Involvement of Chromosomally Encoded Homologs of the RRNPP Protein Family in Enterococcus faecalis Biofilm Formation and Urinary Tract Infection Pathogenesis

ABSTRACT Enterococcus faecalis is an opportunistic pathogen capable of causing infections, including endocarditis and urinary tract infections (UTI). One of the well-characterized quorum-sensing pathways in E. faecalis involves coordination of the conjugal transfer of pheromone-responsive plasmids by PrgX, a member of the RRNPP protein family. Members of this protein family in various Firmicutes have also been shown to contribute to numerous cellular processes, including sporulation, competence, conjugation, nutrient sensing, biofilm formation, and virulence. As PrgX is a plasmid-encoded RRNPP family member, we surveyed the genome of the multidrug-resistant strain V583 for additional RRNPP homologs using computational searches and refined those identified hits for predicted structural similarities to known RRNPP family members. This led us to investigate the contribution of the chromosomally encoded RRNPP homologs to biofilm processes and pathogenesis in a catheter-associated urinary tract infection (CAUTI) model. In this study, we identified five such homologs and report that 3 of the 5 homologs, EF0073, EF1599, and EF1316, affect biofilm formation as well as outcomes in the CAUTI model. IMPORTANCE Enterococcus faecalis causes health care-associated infections and displays resistance to a variety of broad-spectrum antibiotics by acquisition of resistance traits as well as the ability to form biofilms. Even though a growing number of factors related to biofilm formation have been identified, mechanisms that contribute to biofilm formation are still largely unknown. Members of the RRNPP protein family regulate a diverse set of biological reactions in low-G+C Gram-positive bacteria (Firmicutes). Here, we identify three predicted structural homologs of the RRNPP family, EF0073, EF1599, and EF1316, which affect biofilm formation and CAUTI pathogenesis.

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(p)ppGpp and CodY Promote Enterococcus faecalis Virulence in a Murine Model of Catheter-Associated Urinary Tract Infection

mSphere ◽

10.1128/msphere.00392-19 ◽

2019 ◽

Vol 4 (4) ◽

Cited By ~ 8

Author(s):

C. Colomer-Winter ◽

A. L. Flores-Mireles ◽

S. Kundra ◽

S. J. Hultgren ◽

J. A. Lemos

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Enterococcus Faecalis ◽

Stringent Response ◽

Nutrient Sensing ◽

Transcriptional Analysis ◽

Content Type ◽

Tract Infections

ABSTRACT In Firmicutes, the nutrient-sensing regulators (p)ppGpp, the effector molecule of the stringent response, and CodY work in tandem to maintain bacterial fitness during infection. Here, we tested (p)ppGpp and codY mutant strains of Enterococcus faecalis in a catheter-associated urinary tract infection (CAUTI) mouse model and used global transcriptional analysis to investigate the relationship of (p)ppGpp and CodY. The absence of (p)ppGpp or single inactivation of codY led to lower bacterial loads in catheterized bladders and diminished biofilm formation on fibrinogen-coated surfaces under in vitro and in vivo conditions. Single inactivation of the bifunctional (p)ppGpp synthetase/hydrolase rel did not affect virulence, supporting previous evidence that the association of (p)ppGpp with enterococcal virulence is not dependent on the activation of the stringent response. Inactivation of codY in the (p)ppGpp0 strain restored E. faecalis virulence in the CAUTI model as well as the ability to form biofilms in vitro. Transcriptome analysis revealed that inactivation of codY restores, for the most part, the dysregulated metabolism of (p)ppGpp0 cells. While a clear linkage between (p)ppGpp and CodY with expression of virulence factors could not be established, targeted transcriptional analysis indicates that a possible association between (p)ppGpp and c-di-AMP signaling pathways in response to the conditions found in the bladder may play a role in enterococcal CAUTI. Collectively, data from this study identify the (p)ppGpp-CodY network as an important contributor to enterococcal virulence in catheterized mouse bladder and support that basal (p)ppGpp pools and CodY promote virulence through maintenance of a balanced metabolism under adverse conditions. IMPORTANCE Catheter-associated urinary tract infections (CAUTIs) are one of the most frequent types of infection found in the hospital setting that can develop into serious and potentially fatal bloodstream infections. One of the infectious agents that frequently causes complicated CAUTI is the bacterium Enterococcus faecalis, a leading cause of hospital-acquired infections that are often difficult to treat due to the exceptional multidrug resistance of some isolates. Understanding the mechanisms by which E. faecalis causes CAUTI will aid in the discovery of new druggable targets to treat these infections. In this study, we report the importance of two nutrient-sensing bacterial regulators, named (p)ppGpp and CodY, for the ability of E. faecalis to infect the catheterized bladder of mice.

