scholarly journals CRISPR-cas system in the acquisition of virulence genes in dental-root canal and hospital-acquired isolates of Enterococcus faecalis

2020 ◽  
Author(s):  
Pourya Gholizadeh ◽  
Mohammad Aghazadeh ◽  
Reza Ghotaslou ◽  
Mohammad Ahangarzadeh Rezaee ◽  
Tahereh Pirzadeh ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Biofilm Formation ◽  
Virulence Genes ◽  
Virulence Determinants ◽  
Root Canals ◽  
Causative Agents ◽  
Life Threatening ◽  
Selective Forces ◽  
Hospital Acquired

Abstract Introduction Enterococcus faecalis is one of the important causative agents of nosocomial and life-threatening infections in human. Several studies have demonstrated that the presence of CRISPR- cas is associated with antibiotic susceptibility and lack of virulence traits. In this study, we aimed to assess the phenotypic and genotypic virulence determinants in relation to CRISPR elements from the dental-root canals and hospital-acquired isolates of E. faecalis . Methods and materials Eighty-eight hospital-acquired and 73 dental-root canal isolates of E. faecalis were assessed in this study. Phenotypic screening of the isolates included biofilm formation, and gelatinase and hemolysis activities. Genotypical screening using PCR was further used to evaluate the presence of CRISPR elements and different virulence-associated genes such as efaA , esp , cylA , hyl , gelE , ace , ebpR , and asa1 . Results Biofilm formation, and gelatinase and hemolysis activity were detected in 93.8%, 29.2% and 19.2% of the isolates, respectively. The most prevalent virulence-associated gene was ace , which was followed by efaA , whereas cylA was the least identified. The presence of CRISPR1- cas , orphan CRISPR2 and CRISPR3- cas was determined in 13%, 55.3% and 17.4% of the isolates, respectively. CRISPR elements were significantly more prevalent in the dental-root canals isolates. An inverse significant correlation was found between CRISPR- cas loci, esp and gelE , while direct correlations were observed in the case of cylA , hyl , gelE (among CRISPR-loci 1 and 3), asa1 , ace , biofilm formation, and hemolysis activity. Conclusion Findings, therefore, indicate that CRISPR- cas might prevent the acquisition of some respective pathogenicity factors in some isolates, though not all; so selective forces could not influence pathogenic traits.

scholarly journals CRISPR-cas system in the acquisition of virulence genes in dental-root canal and hospital-acquired isolates of Enterococcus faecalis

2019 ◽  
Author(s):  
Pourya Gholizadeh ◽  
Mohammad Aghazadeh ◽  
Reza Ghotaslou ◽  
Mohammad Ahangarzadeh Rezaee ◽  
Tahereh Pirzadeh ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Biofilm Formation ◽  
Virulence Genes ◽  
Virulence Determinants ◽  
Root Canals ◽  
Causative Agents ◽  
Life Threatening ◽  
Selective Forces ◽  
Hospital Acquired

Abstract Introduction Enterococcus faecalis is one of the important causative agents of nosocomial and life-threatening infections in human. Several studies have demonstrated that the presence of CRISPR- cas is associated with antibiotic susceptibility and lack of virulence traits. In this study, we aimed to assess the phenotypic and genotypic virulence determinants in relation to CRISPR elements from the dental-root canals and hospital-acquired isolates of E. faecalis .Methods and materials Eighty-eight hospital-acquired and 73 dental-root canal isolates of E. faecalis were assessed in this study. Phenotypic screening of the isolates included biofilm formation, and gelatinase and hemolysis activities. Genotypical screening using PCR was further used to evaluate the presence of CRISPR elements and different virulence-associated genes such as efaA , esp , cylA , hyl , gelE , ace , ebpR , and asa1 .Results Biofilm formation, and gelatinase and hemolysis activity were detected in 93.8%, 29.2% and 19.2% of the isolates, respectively. The most prevalent virulence-associated gene was ace , which was followed by efaA , whereas cylA was the least identified. The presence of CRISPR1- cas , orphan CRISPR2 and CRISPR3- cas was determined in 13%, 55.3% and 17.4% of the isolates, respectively. CRISPR elements were significantly more prevalent in the dental-root canals isolates. An inverse significant correlation was found between CRISPR- cas loci, esp and gelE , while direct correlations were observed in the case of cylA , hyl , gelE (among CRISPR-loci 1 and 3), asa1 , ace , biofilm formation, and hemolysis activity.Conclusion Findings, therefore, indicate that CRISPR- cas might prevent the acquisition of some respective pathogenicity factors in some isolates, though not all; so selective forces could not influence pathogenic traits.

