Enterococcus faecalis in Infected Root Canals in an Ex Vivo Model.

PubMed

Cretella, Gilda; Lajolo, Carlo; Castagnola, Raffaella; Somma, Francesco; Inchingolo, MariaTeresa; Marigo, Luca

2017-04-01

This study examined the bactericidal effect of diode laser irradiation against intracanal Enterococcus faecalis. m total of 128 extracted single-rooted and single-canal teeth were treated with ProTaper instruments (Dentsply Maillefer, Ballaigues, Switzerland). A total of 120 root canals were inoculated with E. faecalis for 21 days, and the samples were randomly divided into five groups: Group 1 (n = 24) samples were irrigated with only saline solution (positive controls); Group 2 (n = 24) was treated with only 5.25% sodium hypochlorite; Group 3 (n = 24) was irrigated with saline solutions activated by diode laser; Group 4 (n = 24) was treated with 5.25% sodium hypochlorite activated by diode laser; and Group 5 (n = 24) was irrigated with saline solution with methylene blue dye activated by the diode laser Fox (Sweden & Martina, Padova, Italy); additionally, eight teeth were not contaminated and their canals were irrigated with saline solution and used as a negative control. The Uro-Quick system was used to determine the microbial residual charge. The data were analyzed using Pearson's chi-square test (p < 0.001). A statistically significant reduction in bacterial count was observed in Group 2 and Group 4 (p < 0.001). There were no statistically significant differences among the other groups (p > 0.001). Evidence indicates that the diode laser was not more effective than sodium hypochlorite in reducing free bacteria.

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.

Hydrogen Sulfide Sensing through Reactive Sulfur Species (RSS) and Nitroxyl (HNO) in Enterococcus faecalis.

PubMed

Shen, Jiangchuan; Walsh, Brenna J C; Flores-Mireles, Ana Lidia; Peng, Hui; Zhang, Yifan; Zhang, Yixiang; Trinidad, Jonathan C; Hultgren, Scott J; Giedroc, David P

2018-05-17

Recent studies of hydrogen sulfide (H 2 S) signaling implicate low molecular weight (LMW) thiol persulfides and other reactive sulfur species (RSS) as signaling effectors. Here, we show that a CstR protein from the human pathogen Enterococcus faecalis ( E. faecalis), previously identified in Staphylococcus aureus ( S. aureus), is an RSS-sensing repressor that transcriptionally regulates a cst-like operon in response to both exogenous sulfide stress and Angeli's salt, a precursor of nitroxyl (HNO). E. faecalis CstR reacts with coenzyme A persulfide (CoASSH) to form interprotomer disulfide and trisulfide bridges between C32 and C61', which negatively regulate DNA binding to a consensus CstR DNA operator. A Δ cstR strain exhibits deficiency in catheter colonization in a catheter-associated urinary tract infection (CAUTI) mouse model, suggesting sulfide regulation and homeostasis is critical for pathogenicity. Cellular polysulfide metabolite profiling of sodium sulfide-stressed E. faecalis confirms an increase in both inorganic polysulfides and LMW thiols and persulfides sensed by CstR. The cst-like operon encodes two authentic thiosulfate sulfurtransferases and an enzyme we characterize here as an NADH and FAD-dependent coenzyme A (CoA) persulfide reductase (CoAPR) that harbors an N-terminal CoA disulfide reductase (CDR) domain and a C-terminal rhodanese homology domain (RHD). Both cysteines in the CDR (C42) and RHD (C508) domains are required for CoAPR activity and complementation of a sulfide-induced growth phenotype of a S. aureus strain lacking cstB, encoding a nonheme Fe II persulfide dioxygenase. We propose that S. aureus CstB and E. faecalis CoAPR employ orthogonal chemistries to lower CoASSH that accumulates under conditions of cellular sulfide toxicity and signaling.

Lipoquinones of some spore-forming rods, lactic-acid bacteria and actinomycetes.

PubMed

Hess, A; Holländer, R; Mannheim, W

1979-11-01

The respiratory quinones of 73 strains of Gram-positive bacteria including spore-forming rods, lactic-acid bacteria and actinomyctes were examined. Menaquinones with seven isoprenoid units (MK-7) were the main quinone type found in representatives of the genus Bacillus and in Sporolactobacillus inulinus. However, a strain of B. thuringiensis produced MK-8 in addition to MK-7, and strains of B. lentus and B. pantothenticus appeared to produce MK-9 and MK-8, respectively, with no MK-7. In the clostridia and lactic-acid bacteria, no quinones were found, except in Pediococcus cerevisiae NCTC 8066 and Lactobacillus casei subsp. rhamnosus ATCC 7469, which contained menaquinones, and Streptococcus faecalis NCTC 775 and HIM 478-1, which contained demethylmenaquinones, in relatively low concentrations. Menaquinones were also found in the actinomycetes (except Actinomyces odontolyticus and Bifidobacterium bifidum which did not produce any quinones) and in Protaminobacter alboflavus ATCC 8458, the so-called Actinobacillus actinoides ATCC 15900 and Noguchia granulosis NCTC 10559.

Effect of a Benzalkonium Chloride Surfactant-Sodium Hypochlorite Combination on Elimination of Enterococcus faecalis.

PubMed

Baron, Aleksandr; Lindsey, Kimberly; Sidow, Stephanie J; Dickinson, Douglas; Chuang, Augustine; McPherson, James C

2016-01-01

The purpose of this investigation was to determine the effect of a sodium hypochlorite-surfactant combination on the removal of Enterococcus faecalis from infected teeth. Sixty-four extracted human single canal anterior teeth were prepared with rotary instrumentation and sterilized. Teeth were divided into 4 groups, N = 16. Three experimental groups were inoculated with E. faecalis and cultured for 21 days before use: positive control group, no irrigation; NaOCl group, irrigated with 5 mL 6% NaOCl; and NaOCl/BAK group, irrigated with 5 mL 6% NaOCl/0.008% benzalkonium chloride (BAK). The negative control group received medium only and no inoculate. Paper point sampling of the canals was obtained before irrigation (S1) for all 4 groups and for 2 groups after irrigation (S2) to determine remaining colony-forming units. After sampling, all teeth were split in half and evaluated for bacterial viability colony-forming units and penetration of dentinal tubules by using fluorescent vital dye staining and confocal laser scanning microscopy. Comparison of pre-irrigation and post-irrigation paper point samples from the 2 irrigated groups showed a significant reduction in bacterial canal load (P < .001, Kruskal-Wallis), with a significantly lower load in the NaOCl/BAK group than in the NaOCl group (P = .001, Mann-Whitney U test); 68.8% of the NaOCl/BAK samples gave no recoverable counts. In contrast, no significant difference between these groups was found for counts recovered from dentin. Confocal laser scanning microscopy showed no differences in tubule penetration. The addition of BAK to NaOCl significantly reduced the number of remaining bacteria within the canal after irrigation compared with NaOCl alone. Published by Elsevier Inc.

Antimicrobial effects of root canal medicaments against Enterococcus faecalis and Streptococcus mutans.

PubMed

Atila-Pektaş, B; Yurdakul, P; Gülmez, D; Görduysus, O

2013-05-01

To compare the antimicrobial activities of Activ Point (Roeko, Langenau, Germany), Calcium Hydroxide Plus Point (Roeko, Langenau, Germany), calcium hydroxide, 1% chlorhexidine gel and bioactive glass (S53P4) against Enterococcus faecalis and Streptococcus mutans. One hundred and twenty extracted single-rooted human teeth were used. After removing the crowns, root canals were prepared by using the Protaper rotary system. Following autoclave sterilization, root canals were incubated at 37 °C with E. faecalis ATCC 29212 and S. mutans RSHM 676 for 1 week. The specimens, which were divided into five treatment groups for each microorganism according to the intracanal medicament used, were tested in 10 experimental runs. In each experimental run, 10 roots were included as treatment, one root as positive control and one root as sterility control. Sterile paper points were utilized to take samples from root canals after the incubation of teeth in thioglycollate medium at 37 °C for 1 week. Samples taken from teeth by sterile paper points were inoculated onto sheep blood agar, and following an overnight incubation, the colonies grown on sheep blood agar were counted and interpreted as colony-forming units. Results were tested statistically by using Kruskal-Wallis and Conover's nonparametric multiple comparison tests. CHX gel (P < 0.001 and P < 0.001), Activ Point (P = 0.003 and P = 0.002) and Ca(OH)₂ (P = 0.010 and P = 0.005) were significantly more effective against E. faecalis than that of Ca(OH)₂ Plus Point and bioactive glass, respectively. On the other hand, compared with Ca(OH)₂ , CHX gel (P < 0.001), and Activ Point (P < 0.001), bioactive glass (P = 0.014) produced significantly lower colony counts of S. mutans. When compared with the positive control, treatment with Ca(OH)₂ Plus Point (P = 0.085 and P = 0.066) did not produce significantly lower colony counts of E. faecalis and S. mutans, respectively. Compared with the medicaments having an antimicrobial

A novel combination approach of human polyclonal IVIG and antibiotics against multidrug-resistant Gram-positive bacteria

PubMed Central

Sallam, Mariam Madkour; Abou-Aisha, Khaled; El-Azizi, Mohamed

2016-01-01

Background Gram-positive bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA) and enterococci, have shown a remarkable ability to develop resistance to antimicrobial agents. Objective We aimed to assess possible enhancement of the antimicrobial activity of vancomycin, amoxicillin, clarithromycin, and azithromycin by human polyclonal intravenous immunoglobulin G (IVIG) against 34 multidrug-resistant (MDR) bacterial isolates, including MRSA, Enterococcus faecium, and Enterococcus faecalis. Materials and methods Double combinations of the antibiotics with the IVIG were assessed by checkerboard assay, where the interaction was evaluated with respect to the minimum inhibitory concentration (MIC) of the antibiotics. The results of the checkerboard assay were verified in vitro using time-kill assay and in vivo using an invasive sepsis murine model. Results The checkerboard assay showed that IVIG enhanced the antimicrobial activity of amoxicillin and clarithromycin against isolates from the three groups of bacteria, which were resistant to the same antibiotics when tested in the absence of IVIG. The efficacy of vancomycin against 15% of the tested isolates was enhanced when it was combined with the antibodies. Antagonism was demonstrated in 47% of the E. faecalis isolates when clarithromycin was combined with the IVIG. Synergism was proved in the time-kill assay when amoxicillin was combined with the antibodies; meanwhile, antagonism was not demonstrated in all tested combinations, even in combinations that showed such response in checkerboard assay. Conclusion The suggested approach is promising and could be helpful to enhance the antimicrobial activity of not only effective antibiotics but also antibiotics that have been proven to be ineffective against MDR bacteria. To our knowledge, this combinatorial approach against MDR bacteria, such as MRSA and enterococci, has not been investigated before. PMID:27994476

Biodiversity of lactic acid bacteria in Moroccan soft white cheese (Jben).

PubMed

Ouadghiri, Mouna; Amar, Mohamed; Vancanneyt, Marc; Swings, Jean

2005-10-15

The bacterial diversity occurring in traditional Moroccan soft white cheese, produced in eight different regions in Morocco, was studied. A total of 164 lactic acid bacteria were isolated, purified and identified by whole-cell protein fingerprinting and rep-PCR genomic fingerprinting. The majority of the strains belonged to the genera Lactobacillus, Lactococcus, Leuconostoc and Enterococcus. Sixteen species were identified: Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus brevis, Lactobacillus buchneri, Lactococcus lactis, Lactococcus garvieae, Lactococcus raffinolactis, Leuconostoc pseudomesenteroides, Leuconostoc mesenteroides, Leuconostoc citreum, Eterococcus durans, Enterococcus faecalis, Enterococcus faecium, Enterococcus saccharominimus and Streptococcus sp.

High-level resistance to gentamicin: genetic transfer between Enterococcus faecalis isolated from food of animal origin and human microbiota.

PubMed

Sparo, M; Urbizu, L; Solana, M V; Pourcel, G; Delpech, G; Confalonieri, A; Ceci, M; Sánchez Bruni, S F

2012-02-01

  To investigate the in vivo gene transfer of high-level gentamicin resistance (HLRG) from Enterococcus faecalis isolated from the food of animal origin to a human isolate, using a mouse model of intestinally colonized human microbiota.   In vitro study: The presence of plasmids involved in HLRG coding was investigated. After the conjugation experiment, the recipient strain, Ent. faecalis JH2-SS, acquired a plasmid responsible for HLRG [minimal inhibitory concentration (MIC) >800 μg ml(-1) ], in a similar position to the donor cells. In vivo study: Seven BALB/c mice were dosed with ceftriaxone (400 mg kg(-1) ) and then inoculated with a dilution of 1/100 of human faeces (HFc). After 72 h, Ent. faecalis JH2-SS (recipient) was inoculated and then, after a further 72 h, the animals were given Ent. faecalis CS19, isolated from the food of animal origin, involved in HLRG (donor). The presence of transconjugant strains in HFc was subsequently recorded on a daily basis until the end of the experiment. The clonal relationship between Ent. faecalis and Escherichia coli in faeces was assessed by RAPD-PCR. Both the in vitro and in vivo studies showed that the receptor strain acquired a plasmid responsible for HLRG (MICs >800 μg ml(-1) ), which migrated with a similar relative mobility value. Transconjugant strains were detected from 24 h after the donor strain inoculation and persisted until the end of the experiment.   The in vivo gene transfer of HLRG from Ent. faecalis strains, isolated from the food of animal origin, to human microbiota has been demonstrated in a mouse model.   The complexity found on the therapeutic responses of invasive infectious diseases caused by Ent. faecalis facilitates the assessment of food of animal origin as a resistant pathogen reservoir. In addition, this study may contribute to the understanding of antimicrobials' resistance gene transfer between Ent. faecalis strains from food and human GI tract. © 2011 The Authors. Letters in

PURIFICATION AND ACTIVITY OF PROTEINASE OF STREPTOCOCCUS FAECALIS VAR. LIQUEFACIENS

PubMed Central

Shugart, Lee R.; Beck, Raymond W.

1964-01-01

Shugart, Lee R. (University of Tennessee, Knoxville) and Raymond W. Beck. Purification and activity of proteinase of Streptococcus faecalis var. liquefaciens. J. Bacteriol. 88:586–590. 1964.—A proteolytic enzyme from Streptococcus faecalis var. liquefaciens was purified 480-fold by ammonium sulfate fractionation and treatment with calcium phosphate gel. Approximately 20% of the original enzyme activity was recovered in the purified fraction. Optimal enzyme activity was found to be at pH 7.6 and 35 C. The enzyme is apparently more susceptible to heat denaturation when complexed with substrate than when heated in the absence of substrate. Michaelis-Menten constants were found to be 0.655% for hemoglobin and 0.133% for casein. Apparent energies of activation on these substrates were calculated to be 9,060 and 12,020 cal, respectively. PMID:14208492

High-purity Nisin Alone or in Combination with Sodium Hypochlorite Is Effective against Planktonic and Biofilm Populations of Enterococcus faecalis.

PubMed

Kajwadkar, Ruma; Shin, Jae M; Lin, Guo-Hao; Fenno, J Christopher; Rickard, Alexander H; Kapila, Yvonne L

2017-06-01

Nisin, a broad-spectrum bacteriocin, has recently been highlighted for its biomedical applications. To date, no studies have examined the antimicrobial and antibiofilm properties of high-purity (>95%) nisin (nisin ZP) on Enterococcus faecalis and biofilms formed by this species. We hypothesize that nisin can inhibit E. faecalis and reduce biofilm biomass, and combinations of nisin and sodium hypochlorite (NaOCl) will enhance the antibiofilm properties against E. faecalis biofilms. Using broth cultures, disc diffusion assays, and biofilm assays, we examined the effects of nisin on various E. faecalis growth parameters and biofilm properties (biovolume, thickness, and roughness). Confocal microscopy was used in conjunction with Imaris and Comstat2 software (Kongens Lyngby, Copenhagen, Denmark) to measure and analyze the biofilm properties. Nisin significantly decreased the growth of planktonic E. faecalis dose dependently. The minimum inhibitory concentrations against E. faecalis strains OG-1 and ATCC 29212 were 15 and 50 μg/mL, and the minimum bactericidal concentrations were 150 and 200 μg/mL, respectively. A reduction in biofilm biovolume and thickness was observed for biofilms treated with nisin at ≥10 μg/mL for 10 minutes. In addition, the combination of nisin with low doses of NaOCl enhanced the antibiofilm properties of both antimicrobial agents. Nisin alone or in combination with low concentrations of NaOCl reduces the planktonic growth of E. faecalis and disrupts E. faecalis biofilm structure. Our results suggest that nisin has potential as an adjunctive endodontic therapeutic agent and as an alternative to conventional NaOCl irrigation. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

In Vitro antibacterial efficacy of 21 Indian timber-yielding plants against multidrug-resistant bacteria causing urinary tract infection.

PubMed

Mishra, Monali P; Padhy, Rabindra N

2013-12-01

To screen methanolic leaf extracts of 21 timber-yielding plants for antibacterial activity against nine species of uropathogenic bacteria isolated from clinical samples of a hospital (Enterococcus faecalis, Staphylococcus aureus, Acinetobacter baumannii, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, and Pseudomonas aeruginosa). Bacterial strains were subjected to antibiotic sensitivity tests by the Kirby-Bauer's disc diffusion method. The antibacterial potentiality of leaf extracts was monitored by the agar-well diffusion method with multidrug-resistant (MDR) strains of nine uropathogens. Two Gram-positive isolates, E. faecalis and S. aureus, were resistant to 14 of the 18 antibiotics used. Gram-negative isolates A. baumannii, C. freundii, E. aerogenes, E. coli, K. pneumoniae, P. mirabilis, and P. aeruginosa were resistant to 10, 12, 9, 11, 11, 10, and 11 antibiotics, respectively, of the 14 antibiotics used. Methanolic leaf extracts of Anogeissus acuminata had the maximum zone of inhibition size-29 mm against S. aureus and 28 mm against E. faecalis and P. aeruginosa. Cassia tora had 29 mm as the zone of inhibition size for E. faecalis, E. aerogenes, and P. aeruginosa. Based on the minimum inhibitory concentration and minimum bactericidal concentration values, the most effective 10 plants against uropathogens could be arranged in decreasing order as follows: C. tora > A. acuminata > Schleichera oleosa > Pterocarpus santalinus > Eugenia jambolana > Bridelia retusa > Mimusops elengi > Stereospermum kunthianum > Tectona grandis > Anthocephalus cadamba. The following eight plants had moderate control capacity: Artocarpus heterophyllus, Azadirachta indica, Dalbergia latifolia, Eucalyptus citriodora, Gmelina arborea, Pongamia pinnata, Pterocarpus marsupium, and Shorea robusta. E. coli, followed by A. baumannii, C. freundii, E. aerogenes, P. mirabilis, and P. aeruginosa were controlled by

In Vitro Antibacterial Efficacy of 21 Indian Timber-Yielding Plants Against Multidrug-Resistant Bacteria Causing Urinary Tract Infection

PubMed Central

Mishra, Monali P.; Padhy, Rabindra N.

2013-01-01

Objectives To screen methanolic leaf extracts of 21 timber-yielding plants for antibacterial activity against nine species of uropathogenic bacteria isolated from clinical samples of a hospital (Enterococcus faecalis, Staphylococcus aureus, Acinetobacter baumannii, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, and Pseudomonas aeruginosa). Methods Bacterial strains were subjected to antibiotic sensitivity tests by the Kirby–Bauer's disc diffusion method. The antibacterial potentiality of leaf extracts was monitored by the agar-well diffusion method with multidrug-resistant (MDR) strains of nine uropathogens. Results Two Gram-positive isolates, E. faecalis and S. aureus, were resistant to 14 of the 18 antibiotics used. Gram-negative isolates A. baumannii, C. freundii, E. aerogenes, E. coli, K. pneumoniae, P. mirabilis, and P. aeruginosa were resistant to 10, 12, 9, 11, 11, 10, and 11 antibiotics, respectively, of the 14 antibiotics used. Methanolic leaf extracts of Anogeissus acuminata had the maximum zone of inhibition size—29 mm against S. aureus and 28 mm against E. faecalis and P. aeruginosa. Cassia tora had 29 mm as the zone of inhibition size for E. faecalis, E. aerogenes, and P. aeruginosa. Based on the minimum inhibitory concentration and minimum bactericidal concentration values, the most effective 10 plants against uropathogens could be arranged in decreasing order as follows: C. tora > A. acuminata > Schleichera oleosa > Pterocarpus santalinus > Eugenia jambolana > Bridelia retusa > Mimusops elengi > Stereospermum kunthianum > Tectona grandis > Anthocephalus cadamba. The following eight plants had moderate control capacity: Artocarpus heterophyllus, Azadirachta indica, Dalbergia latifolia, Eucalyptus citriodora, Gmelina arborea, Pongamia pinnata, Pterocarpus marsupium, and Shorea robusta. E. coli, followed by A. baumannii, C. freundii, E. aerogenes, P. mirabilis, and P

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

Enterococcus faecalis Ebp pili are important for cell-cell aggregation and intraspecies gene transfer

PubMed Central

Leanti La Rosa, Sabina; Camila Montealegre, Maria; Singh, Kavindra V.

2016-01-01

Enterococcus faecalis is an opportunistic pathogen that ranks among the leading causes of biofilm-associated infections. We previously demonstrated that the endocarditis- and biofilm-associated pili (Ebp) of E. faecalis play a major role in biofilm formation, adherence to abiotic surfaces and experimental infections. In this study, derivatives of E. faecalis strain OG1 were engineered to further characterize functions of Ebp pili. Loss of pili resulted in a 36-fold decrease in the number of closely associated cells when OG1RFΔebpABC was mixed with OG1SSpΔebpABC, compared with mixing the Ebp+ parental strains. In addition, using the Ebp+ parental strains as donor and recipient, we found a statistically significant increase (280–360 %, P < 0.05) in the frequency of plasmid transfer versus using Ebp−  mutants in the conjugation experiments. These results demonstrate a previously unrecognized role of Ebp pili, namely, as important contributors to microscale cell aggregation and horizontal spread of genetic material. PMID:26967674

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

The Effect of Er:YAG Laser on Entroccocus faecalis Bacterium in the Pulpectomy of Anterior Primary Teeth

PubMed Central

Bahrololoomi, Zahra; Poursina, Farkhondeh; Birang, Reza; Foroughi, Elnaz; Yousefshahi, Hazhir

2017-01-01

Introduction: Successful root canal therapy depends on the complete elimination of microorganisms such as Entroccocus faecalis, which is impossible to achieve with the traditional methods. Lasers are recently introduced as a new method to solve the problem. The present study is planned and performed to examining the antibacterial effect of Er: YAG laser. Methods: Sixty extracted anterior primary teeth were prepared and sterilized. E. faecalis bacterium was cultured in canals. Samples were randomly divided into two groups. The first group was disinfected by NaOCl 5/25% and Er: YAG laser and the second group just by NaOCl 5/25%. Samples of canal contents were cultured and colony counts were calculated. The results were analyzed statistically by SPSS software and Mann Whitney test. Results: There was no significant difference between colony counts in both groups (P=0.142). But the number of colonies in the first group was lower than in the second group. Conclusion: Although, Er: YAG laser cannot completely eliminate E. faecalis bacterium, its simultaneous use with NaOCl decreases E. faecalis. PMID:29071021

The Effect of Er:YAG Laser on Entroccocus faecalis Bacterium in the Pulpectomy of Anterior Primary Teeth.

PubMed

Bahrololoomi, Zahra; Poursina, Farkhondeh; Birang, Reza; Foroughi, Elnaz; Yousefshahi, Hazhir

2017-01-01

Introduction: Successful root canal therapy depends on the complete elimination of microorganisms such as Entroccocus faecalis , which is impossible to achieve with the traditional methods. Lasers are recently introduced as a new method to solve the problem. The present study is planned and performed to examining the antibacterial effect of Er: YAG laser. Methods: Sixty extracted anterior primary teeth were prepared and sterilized. E. faecalis bacterium was cultured in canals. Samples were randomly divided into two groups. The first group was disinfected by NaOCl 5/25% and Er: YAG laser and the second group just by NaOCl 5/25%. Samples of canal contents were cultured and colony counts were calculated. The results were analyzed statistically by SPSS software and Mann Whitney test. Results: There was no significant difference between colony counts in both groups ( P =0.142). But the number of colonies in the first group was lower than in the second group. Conclusion: Although, Er: YAG laser cannot completely eliminate E. faecalis bacterium, its simultaneous use with NaOCl decreases E. faecalis .

Prevalence, outcome and risk factor associated with vancomycin-resistant Enterococcus faecalis and Enterococcus faecium at a Tertiary Care Hospital in Northern India.

PubMed

Tripathi, A; Shukla, S K; Singh, A; Prasad, K N

2016-01-01

To determine the prevalence, genotype, risk factors and mortality in patients having vancomycin-resistant Enterococcus faecalis (VR E. faecalis) and Enterococcus faecium (VR E. faecium) infection or colonisation. A total of 1488 clinical isolates of E. faecalis and E. faecium were tested for vancomycin resistance by phenotypic (disk diffusion, E-test and broth micro-dilution test) and genotypic polymerase chain reaction methods. Records of all 1488 patients who had E. faecalis or E. faecium infection or colonisation were reviewed for the identification of host, hospital and medication related risk factors associated with VR E. faecalis and VR E. faecium. Of 1488 isolates, 118 (7.9%) were vancomycin-resistant and their distributions were as follows: E. faecalis=72 (61%) and E. faecium=46 (39%). All 118 vancomycin-resistant isolates were vanA genotype (minimum inhibitory concentration [MIC] to vancomycin ≥64 μg/ml and MIC to teicoplanin≥32 μg/ml) and none of the isolates was vanB genotype. Multivariate logistic regression analysis identified ventilator support and hospital stay for ≥48 h as independent risk factors associated with VR E. faecalis and VR E. faecium infection or colonisation. Hospital stay≥48 h was the only independent risk factor for mortality in patients infected with vancomycin-resistant enterococci. Strategies to limit the nosocomial infection especially in patients on ventilator support can reduce VRE incidence and related mortality.

Effectiveness of EDTA and Modified Salt Solution to Detach and Kill Cells from Enterococcus faecalis Biofilm.

PubMed

de Almeida, Josiane; Hoogenkamp, Michel; Felippe, Wilson T; Crielaard, Wim; van der Waal, Suzette V

2016-02-01

Disruption of the matrix of endodontic biofilms will aid in their removal from a root canal. Therefore, the aim of this study was to investigate the efficacy of EDTA and a modified salt solution (MSS) to detach bacteria from biofilms. Forty-eight-hour-old Enterococcus faecalis biofilms were grown on glass coverslips and then treated for 1 hour by immersion in 17% EDTA or MSS. Phosphate-buffered saline served as a negative control. Then, residual biofilm cells on the substrate and the detached cells in the supernatant were collected. Viability was verified by the colony-forming unit (CFU) counting method. Propidium monoazide (PMA) treatment in conjunction with quantitative polymerase chain reaction (qPCR) was also performed to detect the presence of E. faecalis 16S ribonucleic RNA genes. Data were analyzed using 1-way analysis of variance and Tukey or Kruskal-Wallis and Dunn tests. The Pearson R test evaluated the correlation between results from CFU and PMA (α = 5%). qPCR showed that EDTA detached 99% of biofilm cells, and MSS detached 94% of biofilm cells (both P < .001). In contrast to EDTA, MSS was highly antimicrobial. The treatment promoted an ample log 7 reduction of the attached cells (P < .001), and almost no live cells were detected in the supernatant (P < .001). Positive correlations between CFU and qPCR with PMA were observed (r = 0.959 and r = 0.729). EDTA detached cells in biofilms with a minor antimicrobial effect. Besides a great antimicrobial effect, MSS also detached biofilm cells. These dispersals of biofilms give insights into new endodontic biofilm removal strategies. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Enzymes Required for Maltodextrin Catabolism in Enterococcus faecalis Exhibit Novel Activities

PubMed Central

Joyet, Philippe; Mokhtari, Abdelhamid; Riboulet-Bisson, Eliette; Blancato, Víctor S.; Espariz, Martin; Magni, Christian; Sauvageot, Nicolas

2017-01-01

ABSTRACT Maltose and maltodextrins are formed during the degradation of starch or glycogen. Maltodextrins are composed of a mixture of maltooligosaccharides formed by α-1,4- but also some α-1,6-linked glucosyl residues. The α-1,6-linked glucosyl residues are derived from branching points in the polysaccharides. In Enterococcus faecalis, maltotriose is mainly transported and phosphorylated by a phosphoenolpyruvate:carbohydrate phosphotransferase system. The formed maltotriose-6″-phosphate is intracellularly dephosphorylated by a specific phosphatase, MapP. In contrast, maltotetraose and longer maltooligosaccharides up to maltoheptaose are taken up without phosphorylation via the ATP binding cassette transporter MdxEFG-MsmX. We show that the maltose-producing maltodextrin hydrolase MmdH (GenBank accession no. EFT41964) in strain JH2-2 catalyzes the first catabolic step of α-1,4-linked maltooligosaccharides. The purified enzyme converts even-numbered α-1,4-linked maltooligosaccharides (maltotetraose, etc.) into maltose and odd-numbered (maltotriose, etc.) into maltose and glucose. Inactivation of mmdH therefore prevents the growth of E. faecalis on maltooligosaccharides ranging from maltotriose to maltoheptaose. Surprisingly, MmdH also functions as a maltogenic α-1,6-glucosidase, because it converts the maltotriose isomer isopanose into maltose and glucose. In addition, E. faecalis contains a glucose-producing α-1,6-specific maltodextrin hydrolase (GenBank accession no. EFT41963, renamed GmdH). This enzyme converts panose, another maltotriose isomer, into glucose and maltose. A gmdH mutant had therefore lost the capacity to grow on panose. The genes mmdH and gmdH are organized in an operon together with GenBank accession no. EFT41962 (renamed mmgT). Purified MmgT transfers glucosyl residues from one α-1,4-linked maltooligosaccharide molecule to another. For example, it catalyzes the disproportionation of maltotriose by transferring a glucosyl residue to

Highly efficient removal of pathogenic bacteria with magnetic graphene composite.

PubMed

Zhan, Sihui; Zhu, Dandan; Ma, Shuanglong; Yu, Wenchao; Jia, Yanan; Li, Yi; Yu, Hongbing; Shen, Zhiqiang

2015-02-25

Magnetic Fe3O4/graphene composite (abbreviated as G-Fe3O4) was synthesized successfully by solvothermal method to effectively remove both bacteriophage and bacteria in water, which was tested by HRTEM, XRD, BET, XPS, FTIR, CV, magnetic property and zeta-potential measurements. Based on the result of HRTEM, the single-sheet structure of graphene oxide and the monodisperse Fe3O4 nanoparticles on the surface of graphene can be observed obviously. The G-Fe3O4 composite were attractive for removing a wide range of pathogens including not only bacteriophage ms2, but also various bacteria such as S. aureus, E. coli, Salmonella, E. Faecium, E. faecalis, and Shigella. The removal efficiency of E. coli for G-Fe3O4 composite can achieve 93.09%, whereas it is only 54.97% with pure Fe3O4 nanoparticles. Moreover, a detailed verification test of real water samples was conducted and the removal efficiency of bacteria in real water samples with G-Fe3O4 composite can also reach 94.8%.

[Post-marketing surveillance of antibacterial activities of cefozopran against various clinical isolates--I. Gram-positive bacteria].

PubMed

Igari, Jun; Oguri, Toyoko; Hiramatsu, Nobuyoshi; Akiyama, Kazumitsu; Koyama, Tsuneo

2002-02-01

As a post-marketing surveillance, the in vitro antibacterial activities of cefozopran (CZOP), an agent of cephems, against various clinical isolates were yearly evaluated and compared with those of other cephems, oxacephems, penicillins, and carbapenems. Changes in the bacterial sensitivity for CZOP were also evaluated with the resistance ratio calculated with breakpoint MIC. Sixteen species (1,913 strains) of Gram-positive bacteria were isolated from the clinical materials annually collected from 1996 to 2000, and consisted of methicillin-susceptible Staphylococcus aureus (MSSA; n = 178), methicillin-resistant S. aureus (MRSA; n = 199), methicillin-susceptible Staphylococcus epidermidis (MSSE; n = 98), methicillin-resistant S. epidermidis (MRSE; n = 164), Staphylococcus haemolyticus (n = 72), Staphylococcus saprophyticus (n = 28), Enterococcus faecalis (n = 206), Enterococcus faecium (n = 91), Enterococcus avium (n = 72), Streptococcus pyogenes (n = 133), Streptococcus agalactiae (n = 138), penicillin-susceptible Streptococcus pneumoniae (PSSP; n = 133), penicillin-intermediate resistant S. pneumoniae (PISP; n = 100), penicillin-resistant S. pneumoniae (PRSP; n = 29), Streptococcus milleri group (n = 135) and Peptostreptococcus spp. (n = 137). CZOP possessed comparable antibacterial activities against MSSA and MSSE to other cephems, and was also effective on MRSE but not on MRSA. An antibacterial activity of CZOP against S. saprophyticus was comparable to or higher than other cephems. CZOP, however, did not indicate an antibacterial activity against S. haemolyticus, just like other cephems. An antibacterial activity of CZOP against E. faecalis was comparable to cefpirome (CPR) and higher than other cephems. No antibacterial activity of CZOP against E. faecium and E. avium was observed, just like other drugs. An antibacterial activity of CZOP against S. pyogenes was as potent as that of cefotiam (CTM), cefepime (CFPM) and CPR, and that against S. agalactiae was

Comparative Genomics of Enterococci: Variation in Enterococcus faecalis, Clade Structure in E. faecium, and Defining Characteristics of E. gallinarum and E. casseliflavus

PubMed Central

Palmer, Kelli L.; Godfrey, Paul; Griggs, Allison; Kos, Veronica N.; Zucker, Jeremy; Desjardins, Christopher; Cerqueira, Gustavo; Gevers, Dirk; Walker, Suzanne; Wortman, Jennifer; Feldgarden, Michael; Haas, Brian; Birren, Bruce; Gilmore, Michael S.

2012-01-01

ABSTRACT The enterococci are Gram-positive lactic acid bacteria that inhabit the gastrointestinal tracts of diverse hosts. However, Enterococcus faecium and E. faecalis have emerged as leading causes of multidrug-resistant hospital-acquired infections. The mechanism by which a well-adapted commensal evolved into a hospital pathogen is poorly understood. In this study, we examined high-quality draft genome data for evidence of key events in the evolution of the leading causes of enterococcal infections, including E. faecalis, E. faecium, E. casseliflavus, and E. gallinarum. We characterized two clades within what is currently classified as E. faecium and identified traits characteristic of each, including variation in operons for cell wall carbohydrate and putative capsule biosynthesis. We examined the extent of recombination between the two E. faecium clades and identified two strains with mosaic genomes. We determined the underlying genetics for the defining characteristics of the motile enterococci E. casseliflavus and E. gallinarum. Further, we identified species-specific traits that could be used to advance the detection of medically relevant enterococci and their identification to the species level. PMID:22354958

Candida albicans and Enterococcus faecalis in the gut: synergy in commensalism?

PubMed

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.

Bactericidal efficacy of elevated pH on fish pathogenic and environmental bacteria

USGS Publications Warehouse

Starliper, Clifford E.; Watten, Barnaby J.

2013-01-01

Ship ballast water is a recognized medium for transfer and introductions of nonindigenous species. There is a need for new ballast water treatment methods that effectively and safely eliminate or greatly minimize movements of these species. The present study employed laboratory methods to evaluate the bactericidal efficacy of increased pH (pH 10.0–12.0) for exposure durations of up to 72 h to kill a variety of Gram-negative and Gram-positive bacteria including fish pathogens (Aeromonas spp., Yersinia ruckeri, Edwardsiella ictaluri, Serratia liquefaciens, Carnobacterium sp.), other common aquatic-inhabitant bacteria (Serratia marcescens, Pseudomonas fluorescens, Staphylococcus sp., Bacillus sp.) and indicators listed in International Maritime Organization D2 Standards; namely, Vibrio cholera (an environmental isolate from fish), Escherichia coli and Enterococcus faecalis. Volumes of 5 N NaOH were added to tryptic soy broth to obtain desired pH adjustments. Viable cells were determined after 0, 4, 12, 24, 48, and 72 h. Initial (0 h) cell numbers ranged from 3.40 × 104 cfu/mL for Bacillus sp. to 2.44 × 107 cfu/mL for E. faecalis. The effective endpoints of pH and treatment duration necessary to realize 100% bactericidal effect varied; however, all bacteria tested were killed within 72 h at pH 12.0 or lower. The lowest parameters examined, 4 h at pH 10.0, were bactericidal to V. cholera, E. ictaluri, three of four isolates of E. coli, and (three of four) Aeromonas salmonicida subsp. salmonicida. Bactericidal effect was attained at pH 10.0 within 12 h for the other A. salmonicida subsp. salmonicida, and within 24 h for P. fluorescens, and the remaining E. coli.

Evaluation of the antimicrobial effect of super-oxidized water (Sterilox®) and sodium hypochlorite against Enterococcus faecalis in a bovine root canal model

PubMed Central

ROSSI-FEDELE, Giampiero; de FIGUEIREDO, José Antonio Poli; STEIER, Liviu; CANULLO, Luigi; STEIER, Gabriela; ROBERTS, Adam P.

2010-01-01

Ideally root canal irrigants should have, amongst other properties, antimicrobial action associated with a lack of toxicity against periapical tissues. Sodium hypochlorite (NaOCl) is a widely used root canal irrigant, however it has been shown to have a cytotoxic effect on vital tissue and therefore it is prudent to investigate alternative irrigants. Sterilox's Aquatine Alpha Electrolyte® belongs to the group of the super-oxidized waters; it consists of a mixture of oxidizing substances, and has been suggested to be used as root canal irrigant. Super-oxidized waters have been shown to provide efficient cleaning of root canal walls, and have been proposed to be used for the disinfection of medical equipment. Objective To compare the antimicrobial action against Enterococcus faecalis of NaOCl, Optident Sterilox Electrolyte Solution® and Sterilox's Aquatine Alpha Electrolyte® when used as irrigating solutions in a bovine root canal model. Methodology Root sections were prepared and inoculated with E. faecalis JH2-2. After 10 days of incubation the root canals were irrigated using one of three solutions (NaOCl, Optident Sterilox Electrolyte Solution® and Sterilox's Aquatine Alpha Electrolyte®) and subsequently sampled by grinding dentin using drills. The debris was placed in BHI broth and dilutions were plated onto fresh agar plates to quantify growth. Results Sodium hypochlorite was the only irrigant to eliminate all bacteria. When the dilutions were made, although NaOCl was still statistically superior, Sterilox's Aquatine Alpha Electrolyte® solution was superior to Optident Sterilox Electrolyte Solution®. Conclusion Under the conditions of this study Sterilox's Aquatine Alpha Electrolyte® appeared to have significantly more antimicrobial action compared to the Optident Sterilox Electrolyte Solution® alone, however NaOCl was the only solution able to consistently eradicate E. faecalis in the model. PMID:21085808

Incorporation of Exogenous Fatty Acids Protects Enterococcus faecalis from Membrane-Damaging Agents

PubMed Central

Saito, Holly E.; Harp, John R.

2014-01-01

Enterococcus faecalis is a commensal bacterium of the mammalian intestine that can persist in soil and aquatic systems and can be a nosocomial pathogen to humans. It employs multiple stress adaptation strategies in order to survive such a wide range of environments. Within this study, we sought to elucidate whether membrane fatty acid composition changes are an important component for stress adaptation. We noted that E. faecalis OG1RF was capable of changing its membrane composition depending upon growth phase and temperature. The organism also readily incorporated fatty acids from bile, serum, and medium supplemented with individual fatty acids, often dramatically changing the membrane composition such that a single fatty acid was predominant. Growth in either low levels of bile or specific individual fatty acids was found to protect the organism from membrane challenges such as high bile exposure. In particular, we observed that when grown in low levels of bile, serum, or the host-derived fatty acids oleic acid and linoleic acid, E. faecalis was better able to survive the antibiotic daptomycin. Interestingly, the degree of membrane saturation did not appear to be important for protection from the stressors examined here; instead, it appears that a specific fatty acid or combination of fatty acids is critical for stress resistance. PMID:25128342

Pharmacokinetics of enrofloxacin and ceftiofur in plasma, interstitial fluid, and gastrointestinal tract of calves after subcutaneous injection, and bactericidal impacts on representative enteric bacteria.

PubMed

Foster, D M; Jacob, M E; Warren, C D; Papich, M G

2016-02-01

This study's objectives were to determine intestinal antimicrobial concentrations in calves administered enrofloxacin or ceftiofur sodium subcutaneously, and their impact on representative enteric bacteria. Ultrafiltration devices were implanted in the ileum and colon of 12 steers, which received either enrofloxacin or ceftiofur sodium. Samples were collected over 48 h after drug administration for pharmacokinetic/pharmacodynamic analysis. Enterococcus faecalis or Salmonella enterica (5 × 10(5) CFU/mL of each) were exposed in vitro to peak and tail (48 h postadministration) concentrations of both drugs at each location for 24 h to determine inhibition of growth and change in MIC. Enrofloxacin had tissue penetration factors of 1.6 and 2.5 in the ileum and colon, while ciprofloxacin, an active metabolite of enrofloxacin, was less able to cross into the intestine (tissue penetration factors of 0.7 and 1.7). Ceftiofur was rapidly eliminated leading to tissue penetration factors of 0.39 and 0.25. All concentrations of enrofloxacin were bactericidal for S. enterica and significantly reduced E. faecalis. Peak ceftiofur concentration was bactericidal for S. enterica, and tail concentrations significantly reduced growth. E. faecalis experienced growth at all ceftiofur concentrations. The MICs for both organisms exposed to peak and tail concentrations of antimicrobials were unchanged at the end of the study. Enrofloxacin and ceftiofur achieved intestinal concentrations capable of reducing intestinal bacteria, yet the short exposure of ceftiofur in the intestine may select for resistant organisms. © 2015 John Wiley & Sons Ltd.

Effectiveness of Persea major Kopp (Lauraceae) extract against Enterococcus faecalis: a preliminary in vitro study.

PubMed

Volpato, Lusiane; Gabardo, Marilisa Carneiro Leão; Leonardi, Denise Piotto; Tomazinho, Paulo Henrique; Maranho, Leila Teresinha; Baratto-Filho, Flares

2017-03-06

Persea major Kopp (Lauraceae) is a plant with wound healing, antibacterial, and analgesic properties. The aim of this study was to assess the in vitro antibacterial activity of the concentrated crude extract (CCE) and ethyl acetate fraction (EAF) of this plant against Enterococcus faecalis and compare it with calcium hydroxide [Ca(OH) 2 ] paste and 2% chlorhexidine digluconate (CHX). The plant material was collected, and an extract was prepared according to the requirements of the study (CCE and EAF). The minimum inhibitory concentrations (MICs) of CCE, EAF, Ca(OH) 2 , Ca(OH) 2  + CCE, and CHX against E. faecalis were determined using the broth microdilution method RESULTS: The EAF inhibited E. faecalis at concentrations of 166.50, 83.25, and 41.62 mg mL -1 , and 1.00, 0.50, and 0.25% of CHX solutions showed antimicrobial activity. The MICs of Ca(OH) 2 paste were 166.50 and 83.25 mg mL -1 , whereas Ca(OH) 2  + CCE showed antimicrobial activity only at a concentration of 166.50 mg mL -1 . CCE showed no inhibitory effect at any of the concentrations tested CONCLUSIONS: The CCE did not show any antimicrobial activity against E. faecalis; however, the EAF was the most effective among the three highest concentrations tested.