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Enterococcus faecalis Promotes Innate Immune Suppression and Polymicrobial Catheter-Associated Urinary Tract Infection

Infection and Immunity ◽

10.1128/iai.00378-17 ◽

2017 ◽

Vol 85 (12) ◽

Cited By ~ 18

Author(s):

Brenda Yin Qi Tien ◽

Hwee Mian Sharon Goh ◽

Kelvin Kian Long Chong ◽

Soumili Bhaduri-Tagore ◽

Sarah Holec ◽

...

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Enterococcus Faecalis ◽

Opportunistic Pathogen ◽

Gastrointestinal Microbiota ◽

Content Type ◽

E Coli ◽

Raw264.7 Macrophages ◽

Tract Infections

ABSTRACT Enterococcus faecalis, a member of the human gastrointestinal microbiota, is an opportunistic pathogen associated with hospital-acquired wound, bloodstream, and urinary tract infections. E. faecalis can subvert or evade immune-mediated clearance, although the mechanisms are poorly understood. In this study, we examined E. faecalis-mediated subversion of macrophage activation. We observed that E. faecalis actively prevents NF-κB signaling in mouse RAW264.7 macrophages in the presence of Toll-like receptor agonists and during polymicrobial infection with Escherichia coli. E. faecalis and E. coli coinfection in a mouse model of catheter-associated urinary tract infection (CAUTI) resulted in a suppressed macrophage transcriptional response in the bladder compared to that with E. coli infection alone. Finally, we demonstrated that coinoculation of E. faecalis with a commensal strain of E. coli into catheterized bladders significantly augmented E. coli CAUTI. Taken together, these results support the hypothesis that E. faecalis suppression of NF-κB-driven responses in macrophages promotes polymicrobial CAUTI pathogenesis, especially during coinfection with less virulent or commensal E. coli strains.

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Synchrotron X-Ray Fluorescence Microscopy of Gallium in Bladder Tissue following Gallium Maltolate Administration during Urinary Tract Infection

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00616-13 ◽

2013 ◽

Vol 57 (11) ◽

pp. 5197-5201 ◽

Cited By ~ 3

Author(s):

Katherine R. Ball ◽

Francesca Sampieri ◽

Manuel Chirino ◽

Don L. Hamilton ◽

Robert I. R. Blyth ◽

...

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Transitional Epithelium ◽

Bladder Tissue ◽

Content Type ◽

X Ray ◽

E Coli ◽

Tract Infections ◽

X Ray Absorption

ABSTRACTA mouse model of cystitis caused by uropathogenicEscherichia coliwas used to study the distribution of gallium in bladder tissue following oral administration of gallium maltolate during urinary tract infection. The median concentration of gallium in homogenized bladder tissue from infected mice was 1.93 μg/g after daily administration of gallium maltolate for 5 days. Synchrotron X-ray fluorescence imaging and X-ray absorption spectroscopy of bladder sections confirmed that gallium arrived at the transitional epithelium, a potential site of uropathogenicE. coliinfection. Gallium and iron were similarly but not identically distributed in the tissues, suggesting that at least some distribution mechanisms are not common between the two elements. The results of this study indicate that gallium maltolate may be a suitable candidate for further development as a novel antimicrobial therapy for urinary tract infections caused by uropathogenicE. coli.