scholarly journals CRISPR-cas system in the acquisition of virulence genes in dental-root canal and hospital-acquired isolates of Enterococcus faecalis

2019 ◽  
Author(s):  
Pourya Gholizadeh ◽  
Mohammad Aghazadeh ◽  
Reza Ghotaslou ◽  
Mohammad Ahangarzadeh Rezaee ◽  
Tahereh Pirzadeh ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Biofilm Formation ◽  
Virulence Genes ◽  
Virulence Determinants ◽  
Root Canals ◽  
Causative Agents ◽  
Life Threatening ◽  
Selective Forces ◽  
Hospital Acquired

Abstract Introduction Enterococcus faecalis is one of the important causative agents of nosocomial and life-threatening infections in human. Several studies have demonstrated that the presence of CRISPR- cas is associated with antibiotic susceptibility and lack of virulence traits. In this study, we aimed to assess the phenotypic and genotypic virulence determinants in relation to CRISPR elements from the dental-root canals and hospital-acquired isolates of E. faecalis .Methods and materials Eighty-eight hospital-acquired and 73 dental-root canal isolates of E. faecalis were assessed in this study. Phenotypic screening of the isolates included biofilm formation, and gelatinase and hemolysis activities. Genotypical screening using PCR was further used to evaluate the presence of CRISPR elements and different virulence-associated genes such as efaA , esp , cylA , hyl , gelE , ace , ebpR , and asa1 .Results Biofilm formation, and gelatinase and hemolysis activity were detected in 93.8%, 29.2% and 19.2% of the isolates, respectively. The most prevalent virulence-associated gene was ace , which was followed by efaA , whereas cylA was the least identified. The presence of CRISPR1- cas , orphan CRISPR2 and CRISPR3- cas was determined in 13%, 55.3% and 17.4% of the isolates, respectively. CRISPR elements were significantly more prevalent in the dental-root canals isolates. An inverse significant correlation was found between CRISPR- cas loci, esp and gelE , while direct correlations were observed in the case of cylA , hyl , gelE (among CRISPR-loci 1 and 3), asa1 , ace , biofilm formation, and hemolysis activity.Conclusion Findings, therefore, indicate that CRISPR- cas might prevent the acquisition of some respective pathogenicity factors in some isolates, though not all; so selective forces could not influence pathogenic traits.

scholarly journals CRISPR-cas system in the acquisition of virulence genes in the dental-root canal and hospital-acquired isolates of Enterococcus faecalis

2019 ◽  
Author(s):  
Pourya Gholizadeh ◽  
Mohammad Aghazadeh ◽  
Reza Ghotaslou ◽  
Mohammad Ahangarzadeh Rezaee ◽  
Tahereh Pirzadeh ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Biofilm Formation ◽  
Inverse Correlation ◽  
Virulence Determinants ◽  
Gene Encoding ◽  
Virulence Traits ◽  
Causative Agents ◽  
Life Threatening ◽  
Hospital Acquired