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.

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

PubMed

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

1996-06-01

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.

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

Characteristics of a Bacteriocin Derived from Streptococcus faecalis var. zymogenes Antagonistic to Diplococcus peumoniae

PubMed Central

Bottone, Edward; Allerhand, Jona; Pisano, Michael A.

1971-01-01

A bacteriocin-producing strain of Streptococcus faecalis var. zymogenes (E-1) was isolated from clinical material (conjunctiva). The active substance differed from bacteriocins described by other investigators primarily in its spectrum of antibacterial activity, especially by its marked inhibition of Diplococcus pneumoniae. The E-1 bacteriocin also inhibited nonhemolytic strains of enterococci as well as one-third of the Viridans group of streptococcal strains investigated. The degree of inhibition, however, as indicated by the size of the zones against the latter organisms, was significantly reduced. No activity was detected against any of the strains belonging to the following groups of bacteria: hemolytic enterococci, beta-hemolytic streptococci, nonhemolytic streptococci, staphylococci, and various gram-negative species. Similarly, three strains each of Bacillus cereus and Listeria monocytogenes and one strain of Erysipelothrix insidiosa were not inhibited. The bacteriocin was able to diffuse through bacterial membranes as well as cellulose dialyzer tubing. It was inactivated by heating to 80 C for 20 min but resisted inactivation by either trypsin or chloroform. Images PMID:4398532

Antibiotic resistance in bacteria isolated from vegetables with regards to the marketing stage (farm vs. supermarket).

PubMed

Schwaiger, Karin; Helmke, Katharina; Hölzel, Christina Susanne; Bauer, Johann

2011-08-15

The aim of this study was to elucidate whether and to what extent fresh produce from Germany plays a role as a carrier and reservoir of antibiotic resistant bacteria. For this purpose, 1001 vegetables (fruit, root, bulbous vegetables, salads and cereals) were collected from 13 farms and 11 supermarkets in Germany and examined bacteriologically. Phenotypic resistance of Enterobacter cloacae (n=172); Enterobacter gergoviae (n=92); Pantoea agglomerans (n=96); Pseudomonas aeruginosa (n=295); Pseudomonas putida (n=106) and Enterococcus faecalis (n=100) against up to 30 antibiotics was determined by using the microdilution method. Resistance to ß-lactams was most frequently expressed by P. agglomerans and E. gergoviae against cefaclor (41% and 29%). Relatively high resistance rates were also observed for doxycycline (23%), erythromycin (21%) and rifampicin (65%) in E. faecalis, for spectinomycin (28%) and mezlocillin (12%) in E. cloacae, as well as for streptomycin (19%) in P. putida. In P. aeruginosa, relatively low resistance rates were observed for the aminoglycosides amikacin, apramicin, gentamicin, neomycin, netilmicin and tobramycin (<4%); 11% was resistant to streptomycin. No glycopeptide-resistant enterococci were observed. Resistance rates of bacteria isolated from farm samples were higher than those of the retail markets whenever significant differences were observed. This suggests that expressing resistance is at the expense of bacterial viability, since vegetables purchased directly at the farm are probably fresher than at the supermarket, and they have not been exposed to stress factors. However, this should not keep the customer from buying directly at the farm, since the overall resistance rates were not higher than observed in bacteria from human or animal origin. Instead, peeling or washing vegetables before eating them raw is highly recommended, since it reduces not only the risk of contact with pathogens, but also that of ingesting and spreading

Isolation and characterization of potential lactic acid bacteria (LAB) from freshwater fish processing wastes for application in fermentative utilisation of fish processing waste

PubMed Central

R, Jini; HC, Swapna; Rai, Amit Kumar; R, Vrinda; PM, Halami; NM, Sachindra; N, Bhaskar

2011-01-01

Proteolytic and/or lipolytic lactic acid bacteria (LAB) were isolated from visceral wastes of different fresh water fishes. LAB count was found to be highest in case of visceral wastes of Mrigal (5.88 log cfu/g) and lowest in that of tilapia (4.22 log cfu/g). Morphological, biochemical and molecular characterization of the selected LAB isolates were carried out. Two isolates FJ1 (E. faecalis NCIM5367) and LP3 (P. acidilactici NCIM5368) showed both proteolytic and lipolytic properties. All the six native isolates selected for characterization showed antagonistic properties against several human pathogens. All the native isolates were sensitive to antibiotics cephalothin and clindamycin; and, resistant to cotrimoxazole and vancomycin. Considering individually, P. acidilactici FM37, P. acidilactici MW2 and E. faecalis FD3 were sensitive to erythromycin. The two strains FJ1 (E. faecalis NCIM 5367) and LP3 (P. acidilactici NCIM 5368) that had both proteolytic and lipolytic properties have the potential for application in fermentative recovery of lipids and proteins from fish processing wastes. PMID:24031786

A Novel Biomimetic Nanosponge Protects the Retina from the Enterococcus faecalis Cytolysin.

PubMed

LaGrow, Austin L; Coburn, Phillip S; Miller, Frederick C; Land, Craig; Parkunan, Salai Madhumathi; Luk, Brian T; Gao, Weiwei; Zhang, Liangfang; Callegan, Michelle C

2017-01-01

Intraocular infections are a potentially blinding complication of common ocular surgeries and traumatic eye injuries. Bacterial toxins synthesized in the eye can damage intraocular tissue, often resulting in poor visual outcomes. Enteroccocus faecalis causes blinding infections and is responsible for 8 to 17% of postoperative endophthalmitis cases. These infections are increasingly difficult to treat due to the emergence of multidrug-resistant strains. Virulent E. faecalis isolates secrete a pore-forming bicomponent cytolysin that contributes to retinal tissue damage during endophthalmitis. We hypothesized that a biomimetic nanosponge, which mimics erythrocytes, might adsorb subunits of the cytolysin and reduce retinal damage, protecting vision. To test the efficacy of nanosponges in neutralizing the cytolysin in vitro , hemoglobin release assays were performed on culture supernatants from cytolysin-producing E. faecalis with and without preincubation with nanosponges. Treatment with nanosponges for 30 min reduced hemolytic activity by ~70%. To determine whether nanosponges could neutralize the cytolysin in vivo , electroretinography was performed on mice 24 h after intravitreal injection with cytolysin-containing supernatants treated with nanosponges. Pretreatment of cytolysin-containing supernatants with nanosponges increased the A-wave retention from 12.2% to 65.5% and increased the B-wave retention from 21.0% to 77.0%. Histology revealed that in nanosponge-treated eyes, retinas remained intact and attached, with little to no damage. Rabbit nanosponges were also nontoxic and noninflammatory when injected into mouse eyes. In an experimental murine model of E. faecalis endophthalmitis, injection of nanosponges into the vitreous 6 h after infection with a wild-type cytolysin-producing strain increased A-wave retention from 5.9% to 31% and increased B-wave retention from 12.6% to 27.8%. Together, these results demonstrated that biomimetic nanosponges

A Novel Biomimetic Nanosponge Protects the Retina from the Enterococcus faecalis Cytolysin

PubMed Central

LaGrow, Austin L.; Coburn, Phillip S.; Miller, Frederick C.; Land, Craig; Parkunan, Salai Madhumathi; Luk, Brian T.; Gao, Weiwei; Zhang, Liangfang

2017-01-01

ABSTRACT Intraocular infections are a potentially blinding complication of common ocular surgeries and traumatic eye injuries. Bacterial toxins synthesized in the eye can damage intraocular tissue, often resulting in poor visual outcomes. Enteroccocus faecalis causes blinding infections and is responsible for 8 to 17% of postoperative endophthalmitis cases. These infections are increasingly difficult to treat due to the emergence of multidrug-resistant strains. Virulent E. faecalis isolates secrete a pore-forming bicomponent cytolysin that contributes to retinal tissue damage during endophthalmitis. We hypothesized that a biomimetic nanosponge, which mimics erythrocytes, might adsorb subunits of the cytolysin and reduce retinal damage, protecting vision. To test the efficacy of nanosponges in neutralizing the cytolysin in vitro, hemoglobin release assays were performed on culture supernatants from cytolysin-producing E. faecalis with and without preincubation with nanosponges. Treatment with nanosponges for 30 min reduced hemolytic activity by ~70%. To determine whether nanosponges could neutralize the cytolysin in vivo, electroretinography was performed on mice 24 h after intravitreal injection with cytolysin-containing supernatants treated with nanosponges. Pretreatment of cytolysin-containing supernatants with nanosponges increased the A-wave retention from 12.2% to 65.5% and increased the B-wave retention from 21.0% to 77.0%. Histology revealed that in nanosponge-treated eyes, retinas remained intact and attached, with little to no damage. Rabbit nanosponges were also nontoxic and noninflammatory when injected into mouse eyes. In an experimental murine model of E. faecalis endophthalmitis, injection of nanosponges into the vitreous 6 h after infection with a wild-type cytolysin-producing strain increased A-wave retention from 5.9% to 31% and increased B-wave retention from 12.6% to 27.8%. Together, these results demonstrated that biomimetic nanosponges

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. © 2012 Eur J Oral Sci.

Molecular Basis for Lytic Bacteriophage Resistance in Enterococci.

PubMed

Duerkop, Breck A; Huo, Wenwen; Bhardwaj, Pooja; Palmer, Kelli L; Hooper, Lora V

2016-08-30

The human intestine harbors diverse communities of bacteria and bacteriophages. Given the specificity of phages for their bacterial hosts, there is growing interest in using phage therapies to combat the rising incidence of multidrug-resistant bacterial infections. A significant barrier to such therapies is the rapid development of phage-resistant bacteria, highlighting the need to understand how bacteria acquire phage resistance in vivo Here we identify novel lytic phages in municipal raw sewage that kill Enterococcus faecalis, a Gram-positive opportunistic pathogen that resides in the human intestine. We show that phage infection of E. faecalis requires a predicted integral membrane protein that we have named PIPEF (for phage infection protein from E. faecalis). We find that PIPEF is conserved in E. faecalis and harbors a 160-amino-acid hypervariable region that determines phage tropism for distinct enterococcal strains. Finally, we use a gnotobiotic mouse model of in vivo phage predation to show that the sewage phages temporarily reduce E. faecalis colonization of the intestine but that E. faecalis acquires phage resistance through mutations in PIPEF Our findings define the molecular basis for an evolutionary arms race between E. faecalis and the lytic phages that prey on them. They also suggest approaches for engineering E. faecalis phages that have altered host specificity and that can subvert phage resistance in the host bacteria. Bacteriophage therapy has received renewed attention as a potential solution to the rise in antibiotic-resistant bacterial infections. However, bacteria can acquire phage resistance, posing a major barrier to phage therapy. To overcome this problem, it is necessary to understand phage resistance mechanisms in bacteria. We have unraveled one such resistance mechanism in Enterococcus faecalis, a Gram-positive natural resident of the human intestine that has acquired antibiotic resistance and can cause opportunistic infections

D-amino acids reduce Enterococcus faecalis biofilms in vitro and in the presence of antimicrobials used for root canal treatment.

PubMed

Zilm, Peter S; Butnejski, Victor; Rossi-Fedele, Giampiero; Kidd, Stephen P; Edwards, Suzanne; Vasilev, Krasimir

2017-01-01

Enterococcus faecalis is the most frequent species present in post-treatment disease and plays a significant role in persistent periapical infections following root canal treatment. Its ability to persist in stressful environments is inter alia, due to its ability to form biofilms. The presence of certain D-amino acids (DAAs) has previously been shown to reduce formation of Bacillus subtilis biofilms. The aims of this investigation were to determine if DAAs disrupt biofilms in early and late growth stages for clinical E. faecalis strains and to test their efficacy in disrupting E. faecalis biofilms grown in sub-minimum inhibitory concentrations of commonly used endodontic biocides. From thirty-seven E. faecalis strains, the ten "best" biofilm producers were used to test the ability of a mixture containing D-leucine, D-methionine, D-tyrosine and D-tryptophan to reduce biofilm growth over a period of 24, 72 and 144 hours and when compared to their cognate L-Amino Acids (LAAs). We have previously shown that sub-MIC levels of tetracycline and sodium hypochlorite promotes biofilm growth in clinical strains of E. faecalis. DAAs were therefore tested for their effectiveness to reduce biofilm growth in the presence of sub-minimal concentrations of sodium hypochlorite (NaOCl-0.031%) and Odontocide™ (0.25% w/v), and in the presence of Odontopaste™ (0.25% w/v). DAAs significantly reduced biofilm formation for all strains tested in vitro, while DAAs significantly reduced biofilm formation compared to LAAs. The inhibitory effect of DAAs on biofilm formation was concentration dependent. DAAs were also shown to be effective in reducing E. faecalis biofilms in the presence of Odontopaste™ and sub-MIC levels of NaOCl and Odontocide™. The results suggest that the inclusion of DAAs into current endodontic procedures may reduce E. faecalis biofilms.

The acid tolerance response and pH adaptation of Enterococcus faecalis in extract of lime Citrus aurantiifolia from Aceh Indonesia.

PubMed

Mubarak, Zaki; Soraya, Cut

2018-01-01

Background:  The objective of the present study was to evaluate the acid tolerance response and pH adaptation when Enterococcus faecalis interacted with extract of lime ( Citrus aurant iifolia ). Methods : We used E. faecalis ATCC 29212 and lime extract from Aceh, Indonesia. The microbe was analyzed for its pH adaptation, acid tolerance response, and adhesion assay using a light microscope with a magnification of x1000. Further, statistical tests were performed to analyze both correlation and significance of the acid tolerance and pH adaptation as well as the interaction activity. Results : E. faecalis was able to adapt to a very acidic environment (pH 2.9), which was characterized by an increase in its pH (reaching 4.2) at all concentrations of the lime extract (p < 0.05). E. faecalis was also able to provide acid tolerance response to lime extract based on spectrophotometric data (595 nm) (p < 0.05). Also, the interaction activity of E. faecalis in different concentrations of lime extract was relatively stable within 6 up to 12 hours (p < 0.05), but it became unstable within 24-72 hours (p > 0.05) based on the mass profiles of its interaction activity. Conclusions : E. faecalis can adapt to acidic environments (pH 2.9-4.2); it is also able to tolerate acid generated by Citrus auranti ifolia extract, revealing a stable interaction in the first 6-12 hours.

Evaluation of the antibacterial potential of Petroselinum crispum and Rosmarinus officinalis against bacteria that cause urinary tract infections

PubMed Central

Petrolini, Fernanda Villas Boas; Lucarini, Rodrigo; de Souza, Maria Gorete Mendes; Pires, Regina Helena; Cunha, Wilson Roberto; Martins, Carlos Henrique Gomes

2013-01-01

In this study we evaluated the antibacterial activity of the crude hydroalcoholic extracts, fractions, and compounds of two plant species, namely Rosmarinus officinalis and Petroselinum crispum, against the bacteria that cause urinary tract infection. The microdilution method was used for determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The crude hydroalcoholic extract of R. officinalis displayed in vitro activity against Gram-positive bacteria, with satisfactory MBC for the clinical isolate S. saprophyticus. The fractions and the pure compound rosmarinic acid did not furnish promising results for Gram-negative bacteria, whereas fractions 2, 3, and 4 gave encouraging results for Gram-positive bacteria and acted as bactericide against S. epidermidis as well as E. faecalis (ATCC 29212) and its clinical isolate. R. officinalis led to promising results in the case of Gram-positive bacteria, resulting in a considerable interest in the development of reliable alternatives for the treatment of urinary infections. PMID:24516424

Evaluation of the antibacterial potential of Petroselinum crispum and Rosmarinus officinalis against bacteria that cause urinary tract infections.

PubMed

Petrolini, Fernanda Villas Boas; Lucarini, Rodrigo; de Souza, Maria Gorete Mendes; Pires, Regina Helena; Cunha, Wilson Roberto; Martins, Carlos Henrique Gomes

2013-01-01

In this study we evaluated the antibacterial activity of the crude hydroalcoholic extracts, fractions, and compounds of two plant species, namely Rosmarinus officinalis and Petroselinum crispum, against the bacteria that cause urinary tract infection. The microdilution method was used for determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The crude hydroalcoholic extract of R. officinalis displayed in vitro activity against Gram-positive bacteria, with satisfactory MBC for the clinical isolate S. saprophyticus. The fractions and the pure compound rosmarinic acid did not furnish promising results for Gram-negative bacteria, whereas fractions 2, 3, and 4 gave encouraging results for Gram-positive bacteria and acted as bactericide against S. epidermidis as well as E. faecalis (ATCC 29212) and its clinical isolate. R. officinalis led to promising results in the case of Gram-positive bacteria, resulting in a considerable interest in the development of reliable alternatives for the treatment of urinary infections.

Occurrence and Distribution of Proteinase of Streptococcus faecalis var. liquefaciens1

PubMed Central

Shugart, Lee R.; Beck, Raymond W.

1966-01-01

Shugart, Lee R. (University of Tennessee, Knoxville), and Raymond W. Beck. Occurrence and distribution of proteinase of Streptococcus faecalis var. liquefaciens. J. Bacteriol. 92:338–341. 1966.—The proteolytic enzyme produced by Streptococcus faecalis var. liquefaciens (ATCC 13398) was shown to be an exoenzyme. The production of the proteinase was followed in growing cultures, and its distribution was compared with that of the intracellular enzymes reduced nicotinamide adenine dinucleotide (NADH2) peroxidase and lactate dehydrogenase. The proteinase appeared in the culture medium prior to the stationary phase of growth, whereas the other enzymes could be found only in whole cells. Fractionation of whole cells by sonic treatment and by treatment with lysozyme showed the proteinase to be associated primarily with the cell wall and cell membrane, and NADH2 peroxidase to be associated only with the cytoplasmic fractions. PMID:16562116

Synergistic mechanism of Ag+-Zn2+ in anti-bacterial activity against Enterococcus faecalis and its application against dentin infection.

PubMed

Fan, Wei; Sun, Qing; Li, Yanyun; Tay, Franklin R; Fan, Bing

2018-01-31

Ag + and Zn 2+ have already been used in combinations to obtain both enhanced antibacterial effect and low cytotoxicity. Despite this, it is still unclear how the Zn 2+ co-works with Ag + in the synergistic antibacterial activity. The main purposes of this study were to investigate the co-work pattern and optimum ratio between Ag + and Zn 2+ in their synergistic antibacterial activity against E. faecalis, the possible mechanisms behind this synergy and the primary application of optimum Ag + -Zn 2+ co-work pattern against the E. faecalis biofilm on dentin. A serial of Ag + -Zn 2+ atomic combination ratios were tested on both planktonic and biofilm-resident E. faecalis on dentin, their antibacterial efficiency was calculated and optimum ratio determined. And the cytotoxicity of various Ag + -Zn 2+ atomic ratios was tested on MC3T3-E1 Cells. The role of Zn 2+ in Ag + -Zn 2+ co-work was evaluated using a Zn 2+ pretreatment study and membrane potential-permeability measurement. The results showed that the synergistically promoted antibacterial effect of Ag + -Zn 2+ combinations was Zn 2+ amount-dependent with the 1:9 and 1:12 Ag + -Zn 2+ atomic ratios showing the most powerful ability against both planktonic and biofilm-resident E. faecalis. This co-work could likely be attributed to the depolarization of E. faecalis cell membrane by the addition of Zn 2+ . The cytotoxicity of the Ag + -Zn 2+ atomic ratios of 1:9 and 1:12 was much lower than 2% chlorhexidine. The Ag + -Zn 2+ atomic ratios of 1:9 and 1:12 demonstrated similar strong ability against E. faecalis biofilm on dentin but much lower cytotoxicity than 2% chlorhexidine. New medications containing optimum Ag + -Zn 2+ atomic ratios higher than 1:6, such as 1:9 or 1:12, could be developed against E. faecalis infection in root canals of teeth or any other parts of human body.

Antibacterial effect of calcium hydroxide combined with chlorhexidine on Enterococcus faecalis: a systematic review and meta-analysis

PubMed Central

SAATCHI, Masoud; SHOKRANEH, Ali; NAVAEI, Hooman; MARACY, Mohammad Reza; SHOJAEI, Hasan

2014-01-01

Objective Enterococcus faecalis (E. faecalis) is the most frequently isolated strain in failed endodontic therapy cases since it is resistant to calcium hydroxide (CH). Whether a combination of CH and chlorhexidine (CHX) is more effective than CH alone against E. faecalis is a matter of controversy. Thus, the aim of this study was to conduct a systematic review and meta-analysis of the literature. Material and Methods A comprehensive search in PubMed, EMbase, EBSCOhost, The Cochrane Library, SciELO, and BBO databases, Clinical trials registers, Open Grey, and conference proceedings from the earliest available date to February 1, 2013 was carried out and the relevant articles were identified by two independent reviewers. Backward and forward search was performed and then inclusion and exclusion criteria were applied. The included studies were divided into "comparisons" according to the depth of sampling and dressing period of each medicament. Meta-analysis was performed using Stata software 10.0. The level of significance was set at 0.05. Results Eighty-five studies were retrieved from databases and backward/forward searches. Fortyfive studies were considered as relevant (5 in vivo, 18 in vitro, 18 ex vivo, and 4 review articles). Nine studies were included for meta-analysis. Inter-observer agreement (Cohen kappa) was 0.93. The included studies were divided into 21 comparisons for meta-analysis. Chi-square test showed the comparisons were heterogeneous (p<0.001). Random effect model demonstrated no significant difference between CH/CHX mixture and CH alone in their effect on E. faecalis (p=0.115). Conclusions According to the evidence available now, mixing CH with CHX does not significantly increase the antimicrobial activity of CH against E. faecalis. It appears that mixing CH with CHX does not improve its ex vivo antibacterial property as an intracanal medicament against E. faecalis. Further in vivo studies are necessary to confirm and correlate the findings of

Is the mazEF toxin-antitoxin system responsible for vancomycin resistance in clinical isolates of Enterococcus faecalis?

PubMed

Sadeghifard, Nourkhoda; Soheili, Sara; Sekawi, Zamberi; Ghafourian, Sobhan

2014-01-01

The current study was conducted to investigate the relationship between vancomycin-resistant Enterococcus faecalis (VRE) and the presence of mazEF toxin-antitoxin (TA) system, which may be useful as target for novel antimicrobial therapy concepts. The susceptibility of E. faecalis was determined by MIC, and the presence of the mazEF TA system was evaluated by PCR. Among 200 E. faecalis isolates 39.5% showed resistance to vancomycin (VRE), while 60.5% were susceptible strains (VSE). The mazEF TA system was positive in all VRE isolates (100%), but less prevalent (38/121, 31.4%) among the 121 VSE strains. In conclusion, our study demonstrated a positive relationship between the presence of vancomycin resistance and mazEF TA system. This observation may introduce therapeutic options against a novel antimicrobial target in enterococci.

Is the mazEF toxin-antitoxin system responsible for vancomycin resistance in clinical isolates of Enterococcus faecalis?

PubMed Central

Sadeghifard, Nourkhoda; Soheili, Sara; Sekawi, Zamberi; Ghafourian, Sobhan

2014-01-01

The current study was conducted to investigate the relationship between vancomycin-resistant Enterococcus faecalis (VRE) and the presence of mazEF toxin-antitoxin (TA) system, which may be useful as target for novel antimicrobial therapy concepts. The susceptibility of E. faecalis was determined by MIC, and the presence of the mazEF TA system was evaluated by PCR. Among 200 E. faecalis isolates 39.5% showed resistance to vancomycin (VRE), while 60.5% were susceptible strains (VSE). The mazEF TA system was positive in all VRE isolates (100%), but less prevalent (38/121, 31.4%) among the 121 VSE strains. In conclusion, our study demonstrated a positive relationship between the presence of vancomycin resistance and mazEF TA system. This observation may introduce therapeutic options against a novel antimicrobial target in enterococci. PMID:24653969

Effect of photodynamic therapy (PDT) on Enterococcus faecalis biofilm in experimental primary and secondary endodontic infections.

PubMed

Tennert, Christian; Feldmann, Katharina; Haamann, Edwina; Al-Ahmad, Ali; Follo, Marie; Wrbas, Karl-Thomas; Hellwig, Elmar; Altenburger, Markus J

2014-11-04

To determine the antibacterial effect of photodynamic Therapy on Enterococcus faecalis (E. faecalis) biofilms in experimentally infected human root canals in primary infections and endodontic retreatments. One hundred and sixty single-rooted extracted teeth with one root canal were prepared using ProTaper instruments. Seventy specimens were left without root canal filling and autoclaved. The root canals of another 70 specimens were filled with Thermafil and AH Plus and the root canal fillings were removed after 24 hours using ProTaper D files and plasma sterilized. The specimens were infected with a clinical isolate of E. faecalis for 72 hours. Samples were taken using sterile paper points to determine the presence of E. faecalis in the root canals. The specimens were randomly divided into groups according to their treatment with 20 teeth each and a control. In the PDT group the teeth were treated using PDT, consisting of the photosensitizer toluidine blue and the PDT light source at 635 nm. In the NaOCl (sodium hypochlorite) group the root canals were rinsed with 10 mL of 3% NaOCl. In the NaOCl-PDT group the root canals were rinsed with 10 mL of 3% of sodium hypochlorite and then treated with PDT. Samples were taken after treatments using sterile paper points. Additionally, remaining root canal filling material was recovered from the root canal walls. Survival fractions of the samples were calculated by counting colony-forming units. A one-way analysis of variance (ANOVA) was applied to the data to assess the effect of different treatment techniques. Antimicrobial treatment of root canals caused a significant reduction of bacterial load in all groups. NaOCl irrigation eliminated E. faecalis most effectively. PDT alone was less effective compared to NaOCl irrigation and the combination of NaOCl irrigation and PDT. CFU levels recovered from the filling material after NaOCl irrigation of the root canals were 10fold higher compared to PDT and the combination of Na

Identification and catabolic activity of well-derived gasoline-degrading bacteria from a contaminated aquifer.

PubMed Central

Ridgway, H F; Safarik, J; Phipps, D; Carl, P; Clark, D

1990-01-01

Approximately 300 gasoline-degrading bacteria were isolated from well water and core material from a shallow coastal aquifer contaminated with unleaded gasoline. Identification of 244 isolates revealed four genera: Pseudomonas, Alcaligenes, Nocardia, and Micrococcus, with pseudomonads making up 86.9% of bacteria identified. A total of 297 isolates was sorted into 111 catabolic groups on the basis of aerobic growth responses on 15 gasoline hydrocarbons. Each test hydrocarbon was degraded by at least one isolate. Toluene, p-xylene, ethylbenzene, and 1,2,4-trimethylbenzene were most frequently utilized as growth substrates, whereas cyclic and branched alkanes were least utilized. Most isolates were able to grow on 2 or 3 different hydrocarbons, and nearly 75% utilized toluene as a sole source of carbon and energy. Isolates were remarkably specific for hydrocarbon usage, often catabolizing only one of several closely related compounds. A subset of 220 isolates was sorted into 51 groups by polyacrylamide gel electrophoresis. Pseudomonas aeruginosa was partitioned into 16 protein-banding groups (i.e., subspecies) whose catabolic activities were largely restricted to substituted aromatics. Different members of subspecies groups defined by protein-banding pattern analysis often exhibited different growth responses on the same hydrocarbon, implying marked strain diversity. The catabolic activities of well-derived, gasoline-degrading bacteria associated with this contaminated aquifer are consonant with in situ adaptation at the site. PMID:2268163

Identification and catabolic activity of well-derived gasoline-degrading bacteria from a contaminated aquifer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ridgway, H.F.; Safarik, J.; Phipps, D.

1990-11-01

Approximately 300 gasoline-degrading bacteria were isolated from well water and core material from a shallow coastal aquifer contaminated with unleaded gasoline. Identification of 244 isolates revealed four genera: Pseudomonas, Alcaligenes, Nocardia, and Micrococcus, with pseudomonads making up 86.9% of bacteria identified. A total of 297 isolates was sorted into 111 catabolic groups on the basis of aerobic growth responses on 15 gasoline hydrocarbons. Each test hydrocarbon was degraded by at least one isolate. Toluene, p-xylene, ethylbenzene, and 1,2,4-trimethylbenzene were most frequently utilized as growth substrates, whereas cyclic and branched alkanes were least utilized. Most isolates were able to grow onmore » 2 or 3 different hydrocarbons, and nearly 75% utilized toluene as a sole source of carbon and energy. Isolates were remarkably specific for hydrocarbon usage, often catabolizing only one of several closely related compounds. A subset of 220 isolates was sorted into 51 groups by polyacrylamide gel electrophoresis. Pseudomonas aeruginosa was partitioned into 16 protein-banding groups (i.e., subspecies) whose catabolic activities were largely restricted to substituted aromatics. Different members of subspecies groups defined by protein-banding pattern analysis often exhibited different growth responses on the same hydrocarbon, implying marked strain diversity. The catabolic activities of well-derived, gasoline-degrading bacteria associated with this contaminated aquifer are consonant with in situ adaptation at the site.« less

Antimicrobial activity of essential oil from Schinus molle Linn.

PubMed

Gundidza, M

1993-11-01

The essential oil from the fresh leaves of Schinus molle isolated by hydrodistillation was tested for antibacterial activity using the hole plate diffusion method and for antifungal activity using the mycelium or single cell growth inhibition method. Results obtained showed that the volatile oil exhibited significant activity against the following bacterial species: Klebsiella pneumoniae, Alcaligenes faecalis, Pseudomonas aeruginosa, Leuconostoc cremoris, Enterobacter aerogenes, Proteus vulgaris, Clostridium sporogenes, Acinetobacter calcoacetica, Escherichia coli, Beneckea natriegens, Citrobacter freundii, Serratia marcescens, Bacillus subtilis and Brochothrix thermosphacata. The fungal species Aspergillus ochraceus, Aspergillus parasiticus, Fusarium culmorum and Alternaria alternata exhibited significant sensitivity to the volatile oil.

Combined effect of a mixture of tetracycline, acid, and detergent, and Nisin against Enterococcus faecalis and Actinomyces viscosus biofilms

PubMed Central

Balto, Hanan A.; Shakoor, Zahid A.; Kanfar, Maha A.

2015-01-01

Objectives: To evaluate the combined effect of a mixture of tetracycline, acid, and detergent (MTAD) and Nisin against Enterococcus faecalis (E. faecalis) and Actinomyces viscosus (A. viscosus) biofilms. Methods: This study was conducted between June and December 2013 in collaboration with Dental Caries Research Chair, College of Dentistry, King Saud University, Riyadh, Saudi Arabia. Single-species biofilms (n=9/species/observation period) were generated on membrane filter discs and subjected to 5, 10, or 15 minute incubation with MTADN (MTAD with 3% Nisin), 5.25% sodium hypochlorite (NaOCl), or normal saline. The colony forming units were counted using the Dark field colony counter. Results: A 100% bactericidal effect of 5.25% NaOCl was noted during the 3 observation periods; a significant reduction (p=0.000) in mean survival rates of E. faecalis (77.3+13.6) and A. viscosus (39.6+12.6) was noted after 5 minutes exposure to MTADN compared with normal saline (78000000+5291503) declining to almost no growth after 10 and 15 minutes. The survival rates of the E. faecalis and A. viscosus biofilm were no different after treatment with MTADN and 5.25% NaOCl at the 3 observation periods (p=1.000). Conclusion: A combination of MTAD and Nisin was as effective as NaOCl against E. faecalis and A. viscosus biofilms. PMID:25719587

Paenalcaligenes suwonensis sp. nov., isolated from spent mushroom compost.

PubMed

Moon, Ji-Young; Lim, Jun-Muk; Ahn, Jae-Hyung; Weon, Hang-Yeon; Kwon, Soon-Wo; Kim, Soo-Jin

2014-03-01

A bacterial strain, ABC02-12(T), was isolated from spent mushroom compost, a waste product of button mushroom cultivation. Cells of the strain were Gram-stain-negative, catalase- and oxidase-positive, non-spore-forming, aerobic flagellated rods. Optimum growth occurred at 28 °C and pH 7.0. 16S rRNA gene sequence analysis showed that strain ABC02-12(T) shared the highest sequence similarities with Paenalcaligenes hominis CCUG 53761A(T) (96.0 %), Alcaligenes faecalis subsp. parafaecalis G(T) (95.7 %), Alcaligenes faecalis subsp. faecalis IAM 12369(T) (95.4 %) and Pusillimonas noertemannii BN9(T) (95.3 %). According to the phylogenetic tree, strain ABC02-12(T) formed a robust cluster with Paenalcaligenes hominis CCUG 53761A(T) and Paenalcaligenes hermetiae KBL009(T). The quinone system was ubiquinone Q-8 with minor amounts of Q-7. The major fatty acids (>5 % of total fatty acids) were C16 : 0, C16 : 1ω6c and/or C16 : 1ω7c (summed feature 3), C18 : 1ω7c and/or C18 : 1ω6c (summed feature 8), C17 : 0 cyclo, and iso-C16 : 1 I, C14 : 0 3-OH and/or an unknown fatty acid (summed feature 2). The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unknown aminolipid. Putrescine was the principal polyamine, with small amounts of 2-hydroxyputrescine and cadaverine. On the basis of the evidence presented in this study, strain ABC02-12(T) is a representative of a novel species within the genus Paenalcaligenes, for which the name Paenalcaligenes suwonensis sp. nov. is proposed. The type strain is ABC02-12(T) ( = KACC 16537(T) = NBRC 108927(T)).

SNP diversity of Enterococcus faecalis and Enterococcus faecium in a South East Queensland waterway, Australia, and associated antibiotic resistance gene profiles

PubMed Central

2011-01-01

Background Enterococcus faecalis and Enterococcus faecium are associated with faecal pollution of water, linked to swimmer-associated gastroenteritis and demonstrate a wide range of antibiotic resistance. The Coomera River is a main water source for the Pimpama-Coomera watershed and is located in South East Queensland, Australia, which is used intensively for agriculture and recreational purposes. This study investigated the diversity of E. faecalis and E. faecium using Single Nucleotide Polymorphisms (SNPs) and associated antibiotic resistance profiles. Results Total enterococcal counts (cfu/ml) for three/six sampling sites were above the United States Environmental Protection Agency (USEPA) recommended level during rainfall periods and fall into categories B and C of the Australian National Health and Medical Research Council (NHMRC) guidelines (with a 1-10% gastrointestinal illness risk). E. faecalis and E. faecium isolates were grouped into 29 and 23 SNP profiles (validated by MLST analysis) respectively. This study showed the high diversity of E. faecalis and E. faecium over a period of two years and both human-related and human-specific SNP profiles were identified. 81.8% of E. faecalis and 70.21% of E. faecium SNP profiles were associated with genotypic and phenotypic antibiotic resistance. Gentamicin resistance was higher in E. faecalis (47% resistant) and harboured the aac(6')-aph(2') gene. Ciprofloxacin resistance was more common in E. faecium (12.7% resistant) and gyrA gene mutations were detected in these isolates. Tetracycline resistance was less common in both species while tet(L) and tet(M) genes were more prevalent. Ampicillin resistance was only found in E. faecium isolates with mutations in the pbp5 gene. Vancomycin resistance was not detected in any of the isolates. We found that antibiotic resistance profiles further sub-divided the SNP profiles of both E. faecalis and E. faecium. Conclusions The distribution of E. faecalis and E. faecium

Exploring different photodynamic therapy parameters to optimize elimination of Enterococcus faecalis in planktonic form.

PubMed

Soares, Janir Alves; Soares, Suelleng Maria Cunha Santos; de Jesus Tavarez, Rudys Rodolfo; de Castro Rizzi, Claudia; Vaz Rodrigues, Silvana Cristina Gama; Maia Filho, Etevaldo Matos; Brito-Júnior, Manoel; Pereira, Rodrigo Dantas; Magalhães, Paula Prazeres; de Macêdo Farias, Luiz

2018-06-01

The failure of endodontic treatment is linked to the presence of Enterococcus faecalis in the root canals. The scope of this study was to evaluate the influence of the energy dose and frequency of photodynamic therapy (PDT cycles), as well as the volume of bacterial suspensions (BS) in the elimination of Enterococcus faecalis in planktonic form. In four successive assays BS of Enterococcus faecalis ATCC 19433 were irradiated with a diode laser (40 mW) using the photosensitizer (PS) methylene blue (MB) (0.005 μg/mL). Group 1 - Effect of energy dose: 100 μ L of BS and 100 μ L of PS were irradiated by 1, 2.5, 5, 7.5 and 10 minute s. Group 2 - Effect of PDT cycles: The BS received 1, 2, 3 or 4 PDT cycles (in each cycle 100 m L of PS was added and irradiated by 2.5 minutes). Group 3 - Effect of energy dose and bacterial suspension volume: 10 μ L of BS and 10 μ L of PS were irradiated similar to group 1. Group 4 - Effect of energy dose, bacterial suspension volume and PDT cycles: 10 μ L of BS and 10 μL of PS were irradiated according to group 2. The laser source and MB isolated represented the controls. The mean log reduction after separate applying laser light and MB were 0.01 and 0.07, respectively. It was found that wells with 100 μ L of BS irradiated with 2.4 to 24 J of energy did not cause significant bacterial elimination (p > 0.05), on the other hand PDT cycles above 12 J increased significantly bacterial elimination (p < 0.05). In 10 μ L wells irradiation from 12 J of energy provided higher bacterial elimination (p < 0.05) which combined with PDT cycle resulted in the logarithmic elimination of E. faecalis (p < 0.05). The energy dose, the volume of the bacterial suspension and, especially, the PDT cycles optimized the bacterial elimination of Enterococcus faecalis in planktonic form. Copyright © 2018 Elsevier B.V. All rights reserved.

Comparative Analysis of Protaper and Waveone Systems to Reduce Enterococcus Faecalis from Root Canal System in Primary Molars--An in Vitro Study.

PubMed

Pinheiro, Sérgio Luiz; Pessoa, Carolina; da Silva, Josianne Neres; Gonçalves, Rafael Orro; Duarte, Danilo Antonio; da Silveira Bueno, Carlos Eduardo

2016-01-01

To assess, in vitro, the ability of the ProTaper(™) and WaveOne(™) systems to reduce Enterococcus faecalis contamination in primary molars. Sixty roots of primary molars were contaminated with E. faecalis. Roots were randomly allocated to one of four groups (n=20): ProTaper(™), WaveOne(™), control A, or control B. The files used were S1 and S2/F1 and F2 (ProTaper(™) system) and 25.08 (WaveOne(™) system). In control group A, the root canal was left uninstrumented, whereas in control group B, the root canal was irrigated with NaCl 0.9%. E. faecalis was sampled from the root canal system before and after instrumentation and the Wilcoxon test and Mann-Whitney U were used. There were no differences in E. faecalis counts between pre-instrumentation counts in the ProTaper™ and WaveOne(™) (p>0.05). The ProTaper(™) system led to an 89.36% reduction in E. faecalis burden, versus 78.10% with the WaveOne(™) system (p>0.05). Instrumentation time was shorter with WaveOne(™) (p<0.0001). The ProTaper(™) and WaveOne™ systems were equally effective in reducing Enterococcus faecalis in primary molars. The WaveOne(™) system was associated with shorter instrumentation time.

Cloning of the Alcaligenes latus polyhydroxyalkanoate biosynthesis genes and use of these genes for enhanced production of Poly(3-hydroxybutyrate) in Escherichia coli.

PubMed

Choi, J I; Lee, S Y; Han, K

1998-12-01

Polyhydroxyalkanoates (PHAs) are microbial polyesters that can be used as completely biodegradable polymers, but the high production cost prevents their use in a wide range of applications. Recombinant Escherichia coli strains harboring the Ralstonia eutropha PHA biosynthesis genes have been reported to have several advantages as PHA producers compared with wild-type PHA-producing bacteria. However, the PHA productivity (amount of PHA produced per unit volume per unit time) obtained with these recombinant E. coli strains has been lower than that obtained with the wild-type bacterium Alcaligenes latus. To endow the potentially superior PHA biosynthetic machinery to E. coli, we cloned the PHA biosynthesis genes from A. latus. The three PHA biosynthesis genes formed an operon with the order PHA synthase, beta-ketothiolase, and reductase genes and were constitutively expressed from the natural promoter in E. coli. Recombinant E. coli strains harboring the A. latus PHA biosynthesis genes accumulated poly(3-hydroxybutyrate) (PHB), a model PHA product, more efficiently than those harboring the R. eutropha genes. With a pH-stat fed-batch culture of recombinant E. coli harboring a stable plasmid containing the A. latus PHA biosynthesis genes, final cell and PHB concentrations of 194.1 and 141.6 g/liter, respectively, were obtained, resulting in a high productivity of 4.63 g of PHB/liter/h. This improvement should allow recombinant E. coli to be used for the production of PHB with a high level of economic competitiveness.

The Impact of a 940 nm Diode Laser with Radial Firing Tip and Bare End Fiber Tip on Enterococcus faecalis in the Root Canal Wall Dentin of Bovine Teeth: An In Vitro Study.

PubMed

Schulte-Lünzum, Ruth; Gutknecht, Norbert; Conrads, Georg; Franzen, Rene

2017-07-01

This in vitro study aimed to compare the bactericidal effect of two different laser delivery systems, a radial firing tip (RFT) and bare end fiber tip (BFT) used with the 940 nm diode laser on Enterococcus faecalis inoculated onto bovine radicular dentin. A total of 100 bovine dentin slices with a defined thickness of 500 and 1000 μm were prepared. They were assigned into four test groups together with untreated samples served as control for each slice thickness. The slices were inoculated on one side with 1 μL E. faecalis suspension and laser irradiation was performed indirectly on the opposite side with the 940 nm diode laser delivered with a 200 μm RFT and a BFT at 1 and 1.5 W in continuous wave mode for 8 sec per cycle and repeated four times. After irradiation, the remaining bacteria were detached and the produced suspension was diluted and plated onto blood agar plates with 5% sheep blood and incubated overnight at 37°C in a CO 2 -rich atmosphere. The colony-forming units of E. faecalis were counted and the bacterial reduction was analyzed. The diode laser equipped with RFT fiber design further reduced the number of vital E. faecalis cells significantly compared with BFT design, regardless of the used power and dentin thickness (p < 0.0001). The highest average value of 4 log kills was observed in 500 μm slice thickness irradiated with RFT at 1.5 W. Temperature measurements on the external root surface at 1 mm from the apex did not elicit a harmful temperature elevation in both power settings and fiber designs. Within the studied parameters, 940 nm diode laser in conjugation with RFT showed a satisfactory bactericidal effect without any thermal side effect to the tooth-supporting tissues.