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Draft Genome Sequence of Staphylococcus epidermidis UMB7765, Isolated from the Urobiome of a Woman with Recurrent Urinary Tract Infection

Microbiology Resource Announcements ◽

10.1128/mra.00417-20 ◽

2020 ◽

Vol 9 (20) ◽

Author(s):

Lucy Kemper ◽

Taylor Miller-Ensminger ◽

Adelina Voukadinova ◽

Alan J. Wolfe ◽

Catherine Putonti

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Urinary Tract Infections ◽

Staphylococcus Epidermidis ◽

Recurrent Urinary Tract Infection ◽

Draft Genome ◽

Positive Bacterium ◽

Content Type ◽

Tract Infections

Staphylococcus epidermidis is a Gram-positive bacterium that is resistant to many antibiotics. Here, we present the 2.5-Mb draft genome of S. epidermidis UMB7765, isolated from a voided urine sample from a female with recurrent urinary tract infections.

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Paraoxonase and acylated homoserine lactones in urine from patients with urinary tract infections

10.1101/641688 ◽

2019 ◽

Author(s):

John Lafleur ◽

Richard L. Amdur

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Biofilm Formation ◽

Innate Immune ◽

Western Blot Assay ◽

Western Blots ◽

Homoserine Lactones ◽

Tract Infections ◽

Acylated Homoserine Lactones

AbstractParaoxonases are mammalian enzymes that have a number of roles including the inhibition of bacterial virulence and biofilm formation by microorganisms that quorum sense with acylated homoserine lactones. Paraoxonases have previously been reported to inhibit P. aeruginosa biofilm formation in mammalian airways and skin. An innate immune role for paraoxonases in urinary tract infection has not previously been reported. We performed western blots for paraoxonase1 in urine from patients with urinary tract infection; we also tested urinary tract infection urine for the presence of acylated homoserine lactones using a cellular reporter system. We report here that paraoxonase1 was not found with our western blot assay in the urine of normal control patients; in those with urinary tract infection, paraoxonase1 was associated with E. coli UTI. Acylated homoserine lactones, but not paraoxonases, were found in the bulk urine of those with P. aeruginosa urinary tract infection. We hypothesize that paraoxonase may play a similar innate immune role in infected urine as has previously described in skin and airways.

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Spread of Enterococcal Surface Protein in Antibiotic Resistant Entero-coccus faecium and Enterococcus faecalis isolates from Urinary Tract Infections

The Open Microbiology Journal ◽

10.2174/1874285801509010014 ◽

2015 ◽

Vol 9 (1) ◽

pp. 14-17 ◽

Cited By ~ 21

Author(s):

