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

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.

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

10.21203/rs.2.13402/v3 ◽

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.

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

10.21203/rs.2.13402/v2 ◽

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

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

10.21203/rs.2.13402/v4 ◽

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.

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

Microorganisms ◽

10.3390/microorganisms8111771 ◽

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.

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

Vojnosanitetski pregled ◽

10.2298/vsp131228062d ◽

2015 ◽

Vol 72 (4) ◽

pp. 379-382 ◽

Cited By ~ 1

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.

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

Infection and Immunity ◽

10.1128/iai.72.10.6032-6039.2004 ◽

2004 ◽

Vol 72 (10) ◽

pp. 6032-6039 ◽

Cited By ~ 140

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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Incidence of Virulence Factors and Antibiotic Resistance among Enterococci Isolated from Food

Applied and Environmental Microbiology ◽

10.1128/aem.67.9.4385-4389.2001 ◽

2001 ◽

Vol 67 (9) ◽

pp. 4385-4389 ◽

Cited By ~ 227

Author(s):

Charles M. A. P. Franz ◽

Albrecht B. Muscholl-Silberhorn ◽

Nuha M. K. Yousif ◽

Marc Vancanneyt ◽

Jean Swings ◽

...

Keyword(s):

Antibiotic Resistance ◽

Enterococcus Faecalis ◽

Virulence Factors ◽

Enterococcus Faecium ◽

Antibiotic Susceptibility ◽

Virulence Determinants ◽

Virulence Traits

ABSTRACT The incidence of virulence factors among 48 Enterococcus faecium and 47 Enterococcus faecalis strains from foods and their antibiotic susceptibility were investigated. No strain was resistant to all antibiotics, and for some strains, multiple resistances were observed. Of E. faecium strains, 10.4% were positive for one or more virulence determinants, compared to 78.7% of E. faecalis strains. Strains exhibiting virulence traits were not necessarily positive for all traits; thus, the incidence of virulence factors may be considered to be strain specific.

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Cardiac Microlesions Form During Severe Bacteremic Enterococcus faecalis Infection

10.1101/2020.01.14.906669 ◽

2020 ◽

Author(s):

Armand O. Brown ◽

Kavindra V. Singh ◽

Melissa R. Cruz ◽

Karan Gautam Kaval ◽

Liezl E. Francisco ◽

...

Keyword(s):

Immune Response ◽

Enterococcus Faecalis ◽

Pneumococcal Disease ◽

Blood Stream Infection ◽

Blood Stream ◽

Virulence Determinants ◽

C Elegans ◽

Bond Forming ◽

Hospital Acquired ◽

Pro Inflammatory Cytokines

AbstractEnterococcus faecalis is a significant cause of hospital-acquired bacteremia. Herein, the discovery is reported that cardiac microlesions form during severe bacteremic E. faecalis infection in mice. The cardiac microlesions were identical in appearance to those formed by Streptococcus pneumoniae during invasive pneumococcal disease (IPD). However, E. faecalis does not encode the virulence determinants implicated in pneumococcal microlesion formation. Rather, disulfide bond forming protein DsbA was found to be required for E. faecalis virulence in a C. elegans model and was necessary for efficient cardiac microlesion formation. Furthermore, E. faecalis promoted cardiomyocyte apoptotic and necroptotic cell death at sites of microlesion formation. Additionally, loss of DsbA caused an increase in pro-inflammatory cytokines unlike the wild-type strain, which suppressed the immune response. In conclusion, we establish that E. faecalis is capable of forming cardiac microlesions and identify features of both the bacterium and the host response that are mechanistically involved.SUMMARYThis work presents the observation of cardiac microlesion formation during severe blood stream infection with Enterococcus faecalis in mice. Moreover, we identify the contribution of a novel enterococcal virulence determinant in modulating microlesion formation and the host immune response.

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Pathogenicity Biofilm formation of Enterococcus faecalis

Journal of Syiah Kuala Dentistry Society ◽

10.24815/jds.v5i1.20011 ◽

2020 ◽

Vol 5 (1) ◽

pp. 47-50

Author(s):

Nur Asmah

Keyword(s):

Enterococcus Faecalis ◽

Root Canal ◽

Biofilm Formation ◽

Laboratory Diagnosis ◽

Toxin Production ◽

Limited Information ◽

Periapical Lesions ◽

Alkaline Conditions ◽

Autolytic Activity ◽

Acid Reaction

Biofilm formation is closely related to the pathogenicity of E. faecalis in persistent root canal infections. Limited information is available about the ability and characteristics of E. faecalis biofilm-forming in the persistent pathogenicity of root canal infections. Based on these problems, the purpose of this paper is to improve the molecular understanding of E. faecalis on virulence factors associated with biofilm formation against persistent root canal infections to support laboratory diagnosis and therapy of oral E. faecalis. In conclusion, the release of cytokines triggers the dlt gene of LTA to improving: homeostasis, autolytic activity, and bacterial envelope properties. The role of cytolysin activated by the cylLL and cylLS genes improving the survival ability of E. Faecalis. Hyaluronidase will facilitate other bacteria to migrate from the root canal to periapical lesions resulting in the triggering of toxin production, which will increase damage and inflammation in the host. Enterococcus faecalis, through dentine matrix formation, hydrolyze collagen and induce apatite deposition in developing biofilms. Besides, these bacteria can also increase their tolerance to antimicrobials by blocking the inflammatory response's acid reaction. Alkaline conditions will neutralize the lactic acid secreted by osteoclasts to absorb hard tissue

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