Biphenyl-metabolizing bacteria in the rhizosphere of horseradish and bulk soil contaminated by polychlorinated biphenyls as revealed by stable isotope probing.

PubMed

Uhlik, Ondrej; Jecna, Katerina; Mackova, Martina; Vlcek, Cestmir; Hroudova, Miluse; Demnerova, Katerina; Paces, Vaclav; Macek, Tomas

2009-10-01

DNA-based stable isotope probing in combination with terminal restriction fragment length polymorphism was used in order to identify members of the microbial community that metabolize biphenyl in the rhizosphere of horseradish (Armoracia rusticana) cultivated in soil contaminated with polychlorinated biphenyls (PCBs) compared to members of the microbial community in initial, uncultivated bulk soil. On the basis of early and recurrent detection of their 16S rRNA genes in clone libraries constructed from [(13)C]DNA, Hydrogenophaga spp. appeared to dominate biphenyl catabolism in the horseradish rhizosphere soil, whereas Paenibacillus spp. were the predominant biphenyl-utilizing bacteria in the initial bulk soil. Other bacteria found to derive carbon from biphenyl in this nutrient-amended microcosm-based study belonged mostly to the class Betaproteobacteria and were identified as Achromobacter spp., Variovorax spp., Methylovorus spp., or Methylophilus spp. Some bacteria that were unclassified at the genus level were also detected, and these bacteria may be members of undescribed genera. The deduced amino acid sequences of the biphenyl dioxygenase alpha subunits (BphA) from bacteria that incorporated [(13)C]into DNA in 3-day incubations of the soils with [(13)C]biphenyl are almost identical to that of Pseudomonas alcaligenes B-357. This suggests that the spectrum of the PCB congeners that can be degraded by these enzymes may be similar to that of strain B-357. These results demonstrate that altering the soil environment can result in the participation of different bacteria in the metabolism of biphenyl.

Quantifying the extruded bacteria following use of two rotary instrumentation systems.

PubMed

Mohammadi, Zahed; Khademi, Abbasali

2007-01-01

All instrumentation techniques have been reported to be associated with extrusion of infected debris. The aim of this study was to evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using the two engine-driven techniques utilizing nickel-titanium instruments (Flex Master and Mtwo). Seventy extracted maxillary central incisor teeth were used. Access cavities were prepared and root canals were then contaminated with a suspension of Enterococcus faecalis and dried. The contaminated roots were divided into two experimental groups of 30 teeth each and one control group of 10 teeth. Group 1, Flex Master; Group2, Mtwo; Group 3, control group: no instrumentation was attempted. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 h. Colonies of bacteria were counted and the results were given as number of colony-forming units. The obtained data were analyzed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with α = 0.05 as the level for statistical significance. Findings showed that there was no significant difference as to the number of extruded bacteria between two engine-driven systems (P>0.05). Both engine-driven Nickel-Titanium systems extruded bacteria through the apical foramen.

Evaluation of Enterococcus faecalis clinical isolates with 'penicillin-resistant, ampicillin-susceptible' phenotype as reported by Vitek-2 Compact system.

PubMed

Tan, Yen Ee; Ng, Lily S Y; Tan, Thean Yen

2014-10-01

It has been recently reported that ampicillin susceptibility cannot accurately predict piperacillin and imipenem susceptibilities in penicillin-resistant, ampicillin-susceptible (Pen-R, Amp-S) Enterococcus faecalis isolates, contrary to the current Clinical and Laboratory Standards Institute (CLSI) recommendations. This has important therapeutic implications. Such isolates were noted after the use of Vitek-2 Compact system AST-GP67 susceptibility cards in a Singapore general hospital and they were increasing in numbers. The primary aim of this study was to evaluate these clinical isolates against microbroth dilution (MBD) technique and other commonly used antimicrobial susceptibility test (AST) methods for penicillin and ampicillin. The secondary aim was to evaluate whether ampicillin susceptibility could indeed be a reliable surrogate marker for piperacillin and imipenem susceptibilities in E. faecalis isolates that were confirmed Pen-R, Amp-S.From 2009 to 2013, a total of 49 isolates (5%) of 983 non-duplicate E. faecalis tested by Vitek-2 displayed the 'Pen-R, Amp-S' phenotype in a general hospital in Singapore. These were tested against MBD which was the reference method, Etest and disc diffusion for penicillin and ampicillin. Susceptibilities to piperacillin and imipenem were also tested using MBD. In addition, β-lactamase production test was performed. Forty E. faecalis isolates with penicillin-susceptible, ampicillin-susceptible (Pen-S, Amp-S) phenotype were included for comparative purposes.The categorical agreement rate was 100% for all AST methods in ampicillin reporting for the 'Pen-R, Amp-S' group of E. faecalis isolates. However, a large number of isolates (46 isolates, 93.9%) fell into the major error category for penicillin testing by the Vitek-2 system. Penicillin minimum inhibitory concentrations (MICs) generated by the Vitek-2 system for the majority of these isolates were two doubling dilutions higher compared to those obtained by the reference

In vitro evaluation of apical extrusion of bacteria following use of new rotary instrumentation system.

PubMed

Mohammadi, Zahed

2009-04-01

The aim of this study was to evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using two engine-driven nickel-titanium instruments (Flex Master and V-Taper). Seventy extracted maxillary central incisor teeth were used. After preparing access cavities, root canals were contaminated with a suspension of Enterococcus faecalis, then dried. The contaminated roots were divided into two experimental groups of 30 teeth each and one control group of 10 teeth. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 hours. Colonies of bacteria were counted, and the results were given as number of colony-forming units. The data obtained were analyzed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with alpha = 0.05 as the level for statistical significance. Results showed that there was no significant difference as to the number of extruded bacteria between the two engine-driven systems (P > 0.05). Both engine-driven nickel-titanium systems extruded bacteria through the apical foramen.

Proteomic Investigation of the Response of Enterococcus faecalis V583 when Cultivated in Urine.

PubMed

Arntzen, Magnus Øverlie; Karlskås, Ingrid Lea; Skaugen, Morten; Eijsink, Vincent G H; Mathiesen, Geir

2015-01-01

Enterococcus faecalis is a robust bacterium, which is able to survive in and adapt to hostile environments such as the urinary tract and bladder. In this label-free quantitative proteomic study based on MaxQuant LFQ algorithms, we identified 127 proteins present in the secretome of the clinical vancomycin-resistant isolate E. faecalis V583 and we compared proteins secreted in the initial phase of cultivation in urine with the secretome during cultivation in standard laboratory medium, 2xYT. Of the 54 identified proteins predicted to be secreted, six were exclusively found after cultivation in urine including the virulence factor EfaA ("endocarditis specific antigen") and its homologue EF0577 ("adhesion lipoprotein"). These two proteins are both involved in manganese transport, known to be an important determinant of colonization and infection, and may additionally function as adhesins. Other detected urine-specific proteins are involved in peptide transport (EF0063 and EF3106) and protease inhibition (EF3054). In addition, we found an uncharacterized protein (EF0764), which had not previously been linked to the adaptation of V583 to a urine environment, and which is unique to E. faecalis. Proteins found in both environments included a histone-like protein, EF1550, that was up-regulated during cultivation in urine and that has a homologue in streptococci (HlpA) known to be involved in bacterial adhesion to host cells. Up-regulated secreted proteins included autolysins. These results from secretome analyses are largely compatible with previously published data from transcriptomics studies. All in all, the present data indicate that transport, in particular metal transport, adhesion, cell wall remodelling and the unknown function carried out by the unique EF0764 are important for enterococcal adaptation to the urine environment. These results provide a basis for a more targeted exploration of novel proteins involved in the adaptability and pathogenicity of E. faecalis.

[The frequency of bacteria in human gallstones].

PubMed

Lévay, Bernadett; Szabó, Györgyi; Szijártó, Attila; Gamal, Eldin Mohamed

2013-12-01

Complications caused by lost gallstones within the abdominal cavity are well known. Abscesses, perforation of the gastro-intestinal tract were all described in the literature, but gallstones were found in hernial sac, or even in sputum after it penetrated through the diaphragm into the respiratory tract. These complications can develop between several weeks to several years postoperatively. Most complications can be treated surgically only. Fifty gallstones and bile samples were collected from 50 patients who underwent cholecystectomy (36 female / 14 male, avarge age: 60.8 ± 6.8 years). All samples were sent for microbiological examination. bacterial colonization of the gallstone and the bile were found in 16 cases. Four of them showed acute inflammation in the gallbladder while pathological signs of chronic inflammation in the gallbladder wall were detected in eight cases. Empyema was found in four cases. Bacteria from enteral origin (Esherichia coli, Enterococcus faecalis, Enterobacter cloacae) was detected in 13 cases, while non-enteral (Klebsiella penumoniae, Streptococcus alfa-haemoliticus) colony were detected in three cases. Positive bacterial cultures were identified in twelve female and fourmale patients. Different types of bacteria can be found in the gallstones, which may cause various complications.

First evidence of a membrane-bound, tyramine and beta-phenylethylamine producing, tyrosine decarboxylase in Enterococcus faecalis: a two-dimensional electrophoresis proteomic study.

PubMed

Pessione, Enrica; Pessione, Alessandro; Lamberti, Cristina; Coïsson, Daniel Jean; Riedel, Kathrin; Mazzoli, Roberto; Bonetta, Silvia; Eberl, Leo; Giunta, Carlo

2009-05-01

The soluble and membrane proteome of a tyramine producing Enterococcus faecalis, isolated from an Italian goat cheese, was investigated. A detailed analysis revealed that this strain also produces small amounts of beta-phenylethylamine. Kinetics of tyramine and beta-phenylethylamine accumulation, evaluated in tyrosine plus phenylalanine-enriched cultures (stimulated condition), suggest that the same enzyme, the tyrosine decarboxylase (TDC), catalyzes both tyrosine and phenylalanine decarboxylation: tyrosine was recognized as the first substrate and completely converted into tyramine (100% yield) while phenylalanine was decarboxylated to beta-phenylethylamine (10% yield) only when tyrosine was completely depleted. The presence of an aspecific aromatic amino acid decarboxylase is a common feature in eukaryotes, but in bacteria only indirect evidences of a phenylalanine decarboxylating TDC have been presented so far. Comparative proteomic investigations, performed by 2-DE and MALDI-TOF/TOF MS, on bacteria grown in conditions stimulating tyramine and beta-phenylethylamine biosynthesis and in control conditions revealed 49 differentially expressed proteins. Except for aromatic amino acid biosynthetic enzymes, no significant down-regulation of the central metabolic pathways was observed in stimulated conditions, suggesting that tyrosine decarboxylation does not compete with the other energy-supplying routes. The most interesting finding is a membrane-bound TDC highly over-expressed during amine production. This is the first evidence of a true membrane-bound TDC, longly suspected in bacteria on the basis of the gene sequence.

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

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

Optimizing supercritical carbon dioxide in the inactivation of bacteria in clinical solid waste by using response surface methodology.

PubMed

Hossain, Md Sohrab; Nik Ab Rahman, Nik Norulaini; Balakrishnan, Venugopal; Alkarkhi, Abbas F M; Ahmad Rajion, Zainul; Ab Kadir, Mohd Omar

2015-04-01

Clinical solid waste (CSW) poses a challenge to health care facilities because of the presence of pathogenic microorganisms, leading to concerns in the effective sterilization of the CSW for safe handling and elimination of infectious disease transmission. In the present study, supercritical carbon dioxide (SC-CO2) was applied to inactivate gram-positive Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis, and gram-negative Escherichia coli in CSW. The effects of SC-CO2 sterilization parameters such as pressure, temperature, and time were investigated and optimized by response surface methodology (RSM). Results showed that the data were adequately fitted into the second-order polynomial model. The linear quadratic terms and interaction between pressure and temperature had significant effects on the inactivation of S. aureus, E. coli, E. faecalis, and B. subtilis in CSW. Optimum conditions for the complete inactivation of bacteria within the experimental range of the studied variables were 20 MPa, 60 °C, and 60 min. The SC-CO2-treated bacterial cells, observed under a scanning electron microscope, showed morphological changes, including cell breakage and dislodged cell walls, which could have caused the inactivation. This espouses the inference that SC-CO2 exerts strong inactivating effects on the bacteria present in CSW, and has the potential to be used in CSW management for the safe handling and recycling-reuse of CSW materials. Copyright © 2015 Elsevier Ltd. All rights reserved.

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.

21 CFR 172.809 - Curdlan.

Code of Federal Regulations, 2011 CFR

2011-04-01

...) produced by pure culture fermentation from the nonpathogenic and nontoxicogenic bacterium Alcaligenes...) Coliform bacteria, not more than 3 per gram. (c) Curdlan is used or intended for use in accordance with...

21 CFR 172.809 - Curdlan.

Code of Federal Regulations, 2010 CFR

2010-04-01

...) produced by pure culture fermentation from the nonpathogenic and nontoxicogenic bacterium Alcaligenes...) Coliform bacteria, not more than 3 per gram. (c) Curdlan is used or intended for use in accordance with...

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Meining, Winfried, E-mail: wim@csb.ki.se; Scheuring, Johannes; Fischer, Markus

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,more » α = γ = 90.0, β = 119.1°. A selenomethionine derivative was prepared and is currently being tested in crystallization trials.« less

Comparative Genomic Analysis of Pathogenic and Probiotic Enterococcus faecalis Isolates, and Their Transcriptional Responses to Growth in Human Urine

PubMed Central

Snipen, Lars; Nes, Ingolf F.; Brede, Dag A.

2010-01-01

Urinary tract infection (UTI) is the most common infection caused by enterococci, and Enterococcus faecalis accounts for the majority of enterococcal infections. Although a number of virulence related traits have been established, no comprehensive genomic or transcriptomic studies have been conducted to investigate how to distinguish pathogenic from non-pathogenic E. faecalis in their ability to cause UTI. In order to identify potential genetic traits or gene regulatory features that distinguish pathogenic from non-pathogenic E. faecalis with respect to UTI, we have performed comparative genomic analysis, and investigated growth capacity and transcriptome profiling in human urine in vitro. Six strains of different origins were cultivated and all grew readily in human urine. The three strains chosen for transcriptional analysis showed an overall similar response with respect to energy and nitrogen metabolism, stress mechanism, cell envelope modifications, and trace metal acquisition. Our results suggest that citrate and aspartate are significant for growth of E. faecalis in human urine, and manganese appear to be a limiting factor. The majority of virulence factors were either not differentially regulated or down-regulated. Notably, a significant up-regulation of genes involved in biofilm formation was observed. Strains from different origins have similar capacity to grow in human urine. The overall similar transcriptional responses between the two pathogenic and the probiotic strain suggest that the pathogenic potential of a certain E. faecalis strain may to a great extent be determined by presence of fitness and virulence factors, rather than the level of expression of such traits. PMID:20824220

An In Vitro Study on the Effects of Nisin on the Antibacterial Activities of 18 Antibiotics against Enterococcus faecalis

PubMed Central

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

Bactericidal effect of the 908 nm diode laser on Enterococcus faecalis in infected root canals

PubMed Central

Preethee, Thomas; Kandaswamy, Deivanayagam; Arathi, Ganesh; Hannah, Rosaline

2012-01-01

Aim: The aim of this study is to evaluate the bactericidal effect of 908 nm diode laser in conjunction with various irrigation regimes in disinfection of apical third of root dentin. Materials and Methods: Sixty prepared teeth with single canals were contaminated with Enterococcus faecalis. The specimens were divided into 6 groups (n = 10): Group 1 and 3 and 5 were subjected to chemo-mechanical preparation using 5.25% sodium hypochlorite (NaOCl), 17% Ethylenediaminetetraacetic acid (EDTA); 1.3% NaOCl, MTAD (mixture of doxycycline, citric acid and a detergent (Tween 80); and, 8.5% saline, respectively followed by 908 nm diode laser irradiation; Group 2 and 4, followed the same procedure as Group1 and 3, however without laser irradiation; and, Group 6, rinsed with saline solution (control). Dentin shavings from apical third were analyzed for the presence of E. faecalis using culture method and Polymerase Chain reaction (PCR). Results: One-way Analysis of variance showed statistically significant differences between the laser irradiated groups, non irradiated groups and the control group. Conclusion: 908 nm diode used in conjunction with conventional chemomechanical techniques demonstrated a significant elimination of E. faecalis in the apical third of root dentin. PMID:22368335

Chemical Compositions and Aroma Evaluation of Volatile Oil from the Industrial Cultivation Medium of Enterococcus faecalis.

PubMed

Ono, Toshirou; Usami, Atsushi; Nakaya, Satoshi; Maeba, Keisuke; Yonejima, Yasunori; Toyoda, Masanori; Ikeda, Atsushi; Miyazawa, Mitsuo

2015-01-01

Enterococcus faecalis is one of the major lactic acid bacterium (LAB) species colonizing the intestines of animals and humans. The characteristic odor of the volatile oils obtained from both the liquid medium after incubation (MAI) and liquid medium before incubation (MBI) in the cultivation process of E. faecalis was investigated to determine the utility of the liquid medium. In total, fifty-six and thirty-two compounds were detected in the volatile oils from the MAI (MAI oil) and MBI (MBI oil), respectively. The principle components of MAI oil were 2,5-dimethylpyrazine (19.3%), phenylacetaldehyde (19.3%), and phenylethyl alcohol (9.3%). The aroma extract dilution analysis (AEDA) method was performed using gas chromatography-olfactometry (GC-O). The total number of aroma-active compounds identified in the volatile oil from MBI and MAI was thirteen compounds; in particular, 5-methyl-2-furanmethanol, phenylacetaldehyde, and phenylethyl alcohol were the most primary aroma-active compounds in MAI oil. These results imply that the industrial cultivation medium after incubation of E. faecalis may be utilized as a source of volatile oils.

Comparison of the gentamicin resistance transposon Tn5281 with regions encoding gentamicin resistance in Enterococcus faecalis isolates from diverse geographic locations.

PubMed Central

Hodel-Christian, S L; Murray, B E

1992-01-01

The genetic determinant encoding gentamicin resistance (Gmr) on the beta-lactamase encoding plasmid pBEM10 of Enterococcus faecalis HH22 is carried on a transposon, termed Tn5281, that is highly related to the staphylococcal Gmr transposons Tn4001 found in Australian isolates of Staphylococcus aureus and Tn4031 found in United States isolates of Staphylococcus epidermidis. We have now studied plasmid DNA from Gmr strains of E. faecalis isolated from diverse geographical locations (Houston, Pennsylvania, Thailand, and Chile) by using restriction endonuclease analysis and DNA-DNA hybridization to determine whether other Gmr E. faecalis carry Tn5281 or a similar type of element. We also compared these enterococci to several United States isolates of Staphylococcus aureus with nonmobile Gmr determinants. Three E. faecalis isolates (from Houston and Chile) carried Tn5281-like elements, whereas two isolates (from Houston and Pennsylvania) had restriction endonuclease and DNA-DNA hybridization patterns more similar to those of the Tn4001-IS257 hybrid found in the nonmobile Gmr determinants in United States isolates of S. aureus. A strain from Thailand had a third pattern unrelated to either Tn5281 or the nonmobile Gmr determinants present in United States isolates of S. aureus. Our results demonstrate that there is both similarity and diversity between the Gmr determinant of strains of E. faecalis isolated in diverse geographic locations. Images PMID:1332593

ANALYSIS OF ENTEROCOCCUS FAECALIS IN SAMPLES FROM TURKISH PATIENTS WITH PRIMARY ENDODONTIC INFECTIONS AND FAILED ENDODONTIC TREATMENT BY REAL-TIME PCR SYBR GREEN METHOD

PubMed Central

Ozbek, Selcuk M.; Ozbek, Ahmet; Erdogan, Aziz S.

2009-01-01

Objective: The aims of this study were to investigate the presence of Enterococcus faecalis in primary endodontic infections and failed endodontic treatments using real-time PCR and to determine the statistical importance of the presence of E. faecalis in a Turkish population with endodontic infections. Material and Methods: E. faecalis was investigated from 79 microbial samples collected from patients who were treated at the Endodontic Clinic of the Dental School of Atatürk University (Erzurum, Turkey). Microbial samples were taken from 43 patients (Group 1) with failed endodontic treatments and 36 patients (Group 2) with chronic apical periodontitis (primary endodontic infections). DNA was extracted from the samples by using a QIAamp® DNA mini-kit and analyzed with real-time PCR SYBR Green. Results: E. faecalis was detected in 41 out of 79 patients, suggesting that it exists in not less than 61% of all endodontic infections when the proportion test (z= -1.645, faecalis in 32 out of 43 (74.4%) in Group 1, and in 9 out of 36 (25%) in Group 2. Conclusions: These results suggest that E. faecalis is a frequent isolate for endodontic infections in Turkish patients, and is more often associated with failed endodontic treatments than primary endodontic infections. PMID:19936510

Optimizing supercritical carbon dioxide in the inactivation of bacteria in clinical solid waste by using response surface methodology

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hossain, Md. Sohrab; Nik Ab Rahman, Nik Norulaini; Balakrishnan, Venugopal

2015-04-15

Highlights: • Supercritical carbon dioxide sterilization of clinical solid waste. • Inactivation of bacteria in clinical solid waste using supercritical carbon dioxide. • Reduction of the hazardous exposure of clinical solid waste. • Optimization of the supercritical carbon dioxide experimental conditions. - Abstract: Clinical solid waste (CSW) poses a challenge to health care facilities because of the presence of pathogenic microorganisms, leading to concerns in the effective sterilization of the CSW for safe handling and elimination of infectious disease transmission. In the present study, supercritical carbon dioxide (SC-CO{sub 2}) was applied to inactivate gram-positive Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis,more » and gram-negative Escherichia coli in CSW. The effects of SC-CO{sub 2} sterilization parameters such as pressure, temperature, and time were investigated and optimized by response surface methodology (RSM). Results showed that the data were adequately fitted into the second-order polynomial model. The linear quadratic terms and interaction between pressure and temperature had significant effects on the inactivation of S. aureus, E. coli, E. faecalis, and B. subtilis in CSW. Optimum conditions for the complete inactivation of bacteria within the experimental range of the studied variables were 20 MPa, 60 °C, and 60 min. The SC-CO{sub 2}-treated bacterial cells, observed under a scanning electron microscope, showed morphological changes, including cell breakage and dislodged cell walls, which could have caused the inactivation. This espouses the inference that SC-CO{sub 2} exerts strong inactivating effects on the bacteria present in CSW, and has the potential to be used in CSW management for the safe handling and recycling-reuse of CSW materials.« less

A comparison of three rotary systems and hand instrumentation technique for the elimination of Enterococcus faecalis from the root canal.

PubMed

Gorduysus, Melahat; Nagas, Emre; Torun, Ozgur Yildirim; Gorduysus, Omer

2011-12-01

The aim of this study was to compare the in vitro reduction of a bacterial population in a root canal by mechanical instrumentation using three rotary systems and hand instrumentation technique. The root canals contaminated with a suspension of Enterococcus faecalis were instrumented using ProTaper, K3, HeroShaper and K-file hand instrumentation technique. Later the root canals were sampled. After serial dilutions, samples were incubated in culture media for 24 h. Bacterial colonies were counted and the results were given as number of colony-forming units per millilitre. The results showed that all the canal instrumentation systems reduced the number of bacterial cells in the root canals. Statistically, ProTaper instruments were more effective in reducing the number of bacteria than the other rotary files or hand instruments. © 2010 The Authors. Australian Endodontic Journal © 2010 Australian Society of Endodontology.

In vitro evaluation of antibacterial effect of AH Plus incorporated with quaternary ammonium epoxy silicate against Enterococcus faecalis.

PubMed

Gong, Shi-Qiang; Huang, Zhi-Bin; Shi, Wei; Ma, Bo; Tay, Franklin R; Zhou, Bin

2014-10-01

The purpose of this study was to evaluate the in vitro antibacterial effect of AH Plus (Dentsply, DeTrey, Konstanz, Germany) incorporated with quaternary ammonium epoxy silicate (QAES) against Enterococcus faecalis. QAES particles were synthesized by the cocondensation of tetraethoxysilane with 2 trialkoxysilanes (3-[trimethoxysilyl]propyldimethyloctadecyl ammonium chloride and 3-glycidyloxypropyltrimethoxysilane) through a 1-pot sol-gel route. Dried QAES particles were then characterized by attenuated total reflection Fourier transform infrared spectroscopy and scanning electron microscopy. AH Plus sealers incorporated with 0-8 wt% QAES were tested after 4 weeks of water aging to assess the in vitro antibacterial activity against E. faecalis by the direct contact test (DCT) and 3-dimensional image analysis of live/dead-stained E. faecalis biofilms using confocal laser scanning microscopy. The Fourier transform infrared spectroscopy spectrum of QAES particles revealed the coexistence of the characteristic absorbance band of the siloxane backbone (Si-O-Si) from 1,000-1,100 cm(-1), epoxide band peaking at ∼916 cm(-1), and C-N stretching vibration peaking at 1,373 cm(-1). The scanning electron microscopic image showed the spherical morphology of QAES particles with ∼120 nm in diameter and a rough surface. DCT results revealed that AH Plus alone (0 wt% QAES) after 4 weeks of water aging had no inhibitory effect on E. faecalis growth (P = .569). AH Plus incorporated with QAES (2-8 wt%) showed antibacterial activity against E. faecalis as shown in DCT and biofilm viability results (P < .001). The incorporation of QAES into epoxy resin-based AH Plus may be a promising approach for controlling endodontic infection at the time of canal filling and preventing subsequent reinfection. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Genetic Variability of Vancomycin-Resistant Enterococcus faecium and Enterococcus faecalis Isolates from Humans, Chickens, and Pigs in Malaysia

PubMed Central

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

2013-01-01

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. PMID:23666337

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

Ab initio single and multideterminant methods used in the determination of reduction potentials and magnetic properties of Rieske ferredoxins

NASA Astrophysics Data System (ADS)

Powers, Nathan Lee

2008-10-01

The [Fe2S2]2+/[Fe2S 2]+ electronic structure of seven Rieske protein active sites (bovine mitochondrial cytochrome bc1 complex, spinach chloroplast cytochrome b6f complex, Rieske-type ferredoxin associated with biphenyl dioxygenase from Burkholderia cepacia, yeast cytochrome bcl complex from Saccharomyces cerevisiae, Rieske subunit of arsenite oxidase from Alcaligenes faecalis, respiratory-type Rieske protein from Thermus thermophilus, and Rieske protein II (soxF) from Sulfolobus acidocaldarius), which lie in a reduction potential range from -150 mV to 375 mV, have been studied by both single and multi-determinant quantum mechanical methods. Calculated reduction potentials and magnetic properties are found comparable to experimental values.

Ultrasonic activation and chemical modification of photosensitizers enhances the effects of photodynamic therapy against Enterococcus faecalis root-canal isolates.

PubMed

Tennert, C; Drews, A M; Walther, V; Altenburger, M J; Karygianni, L; Wrbas, K T; Hellwig, E; Al-Ahmad, A

2015-06-01

The aim of this study was to evaluate the effect of photodynamic therapy (PDT) on Enterococcus faecalis biofilms in artificially infected root canals using modified photosensitizers and passive ultrasonic activation. Two hundred and seventy extracted human teeth with one root canal were instrumented utilizing ProTaper files, autoclaved, infected with E. faecalis T9 for 72 h and divided into different groups: irrigation with 3% sodium hypochlorite (NaOCl), 20% ethylenediaminetetraacetic acid (EDTA), or 20% citric acid, PDT without irrigation, PDT accompanied by irrigation with NaOCl, EDTA, or citric acid, PDT using an EDTA-based photosensitizer or a citric-acid-based photosensitizer and PDT with ultrasonic activation of the photosensitizer. A 15 mg/ml toluidine blue served as the photosensitizer, activated by a 100 mW LED light source. Sterile paper points were used for sampling the root canals and dentin chips were collected to assess the remaining contamination after treatment. Samples were cultured on blood agar plates and colony forming units were quantified. PDT alone achieved a reduction in E. faecalis counts by 92.7%, NaOCl irrigation alone and combined with PDT by 99.9%. The antibacterial effects increased by the combination of irrigation using EDTA or citric acid and PDT compared to irrigation alone. More than 99% of E. faecalis were killed using PDT with the modified photosensitizers and ultrasonic activation. NaOCl based disinfection achieved the highest antimicrobial effect. Using PDT with an EDTA-based or citric-acid-based phozosensitizer or activating the photosensitizer with ultrasound resulted in a significantly higher reduction in E. faecalis counts compared to conventional PDT. Copyright © 2015 Elsevier B.V. All rights reserved.

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

Antimicrobial activity of polyphenol-rich fractions from Sida alba L. (Malvaceae) against co-trimoxazol-resistant bacteria strains.

PubMed

Konaté, Kiessoun; Hilou, Adama; Mavoungou, Jacques François; Lepengué, Alexis Nicaise; Souza, Alain; Barro, Nicolas; Datté, Jacques Y; M'batchi, Bertrand; Nacoulma, Odile Germaine

2012-02-24

The increased resistance of microorganisms to the currently used antimicrobials has lead to the evaluation of other agents that might have antimicrobial activity. Medicinal plants are sources of phytochemicals which are able to initiate different biological activities including antimicrobials In vitro antibacterial (MIC, MBC and time-kill studies) of polyphenol-rich fractions from Sida alba L. (Malvaceae) was assessed using ten bacteria strains (Gram-negative and Gram-positive). All test bacteria were susceptible to the polyphenol-rich fractions. Time-kill results showed that after 5 h exposition there was no viable microorganism in the initial inoculum and the effect of polyphenol-rich fractions was faster on Enterococcus faecalis (Gram-positive bacterium) comparatively to the other bacteria strains. The data analysis indicates that the tested of polyphenol-rich fractions has significant effects when compared with the standard antibiotic. These results therefore justify the traditional use of sida alba L., alone or in combination with other herbs to treat bacterial infections.

Antimicrobial activity of polyphenol-rich fractions from Sida alba L. (Malvaceae) against co-trimoxazol-resistant bacteria strains

PubMed Central

2012-01-01

Background The increased resistance of microorganisms to the currently used antimicrobials has lead to the evaluation of other agents that might have antimicrobial activity. Medicinal plants are sources of phytochemicals which are able to initiate different biological activities including antimicrobials Materials and methods In vitro antibacterial (MIC, MBC and time-kill studies) of polyphenol-rich fractions from Sida alba L. (Malvaceae) was assessed using ten bacteria strains (Gram-negative and Gram-positive). Results All test bacteria were susceptible to the polyphenol-rich fractions. Time-kill results showed that after 5 h exposition there was no viable microorganism in the initial inoculum and the effect of polyphenol-rich fractions was faster on Enterococcus faecalis (Gram-positive bacterium) comparatively to the other bacteria strains. Conclusion The data analysis indicates that the tested of polyphenol-rich fractions has significant effects when compared with the standard antibiotic. These results therefore justify the traditional use of sida alba L., alone or in combination with other herbs to treat bacterial infections. PMID:22364123

Proteomic Investigation of the Response of Enterococcus faecalis V583 when Cultivated in Urine

PubMed Central

Arntzen, Magnus Øverlie; Karlskås, Ingrid Lea; Skaugen, Morten; Eijsink, Vincent G. H.; Mathiesen, Geir

2015-01-01

Enterococcus faecalis is a robust bacterium, which is able to survive in and adapt to hostile environments such as the urinary tract and bladder. In this label-free quantitative proteomic study based on MaxQuant LFQ algorithms, we identified 127 proteins present in the secretome of the clinical vancomycin-resistant isolate E. faecalis V583 and we compared proteins secreted in the initial phase of cultivation in urine with the secretome during cultivation in standard laboratory medium, 2xYT. Of the 54 identified proteins predicted to be secreted, six were exclusively found after cultivation in urine including the virulence factor EfaA (“endocarditis specific antigen”) and its homologue EF0577 (“adhesion lipoprotein”). These two proteins are both involved in manganese transport, known to be an important determinant of colonization and infection, and may additionally function as adhesins. Other detected urine-specific proteins are involved in peptide transport (EF0063 and EF3106) and protease inhibition (EF3054). In addition, we found an uncharacterized protein (EF0764), which had not previously been linked to the adaptation of V583 to a urine environment, and which is unique to E. faecalis. Proteins found in both environments included a histone-like protein, EF1550, that was up-regulated during cultivation in urine and that has a homologue in streptococci (HlpA) known to be involved in bacterial adhesion to host cells. Up-regulated secreted proteins included autolysins. These results from secretome analyses are largely compatible with previously published data from transcriptomics studies. All in all, the present data indicate that transport, in particular metal transport, adhesion, cell wall remodelling and the unknown function carried out by the unique EF0764 are important for enterococcal adaptation to the urine environment. These results provide a basis for a more targeted exploration of novel proteins involved in the adaptability and pathogenicity of E

Functional analysis of AtlA, the major N-acetylglucosaminidase of Enterococcus faecalis.

PubMed

Eckert, Catherine; Lecerf, Maxime; Dubost, Lionel; Arthur, Michel; Mesnage, Stéphane

2006-12-01

The major peptidoglycan hydrolase of Enterococcus faecalis, AtlA, has been identified, but its enzyme activity remains unknown. We have used tandem mass spectrometry analysis of peptidoglycan hydrolysis products obtained using the purified protein to show that AtlA is an N-acetylglucosaminidase. To gain insight into the regulation of its enzyme activity, the three domains of AtlA were purified alone or in combination following expression of truncated forms of the atlA gene in Escherichia coli or partial digestion of AtlA by proteinase K. The central domain of AtlA was catalytically active, but its activity was more than two orders of magnitude lower than that of the complete protein. Partial proteolysis of AtlA was detected in vivo: zymograms of E. faecalis extracts revealed two catalytically active protein bands of 62 and 72 kDa that were both absent in extracts from an atlA null mutant. Limited digestion of AtlA by proteinase K in vitro suggested that the proteolytic cleavage of AtlA in E. faecalis extracts corresponds to the truncation of the N-terminal domain, which is rich in threonine and glutamic acid residues. We show that the truncation of the N-terminal domain from recombinant AtlA has no impact on enzyme activity. The C-terminal domain of the protein, which contains six LysM modules bound to highly purified peptidoglycan, was required for optimal enzyme activity. These data indicate that AtlA is not produced as a proenzyme and that control of the AtlA glucosaminidase activity is likely to occur at the level of LysM-mediated binding to peptidoglycan.

Apical extrusion of intracanal bacteria following use of two engine-driven instrumentation techniques.

PubMed

Er, K; Sümer, Z; Akpinar, K E

2005-12-01

To evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using the two engine-driven techniques utilizing nickel-titanium instruments (ProTaper and System GT). Forty extracted single-rooted human mandibular premolar teeth were used. Access cavities were prepared and root canals were then contaminated with a suspension of Enterococcus faecalis and dried. The contaminated roots were divided into two experimental groups of 15 teeth each and one control group of 10 teeth. Group 1, ProTaper group: the root canals were instrumented using ProTaper instruments. Group 2, System GT group: the root canals were instrumented using System GT instruments. Group 3, control group: no instrumentation was attempted. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 h. Colonies of bacteria were counted and the results were given as number of colony-forming units. The data obtained were analysed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with alpha = 0.05 as the level for statistical significance. There was no significant difference as to the number of extruded bacteria between the ProTaper and System GT engine-driven systems (P > 0.05). Both engine-driven nickel-titanium systems extruded bacteria through the apical foramen.

Effectiveness of repeated photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm: an in vitro study.

PubMed

Prażmo, Ewa Joanna; Godlewska, Renata Alicja; Mielczarek, Agnieszka Beata

2017-04-01

The study aimed to investigate the effectiveness of photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm and to analyse how a repeated light irradiation, replenishment of oxygen and photosensitiser affect the results of the photodynamic disinfecting protocol. After chemomechanical preparation, 46 single-rooted human teeth were infected with a clinical strain of E. faecalis and incubated for a week in microaerobic conditions. The experimental procedures included groups of single application of photodynamic therapy, two cycles of PDT, irrigation with 5.25% NaOCl solution and negative and positive control. The number of residing bacterial colonies in the root canals was determined based on the CFU/ml method. In the group of preparations irrigated with NaOCl, bacterial colonies were not observed. A single PDT eliminated 45% of the initial CFU/ml. Repeated PDT eradicated 95% of the intracanal bacterial biofilm. Photodynamic therapy has a high potential for the elimination of E. faecalis biofilm. There is a safe therapeutic window where photoinduced disinfection can be used as an adjuvant to conventional endodontic treatment, which remains the most effective.

Effect of lipoteichoic acid and lipids on lysis of intact cells of Streptococcus faecalis.

PubMed Central

Cleveland, R F; Daneo-Moore, L; Wicken, A J; Shockman, G D

1976-01-01

Autolysis of intact cells of Streptococcus faecalis was inhibited to a greater extent by phospholipids than by lipoteichoic acid, suggesting a possible difference in the accessibility of native autolysin to these substances. PMID:821938

Preliminary survey of antibiotic-resistant fecal indicator bacteria and pathogenic Escherichia coli from river-water samples collected in Oakland County, Michigan, 2003

USGS Publications Warehouse

Fogarty, Lisa R.; Duris, Joseph W.; Aichele, Stephen S.

2005-01-01

A preliminary study was done in Oakland County, Michigan, to determine the concentration of fecal indicator bacteria (fecal coliform bacteria and enterococci), antibiotic resistance patterns of these two groups, and the presence of potentially pathogenic Escherichia coli (E. coli). For selected sites, specific members of these groups [E. coli, Enterococcus faecium (E. faecium) and Enterococcus faecalis (E. faecalis)] were isolated and tested for levels of resistance to specific antibiotics used to treat human infections by pathogens in these groups and for their potential to transfer these resistances. In addition, water samples from all sites were tested for indicators of potentially pathogenic E. coli by three assays: a growth-based assay for sorbitol-negative E. coli, an immunological assay for E. coli O157, and a molecular assay for three virulence and two serotype genes. Samples were also collected from two non-urbanized sites outside of Oakland County. Results from the urbanized Oakland County area were compared to those from these two non-urbanized sites. Fecal indicator bacteria concentrations exceeded State of Michigan recreational water-quality standards and (or) recommended U.S. Environmental Protection Agency (USEPA) standards in samples from all but two Oakland County sites. Multiple-antibiotic-resistant fecal coliform bacteria were found at all sites, including two reference sites from outside the county. Two sites (Stony Creek and Paint Creek) yielded fecal coliform isolates resistant to all tested antibiotics. Patterns indicative of extended-spectrum-β-lactamase (ESBL)- producing fecal coliform bacteria were found at eight sites in Oakland County and E. coli resistant to clinically significant antibiotics were recovered from the River Rouge, Clinton River, and Paint Creek. Vancomycin-resistant presumptive enterococci were found at six sites in Oakland County and were not found at the reference sites. Evidence of acquired antibiotic resistances was

Biphenyl-Metabolizing Bacteria in the Rhizosphere of Horseradish and Bulk Soil Contaminated by Polychlorinated Biphenyls as Revealed by Stable Isotope Probing▿ †

PubMed Central

Uhlik, Ondrej; Jecna, Katerina; Mackova, Martina; Vlcek, Cestmir; Hroudova, Miluse; Demnerova, Katerina; Paces, Vaclav; Macek, Tomas

2009-01-01

DNA-based stable isotope probing in combination with terminal restriction fragment length polymorphism was used in order to identify members of the microbial community that metabolize biphenyl in the rhizosphere of horseradish (Armoracia rusticana) cultivated in soil contaminated with polychlorinated biphenyls (PCBs) compared to members of the microbial community in initial, uncultivated bulk soil. On the basis of early and recurrent detection of their 16S rRNA genes in clone libraries constructed from [13C]DNA, Hydrogenophaga spp. appeared to dominate biphenyl catabolism in the horseradish rhizosphere soil, whereas Paenibacillus spp. were the predominant biphenyl-utilizing bacteria in the initial bulk soil. Other bacteria found to derive carbon from biphenyl in this nutrient-amended microcosm-based study belonged mostly to the class Betaproteobacteria and were identified as Achromobacter spp., Variovorax spp., Methylovorus spp., or Methylophilus spp. Some bacteria that were unclassified at the genus level were also detected, and these bacteria may be members of undescribed genera. The deduced amino acid sequences of the biphenyl dioxygenase α subunits (BphA) from bacteria that incorporated [13C]into DNA in 3-day incubations of the soils with [13C]biphenyl are almost identical to that of Pseudomonas alcaligenes B-357. This suggests that the spectrum of the PCB congeners that can be degraded by these enzymes may be similar to that of strain B-357. These results demonstrate that altering the soil environment can result in the participation of different bacteria in the metabolism of biphenyl. PMID:19700551

Characterization of Multi-Drug Resistant Enterococcus faecalis Isolated from Cephalic Recording Chambers in Research Macaques (Macaca spp.).

PubMed

Woods, Stephanie E; Lieberman, Mia T; Lebreton, Francois; Trowel, Elise; de la Fuente-Núñez, César; Dzink-Fox, Joanne; Gilmore, Michael S; Fox, James G

2017-01-01

Nonhuman primates are commonly used for cognitive neuroscience research and often surgically implanted with cephalic recording chambers for electrophysiological recording. Aerobic bacterial cultures from 25 macaques identified 72 bacterial isolates, including 15 Enterococcus faecalis isolates. The E. faecalis isolates displayed multi-drug resistant phenotypes, with resistance to ciprofloxacin, enrofloxacin, trimethoprim-sulfamethoxazole, tetracycline, chloramphenicol, bacitracin, and erythromycin, as well as high-level aminoglycoside resistance. Multi-locus sequence typing showed that most belonged to two E. faecalis sequence types (ST): ST 4 and ST 55. The genomes of three representative isolates were sequenced to identify genes encoding antimicrobial resistances and other traits. Antimicrobial resistance genes identified included aac(6')-aph(2"), aph(3')-III, str, ant(6)-Ia, tetM, tetS, tetL, ermB, bcrABR, cat, and dfrG, and polymorphisms in parC (S80I) and gyrA (S83I) were observed. These isolates also harbored virulence factors including the cytolysin toxin genes in ST 4 isolates, as well as multiple biofilm-associated genes (esp, agg, ace, SrtA, gelE, ebpABC), hyaluronidases (hylA, hylB), and other survival genes (ElrA, tpx). Crystal violet biofilm assays confirmed that ST 4 isolates produced more biofilm than ST 55 isolates. The abundance of antimicrobial resistance and virulence factor genes in the ST 4 isolates likely relates to the loss of CRISPR-cas. This macaque colony represents a unique model for studying E. faecalis infection associated with indwelling devices, and provides an opportunity to understand the basis of persistence of this pathogen in a healthcare setting.