Hossein Samadi Kafil ◽

Ashraf Mohabati Mobarez

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Urinary Tract Infections ◽

Biofilm Formation ◽

Urinary Infection ◽

Surface Protein ◽

Antibiotic Resistant ◽

Specific Pcr ◽

Tract Infections

Enterococci rank among leading cause of nosocomial bacteremia and urinary tract infection in hospital and community acquired infections. Several traits that may contribute to enhanced virulence have been identified in Enterococci. Extracellular surface protein (Esp) is a virulence factor that contributes in biofilm formation and resistance to environmental stresses. In this study we aimed to determine occurrence ofespinE. faeciumandE. faecalisisolates isolated from urinary tract infections and to investigate whether there is any correlation between presence ofespand antibiotic resistance. One hundred and sixty six isolates were collected from patients with UTI and after identification by biochemical and PCR, antibiotic resistances were examined. The presence ofespwas investigated by primer-specific PCR. 43.3% of isolates identified asE. faeciumand 56.7% asE. faecalis. Theespgene was found in 76.1% ofE. faeciumisolates and 77.9% ofE. faecalisisolate. There were significant correlation betweenesppositiveE. faeciumand resistance to Vancomycin (p<0.01), also inE.faecaliswe found correlation betweenesppositive and resistance to Ampicillin, Chloramphenicol and Tetracycline (p<0.01, p<0.01, p<0.01 respectively). Occurrence ofespin our isolates from urinary tract infection was high that indicates importance of this gene in urinary tract infections and shows importance of ability to forming biofilm and hydrophobicity of surface of Enterococci for causing urinary infection by Enterococci. Also, our finding showed significant correlation between resistance to antibiotics and presence ofespin Enterococci.

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Contribution of Individual Ebp Pilus Subunits of Enterococcus faecalis OG1RF to Pilus Biogenesis, Biofilm Formation and Urinary Tract Infection

PLoS ONE ◽

10.1371/journal.pone.0068813 ◽

2013 ◽

Vol 8 (7) ◽

pp. e68813 ◽

Cited By ~ 43

Author(s):

Jouko Sillanpää ◽

Chungyu Chang ◽

Kavindra V. Singh ◽

Maria Camila Montealegre ◽

Sreedhar R. Nallapareddy ◽

...

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Enterococcus Faecalis ◽

Biofilm Formation ◽

Pilus Biogenesis

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Efficacy of Ceftobiprole Medocaril against Enterococcus faecalis in a Murine Urinary Tract Infection Model

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.06102-11 ◽

2012 ◽

Vol 56 (6) ◽

pp. 3457-3460 ◽

Cited By ~ 6

Author(s):

Kavindra V. Singh ◽

Barbara E. Murray

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Enterococcus Faecalis ◽

Infection Model ◽

Content Type ◽

The Stability

ABSTRACTWe evaluated ceftobiprole against the well-characterizedEnterococcus faecalisstrain OG1RF (with and without the β-lactamase [Bla] plasmid pBEM10) in a murine urinary tract infection (UTI) model. Ceftobiprole was equally effective for Bla+and Bla−OG1 strains, while ampicillin was moderately to markedly (depending on the inoculum) less effective against Bla+than Bla−OG1 strains. These data illustrate anin vivoeffect on ampicillin of Bla production byE. faecalisand the stability and efficacy of ceftobiprole in experimental UTI.

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Resistance to different antibiotics in clinical isolates of Enterococcus faecalis obtained from urine cultures of patients with urinary tract infection

10.1101/2020.09.20.305623 ◽

2020 ◽

Author(s):

Mariana Islas Rodríguez ◽

José Carlos Valencia Esquivel ◽

Silvia Patricia Rodríguez Peña ◽

Elisangela Oliveira de Freitas ◽

Jorge Angel Almeida Villegas

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Enterococcus Faecalis ◽

Exclusion Criteria ◽

Sensitivity Tests ◽

Resistance Patterns ◽

Toluca Valley ◽

Tract Infections ◽

Bacitracin A

AbstractObjectiveTo identify patterns of resistance against various antibiotics in Enterococcus faecalis in urinary tract infections in a population of the Toluca valley, MexicoMethods155 samples were collected from patients with suspected urinary tract infection without exclusion criteria such as age or gender. Automated equipment was used for the identification of the etiological agent and sensitivity tests.Results80 positive cultures were obtained, of which 20 strains belong to Enterococcus faecalis, which show 100% sensitivity for penicillins, linezolid, vancomycin, bacitracin, a high pattern of sensitivity for quinolones, and a high pattern of resistance to rifampicin, erythromycin and 100% resistance in tetracyclineConclusionIt shows 100% sensitivity to penicillins, vancomycin and linezolid, first-line treatments and for cases of infection complicated by Enterococci. And 100% resistance for tetracycline and high resistance patterns for erythromycin and rifampin.

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