Identification Of Enterococcus Faecalis Research Articles

Relationship between the Presence of Human Papillomavirus in the Female Urethra and Recurrent Urinary Tract Infections.

Introduction: Recurrent urinary tract infections (rUTIs) are highly prevalent health issues among women, significantly impacting their quality of life. Urethral pain or urethritis can arise from infectious or non-infectious origins. The presence of Human Papillomavirus (HPV) in the urogenital tract has been associated with high-risk sexual behaviors, but its presence in the female urethra without such behaviors has not been thoroughly investigated. Objectives: The study aims to determine the presence of HPV in the urethra of women with recurrent urinary tract infections (rUTIs) and concomitant urethral syndrome and to compare the clinical and microbiological characteristics of women with and without urethral HPV, specifically focusing on those without high-risk sexual behaviors. Methods: This prospective multicenter study included 138 women over 18 years old with rUTIs and concomitant urethral pain syndrome. High-risk sexual behaviors, sexually transmitted infections, and vaginitis were set as exclusion criteria. Participants were divided into two groups: NHPV (n = 72) with no urethral HPV and HPV (n = 66) with urethral HPV presence. Variables analyzed included age, body mass index (BMI), follow-up duration, comorbidities, treatments, toxic habits, surgical history, main symptoms, urine sediment findings, and cultures from urine and vaginal exudate. HPV genotyping was also performed. Descriptive statistics were used, along with Student's t-test, Chi-square, Fisher's exact test, ANOVA, and multivariate cluster analysis. Results: The NHPV group was older on average (48.75 years) compared to the HPV group (39.09 years). The HPV group had a longer follow-up period (2634 days vs. 1975 days in NHPV). Urinary incontinence was significantly more common in NHPV (63.89%) compared to HPV (18.18%) (p = 0.001). HPV-positive women had a higher prevalence of verrucous lesions in the vaginal introitus (64% vs. 8% in NHPV). Additionally, the HPV group showed higher rates of pyuria (27.27%), vaginal Candida albicans (36.26%), and positive urine cultures for Escherichia coli (47.83%), Enterococcus faecalis (36.36%), and Klebsiella pneumoniae (21.74%). No significant differences were observed between the groups concerning BMI, smoking habits, diabetes, or the main symptom at consultation. The most common HPV genotypes identified were G35, G42, and G66 (each 27%). Multivariate analysis revealed that sensitivity to nitrofurantoin was the most significant variable in the HPV group (importance of 0.96), followed by fosfomycin (0.79), trimethoprim (0.79), and amoxicillin (0.71). Conclusions: HPV was present in the urethra of 47.8% of women with rUTIs and concomitant urethral syndrome who did not exhibit high-risk sexual behaviors. These women were younger and had a longer duration of symptoms compared to those without urethral HPV. The identification of Enterococcus faecalis, Escherichia coli, and Klebsiella pneumoniae was more common in the HPV group. The sensitivity of bacteria to nitrofurantoin and fosfomycin is crucial for the clinical management of these patients. The presence of urethral HPV should be considered in the evaluation and treatment of women with rUTIs and urethral syndrome.

Investigation of some changes and clonal relationship in enterococci isolates due to relocation of a hospital.

To investigate the isolation rates, antimicrobial resistance rates, minimum inhibitory concentration values of antimicrobial agents, and clonal relationships of Enterococcus faecalis and Enterococcus faeciumdue to the relocation of a hospital to a newly constructed building. The comparative, prospective study was conducted at adult general intensive care units of the Mus State Hospital, Mus, Turkey, in two phases; before the relocation from January 25 to December 1, 2014, and after the relocation from February 10 to May 24, 2015. Rectal swab samples were collected 72 hours post-hospitalisation. Identification of Enterococcus faecalis and Enterococcus faeciumisolates was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and antimicrobial resistance with minimum inhibitory concentration values was detected with Vitek 2 system. The clonal relatedness among the strains was investigated by pulsed-field gel electrophoresis. Data was analysed using SPSS 23. Of the 69 patients, 37(53.62%) were related to pre-relocation phase; 20(54.1%) females and 17(45.9%) males with mean age 62.81±21.71 years. There were 32(46.37%) patients in the post-relocation phase; 13(40.6%) females and 19(59.4%) males with mean age 62.69±21.35 years (p>0.05). Of the 84 enterococci strains isolated, 51(60.7%) were Enterococcus faecium; 28(55%) before relocation and 23(45%) after relocation (p=0.77). The remaining 33(39.3%) isolates were Enterococcus faecalis; 16(48.5%) before relocation and 17(51.5%) after relocation (p=0.73). Multiple strains were located in 7(18.9%) patients before relocation and in 7(21.9%) after relocation. In 1(3.1%) patient after relocation, 2(8.7%) Enterococcus faecium isolates with different resistance and pulsed-field gel electrophoresis patterns were detected. There were no significant differences between the isolation and antibiotic resistance rates before and after relocation (p>0.05), and a clonal relation between the isolates was not detected (p>0.05). Decreased minimum inhibitory concentration values were noted for some antibiotics. Clonal relationship between the isolates and change in the rates of isolation and antimicrobial resistance of Enterococcus faecalis and Enterococcus faecium was not detected due to relocation. Minimum inhibitory concentration values could be used to reveal relocation-related changes in isolates obtained from patients hospitalised in intensive care units.