Bacterial size matters: Multiple mechanisms controlling septum cleavage and diplococcus formation are critical for the virulence of the opportunistic pathogen Enterococcus faecalis

PubMed Central

Salamaga, Bartłomiej; Prajsnar, Tomasz K.; Willemse, Joost; Bewley, Martin A.; Chau, Françoise

2017-01-01

Enterococcus faecalis is an opportunistic pathogen frequently isolated in clinical settings. This organism is intrinsically resistant to several clinically relevant antibiotics and can transfer resistance to other pathogens. Although E. faecalis has emerged as a major nosocomial pathogen, the mechanisms underlying the virulence of this organism remain elusive. We studied the regulation of daughter cell separation during growth and explored the impact of this process on pathogenesis. We demonstrate that the activity of the AtlA peptidoglycan hydrolase, an enzyme dedicated to septum cleavage, is controlled by several mechanisms, including glycosylation and recognition of the peptidoglycan substrate. We show that the long cell chains of E. faecalis mutants are more susceptible to phagocytosis and are no longer able to cause lethality in the zebrafish model of infection. Altogether, this work indicates that control of cell separation during division underpins the pathogenesis of E. faecalis infections and represents a novel enterococcal virulence factor. We propose that inhibition of septum cleavage during division represents an attractive therapeutic strategy to control infections. PMID:28742152

Fitness costs of various mobile genetic elements in Enterococcus faecium and Enterococcus faecalis

PubMed Central

Starikova, Irina; Al-Haroni, Mohammed; Werner, Guido; Roberts, Adam P.; Sørum, Vidar; Nielsen, Kaare M.; Johnsen, Pål J.

2013-01-01

Objectives To determine the fitness effects of various mobile genetic elements (MGEs) in Enterococcus faecium and Enterococcus faecalis when newly acquired. We also tested the hypothesis that the biological cost of vancomycin resistance plasmids could be mitigated during continuous growth in the laboratory. Methods Different MGEs, including two conjugative transposons (CTns) of the Tn916 family (18 and 33 kb), a pathogenicity island (PAI) of 200 kb and vancomycin-resistance (vanA) plasmids (80–200 kb) of various origins and classes, were transferred into common ancestral E. faecium and E. faecalis strains by conjugation assays and experimentally evolved (vanA plasmids only). Transconjugants were characterized by PFGE, S1 nuclease assays and Southern blotting hybridization analyses. Single specific primer PCR was performed to determine the target sites for the insertion of the CTns. The fitness costs of various MGEs in E. faecium and E. faecalis were estimated in head-to-head competition experiments, and evolved populations were generated in serial transfer assays. Results The biological cost of a newly acquired PAI and two CTns were both host- and insertion-locus-dependent. Newly acquired vanA plasmids may severely reduce host fitness (25%–27%), but these costs were rapidly mitigated after only 400 generations of continuous growth in the absence of antibiotic selection. Conclusions Newly acquired MGEs may impose an immediate biological cost in E. faecium. However, as demonstrated for vanA plasmids, the initial costs of MGE carriage may be mitigated during growth and beneficial plasmid–host association can rapidly emerge. PMID:23833178

Nonclinical and Clinical Enterococcus faecium Strains, but Not Enterococcus faecalis Strains, Have Distinct Structural and Functional Genomic Features

PubMed Central

Kim, Eun Bae

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. PMID:24141120

Isolation and identification of lactic acid bacteria from fermented red dragon fruit juices.

PubMed

Ong, Yien Yien; Tan, Wen Siang; Rosfarizan, Mohamad; Chan, Eng Seng; Tey, Beng Ti

2012-10-01

Red dragon fruit or red pitaya is rich in potassium, fiber, and antioxidants. Its nutritional properties and unique flesh color have made it an attractive raw material of various types of food products and beverages including fermented beverages or enzyme drinks. In this study, phenotypic and genotypic methods were used to confirm the identity of lactic acid bacteria (LAB) appeared in fermented red dragon fruit (Hylocereus polyrhizus) beverages. A total of 21 isolates of LAB were isolated and characterized. They belonged to the genus of Enterococcus based on their biochemical characteristics. The isolates can be clustered into two groups by using the randomly amplified polymorphic DNA method. Nucleotide sequencing and restriction fragment length polymorphism of the 16S rRNA region suggested that they were either Enterococcus faecalis or Enterococcus durans. Current research revealed the use of biochemical analyses and molecular approaches to identify the microbial population particularly lactic acid bacteria from fermented red dragon fruit juices. © 2012 Institute of Food Technologists®

Antibacterial effect of triantibiotic mixture, chlorhexidine gel, and two natural materials Propolis and Aloe vera against Enterococcus faecalis: An ex vivo study.

PubMed

Bazvand, Leila; Aminozarbian, Mohammad Ghasem; Farhad, Alireza; Noormohammadi, Hamid; Hasheminia, Seyed Mohsen; Mobasherizadeh, Sina

2014-07-01

The aim of this ex vivo study was to compare the antimicrobial effect of triantibiotic paste, 0.2% chlorhexidine gel, Propolis and Aloe vera on Enterococcus faecalis in deep dentin. Ninety fresh extracted single-rooted teeth were used in a dentin block model. Seventy-five teeth were infected with E. faecalis and divided into four experimental groups (n = 15). Experimental groups were treated with triantibiotic mixture with distilled water, 0.2% chlorhexidine gel, 70% ethanol + Propolis and Aloe vera. Fifteen teeth treated with distilled water as the positive control and 15 samples, free of bacterial contamination, were considered as the negative control. Gates-Glidden drill #4 was used for removal of surface dentin and Gates-Glidden drill #5 was used to collect samples of deep dentin. The samples were prepared and colony-forming units were counted. Data were analyzed by one-way ANOVA and post hoc Tukey tests. Statistical significance was defined at P < 0.05. Triantibiotic mixture group exhibited the least bacterial growth. However, the rate of bacterial growth showed no significant differences between chlorhexidine and Propolis groups (P > 0.05). Aloe vera had antibacterial effects on E. faecalis, but in comparison with other medicaments, it was less effective (P < 0.05). This experimental study showed that triantibiotic mixture, 0.2% chlorhexidine gel, Propolis and Aleo vera were relatively effective against E. faecalis. All the intracanal medicements had similar effects on E. faecalis in deep dentin except for Aloe vera.

An ex vivo quantification of the apically extruded bacteria following use of nickel-titanium rotary instruments.

PubMed

Mohammadi, Zahed

2007-01-01

The aim of this study was to evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using the five different engine-driven techniques utilizing nickel-titanium instruments. One-hundred and ten extracted maxillary central incisor teeth were used. Access cavities were prepared, and root canals were then contaminated with a suspension of Enterococcus faecalis and dried. The contaminated roots were divided into five experimental groups of 20 teeth each and one control group of 10 teeth: Group 1, HERO 642; Group 2, Flex Master; Group 3, Profile GT; Group 4, RoCe; Group 5, K3; and Group 6, control group. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 hours. Colonies of bacteria were counted, and the results were given as the number of colony-forming units. The data obtained were analyzed using one-way ANOVA, with p = 0.05 as the level for statistical significance. There was no significant difference as to the number of extruded bacteria between five engine-driven systems (p > 0.05). All engine-driven nickel-titanium systems extruded bacteria through the apical foramen.

Comparison of Antibacterial Effects of 810 and 980- nanometer Diode Lasers on Enterococcus Faecalis in the Root Canal System -An in vitro study.

PubMed

Asnaashari, Mohamad; Ebad, Leila Tahmasebi; Shojaeian, Shiva

2016-10-01

Background and aim: Use of laser technology in endodontics has greatly increased in the recent years due to the introduction of new wavelengths and methods and optimal antimicrobial and smear layer removal properties of lasers. This in vitro study aimed to compare the antibacterial effects of diode lasers of 810 nm and 980 nm wavelength on Enterococcus faecalis (E. faecalis) biofilm in the root canal system. Materials and methods: Fifty single-canal human anterior teeth were cleaned, shaped, sterilized and randomly divided into four groups namely two experimental, one positive and one negative control group. The experimental and positive control groups were inoculated with E. faecalis and incubated for two weeks. The experimental group one (n=20) received 810 nm diode laser irradiation (1.5W) while the experimental group two (n=20) was subjected to 980 nm diode laser irradiation (1.5W). The E. faecalis colony forming units (CFUs) were counted in each root canal before and after laser irradiation. Results: Laser irradiation significantly decreased the bacterial colony count in both experimental groups. The reduction in microbial count was significantly greater in 810 nm laser group compared to 980 nm laser group. Conclusion: Irradiation of both 810 and 980 nm lasers significantly decreased the E. faecalis count in the root canal system; 810 nm laser was more effective in decreasing the intracanal microbial load.

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

PubMed Central

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

2016-01-01

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 Phe45 in helix II and Phe18 in the α1α2 loop and a hydrogen bonding between Ser15 in helix I and Ile20 in the α1α2 loop, resulting in its high thermal stability. Phe45-mediated hydrophobic packing may block acyl chain binding subpocket II entry. Furthermore, Ser58 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. PMID:26631734

Pseudomonas aeruginosa Las quorum sensing autoinducer suppresses growth and biofilm production in Legionella species.

PubMed

Kimura, Soichiro; Tateda, Kazuhiro; Ishii, Yoshikazu; Horikawa, Manabu; Miyairi, Shinichi; Gotoh, Naomasa; Ishiguro, Masaji; Yamaguchi, Keizo

2009-06-01

Bacteria commonly communicate with each other by a cell-to-cell signalling mechanism known as quorum sensing (QS). Recent studies have shown that the Las QS autoinducer N-(3-oxododecanoyl)-l-homoserine lactone (3-oxo-C(12)-HSL) of Pseudomonas aeruginosa performs a variety of functions not only in intraspecies communication, but also in interspecies and interkingdom interactions. In this study, we report the effects of Pseudomonas 3-oxo-C(12)-HSL on the growth and suppression of virulence factors in other bacterial species that frequently co-exist with Ps. aeruginosa in nature. It was found that 3-oxo-C(12)-HSL, but not its analogues, suppressed the growth of Legionella pneumophila in a dose-dependent manner. However, 3-oxo-C(12)-HSL did not exhibit a growth-suppressive effect on Serratia marcescens, Proteus mirabilis, Escherichia coli, Alcaligenes faecalis and Stenotrophomonas maltophilia. A concentration of 50 microM 3-oxo-C(12)-HSL completely inhibited the growth of L. pneumophila. Additionally, a significant suppression of biofilm formation was demonstrated in L. pneumophila exposed to 3-oxo-C(12)-HSL. Our results suggest that the Pseudomonas QS autoinducer 3-oxo-C(12)-HSL exerts both bacteriostatic and virulence factor-suppressive activities on L. pneumophila alone.

What about Urinary Tract Infections and its Antibiotic Resistance Bacteria in Ilam, Iran?

PubMed

Mohebi, Reza; Esmaili, Khadijeh; Sadeghifard, Nourkhoda; Pakzad, Iraj; Ghafourian, Sobhan

2018-06-22

Because of unknown situation of antibiotic resistance pattern in main hospital in Ilam, Iran, in an attempt we aimed to evaluate the antibiotic resistance pattern of uropathogenic bacteria obtained from referred patients to Imam Khomaini Hospital, Ilam, Iran. For this reason, 114 bacteria were collected during 9 month period and evaluated for their antibiotic resistance pattern. Our results demonstrated that E. coli as the dominant responsible for urinary tract infection. Our results demonstrated that 61.4 % (n = 70) of isolates were positive for E.coli, while the lowest prevalence observed for Staphylococcus aureus and Acinetobacter baumannii. The results demonstrated that 6.4% (n = 7) were MBL producer. Despite, only 4 gram positive bacteria were obtained from patients with urinary tract infections but 100% (n = 1) of S. aureus were methicillin resistant S. aureus (MRSA) and 66.7% (n = 2) of E. faecalis were positive for resistance to vancomycin. In conclusion, we strongly recommended doing the cohort study among all hospital in Iran to evaluate the situation of antibiotic resistance and make a real panel to control this issue. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Antimicrobial activity of Chlorhexidine, Peracetic acid and Sodium hypochlorite/etidronate irrigant solutions against Enterococcus faecalis biofilms.

PubMed

Arias-Moliz, M T; Ordinola-Zapata, R; Baca, P; Ruiz-Linares, M; García García, E; Hungaro Duarte, M A; Monteiro Bramante, C; Ferrer-Luque, C M

2015-12-01

To evaluate the antimicrobial effect of 2.5% sodium hypochlorite alone (NaOCl) and associated with 9% HEBP (NaOCl/HEBP), 2% peracetic acid (PAA) and 2% chlorhexidine (CHX), on the viability of Enterococcus faecalis biofilms attached to dentine. Biofilms of E. faecalis were grown on the surface of dentine blocks for 5 days and then exposed to the irrigating solutions for 3 min. Distilled water was used as the control. The total biovolume and the percentage of dead cells of the infected dentine were measured by means of confocal microscopy and the live/dead technique. Nonparametric tests were used to determine statistical differences (P < 0.05). NaOCl and the NaOCl/HEBP mixture were associated with a significantly greater percentage of dead cells, followed by PAA (P < 0.05). No significant antimicrobial effect of CHX was observed in comparison with the control group. Total biovolume decreased significantly in NaOCl, NaOCl/HEBP and PAA solutions in comparison with the CHX and control groups. NaOCl alone or associated with HEBP were the most effective irrigant solutions in dissolving and killing E. faecalis biofilms. © 2014 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Biochemical and Structural Basis for Inhibition of Enterococcus faecalis Hydroxymethylglutaryl-CoA Synthase, mvaS, by Hymeglusin

DOE Office of Scientific and Technical Information (OSTI.GOV)

Skaff, D. Andrew; Ramyar, Kasra X.; McWhorter, William J.

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;more » 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.« less

Demetallization of Enterococcus faecalis biofilm: a preliminary study

PubMed Central

ESTRELA, Carlos; COSTA E SILVA, Rodrigo; URBAN, Roberta Cerasi; GONÇALVES, Pablo José; SILVA, Júlio A.; ESTRELA, Cyntia R.A.; PECORA, Jesus Djalma; PETERS, Ove A.

2018-01-01

Abstract Objectives To determine the concentration of calcium, iron, manganese and zinc ions after the application of chelator to Enterococcus faecalis biofilms. Material and Methods Fifty bovine maxillary central incisors were prepared and inoculated with E. faecalis for 60 days. The following were used as irrigation solutions: 17% EDTA (pH 3, 7 and 10), 2.5% sodium hypochlorite (NaOCl) combined with 17% EDTA (pH 3, 7 and 10), distilled water (pH 3, 7 and 10), and 2.5% NaOCl. Each solution was kept in the root canal for five minutes. Fifteen uncontaminated root canals were irrigated with 17% EDTA (pH 3, 7 and 10). Six teeth were used as bacterial control. The number of calcium, iron, manganese and zinc ions was determined using flame atomic absorption spectrometry. Mean ± standard deviation (SD) values were used for descriptive statistics. Results Calcium chelation using 17% EDTA at pH 7 was higher than at pH 3 and 10, regardless of whether bacterial biofilm was present. The highest concentration of iron occurred at pH 3 in the presence of bacterial biofilm. The highest concentration of manganese found was 2.5% NaOCl and 17% EDTA at pH 7 in the presence of bacterial biofilm. Zinc levels were not detectable. Conclusions The pH of chelating agents affected the removal of calcium, iron, and manganese ions. The concentration of iron ions in root canals with bacterial biofilm was higher after the use of 17% EDTA at pH 3 than after the use of the other solutions at all pH levels. PMID:29451651

FORMATE—PYRUVATE EXCHANGE REACTION IN STREPTOCOCCUS FAECALIS II.

PubMed Central

Oster, M. O.; Wood, N. P.

1964-01-01

Oster, M. O. (A. & M. College of Texas, College Station), and N. P. Wood. Formate-pyruvate exchange reaction in Streptococcus faecalis. II. Reaction conditions for cell extracts. J. Bacteriol. 87:104–113. 1964.—In contrast to intact cells of Streptococcus faecalis, no stimulation of the formate-pyruvate exchange reaction was observed in cell extracts when yeast extract was added to the reaction mixture. A heated extract of Micrococcus lactilyticus, vitamin K5, ferrous sulfate, and ferrous ammonium sulfate stimulated an active exchange by protecting the system from oxygen. Tetrahydrofolate, 2,3-dimercaptopropanol, and sodium sulfide provided partial protection, whereas ascorbate, glutathione, sodium hydrosulfite, ammonium sulfide, and sodium bisulfite gave insufficient protection or were inhibitory. Oxidation-reduction (O-R) indicators were not inhibitory and were used to estimate the O-R potentials of reaction mixtures. A potential at least as negative as −125 mv was estimated to be necessary to preserve or initiate formate-pyruvate exchange activity. The reaction operated over a narrow pH range when strict anaerobic conditions were not maintained but, when the system was suitably poised, the pH range was broader. The influence of high phosphate concentrations was less under strictly anaerobic conditions, and orthophosphate could be replaced by small amounts of pyrophosphate. Effect of temperature, time, and amount of extract is presented. Addition of reduced benzyl viologen and hydrogen-saturated palladium in the buffer during 8 hr of dialysis prevented inactivation of extracts. Recovery of activity could be obtained after ammonium sulfate treatment when a combination of palladium chloride, neutral red, and hydrogen bubbling were used. PMID:14102842

Relevance of the clustered regularly interspaced short palindromic repeats of Enterococcus faecalis strains isolated from retreatment root canals on periapical lesions, resistance to irrigants and biofilms

PubMed Central

Tong, Zhongchun; Du, Yu; Ling, Junqi; Huang, Lijia; Ma, Jinglei

2017-01-01

A high prevalence of Enterococcus faecalis (E. faecalis) is observed in teeth with root canal treatment failures. Clustered regularly interspaced short palindromic repeats (CRISPR) are widely distributed in prokaryotes that have adaptive immune systems against mobile elements, including pathogenic genes. The present study investigated the relevance of the CRISPR in E. faecalis strains isolated from retreated root canals on biofilms, periapical lesions and drug resistance. A total of 20 E. faecalis strains were extracted from the root canals of teeth referred for root canal retreatment. CRISPR-Cas loci were identified by two pairs of relevant primers and polymerase chain reaction. The susceptibility of the 20 isolated strains to intracanal irrigants was evaluated by 1- and 5-minute challenges with a mixture of a tetracycline isomer, an acid and a detergent (MTAD), 2% chlorhexidine (CHX) and 5.25% sodium hypochlorite (NaOCl). The microtiter plate assay and crystal violet staining were used to compare the biofilm formation of the E. faecalis isolate strains. Out of the 20 E. faecalis isolate strains, 5 strains that lacked CRISPR-cas determinants exhibited significant periapical lesions. Among the 15 strains containing CRISPR-cas determinants, 8 were isolated from root canals with inadequate fillings and 7 were isolated from root canals without any fillings. The five strains lacking CRISPR-cas loci were observed to be more resistant to MTAD and 2% CHX than the 15 strains that had CRISPR-cas loci. All of the strains exhibited the same susceptibility to 5.25% NaOCl. Furthermore, the 5 strains lacking CRISPR-cas determinants generated more biofilm than the other 15 strains. Thus, the results of the present study suggested that E. faecalis root canal isolates lacking CRISPR-cas exhibit higher resistance to intracanal irrigants, stronger biofilm formation and generate significant periapical lesions. PMID:29285081

Relevance of the clustered regularly interspaced short palindromic repeats of Enterococcus faecalis strains isolated from retreatment root canals on periapical lesions, resistance to irrigants and biofilms.

PubMed

Tong, Zhongchun; Du, Yu; Ling, Junqi; Huang, Lijia; Ma, Jinglei

2017-12-01

A high prevalence of Enterococcus faecalis ( E. faecalis ) is observed in teeth with root canal treatment failures. Clustered regularly interspaced short palindromic repeats (CRISPR) are widely distributed in prokaryotes that have adaptive immune systems against mobile elements, including pathogenic genes. The present study investigated the relevance of the CRISPR in E. faecalis strains isolated from retreated root canals on biofilms, periapical lesions and drug resistance. A total of 20 E. faecalis strains were extracted from the root canals of teeth referred for root canal retreatment. CRISPR-Cas loci were identified by two pairs of relevant primers and polymerase chain reaction. The susceptibility of the 20 isolated strains to intracanal irrigants was evaluated by 1- and 5-minute challenges with a mixture of a tetracycline isomer, an acid and a detergent (MTAD), 2% chlorhexidine (CHX) and 5.25% sodium hypochlorite (NaOCl). The microtiter plate assay and crystal violet staining were used to compare the biofilm formation of the E. faecalis isolate strains. Out of the 20 E. faecalis isolate strains, 5 strains that lacked CRISPR-cas determinants exhibited significant periapical lesions. Among the 15 strains containing CRISPR-cas determinants, 8 were isolated from root canals with inadequate fillings and 7 were isolated from root canals without any fillings. The five strains lacking CRISPR-cas loci were observed to be more resistant to MTAD and 2% CHX than the 15 strains that had CRISPR-cas loci. All of the strains exhibited the same susceptibility to 5.25% NaOCl. Furthermore, the 5 strains lacking CRISPR-cas determinants generated more biofilm than the other 15 strains. Thus, the results of the present study suggested that E. faecalis root canal isolates lacking CRISPR-cas exhibit higher resistance to intracanal irrigants, stronger biofilm formation and generate significant periapical lesions.

[The antagonistic properties of microaerophilic bacteria isolated from the human and mink digestive tracts].

PubMed

Sudenko, V I; Groma, L I; Podgorskiĭ, V S

1996-01-01

Study of antagonistic properties of microaerophilic bacteria isolated from human and mink gastroenteric tract have helped to establish differences in species composition, quantity and level of antagonistic activity of the studied microorganisms in respect to pathogenic microflora. It is shown that lactic acid bacteria identified as Lactobacillus fermentum and L. reuteri prevail among the strains isolated from the stomach and thin intestine of minks kept in the 30-km zone of Chernobyl NPP. Species composition of microaerophilic bacteria isolated from the digestive tract of the control minks is more variable. Antagonistically active bifidobacteria prevail in large intestine of experimental and control animals. Strains of lactic acid bacteria with the expressed antagonistic activity belonging to L. bavaricus, L. reuteri, L. coryniformis and L. maltaromicus have been found parallel with such known producers of antibiotic-like substances as L. fermentum. L. acidophilum. Streptococcus faecalis and bifidobacteria. L. maltaromicus most frequently occurred among antagonistically active strains revealed in feces of people which stayed in the zone of liquidation of the Chernobyl accident. Microaerophilic strains of bacteria (lactic acid, bifidobacteria and enterococci) manifest the expressed antagonistic activity connected with the capacity to not only acid formation but also to accumulation of antibiotic products of unknown nature. A strain of lactic acid bacteria L. fermentum 91 has been isolated from the contents of human gastroenteric tract. These bacteria are distinguished by most expressed and stable antagonism and characterized by the lack of pathogenicity in respect of albino mice that may be used to raise the microorganism resistance to gastric diseases.

Role of N-acetylglucosaminidase and N-acetylmuramidase activities in Enterococcus faecalis peptidoglycan metabolism.

PubMed

Mesnage, Stéphane; Chau, Françoise; Dubost, Lionel; Arthur, Michel

2008-07-11

Identification of the full complement of peptidoglycan hydrolases detected by zymogram in Enterococcus faecalis extracts led to the characterization of two novel hydrolases that we named AtlB and AtlC. Both enzymes have a similar modular organization comprising a central catalytic domain fused to two LysM peptidoglycan-binding modules. AtlB and AtlC displayed N-acetylmuramidase activity, as demonstrated by tandem mass spectrometry analyses of peptidoglycan fragments generated by the purified enzymes. The genes encoding AtlB and AtlC were deleted either alone or in combination with the gene encoding AtlA, a previously described N-acetylglucosaminidase. No autolytic activity was detected in the triple mutant indicating that AtlA, AtlB, and AtlC account for the major hydrolytic activities in E. faecalis. Analysis of cell size distribution by flow cytometry showed that deletion of atlA resulted in the formation of long chains. Thus, AtlA digests the septum and is required for cell separation after cell division. We found that AtlB could act as a surrogate for AtlA, although the enzyme was less efficient at septum digestion. Deletion of atlC had no impact on cell morphology. Labeling of the peptidoglycan with N-[14C]acetylglucosamine revealed an unusually slow turnover as compared with model organisms, almost completely dependent upon the combined activities of AtlA and AtlB. In contrast to atlA, the atlB and atlC genes are located in putative prophages. Because AtlB and AtlC were produced in the absence of cell lysis or production of phage progeny, these enzymes may have been hijacked by E. faecalis to contribute to peptidoglycan metabolism.

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.

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

PubMed Central

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. PMID:26005332

Comparison of Antibacterial Effects of 810 and 980- nanometer Diode Lasers on Enterococcus Faecalis in the Root Canal System —An in vitro study

PubMed Central

Asnaashari, Mohamad; Ebad, Leila Tahmasebi

2016-01-01

Background and aim: Use of laser technology in endodontics has greatly increased in the recent years due to the introduction of new wavelengths and methods and optimal antimicrobial and smear layer removal properties of lasers. This in vitro study aimed to compare the antibacterial effects of diode lasers of 810 nm and 980 nm wavelength on Enterococcus faecalis (E. faecalis) biofilm in the root canal system. Materials and methods: Fifty single-canal human anterior teeth were cleaned, shaped, sterilized and randomly divided into four groups namely two experimental, one positive and one negative control group. The experimental and positive control groups were inoculated with E. faecalis and incubated for two weeks. The experimental group one (n=20) received 810 nm diode laser irradiation (1.5W) while the experimental group two (n=20) was subjected to 980 nm diode laser irradiation (1.5W). The E. faecalis colony forming units (CFUs) were counted in each root canal before and after laser irradiation. Results: Laser irradiation significantly decreased the bacterial colony count in both experimental groups. The reduction in microbial count was significantly greater in 810 nm laser group compared to 980 nm laser group. Conclusion: Irradiation of both 810 and 980 nm lasers significantly decreased the E. faecalis count in the root canal system; 810 nm laser was more effective in decreasing the intracanal microbial load. PMID:27853346

Biochemical and genetic analyses of acetoin catabolism in Alcaligenes eutrophus.

PubMed Central

Fründ, C; Priefert, H; Steinbüchel, A; Schlegel, H G

1989-01-01

In genetic studies on the catabolism of acetoin in Alcaligenes eutrophus, we used Tn5::mob-induced mutants which were impaired in the utilization of acetoin as the sole carbon source for growth. The transposon-harboring EcoRI restriction fragments from 17 acetoin-negative and slow-growing mutants (class 2a) and from six pleiotropic mutants of A. eutorphus, which were acetoin-negative and did not grow chemolithoautotrophically (class 2b), were cloned from pHC79 gene banks. The insertions of Tn5 were mapped on four different chromosomal EcoRI restriction fragments (A, C, D, and E) in class 2a mutants. The native DNA fragments were cloned from a lambda L47 or from a cosmid gene bank. Evidence is provided that fragments A (21 kilobase pairs [kb]) and C (7.7 kb) are closely linked in the genome; the insertions of Tn5 covered a region of approximately 5 kb. Physiological experiments revealed that this region encodes for acetoin:dichlorophenol-indophenol oxidoreductase, a fast-migrating protein, and probably for one additional protein that is as yet unknown. In mutants which were not completely impaired in growth on acetoin but which grew much slower and after a prolonged lag phase, fragments D (7.2 kb) and E (8.1 kb) were inactivated by insertion of Tn5::mob. No structural gene could be assigned to the D or E fragments. In class 2b mutants, insertions of Tn5 were mapped on fragment B (11.3 kb). This fragment complemented pleiotropic hno mutants in trans; these mutants were impaired in the formation of a rpoN-like protein. The expression of the gene cluster on fragments A and C seemed to be rpoN dependent. PMID:2556366

Production of poly-beta-hydroxybutyrate (PHB) by Alcaligenes latus from maple sap.

PubMed

Yezza, Abdessalem; Halasz, Annamaria; Levadoux, Wayne; Hawari, Jalal

2007-11-01

Maple sap, an abundant natural product especially in Canada, is rich in sucrose and thus may represent an ideal renewable feedstock for the production of a wide variety of value-added products. In the present study, maple sap or sucrose was employed as a carbon source to Alcaligenes latus for the production of poly-beta-hydroxybutyrate (PHB). In shake flasks, the biomass obtained from both the sap and sucrose were 4.4 +/- 0.5 and 2.9 +/- 0.3 g/L, and the PHB contents were 77.6 +/- 1.5 and 74.1 +/- 2.0%, respectively. Subsequent batch fermentation (10 L sap) resulted in the formation of 4.2 +/- 0.3 g/L biomass and a PHB content of 77.0 +/- 2.6%. The number average molecular weights of the PHB produced by A. latus from maple sap and pure sucrose media were 300 +/- 66 x 10(3) and 313 +/- 104 x 10(3) g/mol, respectively. Near-infrared, (1)H magnetic resonance imaging (MRI), and (13)C-MRI spectra of the microbially produced PHB completely matched those obtained with a reference material of poly[(R)-3-hydroxybutyric acid]. The polymer was found to be optically active with [alpha](25) (D) equaled to -7.87 in chloroform. The melting point (177.0 degrees C) and enthalpy of fusion (77.2 J/g) of the polymer were also in line with those reported, i.e., 177 degrees C and 81 J/g, respectively.

Thermodynamics of ligand binding to histone deacetylase like amidohydrolase from Bordetella/Alcaligenes.

PubMed

Meyners, Christian; Baud, Matthias G J; Fuchter, Matthew J; Meyer-Almes, Franz-Josef

2014-03-01

Thermodynamic studies on ligand-protein binding have become increasingly important in the process of drug design. In combination with structural data and molecular dynamics simulations, thermodynamic studies provide relevant information about the mode of interaction between compounds and their target proteins and therefore build a sound basis for further drug optimization. Using the example of histone deacetylases (HDACs), particularly the histone deacetylase like amidohydrolase (HDAH) from Bordetella/Alcaligenes, a novel sensitive competitive fluorescence resonance energy transfer-based binding assay was developed and the thermodynamics of interaction of both fluorescent ligands and inhibitors to histone deacetylase like amidohydrolase were investigated. The assay consumes only small amounts of valuable target proteins and is suitable for fast kinetic and mechanistic studies as well as high throughput screening applications. Binding affinity increased with increasing length of aliphatic spacers (n = 4-7) between the hydroxamate moiety and the dansyl head group of ligand probes. Van't Hoff plots revealed an optimum in enthalpy contribution to the free energy of binding for the dansyl-ligand with hexyl spacer. The selectivity in the series of dansyl-ligands against human class I HDAC1 but not class II HDACs 4 and 6 increased with the ratio of ΔH(0)/ΔG(0). The data clearly emphasize the importance of thermodynamic signatures as useful general guidance for the optimization of ligands or rational drug design. Copyright © 2014 John Wiley & Sons, Ltd.

Development of eco-friendly bioplastic like PHB by distillery effluent microorganisms.

PubMed

Gangurde, Nilesh S; Sayyed, Riyaz Z; Kiran, Shashi; Gulati, Arvind

2013-01-01

During screening for poly-β-hydroxybutyrate (PHB) producing bacteria from distillery effluent sample, six out of 30 isolates comprising of three strains of Alcaligenes sp., two strains of Bacillus sp., and one strain of Pseudomonas sp. were found to accumulate varying levels of intracellular PHB. Amongst the various isolates, Alcaligenes sp. RZS4 was found as the potent PHB-producing organism, accumulating higher amounts of PHB. PHB productivity was further enhanced in the presence of oxygen, nitrogen-limiting conditions, and cloning of PHB synthesizing genes of Alcaligenes sp. RZS 4 into Escherichia coli. A twofold increase in PHB yield was obtained from recombinant E. coli vis-à-vis Alcaligenes sp.; the recombinant E. coli accumulated more PHB in NDMM, produced good amount of PHB in a single-stage cultivation process under both nutrient-rich and nutrient-deficient conditions. Extraction of PHB with acetone-alcohol (1:1) was found as suitable method for optimum extraction of PHB as this mixture selectively extracted PHB without affecting the non-PHB cell mass. PHB extract from recombinant E. coli showed the presence of C-H, =O stretching, =C-H deformation, =C-H, =CH, and =C-O functional groups characteristic of PHB.

Antibacterial Effect of Azadirachta indica (Neem) or Curcuma longa (Turmeric) against Enterococcus faecalis Compared with That of 5% Sodium Hypochlorite or 2% Chlorhexidine in vitro.

PubMed

Joy Sinha, Dakshita; D S Nandha, Kanwar; Jaiswal, Natasha; Vasudeva, Agrima; Prabha Tyagi, Shashi; Pratap Singh, Udai

2017-01-01

The purpose of this study was to compare the antibacterial properties of Azadirachta indica (neem) or Curcuma longa (turmeric) against Enterococcus faecalis with those of 5% sodium hypochlorite or 2% chlorhexidine as root canal irrigants in vitro. The activity of neem, chlorhexidine, sodium hypochlorite, or turmeric against E. faecalis was measured on agar plates using the agar diffusion method. The tube dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the irrigants used. Chlorhexidine or neem exhibited the greatest antibacterial activity when used as endodontic irrigants against E. faecalis, followed by sodium hypochlorite. No statistically significant difference was observed between neem, sodium hypochlorite, or chlorhexidine. The MIC of neem was 1: 128, which was similar to that of chlorhexidine. The MBC for each of these irrigants was 1: 16. Neem yielded antibacterial activity equivalent to 2% chlorhexidine or sodium hypochlorite against E. faecalis, suggesting that it offers a promising alternative to the other root canal irrigants tested.

Comparison of apical extrusion of intracanal bacteria by various glide-path establishing systems: an in vitro study.

PubMed

Dagna, Alberto; El Abed, Rashid; Hussain, Sameeha; Abu-Tahun, Ibrahim H; Visai, Livia; Bertoglio, Federico; Bosco, Floriana; Beltrami, Riccardo; Poggio, Claudio; Kim, Hyeon-Cheol

2017-11-01

This study compared the amount of apically extruded bacteria during the glide-path preparation by using multi-file and single-file glide-path establishing nickel-titanium (NiTi) rotary systems. Sixty mandibular first molar teeth were used to prepare the test apparatus. They were decoronated, blocked into glass vials, sterilized in ethylene oxide gas, infected with a pure culture of Enterococcus faecalis, randomly assigned to 5 experimental groups, and then prepared using manual stainless-steel files (group KF) and glide-path establishing NiTi rotary files (group PF with PathFiles, group GF with G-Files, group PG with ProGlider, and group OG with One G). At the end of canal preparation, 0.01 mL NaCl solution was taken from the experimental vials. The suspension was plated on brain heart infusion agar and colonies of bacteria were counted, and the results were given as number of colony-forming units (CFU). The manual instrumentation technique tested in group KF extruded the highest number of bacteria compared to the other 4 groups ( p < 0.05). The 4 groups using rotary glide-path establishing instruments extruded similar amounts of bacteria. All glide-path establishment instrument systems tested caused a measurable apical extrusion of bacteria. The manual glide-path preparation showed the highest number of bacteria extruded compared to the other NiTi glide-path establishing instruments.

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.

Using Phage Lytic Enzymes to Destroy Pathogenic and BW Bacteria

DTIC Science & Technology

2005-07-14

against antibiotic resistant Enterococcus faecalis and Enterococcus faecium . J Bacteriol. 186:4808-12. Cheng, Q., D. Nelson, S. Zhu, and V.A...Lysins from Enterococcus faecalis RU-654 3. Fischetti, Vincent A. Schuch, Raymond Lytic Enzymes and spore surface antigens for detection and

Streptococcus gordonii pheromone s.g.cAM373 may influence the reservoir of antibiotic resistance determinants of Enterococcus faecalis origin in the oral metagenome.

PubMed

Mansfield, Jillian M; Herrmann, Paul; Jesionowski, Amy M; Vickerman, M Margaret

2017-11-01

Streptococcus gordonii produces a pheromone heptapeptide, s.g.cAM373, which induces a conjugative mating response in Enterococcus faecalis cells carrying the responsive plasmid, pAM373. We investigated the extent of this intergeneric signaling on DNA acquisition by streptococcal species likely to cohabit oral biofilms. E. faecalis/pAM373/pAMS470 cells were incubated with synthetic s.g.cAM373, reverse peptide s.g.cAM373-R, or peptide-free medium and examined for their abilities to transfer plasmid DNA to streptococcal species in the presence of DNase. Preinduction of E. faecalis donors with s.g.cAM373 resulted in transconjugation frequencies in non-pheromone producing strains of Streptococcus mutans, Streptococcus sanguinis, Streptococcus anginosus, and Streptococcus suis that were significantly higher than frequencies when donors were preincubated with s.g.cAM373-R or medium alone. Peptide-mediated communication between commensal streptococci and E. faecalis carrying pheromone-responsive plasmids may facilitate conjugative DNA transfer to bystander species, and influence the reservoir of antibiotic resistance determinants of enterococcal origin in the oral metagenome.

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

Antimicrobial activity of sodium hypochlorite associated with intracanal medication for Candida albicans and Enterococcus faecalis inoculated in root canals.

PubMed

Valera, Marcia Carneiro; Silva, Katy Costa Godinho da; Maekawa, Lilian Eiko; Carvalho, Cláudio Antonio Talge; Koga-Ito, Cristiane Yumi; Camargo, Carlos Henrique Ribeiro; Lima, Raphael Silva e

2009-01-01

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. 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. Both 1% NaOCl irrigation and the intracanal medications were effective in eliminating E. faecalis and C. albicans inoculated in root canals.

Identification of bacteria in drinking and purified water during the monitoring of a typical water purification system

PubMed Central

Penna, Vessoni Thereza Christina; Martins, Silva Alzira Maria; Mazzola, Priscila Gava

2002-01-01

Background A typical purification system that provides purified water which meets ionic and organic chemical standards, must be protected from microbial proliferation to minimize cross-contamination for use in cleaning and preparations in pharmaceutical industries and in health environments. Methodology Samples of water were taken directly from the public distribution water tank at twelve different stages of a typical purification system were analyzed for the identification of isolated bacteria. Two miniature kits were used: (i) identification system (api 20 NE, Bio-Mérieux) for non-enteric and non-fermenting gram-negative rods; and (ii) identification system (BBL crystal, Becton and Dickson) for enteric and non-fermenting gram-negative rods. The efficiency of the chemical sanitizers used in the stages of the system, over the isolated and identified bacteria in the sampling water, was evaluated by the minimum inhibitory concentration (MIC) method. Results The 78 isolated colonies were identified as the following bacteria genera: Pseudomonas, Flavobacterium and Acinetobacter. According to the miniature kits used in the identification, there was a prevalence of isolation of P. aeruginosa 32.05%, P. picketti (Ralstonia picketti) 23.08%, P. vesiculares 12.82%,P. diminuta 11.54%, F. aureum 6.42%, P. fluorescens 5.13%, A. lwoffi 2.56%, P. putida 2.56%, P. alcaligenes 1.28%, P. paucimobilis 1.28%, and F. multivorum 1.28%. Conclusions We found that research was required for the identification of gram-negative non-fermenting bacteria, which were isolated from drinking water and water purification systems, since Pseudomonas genera represents opportunistic pathogens which disperse and adhere easily to surfaces, forming a biofilm which interferes with the cleaning and disinfection procedures in hospital and industrial environments. PMID:12182763

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

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. Copyright © 2015 Elsevier B.V. All rights reserved.

An in vitro evaluation of the antibacterial efficacy of chlorine dioxide on E. faecalis in bovine incisors.

PubMed

Eddy, Russell S; Joyce, Anthony P; Roberts, Steven; Buxton, Thomas B; Liewehr, Frederick

2005-09-01

This study investigated the ability of chlorine dioxide to eliminate Enterococcus faecalis from dentinal tubules of bovine incisors. Thirty-seven extracted bovine incisor roots were sectioned into seventy-four 5 mm disks. Standardized lumens were filled with either sterile Brain Heart Infusion Broth (contamination controls, n = 10) or BHI containing E. faecalis (1.0 x 10 cfu/ml). Disks were incubated in 5% CO2 at 37 degrees C for 72 h. To simulate endodontic instrumentation the lumens were again enlarged. Sixty disks were randomly divided into four experimental groups and filled with one of the following irrigants: 10% Clidox-S (chlorine dioxide), 13.8% BioClenz (chlorine dioxide), 5.25% Clorox, or saline. The disks were incubated for 30 min and were then frozen, pulverized, serially diluted in phosphate buffered saline, and plated on BHI plates in triplicate. Total colony forming units were counted macroscopically. Statistical analysis of the data was performed with a Kruskal-Wallis one-way ANOVA on ranks (p < 0.05, n = 60). Bacterial counts, expressed in log10 cfu/disk were as follows (">" denotes significant differences): Saline > Clidox-S = BioClenz > Clorox. All negative controls were sterile. Chlorine dioxide and NaOCL were both effective in eliminating E. faecalis from the dentinal disks within 30 min.

Bacillus subtilis as a Platform for Molecular Characterisation of Regulatory Mechanisms of Enterococcus faecalis Resistance against Cell Wall Antibiotics

PubMed Central

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. PMID:24676422

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.

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. Published by Elsevier B.V.

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

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. Copyright © 2015 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Assessment of microbiological quality of sachet-packaged drinking water in Western Nigeria and its public health significance.

PubMed

Olaoye, O A; Onilude, A A

2009-11-01

To assess the microbiological quality of sachet-packaged drinking water in Western Nigeria and its impact on public health. Cross-sectional microbiological testing. Ninety-two sachet-packaged water samples were analysed for microbiological and metal qualities. Total bacterial and coliform counts were determined, and the presence of Escherichia coli, an important water quality indicator, was tested. The level of conformity of the water processors with the guidelines of Nigeria's quality regulatory agency was also determined. Varying levels of microbial contamination were recorded in samples from the different sampling locations. The total bacteria count ranged between 2.86 and 3.45log colony-forming units (cfu)/ml. The highest coliform count recorded was 1.62log cfu/ml. Faecal coliform E. coli was detected in one sample from Oke-Iho and one sample from Okaka, representing 2.2% of total samples. Lead and manganese were not found in any of the samples. However, iron was detected and the highest iron concentration (0.10mg/l) was detected in samples from Ikorodu. The bacteria that were identified from the water samples included E. coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Klebsiella sp., Proteus vulgaris, Alcaligenes faecalis, Bacillus cereus, Staphylococcus aureus, Streptococcus lactis, Aeromonas sp. and Micrococcus luteum. Many of the water processors did not comply with the guidelines of the quality regulatory agency. Some of the sachet-packaged samples of drinking water were of poor quality. The results indicate a need for Nigeria's quality regulatory agency to take appropriate measures in safeguarding public health.

Microbiological characterization of aquatic microbiomes targeting taxonomical marker genes and antibiotic resistance genes of opportunistic bacteria.