Abstract Introduction: Enterococcus faecalis is one of the important causative agents of nosocomial and life-threatening infections in human. Several studies demonstrated that the presence of CRISPR-cas is associated with the antibiotic susceptibility and lack of virulence traits. In this study, we aimed to assess the relation between the phenotypic and genotypic virulence determinants of E. faecalis with the CRISPR elements in the dental-root canal and hospital-acquired isolates.Methods and materials: Eighty-eight hospital-acquired and 73 dental-root canal isolates of E. faecalis were subjected for study. The phenotype tests including biofilm formation, gelatinase and hemolysis activity were performed and the genotype characteristics including efaA, esp, cylA, hyl, gelE, ace, ebpR, and asa1 were performed by PCR methods. Presences of different types of CRISPR-cas system were determined by PCR.Results: Biofilm formation, gelatinase and hemolysis activity were detected in 93.8%, 29.2% and 19.2% of the isolates, respectively. The most gene encoding virulence traits were ace, followed by efaA and the lowest was cylA. The presence of CRISPR1-cas, orphan CRISPR2 and CRISPR3-cas were determined in 13%, 55.3%, and 17.4% of isolates, respectively, which were present in proportionally more in the dental-root canal. Inverse correlation were found significantly between CRISPR-cas loci, esp and gelE, and direct correlations were found in cylA, hyl, gelE (between some CRISPR-loci), asa1, ace, biofilm formation, gelatinase and hemolysis activitiesConclusion: Findings indicates that CRISPR-cas might prevent the acquisition of some respective pathogenicity factors in some isolates not all, and could not be selective forces to influence pathogenic traits.

scholarly journals CRISPR-cas system in the acquisition of virulence genes in dental-root canal and hospital-acquired isolates of Enterococcus faecalis

Virulence ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 1257-1267
Author(s):  
Pourya Gholizadeh ◽  
Mohammad Aghazadeh ◽  
Reza Ghotaslou ◽  
Mohammad Ahangarzadeh Rezaee ◽  
Tahereh Pirzadeh ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Virulence Genes ◽  
Hospital Acquired

scholarly journals Bacteriophages to Control Multi-Drug Resistant Enterococcus faecalis Infection of Dental Root Canals

2021 ◽  
Vol 9 (3) ◽  
pp. 517
Author(s):  
Mohamed El-Telbany ◽  
Gamal El-Didamony ◽  
Ahmed Askora ◽  
Eman Ariny ◽  
Dalia Abdallah ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Phage Therapy ◽  
Ex Vivo ◽  
Burst Size ◽  
Resistant Bacteria ◽  
Drug Resistant ◽  
Range Analysis ◽  
Root Canals

Phage therapy is an alternative treatment to antibiotics that can overcome multi-drug resistant bacteria. In this study, we aimed to isolate and characterize lytic bacteriophages targeted against Enterococcus faecalis isolated from root canal infections obtained from clinics at the Faculty of Dentistry, Ismalia, Egypt. Bacteriophage, vB_ZEFP, was isolated from concentrated wastewater collected from hospital sewage. Morphological and genomic analysis revealed that the phage belongs to the Podoviridae family with a linear double-stranded DNA genome, consisting of 18,454, with a G + C content of 32.8%. Host range analysis revealed the phage could infect 10 of 13 E. faecalis isolates exhibiting a range of antibiotic resistances recovered from infected root canals with efficiency of plating values above 0.5. One-step growth curves of this phage showed that it has a burst size of 110 PFU per infected cell, with a latent period of 10 min. The lytic activity of this phage against E. faecalis biofilms showed that the phage was able to control the growth of E. faecalis in vitro. Phage vB_ZEFP could also prevent ex-vivo E. faecalis root canal infection. These results suggest that phage vB_ZEFP has potential for application in phage therapy and specifically in the prevention of infection after root canal treatment.

scholarly journals In vitro evaluation of the effectiveness of the chemomechanical preparation against Enterococcus faecalis after single- or multiple-visit root canal treatment

2007 ◽  
Vol 21 (4) ◽  
pp. 308-313 ◽  
Author(s):  
Eduardo Diogo Gurgel-Filho ◽  
Nilton Vivacqua-Gomes ◽  
Brenda Paula Figueiredo de Almeida Gomes ◽  
Caio Cezar Randi Ferraz ◽  
Alexandre Augusto Zaia ◽  
...  
Keyword(s):  
Calcium Hydroxide ◽  
Enterococcus Faecalis ◽  
Root Canal ◽  
Control Groups ◽  
Single Visit ◽  
Root Canals ◽  
Multiple Visit ◽  
Microbial Samples ◽  
Chlorhexidine Gel