PubMed

Alexander, Johannes; Bollmann, Anna; Seitz, Wolfram; Schwartz, Thomas

2015-04-15

The dissemination of medically relevant antibiotic resistance genes (ARGs) (blaVIM-1, vanA, ampC, ermB, and mecA) and opportunistic bacteria (Enterococcus faecium/faecalis, Pseudomonas aeruginosa, Enterobacteriaceae, Staphylococcus aureus, and CNS) was determined in different anthropogenically influenced aquatic habitats in a selected region of Germany. Over a period of two years, four differently sized wastewater treatment plants (WWTPs) with and without clinical influence, three surface waters, four rain overflow basins, and three groundwater sites were analyzed by quantitative Polymerase Chain Reaction (qPCR). Results were calculated in cell equivalents per 100 ng of total DNA extracted from water samples and per 100 mL sample volume, which seems to underestimate the abundance of antibiotic resistance and opportunistic bacteria. High abundances of opportunistic bacteria and ARG were quantified in clinical wastewaters and influents of the adjacent WWTP. The removal capacities of WWTP were up to 99% for some, but not all investigated bacteria. The abundances of most ARG targets were found to be increased in the bacterial population after conventional wastewater treatment. As a consequence, downstream surface water and also some groundwater compartments displayed high abundances of all four ARGs. It became obvious that the dynamics of the ARG differed from the fate of the opportunistic bacteria. This underlines the necessity of an advanced microbial characterization of anthropogenically influenced environments. Copyright © 2015 Elsevier B.V. All rights reserved.

Cellular and Molecular Approaches to Polymer Synthesis by Bacteria

DTIC Science & Technology

1989-03-01

of biodegradable polymers f rom Pseudowonas oleovorans.....L (See appended research summaries pp. 1-4.) 20. DISTRIBUTIONI/AVAILABILITY OF ABSTRACT 21...during the growth of the organism , polymer production is greatest and harvesting gives maximum PHA vield. Other experiments have also been conducted...Functional PHA from Rhodospirillum and Alcaligenes The major emphasis is to produce functional biodegradable polymers and new, totally biodegradable

Study on the biodegradation of crude oil by free and immobilized bacterial consortium in marine environment.

PubMed

Chen, Qingguo; Li, Jingjing; Liu, Mei; Sun, Huiling; Bao, Mutai

2017-01-01

Five strains of bacteria, namely, Exiguobacterium sp. ASW-1, Pseudomonas aeruginosa strain ASW-2, Alcaligenes sp. ASW-3, Alcaligenes sp. ASS-1, and Bacillus sp. ASS-2, were isolated from the Zhejiang coast in China. The mixed flora of the five strains performed well with degrading 75.1% crude oil (1%, w/v) in 7 days. The calcium alginate-activated carbon embedding carrier was used to immobilize bacterial consortium. Immobilized cells performed better than free ones in variations of environmental factors containing incubated temperature, initial pH, salinity of the medium and crude oil concentration. The degradation process of crude oil by immobilized bacteria was accelerated compared with that of the free ones. Bacterial consortium showed better performance on biodegradation of normal alkanes than that of PAHs. Improvement of immobilization on the biodegradation efficiency of normal alkanes (31.9%) was apparently high than that of PAHs (1.9%).

Study on the biodegradation of crude oil by free and immobilized bacterial consortium in marine environment

PubMed Central

Li, Jingjing; Liu, Mei; Sun, Huiling; Bao, Mutai

2017-01-01

Five strains of bacteria, namely, Exiguobacterium sp. ASW-1, Pseudomonas aeruginosa strain ASW-2, Alcaligenes sp. ASW-3, Alcaligenes sp. ASS-1, and Bacillus sp. ASS-2, were isolated from the Zhejiang coast in China. The mixed flora of the five strains performed well with degrading 75.1% crude oil (1%, w/v) in 7 days. The calcium alginate—activated carbon embedding carrier was used to immobilize bacterial consortium. Immobilized cells performed better than free ones in variations of environmental factors containing incubated temperature, initial pH, salinity of the medium and crude oil concentration. The degradation process of crude oil by immobilized bacteria was accelerated compared with that of the free ones. Bacterial consortium showed better performance on biodegradation of normal alkanes than that of PAHs. Improvement of immobilization on the biodegradation efficiency of normal alkanes (31.9%) was apparently high than that of PAHs (1.9%). PMID:28346510

Growth- and Stress-Induced PASTA Kinase Phosphorylation in Enterococcus faecalis.

PubMed

Labbe, Benjamin D; Kristich, Christopher J

2017-11-01

Transmembrane Ser/Thr kinases containing extracellular PASTA domains are ubiquitous among Actinobacteria and Firmicutes Such PASTA kinases regulate critical processes, including antibiotic resistance, cell division, toxin production, and virulence, and are essential for viability in certain organisms. Based on in vitro studies with purified extracellular and intracellular fragments of PASTA kinases, a model for signaling has been proposed, in which the extracellular PASTA domains bind currently undefined ligands (typically thought to be peptidoglycan, or fragments thereof) to drive kinase dimerization, which leads to enhanced kinase autophosphorylation and enhanced phosphorylation of substrates. However, this model has not been rigorously tested in vivo Enterococcus faecalis is a Gram-positive intestinal commensal and major antibiotic-resistant opportunistic pathogen. In E. faecalis , the PASTA kinase IreK drives intrinsic resistance to cell wall-active antimicrobials, suggesting that such antimicrobials may trigger IreK signaling. Here we show that IreK responds to cell wall stress in vivo by enhancing its phosphorylation and that of a downstream substrate. This response requires both the extracellular PASTA domains and specific phosphorylatable residues in the kinase domain. Thus, our results provide in vivo evidence, with an intact full-length PASTA kinase in its native physiological environment, that supports the prevailing model of PASTA kinase signaling. In addition, we show that IreK responds to a signal associated with growth and/or cell division, in the absence of cell wall-active antimicrobials. Surprisingly, the ability of IreK to respond to growth and/or division does not require the extracellular PASTA domains, suggesting that IreK monitors multiple parameters for sensory input in vivo IMPORTANCE Transmembrane Ser/Thr kinases containing extracellular PASTA domains are ubiquitous among Actinobacteria and Firmicutes and regulate critical processes. The

Growth- and Stress-Induced PASTA Kinase Phosphorylation in Enterococcus faecalis

PubMed Central

Labbe, Benjamin D.

2017-01-01

ABSTRACT Transmembrane Ser/Thr kinases containing extracellular PASTA domains are ubiquitous among Actinobacteria and Firmicutes. Such PASTA kinases regulate critical processes, including antibiotic resistance, cell division, toxin production, and virulence, and are essential for viability in certain organisms. Based on in vitro studies with purified extracellular and intracellular fragments of PASTA kinases, a model for signaling has been proposed, in which the extracellular PASTA domains bind currently undefined ligands (typically thought to be peptidoglycan, or fragments thereof) to drive kinase dimerization, which leads to enhanced kinase autophosphorylation and enhanced phosphorylation of substrates. However, this model has not been rigorously tested in vivo. Enterococcus faecalis is a Gram-positive intestinal commensal and major antibiotic-resistant opportunistic pathogen. In E. faecalis, the PASTA kinase IreK drives intrinsic resistance to cell wall-active antimicrobials, suggesting that such antimicrobials may trigger IreK signaling. Here we show that IreK responds to cell wall stress in vivo by enhancing its phosphorylation and that of a downstream substrate. This response requires both the extracellular PASTA domains and specific phosphorylatable residues in the kinase domain. Thus, our results provide in vivo evidence, with an intact full-length PASTA kinase in its native physiological environment, that supports the prevailing model of PASTA kinase signaling. In addition, we show that IreK responds to a signal associated with growth and/or cell division, in the absence of cell wall-active antimicrobials. Surprisingly, the ability of IreK to respond to growth and/or division does not require the extracellular PASTA domains, suggesting that IreK monitors multiple parameters for sensory input in vivo. IMPORTANCE Transmembrane Ser/Thr kinases containing extracellular PASTA domains are ubiquitous among Actinobacteria and Firmicutes and regulate critical

Some Technological Properties of Lactic Acid Bacteria Isolated from Dahi and Datshi, Naturally Fermented Milk Products of Bhutan

PubMed Central

Shangpliang, H. N. J.; Sharma, Sharmila; Rai, Ranjita; Tamang, Jyoti P.

2017-01-01

Dahi and datshi are common naturally fermented milk (NFM) products of Bhutan. Population of lactic acid bacteria (LAB) in dahi (pH 3.7) and datshi (pH 5.2) was 1.4 × 107 and 3.9 × 108 cfu/ml, respectively. Based on 16S rRNA gene sequencing isolates of LAB from dahi and datshi were identified as Enterococcus faecalis, E. faecium, Lactococcus lactis subsp. lactis. LAB strains were tested for some technological properties. All LAB strains except E. faecalis CH2:17 caused coagulation of milk at both 30°C for 48 h. Only E. faecium DH4:05 strain was resistant to pH 3. No significant difference (P > 0.05) of viable counts was observed in MRS broth with and without lysozyme. All LAB strains grew well in 0.3% bile showing their ability to tolerate bile salt. None of the LAB strains showed >70% hydrophobicity. This study, being the first of its microbiological analysis of the NFM of Bhutan, has opened up to an extent of research work that gives a new insight to the products. PMID:28203227

Anaerobic bacteria grow within Candida albicans biofilms and induce biofilm formation in suspension cultures.

PubMed

Fox, Emily P; Cowley, Elise S; Nobile, Clarissa J; Hartooni, Nairi; Newman, Dianne K; Johnson, Alexander D

2014-10-20

The human microbiome contains diverse microorganisms, which share and compete for the same environmental niches. A major microbial growth form in the human body is the biofilm state, where tightly packed bacterial, archaeal, and fungal cells must cooperate and/or compete for resources in order to survive. We examined mixed biofilms composed of the major fungal species of the gut microbiome, Candida albicans, and each of five prevalent bacterial gastrointestinal inhabitants: Bacteroides fragilis, Clostridium perfringens, Escherichia coli, Klebsiella pneumoniae, and Enterococcus faecalis. We observed that biofilms formed by C. albicans provide a hypoxic microenvironment that supports the growth of two anaerobic bacteria, even when cultured in ambient oxic conditions that are normally toxic to the bacteria. We also found that coculture with bacteria in biofilms induces massive gene expression changes in C. albicans, including upregulation of WOR1, which encodes a transcription regulator that controls a phenotypic switch in C. albicans, from the "white" cell type to the "opaque" cell type. Finally, we observed that in suspension cultures, C. perfringens induces aggregation of C. albicans into "mini-biofilms," which allow C. perfringens cells to survive in a normally toxic environment. This work indicates that bacteria and C. albicans interactions modulate the local chemistry of their environment in multiple ways to create niches favorable to their growth and survival. Copyright © 2014 Elsevier Ltd. All rights reserved.

Molecular Viability Testing of UV-Inactivated Bacteria.

PubMed

Weigel, Kris M; Nguyen, Felicia K; Kearney, Moira R; Meschke, John S; Cangelosi, Gerard A

2017-05-15

PCR is effective in detecting bacterial DNA in samples, but it is unable to differentiate viable bacteria from inactivated cells or free DNA fragments. New PCR-based analytical strategies have been developed to address this limitation. Molecular viability testing (MVT) correlates bacterial viability with the ability to rapidly synthesize species-specific rRNA precursors (pre-rRNA) in response to brief nutritional stimulation. Previous studies demonstrated that MVT can assess bacterial inactivation by chlorine, serum, and low-temperature pasteurization. Here, we demonstrate that MVT can detect inactivation of Escherichia coli , Aeromonas hydrophila , and Enterococcus faecalis cells by UV irradiation. Some UV-inactivated E. coli cells transiently retained the ability to synthesize pre-rRNA postirradiation (generating false-positive MVT results), but this activity ceased within 1 h following UV exposure. Viable but transiently undetectable (by culture) E. coli cells were consistently detected by MVT. An alternative viability testing method, viability PCR (vPCR), correlates viability with cell envelope integrity. This method did not distinguish viable bacteria from UV-inactivated bacteria under some conditions, indicating that the inactivated cells retained intact cell envelopes. MVT holds promise as a means to rapidly assess microbial inactivation by UV treatment. IMPORTANCE UV irradiation is increasingly being used to disinfect water, food, and other materials for human use. Confirming the effectiveness of UV disinfection remains a challenging task. In particular, microbiological methods that rely on rapid detection of microbial DNA can yield misleading results, due to the detection of remnant DNA associated with dead microbial cells. This report describes a novel method that rapidly distinguishes living microbial cells from dead microbial cells after UV disinfection. Copyright © 2017 American Society for Microbiology.

[Antibacterial activity of rare Streptomyces species against clinical resistant bacteria].

PubMed

Boughachiche, Faiza; Reghioua, Sihem; Zerizer, Habiba; Boulahrouf, Abderrahmane

2012-01-01

In the search for new antibiotics from Steptomyces, investigating extremes habitats enhances the probability of isolating novel producers. In this context, the antibacterial activity of four Streptomyces strains isolated from Ezzmoul saltpans was studied. Two of them showed antibacterial activity against antibiotic's resistant bacteria (Bacillus cereus: β-lactamines and sulfamides resistant, Streptococcus faecalis: penicillin, tetracycline and cotrimoxazole resistant, and Staphylococcus aureus Mu 50: vancomycine resistant). The most active Streptomyces strain produces one type of polar bioactive molecules that resists to temperature variation and light exposition. Its activity appears in the first culture day and reaches its maximal value in the fourth day. The second strain presents themoresistant activity that reaches its maximal value in the first culture day. It produces two types of bioactive molecules, one is polar and the second is non polar (according to thin layer chromatography technique results).

In vitro antimicrobial activity of various medication preparations on E. faecalis in root canal dentin.

PubMed

Lynne, Richard E; Liewehr, Frederick R; West, Lesley A; Patton, William R; Buxton, Thomas B; McPherson, James C

2003-03-01

The purpose of this study was to evaluate the antimicrobial activity of several medication preparations in root canal dentin infected with Enterococcus faecalis. Roots of extracted bovine incisors were prepared to standardized cylindrical test specimens, 5 mm in height. The smear layer was removed and the samples were autoclaved and then incubated at 37 degrees C/5% CO2 for 24 h in brain-heart infusion (BHI) broth containing 7.0 x 10(4) colony forming units per ml of E. faecalis. The samples were washed in phosphate buffered saline and mounted to individual culture wells with sticky wax. Test medications were applied to fill the canal lumina; medication groups were: (a) sterile H2O (positive control); (b) a 10% mixture of 1.0 g Ca(OH)2 USP in 10 ml sterile H2O; (c) 10% Ca(OH)2 in 0.12% chlorhexidine gluconate (Peridex); (d) Peridex; and (e) uninoculated BHI (negative control). The samples were incubated at 37 degrees C/5% CO2 for 24 h. Dentin samples for quantitative microbiology were then obtained with consecutive sterile burs (ISO 029, 035, 042). All three experimental groups demonstrated significantly greater antimicrobial activity than the positive control (p < 0.001). Group 2 demonstrated significantly greater antimicrobial activity than Group 3 or Group 4 at all dentin depths (p < 0.05). These results suggest that 10% Ca(OH)2 may be more effective than Peridex or 10% Ca(OH)2 in Peridex for the elimination of E. faecalis from dentin tubules.

Evaluation of antimicrobial effect of azadirachtin plant extract (Soluneem (™)) on commonly found root canal pathogenic microorganisms (viz. Enterococcus faecalis) in primary teeth: A microbiological study.

PubMed

Shah, Shanal; Venkataraghavan, Karthik; Choudhary, Prashant; Mohammad, Shameer; Trivedi, Krishna; Shah, Shalin G

2016-01-01

The aim of this study is to evaluate the antimicrobial activity of Soluneem ™ when used as an irrigating solution along with other commonly used irrigating solution sodium hypochlorite (NaOCl) against Enterococcus faecalis. Microorganism used in this study was E. faecalis (Microbial Type Culture Collection 439). Test substance used was Soluneem ™, which was obtained from Vittal Mallya Scientific Research Foundation (VMSRF), Bengaluru. This study was conducted in a microbiology laboratory (Biocare Research India Pvt., Ltd. Laboratory, Ahmedabad, Gujarat) to evaluate the antimicrobial effect of Soluneem ™ (Azadirachtin) on E. faecalis. Antimicrobial activity testing was performed using the macrobroth dilution method according to the Clinical Laboratory Standards Institute guidelines. All determinations were performed thrice. Minimum bactericidal concentration (MBC) was seen as 2.6% for Soluneem ™ while the same was seen at 0.1% for NaOCl. Independent sample t-test was carried out to compare the MBC of Soluneem ™ and NaOCl, which showed that there was no statistically significant difference between them, i.e., 2.6% Soluneem ™ was as effective as 0.1% NaOCl. Soluneem ™ showed antimicrobial activity against E. faecalis at various concentrations. It was also found that the efficacy of Soluneem ™ at 2.6% concentration and above was relatively similar to that of gold standard irrigating solution (NaOCl) on inhibition of E. faecalis.

[Post-marketing surveillance of antibacterial activities of cefozopran against various clinical isolates--I. Gram-positive bacteria].

PubMed

Igari, Jun; Oguri, Toyoko; Hiramatsu, Nobuyoshi; Akiyama, Kazumitsu; Koyama, Tsuneo

2003-10-01

As a post-marketing surveillance, the in vitro antibacterial activities of cefozopran (CZOP), an agent of cephems, against various clinical isolates were yearly evaluated and compared with those of other cephems, oxacephems, penicillins, and carbapenems. Changes in the bacterial susceptibility for CZOP were also evaluated with the resistance ratio calculated with breakpoint MIC. Sixteen species (2,363 strains) of Gram-positive bacteria were isolated from the clinical materials annually collected from 1996 to 2001, and consisted of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), methicillin-susceptible Staphylococcus epidermidis (MSSE), methicillin-resistant Staphylococcus epidermidis (MRSE), Staphylococcus haemolyticus, Staphylococcus saprophyticus, Enterococcus faecalis, Enterococcus faecium, Enterococcus avium, Streptococcus pyogenes, Streptococcus agalactiae, penicillin-susceptible Streptococcus pneumoniae (PSSP), penicillin-intermediate resistant S. pneumoniae (PISP), penicillin-resistant S. pneumoniae (PRSP), Streptococcus milleri group and Peptostreptococcus spp. The antibacterial activity of CZOP either against MSSA or MSSE was preferable (MIC90: 2 or 0.5 micrograms/mL) and comparable to those of other cephems. CZOP was also effective on MRSE (MIC90: 16 micrograms/mL) but not on MRSA. CZOP and other cephems had low antibacterial activity against S. haemolyticus (MIC90: 64 micrograms/mL). The antibacterial activity of CZOP against S. saprophyticus was comparable to or higher than those of other cephems, but the MIC90 of CZOP in 2001 was higher than those in 1996-2000 (32 vs 1-2 micrograms/mL). The antibacterial activity of CZOP against E. faecalis was comparable to that of cefpirome (CPR; MIC90: 16 micrograms/mL) and higher than those of other cephems. No antibacterial activity of CZOP against E. faecium and E. avium was observed, like other drugs. The antibacterial activity of CZOP against S. pyogenes

Human Bile Reduces Antimicrobial Activity of Selected Antibiotics against Enterococcus faecalis and Escherichia coli In Vitro.

PubMed

Wulkersdorfer, Beatrix; Jaros, David; Eberl, Sabine; Poschner, Stefan; Jäger, Walter; Cosentini, Enrico; Zeitlinger, Markus; Schwameis, Richard

2017-08-01

It has been known from previous studies that body fluids, such as cerebrospinal fluid, lung surfactant, and urine, have a strong impact on the bacterial killing of many anti-infective agents. However, the influence of human bile on the antimicrobial activity of antibiotics is widely unknown. Human bile was obtained and pooled from 11 patients undergoing cholecystectomy. After sterilization of the bile fluid by gamma irradiation, its effect on bacterial killing was investigated for linezolid (LZD) and tigecycline (TGC) against Enterococcus faecalis ATCC 29212. Further, ciprofloxacin (CIP), meropenem (MEM), and TGC were tested against Escherichia coli ATCC 25922. Time-kill curves were performed in pooled human bile and Mueller-Hinton broth (MHB) over 24 h. Bacterial counts (in CFU per milliliter after 24 h) of bile growth controls were approximately equal to MHB growth controls for E. coli and approximately 2-fold greater for E. faecalis , indicating a promotion of bacterial growth by bile for the latter strain. Bile reduced the antimicrobial activity of CIP, MEM, and TGC against E. coli as well as the activity of LZD and TGC against E. faecalis This effect was strongest for TGC against the two strains. Degradation of TGC in bile was identified as the most likely explanation. These findings may have important implications for the treatment of bacterial infections of the gallbladder and biliary tract and should be explored in more detail. Copyright © 2017 American Society for Microbiology.

Adsorption of Streptococcus faecalis on diatomite carriers for use in biotransformations.

PubMed

Anderson, W A; Bay, P; Legge, R L; Moo-Young, M

1990-01-01

Adsorption of cells on particulate carriers is potentially one of the most cost-effective immobilization techniques available. Diatomite carriers, such as Celite, have desirable physical properties, are inexpensive, and are suitable for both mycelial and bacterial systems. This work investigated the use of diatomite carriers as a biocatalyst support in a packed-bed reactor where L-tyrosine was enzymatically decarboxylated using adsorbed, non-growing cells of Streptococcus faecalis. Composition of microbial adsorption on different Celite types, with mean pore sizes ranging from 0.55 to 22 microns, showed there was no significant difference in biomass loading capacity under the conditions used. Using Celite 560, biomass loadings in a packed-bed reactor varied from 10 to 30 g dm-3 of reactor volume, which compares favourably with other adsorption methods. When used to decarboxylate L-tyrosine, the reactor was found to have a half-life of 15-20 h. A combination of enzyme activity loss and slow leakage of biomass from the packed-bed reactor was responsible for the decline in conversion. Treatment of the S. faecalis cells with glutaraldehyde significantly reduced the enzyme activity loss and extended the reactor half-life to 65 h, but had little effect on the rate of cell leakage from the reactor. Further work on reduction of cell leakage rate seems necessary for evaluation of the system's practicality.

Antimicrobial resistance profiles of Enterococcus faecalis and Enterococcus faecium isolated from artisanal food of animal origin in Argentina.

PubMed

Delpech, Gastón; Pourcel, Gisela; Schell, Celia; De Luca, María; Basualdo, Juan; Bernstein, Judith; Grenovero, Silvia; Sparo, Mónica

2012-10-01

Enterococci are part of the indigenous microbiota of human gastrointestinal tract and food of animal origin. Enterococci inhabiting non-human reservoirs play a critical role in the acquisition and dissemination of antimicrobial resistance determinants. The aim of this work was to investigate the antimicrobial resistance in Enterococcus faecalis and Enterococcus faecium strains recovered from artisanal food of animal origin. Samples of goat cheese (n = 42), cow cheese (n = 40), artisanal salami (n = 30), and minced meat for the manufacture of hamburgers (n = 60) were analyzed. Phenotypic and genotypic tests for species-level identification of the recovered isolates were carried out. Minimum inhibitory concentration (MIC) study for in vitro quantitative antimicrobial resistance assessment was performed, and 71 E. faecalis and 22 E. faecium were isolated. The recovered enterococci showed different multi-drug resistance patterns that included tretracycline, erythromycin, ciprofloxacin, linezolid, penicillin, ampicillin, vancomycin, teicoplanin, gentamicin (high-level resistance), and streptomycin (high-level resistance). VanA-type E. faecium were detected. β-lactamase activity was not observed. Artisanal foods of animal origin act as a non-human reservoir of E. faecalis and E. faecuim strains, expressing multi-resistance to antimicrobials. In conclusion, the implementation of a continuous antimicrobial resistance surveillance in enterococci isolated from artisanal food of animal origin is important.

Diversity of microbes and potential exoelectrogenic bacteria on anode surface in microbial fuel cells.

PubMed

Sun, Yujiao; Zuo, Jiane; Cui, Longtao; Deng, Qian; Dang, Yan

2010-02-01

Single-chamber microbial fuel cells (MFCs), inoculated with anaerobic sludge and continuously run with two kinds of organic wastewater influents, were systemically investigated. The diversity of microbes, determined by 16S rDNA analysis, was analyzed on three anodes under different conditions. One anode was in a closed circuit in synthetic wastewater containing glucose. The other two anodes, in open or closed circuits, were fed effluent from an anaerobic reactor treating starch wastewater. The chemical oxygen demand (COD) removal efficiency was about 70%, and the exported voltages were about 450 mV. The 16S rDNA molecular clones of microbes on anode surfaces showed significant changes in Eubacterial structure under different conditions. gamma-Proteobacteria and the high G+C gram-positive groups were predominant in the synthetic wastewater, while epsilon-Proteobacteria predominated in the anaerobic reactor effluent. Known exoelectrogenic bacterial species composition also changed greatly depending on substrate. On the artificial substrate, 28% of the bacterial sequences were affiliated with Aeromonas, Pseudomonas, Geobacter, and Desulfobulbus. On the anaerobic effluent, only 6% were affiliated with Geobacter or Clostridium. Because only a few exoelectrogenic bacteria from MFCs have been directly isolated and studied, we compared the community structures of two bacterial anodes, in open and closed circuits, under the same substrate of anaerobic effluent in order to identify additional exoelectrogenic bacterial strains. Alcaligenes monasteriensis, Comamonas denitrificans, and Dechloromonas sp. were found to be potential exoelectrogenic bacteria worthy of further research.

Increased d-lactic Acid intestinal bacteria in patients with chronic fatigue syndrome.

PubMed

Sheedy, John R; Wettenhall, Richard E H; Scanlon, Denis; Gooley, Paul R; Lewis, Donald P; McGregor, Neil; Stapleton, David I; Butt, Henry L; DE Meirleir, Kenny L

2009-01-01

Patients with chronic fatigue syndrome (CFS) are affected by symptoms of cognitive dysfunction and neurological impairment, the cause of which has yet to be elucidated. However, these symptoms are strikingly similar to those of patients presented with D-lactic acidosis. A significant increase of Gram positive facultative anaerobic faecal microorganisms in 108 CFS patients as compared to 177 control subjects (p<0.01) is presented in this report. The viable count of D-lactic acid producing Enterococcus and Streptococcus spp. in the faecal samples from the CFS group (3.5 x 10(7) cfu/L and 9.8 x 10(7) cfu/L respectively) were significantly higher than those for the control group (5.0 x 10(6) cfu/L and 8.9 x 10(4) cfu/L respectively). Analysis of exometabolic profiles of Enterococcus faecalis and Streptococcus sanguinis, representatives of Enterococcus and Streptococcus spp. respectively, by NMR and HPLC showed that these organisms produced significantly more lactic acid (p<0.01) from (13)C-labeled glucose, than the Gram negative Escherichia coli. Further, both E. faecalis and S. sanguinis secrete more D-lactic acid than E. coli. This study suggests a probable link between intestinal colonization of Gram positive facultative anaerobic D-lactic acid bacteria and symptom expressions in a subgroup of patients with CFS. Given the fact that this might explain not only neurocognitive dysfunction in CFS patients but also mitochondrial dysfunction, these findings may have important clinical implications.

Use of photo-Fenton's reaction by 400-nm LED light for endodontic disinfection: A preliminary in vitro study on Enterococcus faecalis.

PubMed

Lagori, Giuseppe; Fornaini, Carlo; Rocca, Jean-Paul; Merigo, Elisabetta

2017-06-01

One of the biggest challenges in endodontics is the complete disinfection of root canals. In addition to mechanical preparation, the technique traditionally also involves channel disinfection with other agents such as sodium hypochlorite, hydrogen peroxide, chlorhexidine, or a combination of these. Some bacterial species are particularly resistant to eradication. Using Enterococcus faecalis in this preliminary study, we tested the bactericidal effectiveness of the Fenton reaction and the photo-Fenton reaction using an LED light with a 400-nm wavelength. Discs of hydroxyapatite were incubated in brain-heart broth contaminated with Enterococcus faecalis. After 4days, they were decontaminated with different bactericidal agents, including some with proven and well-known efficacy (5% sodium hypochlorite and 3% hydrogen peroxide) and other treatments using solutions of 1.5% hydrogen peroxide and 0.15% iron gluconate (Fenton reaction) plus LED light at a Fluence of 4.0J/cm 2 (photo-Fenton reaction). The photo-Fenton reaction demonstrated comparable performance to that of sodium hypochlorite in eliminating Enterococcus faecalis. Copyright © 2017. Published by Elsevier B.V.

Antibacterial Efficacy of Octenisept, Alexidine, Chlorhexidine, and Sodium Hypochlorite against Enterococcus faecalis Biofilms.

PubMed

Bukhary, Sundus; Balto, Hanan

2017-04-01

The purpose of this study was to evaluate the antibacterial effectiveness of Octenisept (OCT; Schülke & Mayr GmBH, Norderstedt, Germany), 1% alexidine (ALX) (Santa Cruz Biotechnology, Inc, Santa Cruz, CA), and 2% chlorhexidine (CHX) against Enterococcus faecalis biofilm using confocal laser scanning microscopy. Root dentin discs were prepared from extracted human teeth, sterilized, and inoculated with E. faecalis strain (ATCC 29212) to establish 3-week-old biofilm model. Infected dentin discs were exposed to OCT (n = 20), 1% ALX (n = 20), and 2% CHX (n = 20) for 10 minutes. Dentin discs (n = 15) exposed to 5.25% sodium hypochlorite (NaOCl) were used as a positive control, whereas specimens exposed to saline (n = 15) were used as a negative control. After exposure, the dentin discs were stained with fluorescent LIVE/DEAD BacLight dye (Invitrogen Molecular Probes, Eugene, OR) and analyzed with confocal laser scanning microscopy to determine the proportion of dead cells in the biofilm. Statistical analysis was performed using the Kruskal-Wallis and Mann-Whitney U tests (P < .05). The highest proportion of dead cells was found in the 5.25% NaOCl group (94.14%; range, 92.30%-98.20%) compared with the experimental groups (P < .05). A significantly greater proportion of dead cells was found in the OCT group (74.14%; range, 70.03%-78.96%) compared with the 1% ALX and 2% CHX groups (P < .05). The proportion of dead cells was 43.89% (range, 24.86%-55.63%) and 42.78% (range, 25.45%-55.06%) in the 1% ALX and 2% CHX groups, respectively, with no statistical significant difference between the 2 groups (P > .05). NaOCl had significantly greater antimicrobial activity against E. faecalis biofilms compared with OCT, CHX, and ALX. OCT was more effective than CHX and ALX. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

An in vivo comparison of antimicrobial efficacy of sodium hypochlorite and Biopure MTAD™ against enterococcus faecalis in primary teeth: A qPCR study.

PubMed

Tulsani, S G; Chikkanarasaiah, N; Bethur, S

2014-01-01

Biopure MTAD™, a new root canal irrigant has shown promising results against the most common resistant microorganism, E. faecalis, in permanent teeth. However, there is lack of studies comparing its antimicrobial effectiveness with NaOCl in primary teeth. The purpose of this study was to compare the in vivo antimicrobial efficacy of NaOCl 2.5% and Biopure MTAD™ against E. faecalis in primary teeth. Forty non vital single rooted primary maxillary anterior teeth of children aged 4-8 years, were irrigated either with NaOCl 2.5% (n=15), Biopure MTAD™ (n=15) and 0.9% Saline (n=10, control group). Paper point samples were collected at baseline (S1) and after chemomechanical preparation (S2) during the pulpectomy procedure. The presence of E. faecalis in S1 & S2 was evaluated using Real time Polymerase Chain Reaction. Statistical significant difference was found in the antimicrobial efficacy of NaOCl 2.5 % and BioPure MTAD™ when compared to saline (p>0.05). However, no statistical significant difference was found between the efficacies of both the irrigants. NaOCl 2.5% and BioPure MTAD™, both irrigants are equally efficient against E. faecalis in necrotic primary anterior teeth. MTAD is a promising irrigant, however clinical studies are required to establish it as ideal root canal irrigant in clinical practice.

Comparison of Antibacterial Efficacy of Turmeric Extract, Morinda Citrifolia and 3% Sodium Hypochlorite on Enterococcus faecalis: An In-vitro Study.

PubMed

Chaitanya, Bathula Vimala; Somisetty, Kusum Valli; Diwan, Abhinav; Pasha, Shiraz; Shetty, Nandaprasad; Reddy, Yashwanth; Nadigar, Shankar

2016-10-01

Sodium hypochlorite (NaOCl), the most commonly used irrigant, has many potential properties like its unique ability to dissolve pulp tissue, excellent antimicrobial activity, but has a cytotoxic effect when injected into periapical tissues. It is also known to produce allergic reactions, foul smell and taste, and potential for corrosion. Facultative organisms such as Enterococcus faecalis and aerobes like Staphylococcus aureus are considered to be the most resistant species and one of the possible causes of root canal treatment failure. So there is a need to find an alternative to sodium hypochlorite to act against these resistant microorganisms. To evaluate and compare the antibacterial efficacy of morinda citrifolia and turmeric extract with 3% NaOCl as a root canal irrigant, against E. faecalis and S.aureus . The antimicrobial efficacy was assessed in vitro using agar well diffusion method. Agar plates were prepared using Brain-Heart Infusion (BHI) agar. Cultures of E.faecalis and S.aureus were grown in nutrient broth at 37°C. Plates were incubated for 24 hours at 37°C and microbial zones of inhibition were recorded. Statistical analysis was performed using ANOVA. NaOCl (3%) showed larger zones of inhibition than herbal irrigants against both the microorganisms. Among the herbal irrigants, morinda citrifolia showed larger zones of inhibition than turmeric hydro-alcoholic extract and turmeric water extract which was statistically significant (p<0.05). NaOCl (3%) showed maximum antibacterial activity against E. faecalis , followed by morinda citrifolia and turmeric extracts. Considering the potential for undesirable properties of NaOCl, use of herbal alternatives in endodontics might prove to be advantageous.

Comparison of Antibacterial Efficacy of Turmeric Extract, Morinda Citrifolia and 3% Sodium Hypochlorite on Enterococcus faecalis: An In-vitro Study

PubMed Central

Somisetty, Kusum Valli; Diwan, Abhinav; Pasha, Shiraz; Shetty, Nandaprasad; Reddy, Yashwanth; Nadigar, Shankar

2016-01-01

Introduction Sodium hypochlorite (NaOCl), the most commonly used irrigant, has many potential properties like its unique ability to dissolve pulp tissue, excellent antimicrobial activity, but has a cytotoxic effect when injected into periapical tissues. It is also known to produce allergic reactions, foul smell and taste, and potential for corrosion. Facultative organisms such as Enterococcus faecalis and aerobes like Staphylococcus aureus are considered to be the most resistant species and one of the possible causes of root canal treatment failure. So there is a need to find an alternative to sodium hypochlorite to act against these resistant microorganisms. Aim To evaluate and compare the antibacterial efficacy of morinda citrifolia and turmeric extract with 3% NaOCl as a root canal irrigant, against E. faecalis and S.aureus. Materials and Methods The antimicrobial efficacy was assessed in vitro using agar well diffusion method. Agar plates were prepared using Brain-Heart Infusion (BHI) agar. Cultures of E.faecalis and S.aureus were grown in nutrient broth at 37°C. Plates were incubated for 24 hours at 37°C and microbial zones of inhibition were recorded. Statistical analysis was performed using ANOVA. Results NaOCl (3%) showed larger zones of inhibition than herbal irrigants against both the microorganisms. Among the herbal irrigants, morinda citrifolia showed larger zones of inhibition than turmeric hydro-alcoholic extract and turmeric water extract which was statistically significant (p<0.05). Conclusion NaOCl (3%) showed maximum antibacterial activity against E. faecalis, followed by morinda citrifolia and turmeric extracts. Considering the potential for undesirable properties of NaOCl, use of herbal alternatives in endodontics might prove to be advantageous. PMID:27891459

Potentiation of the humoral immune response elicited by a commercial vaccine against bovine respiratory disease by Enterococcus faecalis CECT7121.

PubMed

Díaz, A M; Almozni, B; Molina, M A; Sparo, M D; Manghi, M A; Canellada, A M; Castro, M S

2018-04-10

Vaccination against pathogens involved in bovine respiratory disease (BRD) is a useful tool to reduce the risk of this disease however, it has been observed that the commercially available vaccines only partially prevent the infections caused by Pasteurella multocida and Mannheimia haemolytica. Therefore, it is recommended to search for new adjuvant strategies to minimise the economic impact of this respiratory syndrome. A possibility to improve the conventional vaccine response is to modulate the immune system with probiotics, since there is accumulating evidence that certain immunomodulatory strains administered around the time of vaccination can potentiate the immune response. Considering veterinary vaccines are frequently tested in murine models, we have developed an immunisation schedule in BALB/c mice that allows us to study the immune response elicited by BRD vaccine. In order to evaluate a potential strategy to enhance vaccine efficacy, the adjuvant effect of Enterococcus faecalis CECT7121 on the murine specific humoral immune response elicited by a commercial vaccine against BRD was studied. Results indicate that the intragastric administration of E. faecalis CECT7121 was able to induce an increase in the specific antibody titres against the bacterial components of the BRD vaccines (P. multocida and M. haemolytica). The quality of the humoral immune response, in terms of antibody avidity, was also improved. Regarding the cellular immune response, although the BRD vaccination induced a low specific secretion of cytokines in the spleen cell culture supernatants, E. faecalis CECT7121-treated mice showed higher interferon-γ production than immunised control mice. Our results allowed us to conclude that the administration of E. faecalis CECT7121 could be employed as an adjuvant strategy to potentiate humoral immune responses.

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. © 2015 Institute of Food Technologists®

A novel gene, optrA, that confers transferable resistance to oxazolidinones and phenicols and its presence in Enterococcus faecalis and Enterococcus faecium of human and animal origin.

PubMed

Wang, Yang; Lv, Yuan; Cai, Jiachang; Schwarz, Stefan; Cui, Lanqing; Hu, Zhidong; Zhang, Rong; Li, Jun; Zhao, Qin; He, Tao; Wang, Dacheng; Wang, Zheng; Shen, Yingbo; Li, Yun; Feßler, Andrea T; Wu, Congming; Yu, Hao; Deng, Xuming; Xia, Xi; Shen, Jianzhong

2015-08-01

The oxazolidinone-resistant Enterococcus faecalis E349 from a human patient tested negative for the cfr gene and 23S rRNA mutations. Here we report the identification of a novel oxazolidinone resistance gene, optrA, and a first investigation of the extent to which this gene was present in E. faecalis and Enterococcus faecium from humans and food-producing animals. The resistance gene optrA was identified by whole-plasmid sequencing and subsequent cloning and expression in a susceptible Enterococcus host. Transformation and conjugation assays served to investigate the transferability of optrA. All optrA-positive E. faecalis and E. faecium isolates of human and animal origin were analysed for their MICs and their genotype, as well as the location of optrA. The novel plasmid-borne ABC transporter gene optrA from E. faecalis E349 conferred combined resistance or elevated MICs (when no clinical breakpoints were available) to oxazolidinones (linezolid and tedizolid) and phenicols (chloramphenicol and florfenicol). The corresponding conjugative plasmid pE349, on which optrA was located, had a size of 36 331 bp and also carried the phenicol exporter gene fexA. The optrA gene was functionally expressed in E. faecalis, E. faecium and Staphylococcus aureus. It was detected more frequently in E. faecalis and E. faecium from food-producing animals (20.3% and 5.7%, respectively) than from humans (4.2% and 0.6%, respectively). Enterococci with elevated MICs of linezolid and tedizolid should be tested not only for 23S rRNA mutations and the gene cfr, but also for the novel resistance gene optrA. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Quaternized Zn(II) phthalocyanines for photodynamic strategy against resistant periodontal bacteria.

PubMed

Kussovski, Vesselin; Mantareva, Vanya; Durmuş, Mahmut; Angelov, Ivan

2018-04-25

Photodynamic inactivation (PDI) has been featured as an effective strategy in the treatment of acute drug-resistant infections. The efficiency of PDI was evaluated against three periodontal pathogenic bacteria that were tested as drug-resistant strains. In vitro studies were performed with four water-soluble cationic Zn(II) phthalocyanines (ZnPc1-4) and irradiation of a specific light source (light-emitting diode, 665 nm) with three doses (15, 36 and 60 J/cm2). The well detectable fluorescence of ZnPcs allowed the cellular imaging, which suggested relatively high uptakes of ZnPcs into bacterial species. Complete photoinactivation was achieved with all studied ZnPc1-4 for Enterococcus faecalis (E. faecalis) at a light dose of 15 J/cm2. The photodynamic response was high for Prevotella intermedia (P. intermedia) after the application of 6 μM of ZnPc1 and a light dose of 36 J/cm2 and for 6 μM of ZnPc2 at 60 J/cm2. P. intermedia was inactivated with ZnPc3 (4 log) and ZnPc4 (2 log) with irradiation at an optimal dose of 60 J/cm2. Similar photoinactivation results (2 log) were achieved for Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) treated with 6 μM ZnPc1 and ZnPc2 at a light dose of 60 J/cm2. The study suggested that PDI with quaternized Zn(II) phthalocyanines and specific light irradiation appears to be a very useful antimicrobial strategy for effective inactivation of drug-resistant periodontal pathogens.

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. Copyright © 2014 Elsevier B.V. All rights reserved.

Frequency of ace, epa and elrA Genes in Clinical and Environmental Strains of Enterococcus faecalis.

PubMed

Lysakowska, Monika Eliza; Denys, Andrzej; Sienkiewicz, Monika

2012-12-01

Surface proteins play an important role in the pathogenesis of enterococcal infections. Some of them are candidates for a vaccine, e.g., the frequency of endocarditis in rats vaccinated with Ace protein was 75 % as 12 opposed to 100 % in those who weren't. However, there are other components of enterococcal cells, such as Epa antigens or internalin-like proteins, which may be used in the prophylaxis of infections caused by them. However, also other virulence factors and resistance to antibiotics are important during enterococcal infection. Therefore, the relevance of ace, epa, elrA, other virulence genes, as well as resistance to antibiotics was investigated. 161 Enterococcus faecalis strains isolated from teaching hospitals in Lodz, cultured according to standard microbiological methods, were investigated for the presence of genes encoding surface proteins by PCR. Results were analyzed with χ(2) test. The elrA gene was found in all clinical and environmental strains, the ace gene was also widespread among E. faecalis (96.9 %). Both tested epa genes were found in the majority of isolates (83.25 %). There was correlation between the presence of esp and ace genes (p = 0.046) as well as between epa and agg genes (p = 0.0094; χ(2) test). The presence of the genes encoding surface proteins investigated in our study in the great majority of isolates implies that they would appear to be required during E. faecalis infection. Therefore, they could be excellent targets in therapy of enterococcal infections or, as some studies show, candidates for vaccines.

Enterocin C, a class IIb bacteriocin produced by E. faecalis C901, a strain isolated from human colostrum.

PubMed

Maldonado-Barragán, Antonio; Caballero-Guerrero, Belén; Jiménez, Esther; Jiménez-Díaz, Rufino; Ruiz-Barba, José L; Rodríguez, Juan M

2009-07-31

Enterocin C (EntC), a class IIb bacteriocin was purified from culture supernatants of Enterococcus faecalis C901, a strain isolated from human colostrum. Enterocin C consists of two distinct peptides, named EntC1 and EntC2, whose complementary action is required for full antimicrobial activity. The structural genes entC1 and entC2 encoding enterocins EntC1 and EntC2, respectively, and that encoding the putative immunity protein (EntCI) are located in the 9-kb plasmid pEntC, harboured by E. faecalis C901. The N-terminal sequence of both antimicrobial peptides revealed that EntC1 (4284 Da) is identical to Ent1071A, one of the two peptides that form enterocin 1071 (Ent1071), a bacteriocin produced by E. faecalis BFE 1071. In contrast, EntC2 (3867 Da) presents the non-polar alanine residue at position 17 (Ala(17)) instead of the polar threonine residue (Thr(17)) in Ent1071B, the second peptide constituting Ent1071. In spite of peptide similarities, EntC differs from Ent1071 in major aspects, including the complementary activity among its constitutive peptides and its wider inhibitory spectrum of activity. Different amphiphilic alpha-helical conformations between EntC2 and Ent1071B could explain both, acquired complementary activity and increased antimicrobial spectrum.

Isolation and characterization of bacteriocinogenic lactic bacteria from M-Tuba and Tepache, two traditional fermented beverages in México

PubMed Central

de la Fuente-Salcido, Norma M; Castañeda-Ramírez, José Cristobal; García-Almendárez, Blanca E; Bideshi, Dennis K; Salcedo-Hernández, Rubén; Barboza-Corona, José E

2015-01-01

Mexican Tuba (M-Tuba) and Tepache are Mexican fermented beverages prepared mainly with pineapple pulp and coconut palm, respectively. At present, reports on the microbiota and nutritional effects of both beverages are lacking. The purpose of this study was to determine whether M-Tuba and Tepache contain cultivable lactic acid bacteria (LAB) capable of producing bacteriocins. Tepache and M-Tuba contain mesophilic aerobic bacteria, LAB, and yeast. Bacillus subtilis, Listeria monocytogenes, Listeria innocua, Streptococcus agalactiae, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella typhimurium, and Salmonella spp, were the microorganisms most susceptible to metabolites produced by bacterial isolates. M-Tuba and Tepache contain bacteria that harbor genes coding for nisin and enterocin, but not pediocin. The presence of Lactococcus lactis and E. faecium in M-Tuba and Tepache, was identified by 16S rDNA. These bacteria produced bacteriocins of ∼3.5 kDa and 4.0–4.5 kDa, respectively. Partial purified bacteriocins showed inhibitory effect against Micrococcus luteus, L. monocytogenes, L. innocua, Str. agalactiae, S. aureus, Bacillus cereus, B. subtilis, E. faecalis, and K. pneumoniae. We characterized, for the first time, cultivable microbiota of M-Tuba and Tepache, and specifically, identified candidate lactic bacteria (LAB) present in these beverages that were capable of synthesizing antimicrobial peptides, which collectively could provide food preservative functions. PMID:26405529

Heavy metal tolerant halophilic bacteria from Vembanad Lake as possible source for bioremediation of lead and cadmium.

PubMed

Sowmya, M; Rejula, M P; Rejith, P G; Mohan, Mahesh; Karuppiah, Makesh; Hatha, A A Mohamed

2014-07-01

Microorganisms which can resist high concentration of toxic heavy metals are often considered as effective tools of bioremediation from such pollutants. In the present study, sediment samples from Vembanad Lake were screened for the presence of halophilic bacteria that are tolerant to heavy metals. A total of 35 bacterial strains belonging to different genera such as Alcaligenes, Vibrio, Kurthia, Staphylococcus and members of the family Enterobacteriaceae were isolated from 21 sediment samples during February to April, 2008. The salt tolerance and optimum salt concentrations of the isolates revealed that most of them were moderate halophiles followed by halotolerant and extremely halotolerant groups. The minimum inhibitory concentrations (MICs) against cadmium and lead for each isolate revealed that the isolates showed higher MIC against lead than cadmium. Based on the resistance limit concentration, most of them were more tolerant to lead than cadmium at all the three salt concentrations tested. Heavy metal removal efficiency of selected isolates showed a maximum reduction of 37 and 99% against cadmium and lead respectively. The study reveals the future prospects of halophilic microorganisms in the field of bioremediation.

The Relationship among Tyrosine Decarboxylase and Agmatine Deiminase Pathways in Enterococcus faecalis

PubMed Central

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

2017-01-01

Enterococci are considered mainly responsible for the undesirable accumulation of the biogenic amines tyramine and putrescine in cheeses. The biosynthesis of tyramine and putrescine has been described as a species trait in Enterococcus faecalis. Tyramine is formed by the decarboxylation of the amino acid tyrosine, by the tyrosine decarboxylase (TDC) route encoded in the tdc cluster. Putrescine is formed from agmatine by the agmatine deiminase (AGDI) pathway encoded in the agdi cluster. These biosynthesis routes have been independently studied, tyrosine and agmatine transcriptionally regulate the tdc and agdi clusters. The objective of the present work is to study the possible co-regulation among TDC and AGDI pathways in E. faecalis. In the presence of agmatine, a positive correlation between putrescine biosynthesis and the tyrosine concentration was found. Transcriptome studies showed that tyrosine induces the transcription of putrescine biosynthesis genes and up-regulates pathways involved in cell growth. The tyrosine modulation over AGDI route was not observed in the mutant Δtdc strain. Fluorescence analyses using gfp as reporter protein revealed PaguB (the promoter of agdi catabolic genes) was induced by tyrosine in the wild-type but not in the mutant strain, confirming that tdc cluster was involved in the tyrosine induction of putrescine biosynthesis. This study also suggests that AguR (the transcriptional regulator of agdi) was implicated in interaction among the two clusters. PMID:29163401

Block Copolymer Nanoparticles Remove Biofilms of Drug-Resistant Gram-Positive Bacteria by Nanoscale Bacterial Debridement.

PubMed

Li, Jianghua; Zhang, Kaixi; Ruan, Lin; Chin, Seow Fong; Wickramasinghe, Nirmani; Liu, Hanbin; Ravikumar, Vikashini; Ren, Jinghua; Duan, Hongwei; Yang, Liang; Chan-Park, Mary B

2018-06-26

Biofilms and the rapid evolution of multidrug resistance complicate the treatment of bacterial infections. Antibiofilm agents such as metallic-inorganic nanoparticles or peptides act by exerting antibacterial effects and, hence, do not combat biofilms of antibiotics-resistant strains. In this Letter, we show that the block copolymer DA95B5, dextran- block-poly((3-acrylamidopropyl) trimethylammonium chloride (AMPTMA)- co-butyl methacrylate (BMA)), effectively removes preformed biofilms of various clinically relevant multidrug-resistant Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococci (VRE V583), and Enteroccocus faecalis (OG1RF). DA95B5 self-assembles into core-shell nanoparticles with a nonfouling dextran shell and a cationic core. These nanoparticles diffuse into biofilms and attach to bacteria but do not kill them; instead, they promote the gradual dispersal of biofilm bacteria, probably because the solubility of the bacteria-nanoparticle complex is enhanced by the nanoparticle dextran shell. DA95B5, when applied as a solution to a hydrogel pad dressing, shows excellent in vivo MRSA biofilm removal efficacy of 3.6 log reduction in a murine excisional wound model, which is significantly superior to that for vancomycin. Furthermore, DA95B5 has very low in vitro hemolysis and negligible in vivo acute toxicity. This new strategy for biofilm removal (nanoscale bacterial debridement) is orthogonal to conventional rapidly developing resistance traits in bacteria so that it is as effective toward resistant strains as it is toward sensitive strains and may have widespread applications.

Epizootiological characteristics of viable bacteria and fungi in indoor air from porcine, chicken, or bovine husbandry confinement buildings

PubMed Central

Roque, Katharine; Lim, Gyeong-Dong; Jo, Ji-Hoon; Shin, Kyung-Min; Song, Eun-Seob; Gautam, Ravi; Kim, Chang-Yul; Lee, Kyungsuk; Shin, Seungwon; Yoo, Han-Sang; Heo, Yong

2016-01-01

Microorganisms found in bioaerosols from animal confinement buildings not only foster the risk of spreading diseases among livestock buildings, but also pose health hazards to farm workers and nearby residents. This study identified the various microorganisms present in the air of swine, chicken, and cattle farms with different kinds of ventilation conditions in Korea. Microbial air samples were collected onto Petri dishes with bacterial or fungal growth media using a cascade impactor. Endotoxin levels in total dust were determined by the limulus amebocyte lysate kinetic QCL method. Prevalent Gram-positive bacteria were Staphylococcus (S.) lentus, S. chromogenes, Bacillus (B.) cereus, B. licheniformis, and Enterococcus faecalis, while the dominant fungi and Gram-negative bacteria were Candida albicans and Sphingomonas paucimobilis, respectively. Considering no significant relationship between the indoor dust endotoxin levels and the isolation of Gram-negative bacteria from the indoor air, monitoring the indoor airborne endotoxin level was found to be also critical for risk assessment on health for animals or workers. The present study confirms the importance of microbiological monitoring and control on animal husbandry indoor air to ensure animal and worker welfare. PMID:27456779

Genetic modifications to temperate Enterococcus faecalis phage ϕEf11 that abolish the establishment of lysogeny and sensitivity to repressor, and increase host range and productivity of lytic infection

PubMed Central

Zhang, H.; Fouts, D. E.; DePew, J.

2013-01-01

ϕEf11 is a temperate bacteriophage originally isolated by induction from a lysogenic Enterococcus faecalis strain recovered from an infected root canal, and the ϕEf11 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 ϕEf11 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 ϕEf11 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 ϕEf11 and a defective ϕFL1C-like prophage located in the E. faecalis JH2-2 chromosome. The recombinant possessed five ORFs of the defective ϕFL1C-like prophage in place of six ORFs of the ϕEf11 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 ϕEf11 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

The kinetic study of hydrogen bacteria and methanotrophs in pure and defined mixed cultures

DOE Office of Scientific and Technical Information (OSTI.GOV)

Arora, D.K.

The kinetics of pure and mixed cultures of Alcaligenes eutrophus H 16 and Methylobacterium organophilum CRL 26 under double substrate limited conditions were studied. In pure culture growth kinetics, a non-interactive model was found to fit the experimental data best. The yield of biomass on limiting substrate was found to vary with the dilution rate. The variation in the biomass yield may be attributed to the change in metabolic pathways resulting from a shift in the limiting substrates. Both species exhibited wall growth in the chemostat under dark conditions. However, under illuminated conditions, there was significant reduction in wall growth.more » Poly-{beta}-hydroxybutyric acid was synthesized by both species under ammonia and oxygen limiting conditions. The feed gas mixture was optimized to achieve the steady-state coexistence of these two species in a chemostate for the first time. In mixed cultures, the biomass species assays were differentiated on the basis of their selective growth on particular compounds: Sarcosine and D-arabinose were selected for hydrogen bacteria and methylotrophs, respectively. The kinetics parameters estimated from pure cultures were used to predict the growth kinetics of these species in defined mixed cultures.« less

Effect of Passive Ultrasonic Irrigation on Enterococcus faecalis from Root Canals: An Ex Vivo Study.

PubMed

Guerreiro-Tanomaru, Juliane Maria; Chávez-Andrade, Gisselle Moraima; de Faria-Júnior, Norberto Batista; Watanabe, Evandro; Tanomaru-Filho, Mário

2015-01-01

Endodontic irrigation aims to clean and disinfect the root canal system. Passive ultrasonic irrigation (PUI) is based on the use of an ultrasound-activated instrument into the root canal filled with irrigant. The aim of this study was to evaluate, ex vivo, the effectiveness of PUI in eliminating Enterococcus faecalis from root canals. Seventy-five extracted human single-root teeth were used. After root canal preparation, specimens were inoculated with E. faecalis and incubated at 37 °C for 21 days. Specimens were distributed into five groups (n=15), according to the irrigation method: PUI + saline solution (PUI/SS); PUI + 1% NaOCl (PUI/NaOCl); conventional needle irrigation (CNI) + saline solution (CNI/SS); CNI + 1% NaOCl (CNI/NaOCl); No irrigation (control). Microbiological samples were collected at three time points: initial (21 days after inoculation), post-irrigation (immediately after irrigation), and final (7 days after irrigation). Data were obtained in CFU mL-1 and subjected to analysis by ANOVA and Tukey's tests at 5% significance level. The post-irrigation samples did not demonstrate statistical difference between PUI/SS and CNI/SS nor between PUI/NaOCl and CNI/NaOCl (p>0.05), but PUI/NaOCl and CNI/NaOCl had lower CFU mL-1 number than the other groups (p>0.05). Statistically significant difference was observed between the initial and post-irrigation samples and between the post-irrigation and final samples (p<0.05) in all groups, except in the control. The final samples of all groups presented bacterial counts similar to the initial samples. PUI or CNI with 1% NaOCl contribute to disinfection, but are unable to eradicate E. faecalis from the root canal system.

[Effect of compound Chinese traditional medicine on infected root canal bacteria biofilm].

PubMed

Ma, Rui; Huang, Li-li; Xia, Wen-wei; Zhu, Cai-lian; Ye, Dong-xia

2010-08-01

To assess the efficacy of compound Chinese traditional medicine(CTM), which composed of gallic acid, magnolol and polysaccharide of Blettila striata, against the infected root canal bacterial biofilm. Actinomyces viscosus (Av), Enterococcus faecalis (Ef), Fusobacterium nucleatum (Fn) were composed to form biofilm, then confocal laser scan microscope (CLSM) was used to observe and study the bacterial activity. SAS6.12 software package was used for statistical analysis. The biofilm thickness reduced after treatment by both CTM and ZnO (P>0.05),while there was a significant decrease of the percentage of vital bacterias after treatment by CTM (P<0.01). The compound Chinese traditional medicine is effective on biofilm control, so that it would be an effective disinfecting drug for root canal sealers. Supported by Research Fund of Bureau of Traditional Chinese Medicine of Shanghai Municipality (Grant No.2008L008A).

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

Antimicrobial Effect of Ozone Made by KP Syringe of High-Frequency Ozone Generator

PubMed Central

Prebeg, Domagoj; Katunarić, Marina; Budimir, Ana; Šegović, Sanja; Anić, Ivica

2016-01-01

Aim The aim of this study was to evaluate in vitro the antibacterial effect of ozone on suspension of three different bacteria inoculated in prepared canals of extracted human teeth. Material and methods Ozone was produced by special KP syringe of high frequency ozone generator Ozonytron (Biozonix, München, Germany) from aspirated atmospheric air by dielectric barrier discharge and applied through the tip of the syringe to the prepared root canal. The microorganisms used were Enterococcus faecalis, Staphylococcus aureus and Staphylococcus epidermidis. Results However, none of the methods was 100% effective against the three bacterial types in suspension. Application of ozone significantly decreased the absolute count of microorganisms (89.3%), as well as the count of each type of bacteria separately (Staphylococcus aureus 94.0%; Staphylococcus epidermidis 88.6% and Enterococcus faecalis 79.7%). Ozone generated by KP syringe was statistically more effective compared to NaOCl as positive control, for Staphylococcus aureus and Staphylococcus epidermidis. Conclusion The absolute count of Enterococcus faecalis was statistically decreased without a statistically significant difference between the tested group and positive control, respectively. Among the three types of bacteria in suspension, KP probe had the lowest antimicrobial effect against Enterococcus faecalis. PMID:27789911

Electrochemical disinfection of bacteria-laden water using antimony-doped tin-tungsten-oxide electrodes.

PubMed

Ghasemian, Saloumeh; Asadishad, Bahareh; Omanovic, Sasha; Tufenkji, Nathalie

2017-12-01

Electrochemical disinfection has been shown to be an efficient method with a shortrequired contact time for treatment of drinking water supplies, industrial raw water supplies, liquid foodstuffs, and wastewater effluents. In the present work, the electrochemical disinfection of saline water contaminated with bacteria was investigated in chloride-containing solutions using Sb-doped Sn 80% -W 20% -oxide anodes. The influence of current density, bacterial load, initial chloride concentration, solution pH, and the type of bacteria (E. coli D21, E. coli O157:H7, and E. faecalis) on disinfection efficacy was systematically examined. The impact of natural organic matter and a radical scavenger on the disinfection process was also examined. The electrochemical system was highly effective in bacterial inactivation for a 0.1 M NaCl solution contaminated with ∼10 7  CFU/mL bacteria by applying a current density ≥1 mA/cm 2 through the cell.100% inactivation of E. coli D21 was achieved with a contact time of less than 60 s and power consumption of 48 Wh/m 3 , by applying a current density of 6 mA/cm 2 in a 0.1 M NaCl solution contaminated with ∼10 7 CFU/mL. Reactive chlorine species as well as reactive oxygen species (e.g. hydroxyl radicals) generated in situ during the electrochemical process were determined to be responsible for inactivation of bacteria. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kang, Ji Yong; Lee, Hyung Ho; Yoon, Hye Jin

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.more » 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%.« less

Time-dependent antibacterial effects of Aloe vera and Zataria multiflora plant essential oils compared to calcium hydroxide in teeth infected with Enterococcus faecalis.

PubMed

Abbaszadegan, Abbas; Sahebi, Safoora; Gholami, Ahmad; Delroba, Alireza; Kiani, Amin; Iraji, Aida; Abbott, Paul Vincent

2016-02-01

In the present in vitro study, we investigated the time-related antimicrobial efficacy of Aloe vera and Zataria multiflora (Z. multiflora) plant essential oils compared to calcium hydroxide ([Ca[OH]2 ) to eliminate Enterococcus faecalis (E. faecalis) from root canals. A new strain of E. faecalis (Enterococcus spp. AGH04) was isolated from a previously root-filled tooth with persistent apical periodontitis. The 16S rRNA sequence was analyzed and deposited in GeneBank under accession number KF465681. A total of 108 extracted human single-rooted teeth were contaminated with this bacterial strain and treated with Aloe vera essential oil, Z. multiflora essential oil, and Ca(OH)2 for 1, 7, and 14 days. Gas chromatography-mass spectrometry (GC-MS) was used to determine the chemical composition of the oils. The percentage reduction from initial c.f.u./mL counts were calculated and analyzed. Carvacrol, thymol, and linalool were the main constituents of both essential oils. The c.f.u./mL count reductions significantly increased for all three medicaments when the contact time was extended. A statistically-significant difference was observed between the medicaments after 1 and 7 days, but there was no significant difference after 14 days. Both medicinal herbs showed equal antimicrobial efficiency against E. faecalis, comparable to Ca(OH)2 for the prolonged contact time of 14 days. © 2014 Wiley Publishing Asia Pty Ltd.

Diversity and dynamics of antibiotic-resistant bacteria in cheese as determined by PCR denaturing gradient gel electrophoresis.

PubMed

Flórez, Ana Belén; Mayo, Baltasar

2015-12-02

This work reports the composition and succession of tetracycline- and erythromycin-resistant bacterial communities in a model cheese, monitored by polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE). Bacterial 16S rRNA genes were examined using this technique to detect structural changes in the cheese microbiota over manufacturing and ripening. Total bacterial genomic DNA, used as a template, was extracted from cultivable bacteria grown without and with tetracycline or erythromycin (both at 25 μg ml(-1)) on a non-selective medium used for enumeration of total and viable cells (Plate Count agar with Milk; PCA-M), and from those grown on selective and/or differential agar media used for counting various bacterial groups; i.e., lactic acid bacteria (de Man, Rogosa and Sharpe agar; MRSA), micrococci and staphylococci (Baird-Parker agar; BPA), and enterobacteria (Violet Red Bile Glucose agar; VRBGA). Large numbers of tetracycline- and erythromycin-resistant bacteria were detected in cheese samples at all stages of ripening. Counts of antibiotic-resistant bacteria varied widely depending on the microbial group and the point of sampling. In general, resistant bacteria were 0.5-1.0 Log10 units fewer in number than the corresponding susceptible bacteria. The PCR-DGGE profiles obtained with DNA isolated from the plates for total bacteria and the different bacterial groups suggested Escherichia coli, Lactococcus lactis, Enterococcus faecalis and Staphylococcus spp. as the microbial types resistant to both antibiotics tested. This study shows the suitability of the PCR-DGGE technique for rapidly identifying and tracking antibiotic resistant populations in cheese and, by extension, in other foods. Copyright © 2015 Elsevier B.V. All rights reserved.

Efficacy of low concentrations of sodium hypochlorite and low-powered Er,Cr:YSGG laser activated irrigation against an Enterococcus faecalis biofilm.

PubMed

Christo, J E; Zilm, P S; Sullivan, T; Cathro, P R

2016-03-01

To establish the antibacterial efficacy of low concentrations of sodium hypochlorite with and without Er,Cr:YSGG laser activation on Enterococcus faecalis biofilms in extracted teeth. The root canals of 96 decoronated single-rooted extracted human teeth were prepared to a size 40, 0.06 taper 1 mm beyond the apex. They were mounted within a flow cell, which was sterilized before pumping a nutrient media through the root canals. The flow cell was inoculated with E. faecalis (ATCC 700802) and cultivated for 4 weeks. The root-ends were sealed, and the roots were then subjected to one of six treatment groups: group 1: syringe irrigation (SI) with saline (control) using a 27 -gauge Monoject needle 1 mm from the apex for 2 min; group 2: as for group 1 but with 1% NaOCl; group 3: as for group 1 but with 4% NaOCl; group 4: 0.5% NaOCl irrigation for 15 s followed by laser-activated irrigation (LAI) with four 15-s cycles replenishing the irrigant between cycles; group 5: as for group 4 but with 1% NaOCl as the irrigant; group 6: as for group 4 but with 4% NaOCl as the irrigant. Following treatment, teeth were crushed and viable bacteria were quantitated by serial dilution and plating. The colony-forming unit values were compared between groups using one-way anova and Tukey-adjusted post hoc tests. A two-tailed P value of <0.05 was considered statistically significant. The mean number of cells recovered from the 1% NaOCl SI group was significantly higher than that from the 4% NaOCl LAI group (P = 0.02). Within the limitations of this laboratory study, low-powered (0.5 W) Er,Cr:YSGG laser activation did not improve the antibacterial effect of low concentrations of sodium hypochlorite. © 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Biotoxicity of Mars soils: 2. Survival of Bacillus subtilis and Enterococcus faecalis in aqueous extracts derived from six Mars analog soils

NASA Astrophysics Data System (ADS)

Schuerger, Andrew C.; Ming, Doug W.; Golden, D. C.

2017-07-01

The search for an extant microbiota on Mars depends on exploring sites that contain transient or permanent liquid water near the surface. Examples of possible sites for liquid water may be active recurring slope lineae (RSL) and fluid inclusions in ice or salt deposits. The presence of saline fluids on Mars will act to depress the freezing points of liquid water to as low as ‒60 °C, potentially permitting the metabolism and growth of halophilic microorganisms to temperatures significantly below the freezing point of pure water at 0 °C. In order to predict the potential risks of forward contamination by Earth microorganisms to subsurface sites on Mars with liquid brines, experiments were designed to characterize the short-term survival of two bacteria in aqueous soil solutions from six analog soils. The term ''soil'' is used here to denote any loose, unconsolidated matrix with no implications for the presence or absence of organics or biology. The analog soils were previously described (Schuerger et al., 2012, Planetary Space Sci., 72, 91-101), and represented crushed Basalt (benign control), Salt, Acid, Alkaline, Aeolian, and Phoenix analogs on Mars. The survival rates of spores of Bacillus subtilis and vegetative cells of Enterococcus faecalis were tested in soil solutions from each analog at 24, 0, or ‒70 °C for time periods up to 28 d. Survival of dormant spores of B. subtilis were mostly unaffected by incubation in the aqueous extracts of all six Mars analogs. In contrast, survival rates of E. faecalis cells were suppressed by all soil solutions when incubated at 24 °C but improved at 0 and ‒70 °C, except for assays in the Salt and Acid soil solutions in which most cells were killed. Results suggest that Earth microorganisms that form spores may persist in liquid brines on Mars better than non-spore forming species, and thus, spore-forming species may pose a potential forward contamination risk to sites with liquid brines.

Strain-specific inhibition of the adherence of uropathogenic bacteria to bladder cells by probiotic Lactobacillus spp.

PubMed

de Llano, Dolores González; Arroyo, Amalia; Cárdenas, Nivia; Rodríguez, Juan Miguel; Moreno-Arribas, M Victoria; Bartolomé, Begoña

2017-06-01

Urinary tract infections (UTIs), one of most common infections worldwide, face high recurrence rates and increasing antimicrobial resistance. Probiotic bacteria, especially of the genus Lactobacillus, are considered a promising preventive and/or treatment therapy against UTIs. In order to elucidate the mechanisms involved in these beneficial effects, we studied the impact of different Lactobacillus strains (Lactobacillus salivarius UCM572, L. plantarum CLC17 and L. acidophilus 01) in the adherence of reference and clinical uropathogenic strains (Escherichia coli ATCC® 53503, E. coli 10791, Enterococcus faecalis 04-1, En. faecalis 08-1 and Staphylococcus epidermidis 08-3) to T24 epithelial bladder cells. In general, the Lactobacillus strains with previous in vivo evidence of beneficial effects against UTIs (L. salivarius UCM572 and L. acidophilus 01) significantly inhibited the adherence of the five uropathogens to T24 cells, displaying percentages of inhibition ranging between 22.2% and 43.9%, and between 16.5% and 53.7%, respectively. On the other hand, L. plantarum CLC17, a strain with no expected effects on UTIs, showed almost negligible anti-adherence effects.Therefore, these in vitro results suggest that inhibition of the adherence of uropathogens to epithelial bladder cells may be one of the mechanisms involved in the potential beneficial effects of probiotics against UTIs in vivo. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid-bound resistance to heavy metals.

PubMed

Mergeay, M; Nies, D; Schlegel, H G; Gerits, J; Charles, P; Van Gijsegem, F

1985-04-01

Alcaligenes eutrophus strain CH34, which was isolated as a bacterium resistant to cobalt, zinc, and cadmium ions, shares with A. eutrophus strain H16 the ability to grow lithoautotrophically on molecular hydrogen, to form a cytoplasmic NAD-reducing and a membrane-bound hydrogenase, and most metabolic attributes; however, it does not grow on fructose. Strain CH34 contains two plasmids, pMOL28 (163 kilobases) specifying nickel, mercury, and cobalt resistance and pMOL30 (238 kilobases) specifying zinc, cadmium, mercury, and cobalt resistance. The plasmids are self-transmissible in homologous matings, but at low frequencies. The transfer frequency was strongly increased with IncP1 plasmids RP4 and pUZ8 as helper plasmids. The phenotypes of the wild type, cured strains, and transconjugants are characterized by the following MICs (Micromolar) in strains with the indicated phenotypes: Nic+, 2.5; Nic-, 0.6; Cob+A, 5.0; Cob+B, 20.0; Cob-, less than 0.07; Zin+, 12.0; Zin-, 0.6; Cad+, 2.5; and Cad-, 0.6. Plasmid-free cells of strain CH34 are still able to grow lithoautotrophically and to form both hydrogenases, indicating that the hydrogenase genes are located on the chromosome, in contrast to the Hox structural genes of strain H16, which are located on the megaplasmid pHG1 (450 kilobases).

Model-Based Nutrient Feeding Strategies for the Increased Production of Polyhydroxybutyrate (PHB) by Alcaligenes latus.

PubMed

Gahlawat, Geeta; Srivastava, Ashok K

2017-10-01

Polyhydroxyalkanoates (PHAs) are biodegradable polymers which are considered as an effective alternative for conventional plastics due to their mechanical properties similar to the latter. However, the widespread use of these polymers is still hampered due to their higher cost of production as compared to plastics. The production cost could be overcome by obtaining high yields and productivity. The goal of the present research was to enhance the yield of polyhydroxybutyrate (PHB) with the help of two simple fed-batch cultivation strategies. In the present study, average batch kinetic and substrate limitation/inhibition study data of Alcaligenes latus was used for the development of PHB model which was then adopted for designing various off-line nutrient feeding strategies to enhance PHB accumulation. The predictive ability of the model was validated by experimental implementation of two fed-batch strategies. One such dynamic strategy of fed-batch cultivation under pseudo-steady state with respect to nitrogen and simultaneous carbon feeding strategy resulted in significantly high biomass and PHB concentration of 39.17 g/L and 29.64 g/L, respectively. This feeding strategy demonstrated a high PHB productivity and PHB content of 0.6 g/L h and 75%, respectively, which were remarkably high in comparison to batch cultivation. The mathematical model can also be employed for designing various other nutrient feeding strategies.

Impact of peptidoglycan O-acetylation on autolytic activities of the Enterococcus faecalis N-acetylglucosaminidase AtlA and N-acetylmuramidase AtlB.

PubMed

Emirian, Aurélie; Fromentin, Sophie; Eckert, Catherine; Chau, Françoise; Dubost, Lionel; Delepierre, Muriel; Gutmann, Laurent; Arthur, Michel; Mesnage, Stéphane

2009-09-17

Autolysins are potentially lethal enzymes that partially hydrolyze peptidoglycan for incorporation of new precursors and septum cleavage after cell division. Here, we explored the impact of peptidoglycan O-acetylation on the enzymatic activities of Enterococcus faecalis major autolysins, the N-acetylglucosaminidase AtlA and the N-acetylmuramidase AtlB. We constructed isogenic strains with various O-acetylation levels and used them as substrates to assay E. faecalis autolysin activities. Peptidoglycan O-acetylation had a marginal inhibitory impact on the activities of these enzymes. In contrast, removal of cell wall glycopolymers increased the AtlB activity (37-fold), suggesting that these polymers negatively control the activity of this enzyme.

Occurrence of antimicrobial agents, drug-resistant bacteria, and genes in the sewage-impacted Vistula River (Poland).

PubMed

Giebułtowicz, Joanna; Tyski, Stefan; Wolinowska, Renata; Grzybowska, Wanda; Zaręba, Tomasz; Drobniewska, Agata; Wroczyński, Piotr; Nałęcz-Jawecki, Grzegorz

2018-02-01

Antimicrobial agents (antimicrobials) are a group of therapeutic and hygienic agents that either kill microorganisms or inhibit their growth. Their occurrence in surface water may reveal harmful effects on aquatic biota and challenge microbial populations. Recently, there is a growing concern over the contamination of surface water with both antimicrobial agents and multidrug-resistant bacteria. The aim of the study was the determination of the presence of selected antimicrobials at specific locations of the Vistula River (Poland), as well as in tap water samples originating from the Warsaw region. Analysis was performed using the liquid chromatography-electrospray ionization-tandem mass spectrometry method. In addition, the occurrence of drug-resistant bacteria and resistance genes was determined using standard procedures. This 2-year study is the first investigation of the simultaneous presence of antimicrobial agents, drug-resistant bacteria, and genes in Polish surface water. In Poland, relatively high concentrations of macrolides are observed in both surface and tap water. Simultaneous to the high macrolide levels in the environment, the presence of the erm B gene, coding the resistance to macrolides, lincosamides, and streptogramin, was detected in almost all sampling sites. Another ubiquitous gene was int1, an element of the 5'-conserved segment of class 1 integrons that encode site-specific integrase. Also, resistant isolates of Enterococcus faecium and Enterococcus faecalis and Gram-negative bacteria were recovered. Multidrug-resistant bacteria isolates of Gram-negative and Enterococcus were also detected. The results show that wastewater treatment plants (WWTP) are the main source of most antimicrobials, resistant bacteria, and genes in the aquatic environment, probably due to partial purification during wastewater treatment processes.

Phenotypic and genotypic characterization of lactic acid bacteria isolated from raw goat milk and effect of farming practices on the dominant species of lactic acid bacteria.

PubMed

Tormo, Hélène; Ali Haimoud Lekhal, Djamila; Roques, C

2015-10-01

Lactic acid bacteria, in particular Lactococcus lactis, play a decisive role in the cheese making process and more particularly in lactic cheeses which are primarily produced on goat dairy farms. The objective of this study was therefore to identify the main lactic acid bacteria found in raw goats' milk from three different regions in France and evaluate if certain farming practices have an effect on the distribution of species of lactic acid bacteria in the various milk samples. Identification at genus or species level was carried out using phenotypic tests and genotypic methods including repetitive element REP-PCR, species-specific PCR and 16S rRNA gene sequencing. The distribution of the main bacterial species in the milk samples varied depending on farms and their characteristics. Out of the 146 strains identified, L. lactis was the dominant species (60% of strains), followed by Enterococcus (38%) of which Enterococcus faecalis and Enterococcus faecium. Within the species L. lactis, L. lactis subsp lactis was detected more frequently than L. lactis subsp cremoris (74% vs. 26%). The predominance of L. lactis subsp cremoris was linked to geographical area studied. It appears that the animals' environment plays a role in the balance between the dominance of L. lactis and enterococci in raw goats' milk. The separation between the milking parlor and the goat shed (vs no separation) and only straw in the bedding (vs straw and hay) seems to promote L. lactis in the milk (vs enterococci). Copyright © 2015 Elsevier B.V. All rights reserved.

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

Characterization of a TOL-like plasmid from Alcaligenes eutrophus that controls expression of a chromosomally encoded p-cresol pathway.

PubMed Central

Hughes, E J; Bayly, R C; Skurray, R A

1984-01-01

Alcaligenes eutrophus wild-type strain 345 metabolizes m- and p-toluate via a catechol meta-cleavage pathway. DNA analysis, curing studies, and transfer of this phenotype by conjugation and transformation showed that the degradative genes are encoded on a self-transmissible 85-kilobase plasmid, pRA1000. HindIII and XhoI restriction endonuclease analysis of pRA1000 showed it to be similar to the archetypal TOL plasmid, pWWO, differing in the case of HindIII only by the absence of fragments B and D present in pWWO. In strain 345, the presence of pRA1000 prevented the expression of chromosomally encoded enzymes required for the degradation of p-cresol, whereas these enzymes were expressed in strains cured of pRA1000. On the basis of studies with an R68.45-pRA1000 cointegrate plasmid, pRA1001, we conclude that the gene(s) responsible for the effect of p-cresol degradation resides within or near the m- and p-toluate degradative region on pRA1000. Images PMID:6325399

In vitro photodynamic activity of lipid vesicles with zinc phthalocyanine derivative against Enterococcus faecalis.

PubMed

Sobotta, Lukasz; Dlugaszewska, Jolanta; Kasprzycki, Piotr; Lijewski, Sebastian; Teubert, Anna; Mielcarek, Jadwiga; Gdaniec, Maria; Goslinski, Tomasz; Fita, Piotr; Tykarska, Ewa

2018-06-01

Zinc(II) phthalocyanine bearing eight non-peripheral 2-propoxy substituents was subjected to physicochemical study and, after incorporation in lipid vesicles, assessed as a potential photosensitizer for antibacterial photodynamic therapy. The phthalocyanine derivative obtained in the macrocyclization reaction was characterized by MS and NMR techniques. Moreover, its chemical purity was confirmed by HPLC analysis. X-ray structural analysis revealed that overcrowding of the phthalocyanine derivative leads to a strong out-of-plane distortion of the π-system of the macrocycle core. In the UV-Vis absorption spectra of zinc(II) phthalocyanine two characteristic bands were found: the Soret (300-450 nm) and the Q band (600-800 nm). Photophysical properties of mono- and diprotonated forms of phthalocyanine derivative were studied with time-resolved fluorescence spectroscopy. Its tri- and tetraprotonated forms could not be obtained, because compound decomposes in higher acid concentrations. The presented zinc(II) phthalocyanine showed values of singlet oxygen generation Φ Δ  = 0.18 and 0.16, the quantum yield of the photodecomposition Φ P  = 3.06∙10 -4 and 1.23∙10 -5 and the quantum yield of fluorescence Φ FL  = 0.005 and 0.004, designated in DMF and DMSO, respectively. For biological studies, phthalocyanine has been incorporated into modified liposome vesicles containing ethanol. In vitro bacteria photoinactivation study revealed no activity against Escherichia coli and 5.7 log reduction of the Enterococcus faecalis growth. Copyright © 2018 Elsevier B.V. All rights reserved.

Evaluation of polymorphisms in pbp4 gene and genetic diversity in penicillin-resistant, ampicillin-susceptible Enterococcus faecalis from hospitals in different states in Brazil.

PubMed

Infante, Victor Hugo Pacagnelli; Conceição, Natália; de Oliveira, Adriana Gonçalves; Darini, Ana Lúcia da Costa

2016-04-01

The aim of the present study was to verify whether penicillin-resistant, ampicillin-susceptible Enterococcus faecalis (PRASEF) occurred in Brazil prior to the beginning of the 21st century, and to verify whether ampicillin susceptibility can predict susceptibility to other β-lactams in E. faecalis with this inconsistent phenotype. The presence of polymorphisms in the pbp4 gene and genetic diversity among the isolates were investigated. Of 21 PRASEF analyzed, 5 (23.8%) and 4 (19.0%) were imipenem and piperacillin resistant simultaneously by disk diffusion and broth dilution respectively, contradicting the current internationally accepted standards of susceptibility testing. Sequencing of pbp4 gene revealed an amino acid substitution (Asp-573→Glu) in all PRASEF isolates but not in the penicillin-susceptible, ampicillin-susceptible E. faecalis. Most PRASEF (90.5%) had related pulsed-field gel electrophoresis profiles, but were different from other PRASEF described to date. Results demonstrate that penicillin-resistant, ampicillin-susceptible phenotype was already a reality in the 1990s in E. faecalis isolates in different Brazilian states, and some of these isolates were also imipenem- and piperacillin-resistant; therefore, internationally accepted susceptibility criteria cannot be applied to these isolates. According to pbp4 gene sequencing, this study suggests that a specific amino acid substitution in pbp4 gene found in all PRASEF analyzed is associated with penicillin resistance. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Evaluation of pyrrolidonyl arylamidase for the identification of nonfermenting Gram-negative rods.

PubMed

Bombicino, Karina A; Almuzara, Marisa N; Famiglietti, Angela M R; Vay, Carlos

2007-01-01

To evaluate the activity of pyrrolidonyl arylamidase (PYR) for the differentiation and identification of nonfermenting gram negative rods (NFGNR), 293 isolates were tested. A 24 h culture of each test organism was prepared. From this a 108-109 cfu/mL suspension was added to 0.25 mL of sterile physiologic solution. A PYR disk was then added and the test was incubated for 30 minutes at 35-37 degrees C, at environmental atmosphere. Reading was done by adding 1 drop of cinnamaldehyde reagent. Strains of Acinetobacter baumannii, Acinetobacter haemolyticus, Alcaligenes faecalis, Bergeyella zoohelcum, Bordetella bronchiseptica, Bordetella hinzii, Brevundimonas diminuta, Brevundimonas vesicularis, Brucella ovis, Brucella spp., Brucella suis, Burkholderia cepacia complex, Moraxella catarrhalis, Moraxella lacunata, Moraxella nonliquefaciens, Moraxella osloensis, Oligella ureolytica, Pseudomonas alcaligenes, Pseudomonas mendocina, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas stutzeri, Pseudomonas Vb3, Psychrobacter phenylpyruvicus, and Stenotrophomonas maltophilia were PYR negative. On the other hand Achromobacter piechaudii, Achromobacter denitrificans, Achromobacter xylosoxidans, Burkholderia gladioli, Chryseobacterium gleum-indologenes, Comamonas testosroni, Cupriavidus pauculus, Delftia acidovorans, Elizabethkingia meningoseptica, Myroides spp., Ochrobactrum anthropi, Pseudomonas oryzihabitans, Ralstonia pickettii, Rhizobium radiobacter, Shewanella spp., Sphingobacterium multivorum, Sphingobacterium spiritivorum, and Weeksella virosa were PYR positive. Finally, Acinetobacter lwoffii, Pseudomonas aeruginosa, Pseudomonas fluorescens, Roseomonas spp., and Sphingomonas paucimobilis-parapaucimobilis were PYR variable. PYR testing should be considered as a useful tool to facilitate the identification of NFGNR.

Increasing Prevalence of Aminoglycoside-Resistant Enterococcus faecalis Isolates Due to the aac(6')-aph(2") Gene: A Therapeutic Problem in Kermanshah, Iran.

PubMed

Khani, Mitra; Fatollahzade, Mahdie; Pajavand, Hamid; Bakhtiari, Somaye; Abiri, Ramin

2016-03-01

Enterococci are important pathogens in nosocomial infections. Various types of antibiotics, such as aminoglycosides, are used for treatment of these infections. Enterococci can acquire resistant traits, which can lead to therapeutic problems with aminoglycosides. This study was designed to identify the prevalence of, and to compare, the aac(6')-aph(2") and aph(3)-IIIa genes and their antimicrobial resistance patterns among Enterococcus faecalis and E. faecium isolates from patients at Imam Reza hospital in Kermanshah in 2011 - 2012. One hundred thirty-eight clinical specimens collected from different wards of Imam Reza hospital were identified to the species level by biochemical tests. Antimicrobial susceptibility tests against kanamycin, teicoplanin, streptomycin, imipenem, ciprofloxacin, and ampicillin were performed by the disk diffusion method. The minimum inhibitory concentrations of gentamicin, streptomycin, kanamycin, and amikacin were evaluated with the microbroth dilution method. The aminoglycoside resistance genes aac(6')-aph(2") and aph(3")-IIIa were analyzed with multiplex PCR. The prevalence of isolates was 33 (24.1%) for E. faecium and 63 (46%) for E. faecalis. Eighty-nine percent of the isolates were high-level gentamicin resistant (HLGR), and 32.8% of E. faecium isolates and 67.2% of E. faecalis isolates carried aac(6')-aph(2"). The prevalence of aph(3")-IIIa among the E. faecalis and E. faecium isolates was 22.7% and 77.3%, respectively. Remarkably increased incidence of aac(6')-aph(2") among HLGR isolates explains the relationship between this gene and the high level of resistance to aminoglycosides. As the resistant gene among enterococci can be transferred, the use of new-generation antibiotics is necessary.

CATALASE ACTIVITY OF TWO STREPTOCOCCUS FAECALIS STRAINS AND ITS ENHANCEMENT BY AEROBIOSIS AND ADDED CATIONS1

PubMed Central

Jones, Dorothy; Deibel, R. H.; Niven, C. F.

1964-01-01

Jones, Dorothy (American Meat Institute Foundation, Chicago, Ill.), R. H. Deibel, and C. F. Niven, Jr. Catalase activity of two Streptococcus faecalis strains and its enhancement by aerobiosis and added cations. J. Bacteriol. 88:602–610. 1964.—The nature of catalase activity noted in two unusual Streptococcus faecalis strains was determined. Enzyme activity was lost slowly when cultures were maintained by daily transfer in test tubes of broth media. Loss of activity could be prevented by aerobic culture. Supplementation of the growth medium with ferric, manganese, and zinc ions, as well as aerobiosis, enhanced catalase activity. However, addition of these cations to cell suspensions or to cell-free extracts did not increase catalase activity. Although oxygen was observed to be one of the reaction end products, the catalase activity was not inhibited by cyanide or azide, and the iron-porphyrin coenzyme of classical catalase was not detected. The enzyme was purified 185-fold by precipitation with ammonium sulfate, followed by chromotography on a diethylaminoethyl cellulose column. PMID:14208495

Changes in the community structure of free-living heterotrophic bacteria in the open tropical Pacific Ocean in response to microalgal lysate-derived dissolved organic matter.