The purpose was to assess the elimination of Enterococcus faecalis in vitro in human mandibular premolars after chemomechanical preparation with or without the use of a calcium hydroxide dressing. After 60 days of contamination with E. faecalis, the root canals were prepared using the Crown-Down technique combined with 2% chlorhexidine gel irrigation. Then, the specimens were divided into two experimental groups, treated in a single visit or in multiple visits, and two control groups. The multiple-visit group received a dressing with calcium hydroxide for 14 days (CalenTM) and the single-visit group did not receive any medication. In the two control groups, the canals were filled with BHI after chemomechanical preparation with 2% chlorhexidine gel or distilled water. Microbial samples were taken from the root canals for colony forming unit count for each phase of the treatment using sterile paper points inside the root canal lumen. Data were ranked and analyzed by the Kruskal-Wallis statistical test. The residual microbial colonies were then assessed. The results showed that chemomechanical preparation using 2% chlorhexidine gel with no intra-canal dressing reduced by 100% the E. faecalis contamination of the root canal lumen. The calcium-hydroxide group that received the 14-day intra-canal dressing allowed a small number of bacteria to grow between visits, but without statistical differences between groups.

Occurrence of virulence-associated genes in clinical Enterococcus faecalis strains isolated in Londrina, Brazil

2004 ◽  
Vol 53 (11) ◽  
pp. 1069-1073 ◽  
Author(s):  
Elisa Bittencourt de Marques ◽  
Sérgio Suzart
Keyword(s):  
Enterococcus Faecalis ◽  
Virulence Genes ◽  
Epidemiological Studies ◽  
Wide Distribution ◽  
Virulence Determinants ◽  
Simultaneous Presence ◽  
Common Pattern ◽  
Clinical Strains ◽  
Virulence Markers ◽  
Serious Infections

Epidemiological studies have reinforced the importance of Enterococcus faecalis in causing serious infections, and to date, our understanding of how certain virulence factors are involved in the pathogenesis of enterococcal infections is still limited. The aim of the present study was to examine the occurrence of known virulence determinants in a group of E. faecalis strains isolated from different clinical sources in Brazil. A total of 95 E. faecalis strains were investigated for the presence of nine virulence genes including aggA, cylA, cylB, cylM, eep, efaA, enlA, esp and gelE by using PCR. The data showed a relatively wide distribution of the virulence genes among the investigated strains. The clinical strains carried at least one and concomitantly up to as many as eight virulence markers, with two or three being the most common pattern. Most of the strains carried efaA (58.9 %), eep (58.9 %) and esp (57.9 %) genes, whereas the remaining virulence markers were detected in variable percentages ranging from 9.5 to 45 %. Simultaneous presence of virulence markers was observed among clinical strains regardless of their sources. In this study, the efaA + esp + gelE + profile was the virulence genotype most frequently detected among E. faecalis strains. Finally, there was no significant association between virulence markers and clinical sources.

scholarly journals Interactions between Candida albicans and Enterococcus faecalis in an Organotypic Oral Epithelial Model

2020 ◽  
Vol 8 (11) ◽  
pp. 1771
Author(s):  
Akshaya Lakshmi Krishnamoorthy ◽  
Alex A. Lemus ◽  
Adline Princy Solomon ◽  
Alex M. Valm ◽  
Prasanna Neelakantan
Keyword(s):  
Candida Albicans ◽  
Enterococcus Faecalis ◽  
Biofilm Formation ◽  
Opportunistic Pathogen ◽  
Surface Erosion ◽  
Host Immune System ◽  
Microbial Invasion ◽  
Mucosal Surfaces ◽  
Life Threatening ◽  
Hyphal Formation

Candida albicans as an opportunistic pathogen exploits the host immune system and causes a variety of life-threatening infections. The polymorphic nature of this fungus gives it tremendous advantage to breach mucosal barriers and cause oral and disseminated infections. Similar to C. albicans, Enterococcus faecalis is a major opportunistic pathogen, which is of critical concern in immunocompromised patients. There is increasing evidence that E. faecalis co-exists with C. albicans in the human body in disease samples. While the interactive profiles between these two organisms have been studied on abiotic substrates and mouse models, studies on their interactions on human oral mucosal surfaces are non-existent. Here, for the first time, we comprehensively characterized the interactive profiles between laboratory and clinical isolates of C. albicans (SC5314 and BF1) and E. faecalis (OG1RF and P52S) on an organotypic oral mucosal model. Our results demonstrated that the dual species biofilms resulted in profound surface erosion and significantly increased microbial invasion into mucosal compartments, compared to either species alone. Notably, several genes of C. albicans involved in tissue adhesion, hyphal formation, fungal invasion, and biofilm formation were significantly upregulated in the presence of E. faecalis. By contrast, E. faecalis genes involved in quorum sensing, biofilm formation, virulence, and mammalian cell invasion were downregulated. This study highlights the synergistic cross-kingdom interactions between E. faecalis and C. albicans in mucosal tissue invasion.