PubMed

Tada, Yuya; Suzuki, Koji

2016-07-01

Dissolved organic matter derived from phytoplankton (DOMP) can affect the bacterial biomass and community structure in aquatic ecosystems. Here, we examined the community response of free-living heterotrophic bacteria, with respect to cellular nucleic acid levels, to the DOMP lysates derived from three phytoplankton strains in the open tropical Pacific. The free amino acid (FAA) composition of each DOMP lysate differed among the microalgal strains. Terminal restriction fragment-length polymorphism analyses with 16S rRNA genes revealed that the community shifts of high nucleic acid (HNA) and low nucleic acid (LNA) bacteria varied significantly with the different DOMP lysate treatments. Furthermore, the FAA composition in DOMP lysates significantly affected the bacterial community shifts in HNA and LNA. Similarity percentage analysis using 16S rRNA gene deep-sequencing revealed that the DOMP lysates from the pelagophyte Pelagomonas calceolata caused relatively large community shifts with Alcaligenes predominating in the HNA fraction. In contrast, the DOMP lysate from the diatom Thalassiosira oceanica induced a community shift in the LNA fraction with a predominance of uncultured Actinobacteria Thus, the data indicate that the DOMP lysates from different microalgae constitute a primary factor altering the dominant bacterial groups in the open ocean. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Loop-Mediated Isothermal Amplification Label-Based Gold Nanoparticles Lateral Flow Biosensor for Detection of Enterococcus faecalis and Staphylococcus aureus

PubMed Central

Wang, Yi; Li, Hui; Wang, Yan; Zhang, Lu; Xu, Jianguo; Ye, Changyun

2017-01-01

The report describes a simple, rapid and sensitive assay for visual and multiplex detection of Enterococcus faecalis and Staphylococcus aureus based on multiple loop-mediated isothermal amplification (mLAMP) and lateral flow biosensor (LFB). Detection and differentiation of the Ef0027 gene (E. faecalis-specific gene) and nuc gene (S. aureus-specific gene) were determined using fluorescein (FITC)-and digoxin-modified primers in the mLAMP process. In the presence of biotin- and FITC-/digoxin-modified primers, the mLAMP yielded numerous biotin- and FITC-/digoxin-attached duplex products, which were detected by LFB through biotin/streptavidin interaction (biotin on the duplex and streptavidin on the gold nanoparticle) and immunoreactions (FITC/digoxin on the duplex and anti-FITC/digoxin on the LFB test line). The accumulation of gold nanoparticles generated a characteristic red line, enabling visual and multiplex detection of target pathogens without instrumentation. The limit of detection (LoD), analytical specificity and feasibility of LAMP-LFB technique were successfully examined in pure culture and blood samples. The entire procedure, including specimen (blood samples) processing (30 min), isothermal reaction (40 min) and result reporting (within 2 min), could be completed within 75 min. Thus, this assay offers a simple, rapid, sensitive and specific test for multiplex detection of E. faecalis and S. aureus strains. Furthermore, the LAMP-LFB strategy is a universal technique, which can be extended to detect various target sequences by re-designing the specific LAMP primers. PMID:28239371

Characterization of beta-carotene ketolases, CrtW, from marine bacteria by complementation analysis in Escherichia coli.

PubMed

Choi, Seon-kang; Nishida, Yasuhiro; Matsuda, Satoru; Adachi, Kyoko; Kasai, Hiroaki; Peng, Xue; Komemushi, Sadao; Miki, Wataru; Misawa, Norihiko

2005-01-01

A complementation analysis was performed in Escherichia coli to evaluate the efficiency of beta-carotene ketolases (CrtW) from the marine bacteria Brevundimonas sp. SD212, Paracoccus sp. PC1 (Alcaligenes PC-1), and Paracoccus sp. N81106 (Agrobacterium aurantiacum), for astaxanthin production. Each crtW gene was expressed in Escherichia coli synthesizing zeaxanthin due to the presence of plasmid pACCAR25DeltacrtX. Carotenoids that accumulated in the resulting E. coli transformants were examined by chromatographic and spectroscopic analyses. The transformant carrying the Paracoccus sp. PC1 or N81106 crtW gene accumulated high levels of adonixanthin, which is the final astaxanthin precursor for CrtW, and astaxanthin, while the E. coli transformant with crtW from Brevundimonas sp. SD212 did not accumulate any adonixanthin and produced a high level of astaxanthin. These results show efficient conversion by CrtW of Brevundimonas sp. SD212 from adonixanthin to astaxanthin, which is a new-found characteristic of a bacterial CrtW enzyme. The phylogenetic positions between CrtW of the two genera, Brevundimonas and Paracoccus, are distant, although they fall into alpha-Proteobacteria.

Alkaline stress response in Enterococcus faecalis: adaptation, cross-protection, and changes in protein synthesis.

PubMed Central

Flahaut, S; Hartke, A; Giard, J C; Auffray, Y

1997-01-01

The alkaline shock response in Enterococcus faecalis was studied in this work. Cells adapted to an optimum pH of 10.5 were tolerate to pH 11.9 conditions but acquired sensitivity to acid damage. An analysis of stress proteins revealed that 37 polypeptides were amplified. Two of these are DnaK and GroEL. The combined results show that bile salts and alkaline stress responses are closely related. PMID:9023964

Using color histograms and SPA-LDA to classify bacteria.

PubMed

de Almeida, Valber Elias; da Costa, Gean Bezerra; de Sousa Fernandes, David Douglas; Gonçalves Dias Diniz, Paulo Henrique; Brandão, Deysiane; de Medeiros, Ana Claudia Dantas; Véras, Germano

2014-09-01

In this work, a new approach is proposed to verify the differentiating characteristics of five bacteria (Escherichia coli, Enterococcus faecalis, Streptococcus salivarius, Streptococcus oralis, and Staphylococcus aureus) by using digital images obtained with a simple webcam and variable selection by the Successive Projections Algorithm associated with Linear Discriminant Analysis (SPA-LDA). In this sense, color histograms in the red-green-blue (RGB), hue-saturation-value (HSV), and grayscale channels and their combinations were used as input data, and statistically evaluated by using different multivariate classifiers (Soft Independent Modeling by Class Analogy (SIMCA), Principal Component Analysis-Linear Discriminant Analysis (PCA-LDA), Partial Least Squares Discriminant Analysis (PLS-DA) and Successive Projections Algorithm-Linear Discriminant Analysis (SPA-LDA)). The bacteria strains were cultivated in a nutritive blood agar base layer for 24 h by following the Brazilian Pharmacopoeia, maintaining the status of cell growth and the nature of nutrient solutions under the same conditions. The best result in classification was obtained by using RGB and SPA-LDA, which reached 94 and 100 % of classification accuracy in the training and test sets, respectively. This result is extremely positive from the viewpoint of routine clinical analyses, because it avoids bacterial identification based on phenotypic identification of the causative organism using Gram staining, culture, and biochemical proofs. Therefore, the proposed method presents inherent advantages, promoting a simpler, faster, and low-cost alternative for bacterial identification.

Detection and isolation of novel rhizopine-catabolizing bacteria from the environment

PubMed

Gardener; de Bruijn FJ

1998-12-01

Microbial rhizopine-catabolizing (Moc) activity was detected in serial dilutions of soil and rhizosphere washes. The activity observed generally ranged between 10(6) and 10(7) catabolic units per g, and the numbers of nonspecific culture-forming units were found to be approximately 10 times higher. A diverse set of 37 isolates was obtained by enrichment on scyllo-inosamine-containing media. However, none of the bacteria that were isolated were found to contain DNA sequences homologous to the known mocA, mocB, and mocC genes of Sinorhizobium meliloti L5-30. Twenty-one of the isolates could utilize an SI preparation as the sole carbon and nitrogen source for growth. Partial sequencing of 16S ribosomal DNAs (rDNAs) amplified from these strains indicated that five distinct bacterial genera (Arthrobacter, Sinorhizobium, Pseudomonas, Aeromonas, and Alcaligenes) were represented in this set. Only 6 of these 21 isolates could catabolize 3-O-methyl-scyllo-inosamine under standard assay conditions. Two of these, strains D1 and R3, were found to have 16S rDNA sequences very similar to those of Sinorhizobium meliloti. However, these strains are not symbiotically effective on Medicago sativa, and DNA sequences homologous to the nodB and nodC genes were not detected in strains D1 and R3 by Southern hybridization analysis.

Detection and Isolation of Novel Rhizopine-Catabolizing Bacteria from the Environment

PubMed Central

Gardener, Brian B. McSpadden; de Bruijn, Frans J.

1998-01-01

Microbial rhizopine-catabolizing (Moc) activity was detected in serial dilutions of soil and rhizosphere washes. The activity observed generally ranged between 106 and 107 catabolic units per g, and the numbers of nonspecific culture-forming units were found to be approximately 10 times higher. A diverse set of 37 isolates was obtained by enrichment on scyllo-inosamine-containing media. However, none of the bacteria that were isolated were found to contain DNA sequences homologous to the known mocA, mocB, and mocC genes of Sinorhizobium meliloti L5-30. Twenty-one of the isolates could utilize an SI preparation as the sole carbon and nitrogen source for growth. Partial sequencing of 16S ribosomal DNAs (rDNAs) amplified from these strains indicated that five distinct bacterial genera (Arthrobacter, Sinorhizobium, Pseudomonas, Aeromonas, and Alcaligenes) were represented in this set. Only 6 of these 21 isolates could catabolize 3-O-methyl-scyllo-inosamine under standard assay conditions. Two of these, strains D1 and R3, were found to have 16S rDNA sequences very similar to those of Sinorhizobium meliloti. However, these strains are not symbiotically effective on Medicago sativa, and DNA sequences homologous to the nodB and nodC genes were not detected in strains D1 and R3 by Southern hybridization analysis. PMID:9835587

Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid-bound resistance to heavy metals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mergeay, M.; Nies, D.; Schlegel, H.G.

1985-04-01

Alcaligenes eutrophus strain CH34, which was isolated as a bacterium resistant to cobalt, zinc, and cadmium ions, shares with A. eutrophus strain H16 the ability to grow lithoautotrophically on molecular hydrocarbon, to form a cytoplasmic NAD-reducing and a membrane-bound hydrogenase, and most metabolic attributes; however, it does not grow on fructose. Strain CH34 contains two plasmids, pMOL28 (163 kilobases) specifying nickel, mercury, and cobalt resistance and pMOL30 (238 kilobases) specifying zinc, cadmium, mercury, and cobalt resistance. The plasmids are self-transmissible in homologous matings, but at low frequencies. The transfer frequency was strongly increased with IncP1 plasmids RP4 and pUZ8 asmore » helper plasmids. The phenotypes of the wild type, cured strains, and transconjugants are characterized by the following MICs (Micromolar) in strains with the indicated phenotypes: Nic/sup +/, 2.5; Nic/sup -/, 0.6; Cob/sup +/A, 5.0; Cob/sup +/B, 20.0; Cob/sup -/, < 0.07; Zin/sup +/, 12.0; Zin/sup -/, 0.6; Cad/sup +/, 2.5; and Cad/sup -/, 0.6. Plasmid-free cells of strain CH34 are still able to grow lithoautotrophically and to form both hydrogenases, indicating that the hydrogenase genes are located on the chromosome, in contrast to the Hox structural genes of strain H16, which are located on the megaplasmid pHG1 (450 kilobases).« less

Degradation of Chlorophenols by Alcaligenes eutrophus JMP134(pJP4) in Bleached Kraft Mill Effluent

PubMed Central

Valenzuela, J.; Bumann, U.; Cespedes, R.; Padilla, L.; Gonzalez, B.

1997-01-01

The ability of Alcaligenes eutrophus JMP134(pJP4) to degrade 2,4-dichlorophenoxyacetic acid, 2,4,6-trichlorophenol, and other chlorophenols in a bleached kraft mill effluent was studied. The efficiency of degradation and the survival of strain JMP134 and indigenous microorganisms in short-term batch or long-term semicontinuous incubations performed in microcosms were assessed. After 6 days of incubation, 2,4-dichlorophenoxyacetate (400 ppm) or 2,4,6-trichlorophenol (40 to 100 ppm) were extensively degraded (70 to 100%). In short-term batch incubations, indigenous microorganisms were unable to degrade such of compounds. Degradation of 2,4,6-trichlorophenol by strain JMP134 was significantly lower at 200 to 400 ppm of compound. This strain was also able to degrade 2,4-dichlorophenoxyacetate, 2,4,6-trichlorophenol, 4-chlorophenol, and 2,4,5-trichlorophenol when bleached Kraft mill effluent was amended with mixtures of these compounds. On the other hand, the chlorophenol concentration and the indigenous microorganisms inhibited the growth and survival of the strain in short-term incubations. In long-term (>1-month) incubations, strain JMP134 was unable to maintain a large, stable population, although extensive 2,4,6-trichlorophenol degradation was still observed. The latter is probably due to acclimation of the indigenous microorganisms to degrade 2,4,6-trichlorophenol. Acclimation was observed only in long-term, semicontinuous microcosms. PMID:16535488

Evaluation of Antimicrobial and Antifungal efficacy of Chitosan as endodontic irrigant against Enterococcus Faecalis and Candida Albicans Biofilm formed on tooth substrate.

PubMed

Yadav, Pankaj; Chaudhary, Sarika; Saxena, Rajendra K; Talwar, Sangeeta; Yadav, Sudha

2017-03-01

Bacterial biofilms formed on the root canal wall are often difficult to remove. This study aimed to evaluate the cytotoxic effect and antibacterial efficacy of chitosan when used as root canal irrigant against E. Faecalis and Candida albicans biofilm formed on tooth substrate. The present study evaluated antibacterial effect of 0.25% Chitosan, 0.5% Chitosan, 2% chlorhexidine and 3% sodium hypochlorite against Enterococcus faecalis and Candida Albicans . Agar-well diffusion methods, minimal inhibitory concentration tests and biofilm susceptibility assays were used to determine antibacterial activity. Teeth specimens were sectioned to obtain a standardized tooth length of 12mm. Specimens were inoculated with 10 mL of the freshly prepared E. Faecalis suspension and Candida albicans for 4 weeks. The specimens were then instrumented with ProTaper rotary files F3 size. After irrigation with test solution, three sterile paper points were placed into one canal, left for 60 s and transferred to a test tube containing 1 mL of reduced transport fluid. The number of CFU in 1 mL was determined. 3-week biofilm qualitative assay showed complete inhibition of bacterial growth with 3% Sodium hypochlorite, 2% Chlorhexidine and Chitosan except saline, which showed presence of bacterial growth. Significant reduction of colony forming units (CFU)/mL was observed for the chitosan groups and the antibacterial activity of the chitosan groups was at par with 3% NaOCl and 2% Chlorhexidine. It was observed that the chitosan showed no cytotoxicity at 3mg/ml and 10% cytotoxicity at 6mg/ml. The use of chitosan as a root canal irrigant might be an alternative considering the various undesirable properties of NaOCl and chlorhexidine. Key words: Biofilm, Candida albicans, Chitosan, Cytotoxicity, Enterococcus faecalis.

Production and characterization of poly(3-hydroxybutyrate) generated by Alcaligenes latus using lactose and whey after acid protein precipitation process.

PubMed

Berwig, Karina Hammel; Baldasso, Camila; Dettmer, Aline

2016-10-01

Whey after acid protein precipitation was used as substrate for PHB production in orbital shaker using Alcaligenes latus. Statistical analysis determined the most appropriate hydroxide for pH neutralization of whey after protein precipitation among NH4OH, KOH and NaOH 10%w/v. The results were compared to those of commercial lactose. A scale-up test in a 4L bioreactor was done at 35°C, 750rpm, 7L/min air flow, and 6.5 pH. The PHB was characterized through Fourier Transform Infrared Spectroscopy, thermogravimetry and differential scanning calorimetry. NH4OH provided the best results for productivity (p), 0.11g/L.h, and for polymer yield, (YP/S), 1.08g/g. The bioreactor experiment resulted in lower p and YP/S. PHB showed maximum degradation temperature (291°C), melting temperature (169°C), and chemical properties similar to those of standard PHB. The use of whey as a substrate for PHB production did not affect significantly the final product quality. Copyright © 2016 Elsevier Ltd. All rights reserved.

Antimicrobial activity, antibiotic susceptibility and virulence factors of Lactic Acid Bacteria of aquatic origin intended for use as probiotics in aquaculture

PubMed Central

2013-01-01

Background The microorganisms intended for use as probiotics in aquaculture should exert antimicrobial activity and be regarded as safe not only for the aquatic hosts but also for their surrounding environments and humans. The objective of this work was to investigate the antimicrobial/bacteriocin activity against fish pathogens, the antibiotic susceptibility, and the prevalence of virulence factors and detrimental enzymatic activities in 99 Lactic Acid Bacteria (LAB) (59 enterococci and 40 non-enterococci) isolated from aquatic animals regarded as human food. Results These LAB displayed a broad antimicrobial/bacteriocin activity against the main Gram-positive and Gram-negative fish pathogens. However, particular safety concerns based on antibiotic resistance and virulence factors were identified in the genus Enterococcus (86%) (Enterococcus faecalis, 100%; E. faecium, 79%). Antibiotic resistance was also found in the genera Weissella (60%), Pediococcus (44%), Lactobacillus (33%), but not in leuconostocs and lactococci. Antibiotic resistance genes were found in 7.5% of the non-enterococci, including the genera Pediococcus (12.5%) and Weissella (6.7%). One strain of both Pediococcus pentosaceus and Weissella cibaria carried the erythromycin resistance gene mef(A/E), and another two P. pentosaceus strains harboured lnu(A) conferring resistance to lincosamides. Gelatinase activity was found in E. faecalis and E. faecium (71 and 11%, respectively), while a low number of E. faecalis (5%) and none E. faecium exerted hemolytic activity. None enterococci and non-enterococci showed bile deconjugation and mucin degradation abilities, or other detrimental enzymatic activities. Conclusions To our knowledge, this is the first description of mef(A/E) in the genera Pediococcus and Weissella, and lnu(A) in the genus Pediococcus. The in vitro subtractive screening presented in this work constitutes a valuable strategy for the large-scale preliminary selection of putatively safe LAB

Antimicrobial Activity of Kefir against Various Food Pathogens and Spoilage Bacteria

PubMed Central

Kim, Hyunsook

2016-01-01

Kefir is a unique fermented dairy product produced by a mixture of lactic acid bacteria, acetic acid bacteria, and yeast. Here, we compared the antimicrobial spectra of four types of kefirs (A, L, M, and S) fermented for 24, 36, 48, or 72 h against eight food-borne pathogens. Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Enterococcus faecalis, Escherichia coli, Salmonella Enteritidis, Pseudomonas aeruginosa, and Cronobacter sakazakii were used as test strains, and antibacterial activity was investigated by the spot on lawn method. The spectra, potencies, and onsets of activity varied according to the type of kefir and the fermentation time. The broadest and strongest antimicrobial spectrum was obtained after at least 36-48 h of fermentation for all kefirs, although the traditional fermentation method of kefir is for 18-24 h at 25℃. For kefir A, B. cereus, E. coli, S. Enteritidis, P. aeruginosa, and C. sakazakii were inhibited, while B. cereus, S. aureus, E. coli, S. Enteritidis, P. aeruginosa, and C. sakazakii were inhibited to different extents by kefirs L, M, and S. Remarkably, S. aureus, S. Enteritidis, and C. sakazakii were only inhibited by kefirs L, M, and S, and L. monocytogenes by kefir M after fermentation for specific times, suggesting that the antimicrobial activity is attributable not only to a low pH but also to antimicrobial substances secreted during the fermentation. PMID:28115890

Antimicrobial Activity of Kefir against Various Food Pathogens and Spoilage Bacteria.

PubMed

Kim, Dong-Hyeon; Jeong, Dana; Kim, Hyunsook; Kang, Il-Byeong; Chon, Jung-Whan; Song, Kwang-Young; Seo, Kun-Ho

2016-01-01

Kefir is a unique fermented dairy product produced by a mixture of lactic acid bacteria, acetic acid bacteria, and yeast. Here, we compared the antimicrobial spectra of four types of kefirs (A, L, M, and S) fermented for 24, 36, 48, or 72 h against eight food-borne pathogens. Bacillus cereus , Staphylococcus aureus , Listeria monocytogenes , Enterococcus faecalis , Escherichia coli , Salmonella Enteritidis , Pseudomonas aeruginosa , and Cronobacter sakazakii were used as test strains, and antibacterial activity was investigated by the spot on lawn method. The spectra, potencies, and onsets of activity varied according to the type of kefir and the fermentation time. The broadest and strongest antimicrobial spectrum was obtained after at least 36-48 h of fermentation for all kefirs, although the traditional fermentation method of kefir is for 18-24 h at 25℃. For kefir A, B. cereus , E. coli , S . Enteritidis, P. aeruginosa , and C. sakazakii were inhibited, while B. cereus , S. aureus , E. coli , S . Enteritidis, P. aeruginosa , and C. sakazakii were inhibited to different extents by kefirs L, M, and S. Remarkably, S. aureus , S . Enteritidis, and C. sakazakii were only inhibited by kefirs L, M, and S, and L. monocytogenes by kefir M after fermentation for specific times, suggesting that the antimicrobial activity is attributable not only to a low pH but also to antimicrobial substances secreted during the fermentation.

Genetic modifications and introduction of heterologous pdc genes in Enterococcus faecalis for its use in production of bioethanol.

PubMed

Rana, N F; Gente, S; Rincé, A; Auffray, Y; Laplace, J M

2012-09-01

Genetically-modified Enterococcus faecalis has a potential of survival and can be used in ethanolic fermentations. Fermentation profiles of E. faecalis JH2-2 were assessed using glucose and lactose as carbon sources. Deletion of lactate dehydrogenase (ldh) genes increased the ethanol production from 0.25 to 0.82 g/l, which was further increased to 0.96 g/l by the insertion of a pyruvate decarboxylase (pdc) gene (from Sarcina ventriculi or Clostridium acetobutylicum) in place ldh1. When grown on lactose, the pdcSv and pdcCa showed 13.6 and 17.6 U mg(-1) of pdc specific activity, respectively. Highest activity (47 U mg(-1)) and ethanol concentration (2.3 g/l) were obtained with pdcCa using an expression plasmid. Formate and acetate were also produced in high quantities. Transcriptional analysis showed that aldehyde alcohol dehydrogenase gene was upregulated up to 16-fold. Further optimizations are required for higher ethanol production.

Heterologous expression of enterocin AS-48 in several strains of lactic acid bacteria.

PubMed

Fernández, M; Martínez-Bueno, M; Martín, M C; Valdivia, E; Maqueda, M

2007-05-01

Enterococcus faecalis produces a cationic and circular enterocin, AS-48, of 7149 Da, the genetic determinants of which are located within the pMB2 plasmid. We have compared enterocin AS-48 production by different enterococci species with that of other 'safe' lactic acid bacteris (LAB) (GRAS status) and looked into the subsequent application of this enterocin in food production. In an effort to exploit this system for the heterologous expression of enterocin AS-48, a number of vectors containing the as-48 cluster were constructed and used to transform several LAB strains (genera Enterococcus, Lactococcus and Lactobacillus) Heterologous production of enterocin AS-48 failed when bacteria other than those belonging to the genus Enterococcus were used as hosts, although expression of a partial level of resistance against AS-48 were always detected, ruling out the possibility of a lack of recognition of the enterococcal promoters. Our results reveal the special capacity of species from the genus Enterococcus to produce AS-48, an enterocin that requires a post-transcriptional modification to generate a circular peptide with a wide range of inhibitory activity against pathogenic and spoilage bacteria. Preliminary experiments in foodstuffs using nonvirulent enterococci with interesting functional properties reveal the possibility of a biotechnological application of these transformants.

Specificity of the lipase-specific foldases of gram-negative bacteria and the role of the membrane anchor.

PubMed

El Khattabi, M; Ockhuijsen, C; Bitter, W; Jaeger, K E; Tommassen, J

1999-06-01

Folding of lipases that are secreted by Pseudomonads and other gram-negative bacteria via the type II secretion pathway is facilitated by dedicated chaperones, called lipase-specific foldases (Lifs). Lifs are membrane-anchored proteins with a large periplasmic domain. The functional interaction between the Lif and its cognate lipase is specific, since the Pseudomonas aeruginosa Lif was found not to substitute for Lifs from Burkholderia glumae or Acinetobacter calcoaceticus. However, the P. aeruginosa Lif was able to activate the lipase from the closely related species P. alcaligenes. Hybrid proteins constructed from parts of the P. aeruginosa and B. glumae Lifs revealed that the C-terminal 138 amino acids of the B. glumae Lif determine the specificity of the interaction with the cognate lipase. Furthermore, the periplasmic domain of the B. glumae Lif was functional when cloned in frame with a cleavable signal sequence, which demonstrates that the membrane anchor is not essential for Lif function in vivo. However, the recombinant Lif was released into the medium, indicating that the function of the membrane anchor is to prevent secretion of the Lif together with the lipase.

The two-component system GrvRS (EtaRS) regulates ace expression in Enterococcus faecalis OG1RF.

PubMed

Roh, Jung Hyeob; Singh, Kavindra V; La Rosa, Sabina Leanti; Cohen, Ana Luisa V; Murray, Barbara E

2015-01-01

Expression of ace (adhesin to collagen of Enterococcus faecalis), encoding a virulence factor in endocarditis and urinary tract infection models, has been shown to increase under certain conditions, such as in the presence of serum, bile salts, urine, and collagen and at 46 °C. However, the mechanism of ace/Ace regulation under different conditions is still unknown. In this study, we identified a two-component regulatory system GrvRS as the main regulator of ace expression under these stress conditions. Using Northern hybridization and β-galactosidase assays of an ace promoter-lacZ fusion, we found transcription of ace to be virtually absent in a grvR deletion mutant under the conditions that increase ace expression in wild-type OG1RF and in the complemented strain. Moreover, a grvR mutant revealed decreased collagen binding and biofilm formation as well as attenuation in a murine urinary tract infection model. Here we show that GrvR plays a major role in control of ace expression and E. faecalis virulence. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Gentamicin induces efaA expression and biofilm formation in Enterococcus faecalis.

PubMed

Kafil, Hossein Samadi; Mobarez, Ashraf Mohabati; Moghadam, Mehdi Forouzandeh; Hashemi, Zahra Sadat; Yousefi, Mehdi

2016-03-01

Enterococci have been ranked among the leading causes of nosocomial bacteremia and urinary tract infection. This study aimed to investigate the effect of ampicillin, vancomycin, gentamicin and ceftizoxime on biofilm formation and gene expression of colonization factors on Enterococcus faecalis. Twelve clinical isolates of E. faecalis were used to investigate the effect of antibiotics on biofilm formation and gene expression of efaA, asa1, ebpA, esp and ace. Flow system assay and Microtiter plates were used for biofilm assay. Two hundred clinical isolates were used for confirming the effect of antibiotics on biofilm formation. Ampicillin, vancomycin and ceftizoxime did not have any significant effect on biofilm formation, but gentamicin induced biofilm formation in 89% of isolates. In twelve selected isolate gentamicin increased expression of esp (+50.9%) and efaA (+33.9%) genes and reduced or maintained expression of others (asa1:-47.4%, ebpA: 0, ace:-19.2%). Vancomycin increased expression of esp (+89.1%) but reduced the others (asa1: -34.9%, ebpA:-11%, ace:-30%, efaA:-60%). Ceftizoxime increased slightly ebpA (+19.7%) and reduced others (asa1:-66.2%, esp:-35%, ace:-28.1%, efaA:-38.4%). and ampicillin strongly increased expression of ace (+231%), esp (+131%) and ebpA (+83%) but reduced others (asa1:-85.5%, efaA:-47.4%). The findings of the present study showed that antibiotics may have a role in biofilm formation and sustainability of enterococci, especially in case of gentamicin. efaA gene may have an important role, especially in antibiotic induced biofilm formation by gentamicin. Experiments with efaA mutants are needed to investigate the exact effect of efaA on biofilm formation with antibiotic induced cells. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enterococcus faecalis endocarditis following percutaneous manipulation of a biliary tract calculus and ERCP

PubMed Central

Ronald, Allan R; Pattullo, Andrew LS

1990-01-01

A case of Enterococcus faecalis endocarditis followed endoscopic retrograde cholangiopancreatography and percutaneous extraction of a biliary calculus is reported. The most likely cause of endocarditis, though unproven, is the latter procedure, as the bile is often infected during biliary tract obstruction, and bacteremia is frequent during percutaneous manipulations. Initial therapy with vancomycin was unsuccessful in clearing the bacteremia, possibly due to vancomycin tolerance of the isolate and lack of an aminoglycoside in the initial regimen. Cure was obtained when therapy with ampicillin and gentamicin was undertaken. PMID:22553458

An in vitro evaluation of antimicrobial activity of five herbal extracts and comparison of their activity with 2.5% sodium hypochlorite against Enterococcus faecalis.

PubMed

Saxena, Divya; Saha, Suparna Ganguly; Saha, Mainak Kanti; Dubey, Sandeep; Khatri, Margie

2015-01-01

Sodium hypochlorite is the most widely used irrigant in endodontic practice, but it has various disadvantages. Literature has shown that herbal products such as Propolis, Azadirachta indica (AI), Triphala, Curcuma longa, and Morinda citrifolia (MC) possess good antimicrobial properties and thus can be used as potential endodontic irrigants. To evaluate and compare the antimicrobial activity of five herbal extracts, i.e., Propolis, AI, Triphala, C. longa, and MC with that of 2.5% sodium hypochlorite against Enterococcus faecalis. E. faecalis American Type Culture Collection 21292 was inoculated onto brain heart infusion agar plate. Discs impregnated with herbal medicaments were placed on the inoculated plates and incubated at 37°C aerobically for 24 h and growth inhibition zones were measured. Mean zone of inhibition in descending order was found as sodium hypochlorite > Propolis > AI > Triphala > C. longa = MC > ethanol. Statistical analysis was performed using one-way analysis of variance which showed a significant difference in the zone of inhibition of sodium hypochlorite and Propolis (P < 0.001). Propolis showed highest zone of inhibition among all the herbal extracts next to sodium hypochlorite. Propolis and AI have significant antimicrobial activity against E. faecalis.

Enterococcus faecalis Endogenous Endophthalmitis from Valvular Endocarditis

PubMed Central

Barge, Sidnei; Rothwell, Renata; Varandas, Rosário; Agrelos, Luís

2013-01-01

We report a case of a 74-year-old female, with a mitral heart valve, who presented with pain and blurred vision in the right eye for 2 days. Her visual acuity was light perception (LP) in the right eye and 20/40 in the left eye. Slit lamp examination showed corneal edema and hypopyon, and a view of the right fundus was impossible. Echography showed vitreous condensation. One day after presentation, the patient developed acute lung edema requiring hospitalization, so she was not submitted to vitreous tap and intravitreal treatment. The cardiac and systemic evaluations revealed a mitral endocarditis secondary to Enterococcus faecalis. The patient improved systemically with treatment with gentamicin, vancomycin, and linezolid. Her visual acuity remained as no LP, and her intraocular pressure (IOP) has been controlled with brimonidine bid despite developing a total cataract with 360° posterior synechia. A cardiac source for endogenous endophthalmitis should be considered in the presence of a prosthetic cardiac valve. The treatment and followup must be made in cooperation with a cardiologist specialist, but the ophthalmologist can play a key role in the diagnosis. PMID:23936701

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

PubMed

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

2016-10-02

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.

Biofuel Cells Select for Microbial Consortia That Self-Mediate Electron Transfer

PubMed Central

Rabaey, Korneel; Boon, Nico; Siciliano, Steven D.; Verhaege, Marc; Verstraete, Willy

2004-01-01

Microbial fuel cells hold great promise as a sustainable biotechnological solution to future energy needs. Current efforts to improve the efficiency of such fuel cells are limited by the lack of knowledge about the microbial ecology of these systems. The purposes of this study were (i) to elucidate whether a bacterial community, either suspended or attached to an electrode, can evolve in a microbial fuel cell to bring about higher power output, and (ii) to identify species responsible for the electricity generation. Enrichment by repeated transfer of a bacterial consortium harvested from the anode compartment of a biofuel cell in which glucose was used increased the output from an initial level of 0.6 W m−2 of electrode surface to a maximal level of 4.31 W m−2 (664 mV, 30.9 mA) when plain graphite electrodes were used. This result was obtained with an average loading rate of 1 g of glucose liter−1 day−1 and corresponded to 81% efficiency for electron transfer from glucose to electricity. Cyclic voltammetry indicated that the enhanced microbial consortium had either membrane-bound or excreted redox components that were not initially detected in the community. Dominant species of the enhanced culture were identified by denaturing gradient gel electrophoresis and culturing. The community consisted mainly of facultative anaerobic bacteria, such as Alcaligenes faecalis and Enterococcus gallinarum, which are capable of hydrogen production. Pseudomonas aeruginosa and other Pseudomonas species were also isolated. For several isolates, electrochemical activity was mainly due to excreted redox mediators, and one of these mediators, pyocyanin produced by P. aeruginosa, could be characterized. Overall, the enrichment procedure, irrespective of whether only attached or suspended bacteria were examined, selected for organisms capable of mediating the electron transfer either by direct bacterial transfer or by excretion of redox components. PMID:15345423

Biosorption characteristic of Alcaligenes sp. BAPb.1 for removal of lead(II) from aqueous solution.

PubMed

Jin, Yu; Yu, Sumei; Teng, Chunying; Song, Tao; Dong, Liying; Liang, Jinsong; Bai, Xin; Xu, Xiuhong; Qu, Juanjuan

2017-06-01

In this study, strain BAPb.1 was isolated from lead mining area and used as an adsorbent to remove lead(II) ions from aqueous solution. The physicochemical characteristics, heavy metal resistance and antibiotic sensitivity of strain BAPb.1 were investigated. Biosorption capacity was evaluated by batch biosorption experiments, and isothermal characteristics were discussed. Atomic force microscopy (AFM), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX) and Fourier transform infrared spectrometry (FTIR) were conducted to explore the mechanism for lead(II) adsorption. Based on morphological and physiological characteristics as well as the phylogenetic analysis of 16S rDNA sequences, strain BAPb.1 was identified as a member of the genus Alcaligenes. It exhibited high resistances to multiple heavy metals such as lead(II), copper(II), zinc(II), nickel(II) and chromium(VI), and to antibiotics such as kanamycin, ampicillin, streptomycin, chloramphenicol, and tetracycline. The optimum conditions for maximum biosorption rate of 85.2% and maximum capacity of 56.8 mg g -1 were found at pH of 5, adsorbent dosage of 1.5 g L -1 (dry weight), initial lead(II) concentration of 100 mg L -1 , and contact time of 30 min at 30 °C. Biosorption isotherms were well fitted with Langmuir isotherm model. Mechanism analysis reveals that the lead(II) ions may exchange with sodium and potassium ions, and the hydroxyl, carbonyl and phosphate groups on the cell surface can chelate the lead(II) ions, therefore, surface adsorption play significant role in the biosorption process.

Gene transfer of Alcaligenes eutrophus JMP134 plasmid pJP4 to indigenous soil recipients.

PubMed Central

DiGiovanni, G D; Neilson, J W; Pepper, I L; Sinclair, N A

1996-01-01

This study evaluated the potential for gene transfer of a large catabolic plasmid from an introduced organism to indigenous soil recipients. The donor organism Alcaligenes eutrophus JMP134 contained the 80-kb plasmid pJP4, which contains genes that code for mercury resistance. Genes on this plasmid plus chromosomal genes also allow degradation of 2,4-dichloruphenoxyacetic acid (2,4-D). When JMP134 was inoculated into a nonsterile soil microcosm amended with 1,000 micrograms of 2,4-D g-1, significant (10(6) g of soil-1) populations of indigenous recipients or transconjugants arose. These transconjugants all contained an 80-kb plasmid similar in size to pJP4, and all degraded 2,4-D. In addition, all transconjugants were resistant to mercury and contained the tfdB gene of pJP4 as detected by PCR. No mercury-resistant, 2,4-D-degrading organisms with large plasmids or the tfdB gene were found in the 2,4-D-amended but uninoculated control microcosm. These data clearly show that the plasmid pJP4 was transferred to indigenous soil recipients. Even more striking is the fact that not only did the indigenous transconjugant population survive and proliferate but also enhanced rates of 2,4-D degradation occurred relative to microcosms in which no such gene transfer occurred. Overall, these data indicate that gene transfer from introduced organisms is an effective means of bioaugmentation and that survival of the introduced organism is not a prerequisite for biodegradation that utilizes introduced biodegradative genes. PMID:8779592

Quantitative evaluation of apical extrusion of intracanal bacteria using rotary ProTaper, K3XF, twisted and hand K-file system: An ex vivo study.

PubMed

Ghogre, Priyanka; Chourasia, Hemant Ramesh; Agarwal, Manish; Singh, M P; Gurav, Sandeep; Ghogre, Rahul

2015-01-01

The aim of this study was to evaluate the number of intracanal bacteria extruded apically during root canal preparation using rotary ProTaper, K3XF, twisted, and hand K-file system. Seventy extracted single-rooted human mandibular premolar teeth were used. Access cavities were prepared and the teeth were mounted in glass vials. Root canals were then contaminated with a pure culture of Enterococcus faecalis (ATCC 29212) and incubated at 37°C for 24 h. The contaminated roots were divided into four experimental groups of 15 teeth each and one control group of 10 teeth. Group 1: ProTaper; Group 2: K3XF; Group 3: Twisted file; Group 4: Hand K-file; Group 5: Control group. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples were incubated in culture media for 24 h. Colonies of bacteria were counted and the results were given as number of colony-forming units (CFU)/ml. The obtained data were analyzed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests. There was a significant difference between the rotary and hand instrumentation system related to the apically extruded intracanal bacteria. Both the rotary and hand instrumentation systems extruded intracanal bacteria through the apical foramen. K3XF file system showed least bacterial extrusion amongst all instrumentation groups.

Contribution of the autolysin AtlA to the bactericidal activity of amoxicillin against Enterococcus faecalis JH2-2.

PubMed

Bravetti, Anne-Lise; Mesnage, Stéphane; Lefort, Agnès; Chau, Françoise; Eckert, Catherine; Garry, Louis; Arthur, Michel; Fantin, Bruno

2009-04-01

The bactericidal activity of amoxicillin was investigated against Enterococcus faecalis JH2-2 and against an isogenic mutant deficient in the production of the N-acetylglucosaminidase AtlA. Comparison of the two strains indicated that this autolysin contributes to killing by amoxicillin both in vitro and in a rabbit model of experimental endocarditis.

Probiotic Enterococcus faecalis Symbioflor® down regulates virulence genes of EHEC in vitro and decrease pathogenicity in a Caenorhabditis elegans model.

PubMed

Neuhaus, Klaus; Lamparter, Marina C; Zölch, Benjamin; Landstorfer, Richard; Simon, Svenja; Spanier, Britta; Ehrmann, Matthias A; Vogel, Rudi F

2017-03-01

Enterohemorrhagic E. coli O157:H7 (EHEC) shorten the lifespan of Caenorhabditis elegans compared to avirulent bacteria. Co-feeding EHEC with Enterococcus faecalis Symbioflor ® significantly increased the worms' lifespan. The transcriptome of EHEC grown in vitro with or without Symbioflor ® was analyzed using RNA-seq. The analysis revealed downregulation of several virulence-associated genes in the presence of Symbioflor ® , including virulence key genes (e.g., LEE, flagellum, quorum-sensing). The downregulation of the LEE genes was corroborated by lux-transposon mutants. Upregulated genes included acid response genes, due to a decrease in pH exerted by Symbioflor ® . Further genes indicate cellular stress in EHEC (e.g. prophage/mobile elements involved in excision, cell lysis, and cell division inhibition). Thus, the observed protection of C. elegans during an EHEC infection by the probiotic Symbioflor ® is suggested to be caused by triggering concomitant transcriptomic changes. To verify the biological relevance of this modulation, exemplary genes found to be influenced by Symbioflor ® were knocked out (fliD, espB, Z3136, Z3917, and L7052). The lifespan of nematodes changed when using knock-outs as food source and the effect could be complemented in trans. In summary, Symbioflor ® appears to be a protective probiotic in the nematode model.

Molecular characterization of antibiotic resistance in cultivable multidrug-resistant bacteria from livestock manure.

PubMed

Yang, Qingxiang; Tian, Tiantian; Niu, Tianqi; Wang, Panliang

2017-10-01

Diverse antibiotic-resistance genes (ARGs) are frequently reported to have high prevalence in veterinary manure samples due to extensive use of antibiotics in farm animals. However, the characteristics of the distribution and transmission of ARGs among bacteria, especially among different species of multiple antibiotic-resistant bacteria (MARB), have not been well explored. By applying high-throughput sequencing methods, our study uncovered a vast MARB reservoir in livestock manure. The genera Escherichia, Myroides, Acinetobacter, Proteus, Ignatzschineria, Alcaligenes, Providencia and Enterococcus were the predominant cultivable MARB, with compositions of 40.6%-85.7%. From chicken manure isolates, 33 MARB were selected for investigation of the molecular characteristics of antibiotic resistance. A total of 61 ARGs and 18 mobile genetic elements (MGEs) were investigated. We found that 47 ARGs were widely distributed among the 33 MARB isolates. Each isolate carried 27-36 genes responsible for resistance to eight classes of antibiotics frequently used in clinic or veterinary settings. ARGs to the six classes of antibiotics other than streptogramins and vancomycin were present in all 33 MARB isolates with a prevalence of 80%-100%. A total of 12 MGEs were widely distributed among the 33 MARB, with intI1, IS26, ISaba1, and ISEcp1 simultaneously present in 100% of isolates. In addition, 9 gene cassettes within integrons and ISCR1 were detected among MARB isolates encoding resistance to different antibiotic classes. This is the first report revealing the general co-presence of multiple ARGs, various MGEs and ARG cassettes in different species of individual MARB isolates in chicken manure. The results highlight a much higher risk of ARGs spreading through livestock manure to humans than we expected. Copyright © 2017 Elsevier Ltd. All rights reserved.

pHTβ-promoted mobilization of non-conjugative resistance plasmids from Enterococcus faecium to Enterococcus faecalis.

PubMed

Di Sante, Laura; Morroni, Gianluca; Brenciani, Andrea; Vignaroli, Carla; Antonelli, Alberto; D'Andrea, Marco Maria; Di Cesare, Andrea; Giovanetti, Eleonora; Varaldo, Pietro E; Rossolini, Gian Maria; Biavasco, Francesca

2017-09-01

To analyse the recombination events associated with conjugal mobilization of two multiresistance plasmids, pRUM17i48 and pLAG (formerly named pDO1-like), from Enterococcus faecium 17i48 to Enterococcus faecalis JH2-2. The plasmids from two E. faecalis transconjugants (JH-4T, tetracycline resistant, and JH-8E, erythromycin resistant) and from the E. faecium donor (also carrying a pHTβ-like conjugative plasmid, named pHTβ17i48) were investigated by several methods, including PCR mapping and sequencing, S1-PFGE followed by Southern blotting and hybridization, and WGS. Two locations of repApHTβ were detected in both transconjugants, one on a ∼50 kb plasmid (as in the donor) and the other on plasmids of larger sizes. In JH-4T, WGS disclosed an 88.6 kb plasmid resulting from the recombination of pHTβ17i48 (∼50 kb) and a new plasmid, named pLAG (35.3 kb), carrying the tet(M), tet(L), lsa(E), lnu(B), spw and aadE resistance genes. In JH-8E, a 75 kb plasmid resulting from the recombination of pHTβ17i48 and pRUM17i48 was observed. In both cases, the cointegrates were apparently derived from replicative transposition of an IS1216 present in each of the multiresistance plasmids into pHTβ17i48. The cointegrates could resolve to yield the multiresistance plasmids and a pHTβ17i48 derivative carrying an IS1216 (unlike the pHTβ17i48 of the donor). Our results completed the characterization of the multiresistance plasmids carried by the E. faecium 17i48, confirming the role of pHT plasmids in the mobilization of non-conjugative antibiotic resistance elements among enterococci. Results also revealed that mobilization to E. faecalis was associated with the generation of cointegrate plasmids promoted by IS1216-mediated transposition. © The Author 2017. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Isolation and characterization of enterocin W, a novel two-peptide lantibiotic produced by Enterococcus faecalis NKR-4-1.

PubMed

Sawa, Naruhiko; Wilaipun, Pongtep; Kinoshita, Seisuke; Zendo, Takeshi; Leelawatcharamas, Vichien; Nakayama, Jiro; Sonomoto, Kenji

2012-02-01

Enterococcus faecalis NKR-4-1 isolated from pla-ra produces a novel two-peptide lantibiotic, termed enterocin W, comprising Wα and Wβ. The structure of enterocin W exhibited similarity with that of plantaricin W. The two peptides acted synergistically, and their order of binding to the cell membrane was important for their inhibitory activity.

Molecular Epidemiology of Vancomycin-Resistant Enterococcus faecalis and Enterococcus faecium Isolated from Clinical Specimens in the Northwest of Iran.

PubMed

Jahansepas, Ali; Ahangarzadeh Rezaee, Mohammad; Hasani, Alka; Sharifi, Yaeghob; Rahnamaye Farzami, Marjan; Dolatyar, Alireza; Aghazadeh, Mohammad

2018-04-30

This study was conducted to investigate the phenotypic and genotypic characteristics of vancomycin-resistant Enterococcus faecalis and Enterococcus faecium. Antibiotic resistance and virulence genes in the aforementioned resistant isolates were studied using the epsilometer (E)-test and polymerase chain reaction (PCR). These isolates were subjected to typing by pulsed-field gel electrophoresis (PFGE). Thirty vancomycin-resistant enterococci (VRE; 18.75%) were isolated from a total of 160 various clinical specimens cultured for any bacterial growth. Of these, 11 (36.7%) isolates were identified as E. faecalis and 19 (63.3%) as E. faecium. Minimum inhibitory concentrations (MICs) of vancomycin, teicoplanin, and three alternative therapeutic options (linezolid, daptomycin, and quinupristin/dalfopristin) were determined using the E-test. Multiplex PCR was done for confirming species, identification of the resistant genotypes, and the detection of the virulence genes. Finally, the clonal relationship of all VRE strains was studied by PFGE. All VRE strains showed vancomycin MIC ≥256 μg/mL, and 27 (90%) isolates carried the vanA gene, whereas none of the isolates carried vanB. The most common resistance antibiotic pattern observed was toward rifampicin (n = 30 [100%]). Among all virulence genes studied, gelE (n = 28 [93.33%]) was found as the most prevalent virulent gene. VRE isolates exhibited 90%, 46.67%, 100%, and 66.67% resistance to teicoplanin, linezolid, quinupristin/dalfopristin, and daptomycin, respectively. Molecular typing demonstrated 16 PFGE types of VRE isolates (A-P). Although vanA was carried by most of the isolates, PFGE displayed small clonal dissemination among VR E. faecium and VR E. faecalis species.

Recovery of Enterococcus faecalis from cheese in the oral cavity of healthy subjects.

PubMed

Razavi, A; Gmür, R; Imfeld, T; Zehnder, M

2007-08-01

Enterococci are rarely found in the healthy human oral cavity, yet they are strongly associated with filled root canals. The origin of these enterococci remains unknown. Our hypothesis is that they are transient food-born colonizers under healthy conditions. This pilot study reinvestigated the prevalence of enterococci in the oral cavity of healthy volunteers, screened cheese samples for enterococci and investigated colonization of the oral cavity after ingestion of an enterocci-positive cheese. Concentrated oral rinse samples were collected from a cohort of 50 dental students and proved negative for viable enterococci. Twenty cheese samples were obtained from local supermarkets. Enterococci were cultured and identified using standard methods. Viable enterococci were detected in one of five specimens of Swiss Tilsiter, three of five samples of French soft cheese, one of five Mozzarella samples and one of five Feta samples. Eight volunteers from the cohort consumed 10 g of a cheese with high Enterococcus faecalis load. Oral rinse samples were collected before and 1, 10 and 100 min after cheese ingestion. One minute after ingestion, a median of 5,480 E. faecalis colony-forming units was recovered from the oral rinse samples. Bacterial counts were reduced after 10 min, had dropped after 100 min to levels that were significantly (P < 0.005) different from the 1-min and 10-min scores and were below the detection limit after 1 week. These findings suggest that colonization of the healthy oral cavity by enterococci is transitional, but at the same time add weight to our hypothesis that enterococcal root canal infections could be food-borne.

Evaluation of efficacy of combinations of five endodontic sealers with five antibiotics against Enterococcus Faecalis – An in-vitro study

PubMed Central

Sharma, Deepak; Grover, Rohit; Pinnameneni, Prasanth Sai; Dey, Subhra; Raju, P Ramakrishnam

2014-01-01

Background: To evaluate and compare in vitro the antibacterial efficacy of five antibiotics when added individually to five endodontic sealers against Enterococcus faecalis (EF). Materials & Methods: This controlled trial with systematic allocation method was carried out to detect the combined antibacterial activity of five endodontic sealers (Kerr sealer EWT, Endomethasone, AH26, AH Plus, Roekoseal) with five antibiotics regularly used (Amoxicillin, metronidazole, azithromyacin, gatifloxacin, doxycycline) on EF. For each sealerantibiotic combinations, thirty BHI agar plates (15 aerobic and 15 anaerobic) were inoculated with EF, containing five sterile paper discs- three of various sealer- antibiotic combinations, one of sealer alone (positive control) & plain disc as negative control were incubated at 370C for 48 hrs and the zone of inhibition was measured. Data analysis was done by ANOVA and Tukey’s post- hoc test using SPSS( version 17). Results: The findings of this study revealed that sealer-antibiotic combination containing amoxicillin had the significant difference (p<0.001) in the mean zone of inhibition compared to other combinations. Metronidazole showed the minimum zone of inhibition among used antibiotics. The sealers in the decreasing order according to their effectiveness on EF were Kerr sealer endomethasone, AH26, Rockseal, AH plus. Conclusion: Addition of antibiotics to endodontic sealers enhances their antibacterial activity against Enterococcus faecalis. How to cite the article: Sharma D, Grover R, Pinnameneni PS, Dey S, Raju PR. Evaluation of efficacy of combinations of five endodontic sealers with five antibiotics against Enterococcus Faecalis – An in-vitro study. J Int Oral Health 2014;6(2):90-5. PMID:24876708

Increasing Prevalence of Aminoglycoside-Resistant Enterococcus faecalis Isolates Due to the aac(6’)-aph(2”) Gene: A Therapeutic Problem in Kermanshah, Iran

PubMed Central

Khani, Mitra; Fatollahzade, Mahdie; Pajavand, Hamid; Bakhtiari, Somaye; Abiri, Ramin

2016-01-01

Background: Enterococci are important pathogens in nosocomial infections. Various types of antibiotics, such as aminoglycosides, are used for treatment of these infections. Enterococci can acquire resistant traits, which can lead to therapeutic problems with aminoglycosides. Objectives: This study was designed to identify the prevalence of, and to compare, the aac(6’)-aph(2”) and aph(3)-IIIa genes and their antimicrobial resistance patterns among Enterococcus faecalis and E. faecium isolates from patients at Imam Reza hospital in Kermanshah in 2011 - 2012. Patients and Methods: One hundred thirty-eight clinical specimens collected from different wards of Imam Reza hospital were identified to the species level by biochemical tests. Antimicrobial susceptibility tests against kanamycin, teicoplanin, streptomycin, imipenem, ciprofloxacin, and ampicillin were performed by the disk diffusion method. The minimum inhibitory concentrations of gentamicin, streptomycin, kanamycin, and amikacin were evaluated with the microbroth dilution method. The aminoglycoside resistance genes aac(6’)-aph(2”) and aph(3”)-IIIa were analyzed with multiplex PCR. Results: The prevalence of isolates was 33 (24.1%) for E. faecium and 63 (46%) for E. faecalis. Eighty-nine percent of the isolates were high-level gentamicin resistant (HLGR), and 32.8% of E. faecium isolates and 67.2% of E. faecalis isolates carried aac(6’)-aph(2”). The prevalence of aph(3”)-IIIa among the E. faecalis and E. faecium isolates was 22.7% and 77.3%, respectively. Conclusions: Remarkably increased incidence of aac(6’)-aph(2”) among HLGR isolates explains the relationship between this gene and the high level of resistance to aminoglycosides. As the resistant gene among enterococci can be transferred, the use of new-generation antibiotics is necessary. PMID:27217920

The missing link: Bordetella petrii is endowed with both the metabolic versatility of environmental bacteria and virulence traits of pathogenic Bordetellae.

PubMed

Gross, Roy; Guzman, Carlos A; Sebaihia, Mohammed; dos Santos, Vítor A P Martins; Pieper, Dietmar H; Koebnik, Ralf; Lechner, Melanie; Bartels, Daniela; Buhrmester, Jens; Choudhuri, Jomuna V; Ebensen, Thomas; Gaigalat, Lars; Herrmann, Stefanie; Khachane, Amit N; Larisch, Christof; Link, Stefanie; Linke, Burkhard; Meyer, Folker; Mormann, Sascha; Nakunst, Diana; Rückert, Christian; Schneiker-Bekel, Susanne; Schulze, Kai; Vorhölter, Frank-Jörg; Yevsa, Tetyana; Engle, Jacquelyn T; Goldman, William E; Pühler, Alfred; Göbel, Ulf B; Goesmann, Alexander; Blöcker, Helmut; Kaiser, Olaf; Martinez-Arias, Rosa

2008-09-30

Bordetella petrii is the only environmental species hitherto found among the otherwise host-restricted and pathogenic members of the genus Bordetella. Phylogenetically, it connects the pathogenic Bordetellae and environmental bacteria of the genera Achromobacter and Alcaligenes, which are opportunistic pathogens. B. petrii strains have been isolated from very different environmental niches, including river sediment, polluted soil, marine sponges and a grass root. Recently, clinical isolates associated with bone degenerative disease or cystic fibrosis have also been described. In this manuscript we present the results of the analysis of the completely annotated genome sequence of the B. petrii strain DSMZ12804. B. petrii has a mosaic genome of 5,287,950 bp harboring numerous mobile genetic elements, including seven large genomic islands. Four of them are highly related to the clc element of Pseudomonas knackmussii B13, which encodes genes involved in the degradation of aromatics. Though being an environmental isolate, the sequenced B. petrii strain also encodes proteins related to virulence factors of the pathogenic Bordetellae, including the filamentous hemagglutinin, which is a major colonization factor of B. pertussis, and the master virulence regulator BvgAS. However, it lacks all known toxins of the pathogenic Bordetellae. The genomic analysis suggests that B. petrii represents an evolutionary link between free-living environmental bacteria and the host-restricted obligate pathogenic Bordetellae. Its remarkable metabolic versatility may enable B. petrii to thrive in very different ecological niches.

The missing link: Bordetella petrii is endowed with both the metabolic versatility of environmental bacteria and virulence traits of pathogenic Bordetellae

PubMed Central

Gross, Roy; Guzman, Carlos A; Sebaihia, Mohammed; Martins dos Santos, Vítor AP; Pieper, Dietmar H; Koebnik, Ralf; Lechner, Melanie; Bartels, Daniela; Buhrmester, Jens; Choudhuri, Jomuna V; Ebensen, Thomas; Gaigalat, Lars; Herrmann, Stefanie; Khachane, Amit N; Larisch, Christof; Link, Stefanie; Linke, Burkhard; Meyer, Folker; Mormann, Sascha; Nakunst, Diana; Rückert, Christian; Schneiker-Bekel, Susanne; Schulze, Kai; Vorhölter, Frank-Jörg; Yevsa, Tetyana; Engle, Jacquelyn T; Goldman, William E; Pühler, Alfred; Göbel, Ulf B; Goesmann, Alexander; Blöcker, Helmut; Kaiser, Olaf; Martinez-Arias, Rosa

2008-01-01

Background Bordetella petrii is the only environmental species hitherto found among the otherwise host-restricted and pathogenic members of the genus Bordetella. Phylogenetically, it connects the pathogenic Bordetellae and environmental bacteria of the genera Achromobacter and Alcaligenes, which are opportunistic pathogens. B. petrii strains have been isolated from very different environmental niches, including river sediment, polluted soil, marine sponges and a grass root. Recently, clinical isolates associated with bone degenerative disease or cystic fibrosis have also been described. Results In this manuscript we present the results of the analysis of the completely annotated genome sequence of the B. petrii strain DSMZ12804. B. petrii has a mosaic genome of 5,287,950 bp harboring numerous mobile genetic elements, including seven large genomic islands. Four of them are highly related to the clc element of Pseudomonas knackmussii B13, which encodes genes involved in the degradation of aromatics. Though being an environmental isolate, the sequenced B. petrii strain also encodes proteins related to virulence factors of the pathogenic Bordetellae, including the filamentous hemagglutinin, which is a major colonization factor of B. pertussis, and the master virulence regulator BvgAS. However, it lacks all known toxins of the pathogenic Bordetellae. Conclusion The genomic analysis suggests that B. petrii represents an evolutionary link between free-living environmental bacteria and the host-restricted obligate pathogenic Bordetellae. Its remarkable metabolic versatility may enable B. petrii to thrive in very different ecological niches. PMID:18826580

Isolation and Characterization of Enterocin W, a Novel Two-Peptide Lantibiotic Produced by Enterococcus faecalis NKR-4-1

PubMed Central

Sawa, Naruhiko; Wilaipun, Pongtep; Kinoshita, Seisuke; Zendo, Takeshi; Leelawatcharamas, Vichien; Nakayama, Jiro

2012-01-01

Enterococcus faecalis NKR-4-1 isolated from pla-ra produces a novel two-peptide lantibiotic, termed enterocin W, comprising Wα and Wβ. The structure of enterocin W exhibited similarity with that of plantaricin W. The two peptides acted synergistically, and their order of binding to the cell membrane was important for their inhibitory activity. PMID:22138996

Effect of modified nonequilibrium plasma with chlorhexidine digluconate against endodontic biofilms in vitro.

PubMed

Du, Tianfeng; Shi, Qi; Shen, Ya; Cao, Yingguang; Ma, Jingzhi; Lu, Xinpei; Xiong, Zilan; Haapasalo, Markus

2013-11-01

Nonequilibrium plasma has been reported to effectively kill Enterococcus faecalis in endodontic biofilm compared with chlorhexidine digluconate (CHX). The purpose of this study was to evaluate the antimicrobial in vitro activity of modified nonequilibrium plasma with CHX against E. faecalis and multispecies biofilms on bovine dentin discs. Sterile bovine dentin discs were incubated with E. faecalis or a mixture of bacteria from human dental root canal infections to form 1- and 3-week-old biofilms. The specimens were subjected to nonequilibrium plasma, modified nonequilibrium plasma with CHX, and 2% CHX for 2- and 5-minute exposure. After treatment, the biofilms were stained with viability dyes and examined by confocal laser scanning microscopy and 3-dimensional reconstruction analysis. The proportions of bacterial cells killed by the treatments were calculated. The 3-dimensional reconstruction images showed that 1- and 3-week-old biofilms adhered to bovine dentin discs. The proportions of dead cells increased significantly with the longer exposure in each treatment group (P < .05). Modified nonequilibrium plasma was the most effective in killing bacteria in E. faecalis and multispecies biofilms at both 2 and 5 minutes (P < .05). No significant difference was detected between nonequilibrium plasma and CHX groups (P > .05). Significantly more cells were killed in 1-week-old biofilms than in 3-week-old biofilms in all groups (P < .05). The modified nonequilibrium plasma killed more bacteria than conventional nonequilibrium plasma and 2% CHX in E. faecalis and multispecies endodontic biofilms in vitro and thus shows promise as an additional tool in infection control during endodontic treatment. Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Apical extrusion of bacteria when using reciprocating single-file and rotary multifile instrumentation systems.

PubMed

Tinoco, J M; De-Deus, G; Tinoco, E M B; Saavedra, F; Fidel, R A S; Sassone, L M

2014-06-01

To evaluate ex vivo, apical bacterial extrusion associated with two reciprocating single-file systems (WaveOne and Reciproc) compared with a conventional multifile rotary system (BioRace). Forty-five human single-rooted mandibular incisors were used. Endodontic access cavities were prepared, and root canals were contaminated with an Enterococcus faecalis suspension. Following incubation at 37 °C for thirty days, the contaminated teeth were divided into three groups of 15 specimens each (G1 - Reciproc, G2 - WaveOne and G3 - BioRace). Positive and negative controls consisted of 5 infected teeth and 3 uninfected incisors that were instrumented with one of the tested NiTi systems, respectively. Bacteria extruded from the apical foramen during instrumentation were collected into vials containing 0.9% NaCl. The microbiological samples were taken from the vials and incubated in brain heart agar medium for 24 h. The resulting bacterial titre, in colony-forming units (CFU) per mL, was determined, and these data were analysed by Wilcoxon matched-pairs signed rank test and Kruskal-Wallis H-test. The level of significance was set at α = 0.05. No significant difference was found in the number of CFU between the two reciprocating systems (P = 0.41). The conventional multifile rotary system group was associated with significantly higher CFU than both of the two reciprocating groups (P = 0.01). All instrumentation systems extruded bacteria beyond the foramen. However, both reciprocating single-file systems extruded fewer bacteria apically than the conventional multifile rotary system. © 2013 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Physiological and biochemical characterization of the soluble formate dehydrogenase, a molybdoenzyme from Alcaligenes eutrophus.

PubMed Central

Friedebold, J; Bowien, B

1993-01-01

Organoautotrophic growth of Alcaligenes eutrophus on formate was dependent on the presence of molybdate in the medium. Supplementation of the medium with tungstate lead to growth cessation. Corresponding effects of these anions were observed for the activity of the soluble, NAD(+)-linked formate dehydrogenase (S-FDH; EC 1.2.1.2) of the organism. Lack of molybdate or presence of tungstate resulted in an almost complete loss of S-FDH activity. S-FDH was purified to near homogeneity in the presence of nitrate as a stabilizing agent. The native enzyme exhibited an M(r) of 197,000 and a heterotetrameric quaternary structure with nonidentical subunits of M(r) 110,000 (alpha), 57,000 (beta), 19,400 (gamma), and 11,600 (delta). It contained 0.64 g-atom of molybdenum, 25 g-atom of nonheme iron, 20 g-atom of acid-labile sulfur, and 0.9 mol of flavin mononucleotide per mol. The fluorescence spectrum of iodine-oxidized S-FDH was nearly identical to the form A spectrum of milk xanthine oxidase, proving the presence of a pterin cofactor. The molybdenum-complexing cofactor was identified as molybdopterin guanine dinucleotide in an amount of 0.71 mol/mol of S-FDH. Apparent Km values of 3.3 mM for formate and 0.09 mM for NAD+ were determined. The enzyme coupled the oxidation of formate to a number of artificial electron acceptors and was strongly inactivated by formate in the absence of NAD+. It was inhibited by cyanide, azide, nitrate, and Hg2+ ions. Thus, the enzyme belongs to a new group of complex molybdo-flavo Fe-S FDH that so far has been detected in only one other aerobic bacterium. Images PMID:8335630

Efficacy of 4 Irrigation Protocols in Killing Bacteria Colonized in Dentinal Tubules Examined by a Novel Confocal Laser Scanning Microscope Analysis

PubMed Central

Azim, Adham A.; Aksel, Hacer; Zhuang, Tingting; Mashtare, Terry; Babu, Jegdish P.; Huang, George T.-J.

2016-01-01

Introduction The aim of this study was to determine the efficiency of 4 irrigation systems in eliminating bacteria in root canals, particularly in dentinal tubules. Methods Roots of human teeth were prepared to 25/04, autoclaved, and inoculated with Enterococcus faecalis for 3 weeks. Canals were then disinfected by (1) standard needle irrigation, (2) sonically agitating with EndoActivator, (3) XP Endo finisher, or (4) erbium:yttrium aluminum garnet laser (PIPS) (15 roots/group). The bacterial reduction in the canal was determined by MTT assays. For measuring live versus dead bacteria in the dentinal tubules (4 teeth/group), teeth were split open and stained with LIVE/DEAD BackLight. Coronal, middle, and apical thirds of the canal dentin were scanned by using a confocal laser scanning microscope (CLSM) to determine the ratio of dead/total bacteria in the dentinal tubules at various depths. Results All 4 irrigation protocols significantly eliminated bacteria in the canal, ranging from 89.6% to 98.2% reduction (P < .001). XP Endo had the greatest bacterial reduction compared with other 3 techniques (P < .05). CLSM analysis showed that XP Endo had the highest level of dead bacteria in the coronal, middle, and apical segments at 50-μm depth. On the other hand, PIPS had the greatest bacterial killing efficiency at the 150-μm depth in all 3 root segments. Conclusions XP Endo appears to be more efficient than other 3 techniques in disinfecting the main canal space and up to 50 μm deep into the dentinal tubules. PIPS appears to be most effective in killing the bacteria deep in the dentinal tubules. PMID:27130334

Photodynamic therapy for endodontic disinfection.

PubMed

Soukos, Nikolaos S; Chen, Peter Shih-Yao; Morris, Jason T; Ruggiero, Karriann; Abernethy, Abraham D; Som, Sovanda; Foschi, Federico; Doucette, Stephanie; Bammann, Lili Luschke; Fontana, Carla Raquel; Doukas, Apostolos G; Stashenko, Philip P

2006-10-01

The aims of this study were to investigate the effects of photodynamic therapy (PDT) on endodontic pathogens in planktonic phase as well as on Enterococcus faecalis biofilms in experimentally infected root canals of extracted teeth. Strains of microorganisms were sensitized with methylene blue (25 microg/ml) for 5 minutes followed by exposure to red light of 665 nm with an energy fluence of 30 J/cm2. Methylene blue fully eliminated all bacterial species with the exception of E. faecalis (53% killing). The same concentration of methylene blue in combination with red light (222 J/cm2) was able to eliminate 97% of E. faecalis biofilm bacteria in root canals using an optical fiber with multiple cylindrical diffusers that uniformly distributed light at 360 degrees. We conclude that PDT may be developed as an adjunctive procedure to kill residual bacteria in the root canal system after standard endodontic treatment.

Evaluation of bacterial flora during the ripening of Kedong sufu, a typical Chinese traditional bacteria-fermented soybean product.

PubMed

Feng, Zhen; Gao, Wei; Ren, Dan; Chen, Xi; Li, Juan-juan

2013-04-01

Kedong sufu is a typical bacteria-fermented sufu in China. Isolation and identification of the autochthonous bacteria involved would allow the design of specific starters for this speciality. The purpose of the present study was to evaluate the bacterial flora during the ripening of Kedong sufu using polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) and culturing. In terms of bacterial diversity, 22 strains were isolated and identified and 27 strains were detected by DGGE. Regarding bacterial dynamics, the results of culturing and PCR-DGGE exhibited a similar trend towards dominant strains. Throughout the fermentation of sufu, Enterococcus avium, Enterococcus faecalis and Staphylococcus carnosus were the dominant microflora, while the secondary microflora comprised Leuconostoc mesenteroides, Staphylococcus saprophyticus, Streptococcus lutetiensis, Kocuria rosea, Kocuria kristinae, Bacillus pumilus, Bacillus cereus and Bacillus subtilis. This study is the first to reveal the bacterial flora during the ripening of Kedong sufu using both culture-dependent and culture-independent methods. This information will help in the design of autochthonous starter cultures for the production of Kedong sufu with desirable characteristic sensory profiles and shorter ripening times. © 2012 Society of Chemical Industry.

Potent antimicrobial and antibiofilm activities of bacteriogenically synthesized gold-silver nanoparticles against pathogenic bacteria and their physiochemical characterizations.

PubMed

Ramasamy, Mohankandhasamy; Lee, Jin-Hyung; Lee, Jintae

2016-09-01

The objective of this study was to develop a bimetallic nanoparticle with enhanced antibacterial activity that would improve the therapeutic efficacy against bacterial biofilms. Bimetallic gold-silver nanoparticles were bacteriogenically synthesized using γ-proteobacterium, Shewanella oneidensis MR-1. The antibacterial activities of gold-silver nanoparticles were assessed on the planktonic and biofilm phases of individual and mixed multi-cultures of pathogenic Gram negative (Escherichia coli and Pseudomonas aeruginosa) and Gram positive bacteria (Enterococcus faecalis and Staphylococcus aureus), respectively. The minimum inhibitory concentration of gold-silver nanoparticles was 30-50 µM than that of other nanoparticles (>100 µM) for the tested bacteria. Interestingly, gold-silver nanoparticles were more effective in inhibiting bacterial biofilm formation at 10 µM concentration. Both scanning and transmission electron microscopy results further accounted the impact of gold-silver nanoparticles on biocompatibility and bactericidal effect that the small size and bio-organic materials covering on gold-silver nanoparticles improves the internalization and thus caused bacterial inactivation. Thus, bacteriogenically synthesized gold-silver nanoparticles appear to be a promising nanoantibiotic for overcoming the bacterial resistance in the established bacterial biofilms. © The Author(s) 2016.

The Respiratory Arsenite Oxidase: Structure and the Role of Residues Surrounding the Rieske Cluster

PubMed Central

Warelow, Thomas P.; Oke, Muse; Schoepp-Cothenet, Barbara; Dahl, Jan U.; Bruselat, Nicole; Sivalingam, Ganesh N.; Leimkühler, Silke; Thalassinos, Konstantinos; Kappler, Ulrike; Naismith, James H.; Santini, Joanne M.

2013-01-01

The arsenite oxidase (Aio) from the facultative autotrophic Alphaproteobacterium Rhizobium sp. NT-26 is a bioenergetic enzyme involved in the oxidation of arsenite to arsenate. The enzyme from the distantly related heterotroph, Alcaligenes faecalis, which is thought to oxidise arsenite for detoxification, consists of a large α subunit (AioA) with bis-molybdopterin guanine dinucleotide at its active site and a 3Fe-4S cluster, and a small β subunit (AioB) which contains a Rieske 2Fe-2S cluster. The successful heterologous expression of the NT-26 Aio in Escherichia coli has resulted in the solution of its crystal structure. The NT-26 Aio, a heterotetramer, shares high overall similarity to the heterodimeric arsenite oxidase from A. faecalis but there are striking differences in the structure surrounding the Rieske 2Fe-2S cluster which we demonstrate explains the difference in the observed redox potentials (+225 mV vs. +130/160 mV, respectively). A combination of site-directed mutagenesis and electron paramagnetic resonance was used to explore the differences observed in the structure and redox properties of the Rieske cluster. In the NT-26 AioB the substitution of a serine (S126 in NT-26) for a threonine as in the A. faecalis AioB explains a −20 mV decrease in redox potential. The disulphide bridge in the A. faecalis AioB which is conserved in other betaproteobacterial AioB subunits and the Rieske subunit of the cytochrome bc 1 complex is absent in the NT-26 AioB subunit. The introduction of a disulphide bridge had no effect on Aio activity or protein stability but resulted in a decrease in the redox potential of the cluster. These results are in conflict with previous data on the betaproteobacterial AioB subunit and the Rieske of the bc 1 complex where removal of the disulphide bridge had no effect on the redox potential of the former but a decrease in cluster stability was observed in the latter. PMID:24023621

pSK41-Like Plasmid Is Necessary for Inc18-Like vanA Plasmid Transfer from Enterococcus faecalis to Staphylococcus aureus In Vitro

PubMed Central

Clark, Nancye; Patel, Jean B.

2013-01-01

Vancomycin-resistant Staphylococcus aureus (VRSA) is thought to result from the in vivo conjugative transfer of a vanA plasmid from an Enterococcus sp. to S. aureus. We studied bacterial isolates from VRSA cases that occurred in the United States to identify microbiological factors which may contribute to this plasmid transfer. First, vancomycin-susceptible, methicillin-resistant S. aureus (MRSA) isolates from five VRSA cases were tested for their ability to accept foreign DNA by conjugation in mating experiments with Enterococcus faecalis JH2-2 containing pAM378, a pheromone-response conjugative plasmid. All of the MRSA isolates accepted the plasmid DNA with similar transfer efficiencies (∼10−7/donor CFU) except for one isolate, MRSA8, for which conjugation was not successful. The MRSA isolates were also tested as recipients in mating experiments between an E. faecalis isolate with an Inc18-like vanA plasmid that was isolated from a VRSA case patient. Conjugative transfer was successful for 3/5 MRSA isolates. Successful MRSA recipients carried a pSK41-like plasmid, a staphylococcal conjugative plasmid, whereas the two unsuccessful MRSA recipients did not carry pSK41. The transfer of a pSK41-like plasmid from a successful MRSA recipient to the two unsuccessful recipients resulted in conjugal transfer of the Inc18-like vanA plasmid from E. faecalis at a frequency of 10−7/recipient CFU. In addition, conjugal transfer could be achieved for pSK41-negative MRSA in the presence of a cell-free culture filtrate from S. aureus carrying a pSK41-like plasmid at a frequency of 10−8/recipient CFU. These results indicated that a pSK41-like plasmid can facilitate the transfer of an Inc18-like vanA plasmid from E. faecalis to S. aureus, possibly via an extracellular factor produced by pSK41-carrying isolates. PMID:23089754

Comparative analysis of the biological and physical properties of Enterococcus faecalis bacteriophage vB_EfaS_GEC-EfS_3 and Streptococcus mitis bacteriophage vB_SmM_GEC-SmitisM_2.

PubMed

Rigvava, Sophio; Tchgkonia, Irina; Jgenti, Darejan; Dvalidze, Teona; Carpino, James; Goderdzishvili, Marina

2013-01-01

Enterococcus faecalis and Streptococcus mitis are common commensal inhabitants of the human gastrointestinal and genitourinary tracts. However, both species can be opportunistic pathogens and cause disease in nosocomial settings. These infections can be difficult to treat because of the frequency of antibiotic resistance among these strains. Bacteriophages are often suggested as an alternative therapeutic agent against these infections. In this study, E. faecalis and S. mitis strains were isolated from female patients with urinary tract infections. Bacteriophages active against these strains were isolated from sewage water from the Mtkvari River. Two phages, designated vB_EfaS_GEC-EfS_3 (Syphoviridae) and vB_SmM_GEC-SmitisM_2 (Myoviridae), were specific for E. faecalis and S. mitis, respectively. Each phage's growth patterns and adsorption rates were quantified. Sensitivity to ultraviolet light and temperature was determined, as was host range and serology. The S. mitis bacteriophage was found to be more resistant to ultraviolet light and exposure to high temperatures than the E. faecalis bacteriophage, despite having a much greater rate of replication. While each phage was able to infect a broad range of strains of the same species as the host species from which they were isolated, they were unable to infect other host species tested.

Relationships between the antibacterial activity of sodium hypochlorite and treatment time and biofilm age in early Enterococcus faecalis biofilms.

PubMed

Chau, N P T; Chung, N H; Jeon, J G

2015-08-01

To determine the relationships between the antibacterial activity of NaOCl and treatment time and biofilm age in early Enterococcus faecalis biofilms using a linear fitting procedure. Enterococcus faecalis biofilms were formed on hydroxyapatite discs. To investigate the relationship between the antibacterial activity of NaOCl and biofilm age, 22-, 46-, 70- and 94-h-old biofilms were exposed to NaOCl (0-3%) for 5 min. To investigate the relationship between the antibacterial activity of NaOCl and treatment time, 70-h-old biofilms were exposed to NaOCl (0-3%) for 1, 3, 5 and 7 min. After treatment, colony-forming units (CFUs) were counted. To determine the relationships between these variables, linear fitting was performed. The change in the minimum biofilm eradication concentration (MBEC) of NaOCl followed a linear pattern of biofilm age (R = 0.941, R(2)  = 0.886) or treatment time dependence (R = -0.948, R(2)  = 0.898). Below the MBEC, the fitting lines for bacterial CFU count versus NaOCl concentration (R ≤ -0.973, R(2)  ≥ 0.948) in the 22-, 46-, 70- and 94-h-old biofilms implied that the antibacterial activity of NaOCl decreased as the biofilm age increased. The fitting lines for bacterial CFU count versus NaOCl concentration (R ≤ -0.970, R(2)  ≥ 0.942) in the 1-, 3-, 5- and 7-min treatments implied that the antibacterial activity of NaOCl increased with treatment time. These results suggest that the antibacterial activity of NaOCl against early E. faecalis biofilms in root canals may follow a linear pattern depending on biofilm age or treatment time. © 2014 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Cross-Contamination of Residual Emerging Contaminants and Antibiotic Resistant Bacteria in Lettuce Crops and Soil Irrigated with Wastewater Treated by Sunlight/H2O2.

PubMed

Ferro, Giovanna; Polo-López, María I; Martínez-Piernas, Ana B; Fernández-Ibáñez, Pilar; Agüera, Ana; Rizzo, Luigi

2015-09-15

The sunlight/H2O2 process has recently been considered as a sustainable alternative option compared to other solar driven advanced oxidation processes (AOPs) in advanced treatment of municipal wastewater (WW) to be reused for crop irrigation. Accordingly, in this study sunlight/H2O2 was used as disinfection/oxidation treatment for urban WW treatment plant effluent in a compound parabolic collector photoreactor to assess subsequent cross-contamination of lettuce and soil by contaminants of emerging concern (CECs) (determined by QuEChERS extraction and LC-QqLIT-MS/MS analysis) and antibiotic resistant (AR) bacteria after irrigation with treated WW. Three CECs (carbamazepine (CBZ), flumequine (FLU), and thiabendazole (TBZ) at 100 μg L(-1)) and two AR bacterial strains (E. coli and E. faecalis, at 10(5) CFU mL(-1)) were spiked in real WW. A detection limit (DL) of 2 CFU mL(-1) was reached after 120 min of solar exposure for AR E. coli, while AR E. faecalis was more resistant to the disinfection process (240 min to reach DL). CBZ and TBZ were poorly removed after 90 min (12% and 50%, respectively) compared to FLU (94%). Lettuce was irrigated with treated WW for 5 weeks. CBZ and TBZ were accumulated in soil up to 472 ng g(-1) and 256 ng g(-1) and up-taken by lettuce up to 109 and 18 ng g(-1), respectively, when 90 min treated WW was used for irrigation; whereas no bacteria contamination was observed when the bacterial density in treated WW was below the DL. A proper treatment time (>90 min) should be guaranteed in order to avoid the transfer of pathogens from disinfected WW to irrigated crops and soil.

Effects of prolonged exposure to moderate static magnetic field and its synergistic effects with alkaline pH on Enterococcus faecalis.

PubMed

Fan, Wei; Huang, Zhuo; Fan, Bing

2018-02-01

Static magnetic field (SMF) has been shown to biologically affect various microorganisms, but its effects on Enterococcus faecalis, which is associated with multiple dental infections, have not been reported yet. Besides, Enterococcus faecalis was found to be resistant to the alkaline environment provided by a major dental antimicrobial, calcium hydroxide. Therefore, the antibacterial activity of prolonged exposure to moderate SMF (170 mT) and its possible synergistic activity with alkaline pH (pH = 9) were evaluated in the study. The ability to form a biofilm under these conditions was examined by crystal violet assay. Real-time quantitative PCR was performed to evaluate the relative expression of stress (dnaK and groEL) and virulence (efaA, ace, gelE and fsrC) related genes. As the results indicated, cell proliferation was inhibited after 120 h of SMF exposure. What's more, the combined treatment of SMF and alkaline pH showed significantly improved antimicrobial action when compared to single SMF and alkaline pH treatment for more than 24 h and 72 h respectively. However, the ability to form a biofilm was also enhanced under SMF and alkaline pH treatments. SMF can induce stress response by up-regulating the expression of dnaK and elevate virulence gene expression (efaA and ace). These responses were more significant and more genes were up-regulated including groEL, gelE and fsrC when exposed to SMF and alkaline pH simultaneously. Hence, combination of SMF and alkaline pH could be a promising disinfection strategy in dental area and other areas associated with Enterococcus faecalis infections. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production, purification and characterization of halophilic organic solvent tolerant protease from marine crustacean shell wastes and its efficacy on deproteinization.

PubMed

Maruthiah, Thirumalai; Somanath, Beena; Jasmin, Jebamonydhas Vijila; Immanuel, Grasian; Palavesam, Arunachalam

2016-12-01

The quantum of marine fish wastes produced by fish processing industries has necessitated to search new methods for its disposal. Hence, this study is focused on production and purification of halophilic organic solvent tolerant protease (HOSP) from marine Alcaligenes faecalis APCMST-MKW6 using marine shell wastes as substrate. The candidate bacterium was isolated from the marine sediment of Manakudi coast and identified as A. faecalis APCMST-MKW6. The purified protease showed 16.39-fold purity, 70.34 U/mg specific activity with 21.67 % yield. The molecular weight of the purified alkaline protease was 49 kDa. This purified protease registered maximum activity at pH 9 and it was stable between pH 8-9 after 1.30 h of incubation. The optimum temperature registered was 60 °C and it was stable between 50 and 60 °C even after 1.30 h of incubation. This enzyme also showed maximum activity at 20 % NaCl concentration. Further, manganese chloride, magnesium chloride, calcium chloride and barium chloride influenced this enzyme activity remarkably and it was also found to be enhanced by many of the tested surfactants and solvents. The candidate bacterium effectively deproteinized the shrimp shell waste compared to the other tested crustaceans shell wastes and also attained maximum antioxidant activity.

Coupling of bioaugmentation and phytoremediation to improve PCBs removal from a transformer oil-contaminated soil.

PubMed

Salimizadeh, Maryam; Shirvani, Mehran; Shariatmadari, Hossein; Nikaeen, Mahnaz; Leili Mohebi Nozar, Seyedeh

2018-06-07

This study was carried out to assess the dissipation of 17 selected polychlorinated biphenyl (PCB i ) congeners in a transformer oil-contaminated soil using bioaugmentation with 2 PCB-degrading bacterial strains, i.e., Pseudomonas spp. S5 and Alcaligenes faecalis, assisted or not by the maize (Zea mays L.) plantation. After 5 and 10 weeks of treatment, the remaining concentrations of the target PCB i congeners in the soil were extracted and measured using GC-MS. Results showed that the bacterial augmentation treatments with Pseudomonas spp. S5 and A. faecalis led to 21.4% and 20.4% reduction in the total concentration of the target PCBs (ΣPCB i ), respectively, compared to non-bioaugmented unplanted control soil. The ΣPCB i decreased by 35.8% in the non-bioaugmented planted soil compared with the control. The greatest degradation of the PCB congeners was observed over a 10-week period in the soil inoculated with Pseudomonas spp. S5 and cultivated with maize. Under this treatment, the ΣPCB i decreased from 357 to 119 ng g -1 (66.7% lower) and from 1091 to 520 ng g -1 (52.3% lower). Overall, the results suggested that the combined application of phytoremediation and bioaugmentation was an effective technique to remove PCBs and remediate transformer oil-contaminated soils.

Type I and Type II mechanisms of antimicrobial photodynamic therapy: an in vitro study on gram-negative and gram-positive bacteria.

PubMed

Huang, Liyi; Xuan, Yi; Koide, Yuichiro; Zhiyentayev, Timur; Tanaka, Masamitsu; Hamblin, Michael R

2012-08-01

Antimicrobial photodynamic therapy (APDT) employs a non-toxic photosensitizer (PS) and visible light, which in the presence of oxygen produce reactive oxygen species (ROS), such as singlet oxygen ((1) O(2), produced via Type II mechanism) and hydroxyl radical (HO(.), produced via Type I mechanism). This study examined the relative contributions of (1) O(2) and HO(.) to APDT killing of Gram-positive and Gram-negative bacteria. Fluorescence probes, 3'-(p-hydroxyphenyl)-fluorescein (HPF) and singlet oxygen sensor green reagent (SOSG) were used to determine HO(.) and (1) O(2) produced by illumination of two PS: tris-cationic-buckminsterfullerene (BB6) and a conjugate between polyethylenimine and chlorin(e6) (PEI-ce6). Dimethylthiourea is a HO(.) scavenger, while sodium azide (NaN(3)) is a quencher of (1) O(2). Both APDT and killing by Fenton reaction (chemical generation of HO(.)) were carried out on Gram-positive bacteria (Staphylococcus aureus and Enterococcus faecalis) and Gram-negative bacteria (Escherichia coli, Proteus mirabilis, and Pseudomonas aeruginosa). Conjugate PEI-ce6 mainly produced (1) O(2) (quenched by NaN(3)), while BB6 produced HO(.) in addition to (1) O(2) when NaN(3) potentiated probe activation. NaN(3) also potentiated HPF activation by Fenton reagent. All bacteria were killed by Fenton reagent but Gram-positive bacteria needed a higher concentration than Gram-negatives. NaN(3) potentiated Fenton-mediated killing of all bacteria. The ratio of APDT killing between Gram-positive and Gram-negative bacteria was 2 or 4:1 for BB6 and 25:1 for conjugate PEI-ce6. There was a NaN(3) dose-dependent inhibition of APDT killing using both PEI-ce6 and BB6 against Gram-negative bacteria while NaN(3) almost failed to inhibit killing of Gram-positive bacteria. Azidyl radicals may be formed from NaN(3) and HO(.). It may be that Gram-negative bacteria are more susceptible to HO(.) while Gram-positive bacteria are more susceptible to (1) O(2). The differences in Na

Association of red complex, A. actinomycetemcomitans and non-oral bacteria with periodontal diseases.

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