scholarly journals An experience with colistin applied in treatment of imunocompromised patients with peritonitis on peritoneal dialysis

2015 ◽  
Vol 72 (4) ◽  
pp. 379-382 ◽  
Author(s):  
Tatjana Djurdjevic-Mirkovic ◽  
Ljiljana Gvozdenovic ◽  
Gordana Majstorovic-Strazmester ◽  
Violeta Knezevic ◽  
Dejan Celic ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Peritoneal Dialysis ◽  
Complete Recovery ◽  
First Episode ◽  
Exit Site ◽  
Causative Agents ◽  
Life Threatening ◽  
History Of ◽  
Acinetobacter Spp ◽  
Hospital Acquired

Introduction. Immunocompromised patients, such as those with multiple myeloma on peritoneal dialysis, are particularly susceptible to the occurrence of peritonitis. Case report. We presented a 56-year-old female patient with a 10-year history of multiple myeloma. The patient was on peritoneal dialysis since 2010. During 2012 the patient had the first episode of peritonitis that was successfully managed, but in 2013 the second episode of peritonitis occured. Analysis of dialysate culture and exit site swab revealed the presence of multiresistant Acinetobacter spp., which was susceptible only to colistin. Prompt colistin therapy was administered at the doses of 100,000 units/day during six days, which resulted in complete recovery of the patient, as well as improvement of local abdominal findings. Gram-negative bacteria (genus Acinetobacter) are common causative agents in hospital-acquired infections. Studies confirmed susceptibility of Acinetobacter to colistin, which was also the case with the presented patient. Intravenous administration of colistin resulted in a complete remission of this severe, life-threatening peritonitis. Conclusion. Patients with multiple myeloma and renal failure are highly prone to severe life-threatening infections.

scholarly journals Enterococcal Surface Protein, Esp, Enhances Biofilm Formation by Enterococcus faecalis

2004 ◽  
Vol 72 (10) ◽  
pp. 6032-6039 ◽  
Author(s):  
Preeti M. Tendolkar ◽  
Arto S. Baghdayan ◽  
Michael S. Gilmore ◽  
Nathan Shankar
Keyword(s):  
Gastrointestinal Tract ◽  
Cell Surface ◽  
Enterococcus Faecalis ◽  
Human Ecology ◽  
Biofilm Formation ◽  
Molecular Mechanisms ◽  
Surface Protein ◽  
Antibiotic Resistant ◽  
Hospital Acquired Infection ◽  
Hospital Acquired

ABSTRACT Enterococci play a dual role in human ecology. They serve as commensal organisms of the gastrointestinal tract and are also leading causes of multiple antibiotic-resistant hospital-acquired infection. Many nosocomial infections result from the ability of microorganisms to form biofilms. The molecular mechanisms involved in enterococcal biofilm formation are only now beginning to be understood. Enterococcal surface protein, Esp, has been reported to contribute to biofilm formation by Enterococcus faecalis. Recent studies have shown that enterococci form biofilms independently of Esp expression. To precisely determine what role Esp plays in E. faecalis biofilm formation, Esp was expressed on the cell surface of genetically well-defined, natively Esp-deficient strains, and isogenic Esp-positive and Esp-deficient strains were compared for their biofilm-forming ability. The results show that Esp expression leads to a significant increase in biofilm formation, irrespective of the strain tested. The contribution of Esp to biofilm formation was found to be most pronounced in the presence of 0.5% (wt/vol) or greater glucose. These results unambiguously define Esp as a key contributor to the ability of E. faecalis to form biofilms.

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