Identification of Enterococcus faecalis in Different Clonal Lineages with the Pulsed-Field Gel Electrophoresis Method in Imam Khomeini Hospital, Ilam, Iran

Background: Due to the importance of identifying the source of infectious agents, different typing methods have been developed, among which the pulsed-field gel electrophoresis (PFGE) method is known as the gold standard for bacteria. Also, Enterococcus faecalis is classified as a nosocomial infection. Objectives: The current study aimed to identify the source of E. faecalis by the PFGE method. Methods: Bacteria were collected from all cases of urinary tract infections. Then, the identification process was performed. All isolates were evaluated for vancomycin resistance, and then PFGE was carried out. Results: The results of disk diffusion showed that 54% of the isolates showed resistance to vancomycin. Also, 4% of the isolates were intermediate, and 42% showed sensitivity to vancomycin. Afterwards, the PCR of the VanA gene was performed to confirm the results of disk diffusion. Thus, 48 out of 54 (88.8%) isolates had the VanA gene, and none of the four intermediate isolates had the VanA gene. Our results demonstrated that 54 isolates were vancomycin-resistant, and 50 different pulsotypes groups were identified. Conclusions: Our findings showed the isolates of E. faecalis were from different clonal lineages.

Identification of Enterococcus faecalis in a patient with urinary-tract infection based on metagenomic next-generation sequencing: a case report

BackgroundUrinary tract infection (UTI) caused by various pathogenic microorganisms is ubiquitous in the parts of the urinary system such as kidney, ureter, bladder, and urethra. Currently, clinical detection of UTI is mainly focused on urine culture; however, the diagnostic value of urine culture remains limited due to the time-consuming procedure and low detection rate, especially in patients who have used antibiotics. Generally, treatment for UTI relies on empirical medication rather than pathogen diagnosis, which leads to the inappropriate use of antimicrobial agents and a significant increase in resistant strains. Comparatively, metagenomic next-generation sequencing (mNGS) is capable of overcoming the disadvantages of clinical culture, and identifying pathogens for further treatment.Case presentationA 33-year-old male patient was admitted to hospital with a high fever and chills. None of his autoimmune disease or thyroid function related indicators were positive, and he had no risk of endocarditis. His white blood cell count, C-reactive protein, procalcitonin, interleukin 6, and neutrophil proportion were markedly elevated. He was initially diagnosed as having an infection of unknown etiology. Since empirical treatment of Sulperazon and Metronidazole did not relieve his symptoms, both the blood and urine specimens were examined using traditional culture, serological testing, and mNGS assay. Traditional culture and serological testing produced negative results, while the mNGS assay revealed the presence of a potential pathogen, Enterococcus faecalis, in the urine specimen, which was further confirmed by both Sanger sequencing and qPCR analysis. A CT scan of the patient’s whole abdomen showed stones in the right kidney. Once targeted antibiotic therapy was administered, the patient recovered quickly.ConclusionsOur case illustrated that mNGS, as a novel culture-independent approach, demonstrated the capability of rapid, sensitive, and accurate pathogen identification. Furthermore, this technology provides strong support for guiding clinicians to determine appropriate treatment.

Identification of Enterococcus Faecalis and Pseudomonas Aeruginosa on and in Implants in Individuals with Peri-implant Disease: A Cross-Sectional Study.

To verify whether the parts of dental implants can be contaminated by opportunistic pathogens. In this cross-sectional study, 38 individuals (52 implants) were investigated. Samples for microbiologic analysis (for a total of 180 sites) were obtained from each individual, from three types of sites in the following order: (1) the peri-implant sulcus of each implant, (2) the gingival sulcus of adjacent teeth, and (3) inside the implant-abutment connection and the abutment of each implant. Swabs from the oral mucosa (cheeks, tongue, and pharynx) were also collected. Quantitative real-time polymerase chain reaction was carried out for total bacterial counts of Enterococcus faecalis and Pseudomonas aeruginosa. Two-way analysis of variance (sites, species) and Holm-Sidak tests were used for statistical analyses. No opportunistic bacteria were found in the gingival sulcus specimens (38 sites). E faecalis was detected in the peri-implant sulcus (3 of 52 sites) and the inner connection/abutment portion (3 of 52 sites). P aeruginosa was identified only in the oral mucosa swabs (1 of 38 sites) and represented the highest bacterial number (3.5 × 10⁶). Statistically significant differences were only found between species and in the peri-implant sulcus. Within the limitations of this study, significant differences in the presence and levels of nosocomial bacteria were detected in the peri-implant environment of diseased implants. From a clinical point of view, data from this study might suggest that, in patients affected by peri-implantitis, prostheses should be removed and the implant-abutment connection disinfected routinely, along with implant surface decontamination.

Identification of Enterococcus faecalis by PCR in saliva of pediatric patients

Objective: This study aimed to identify E. faecalis in saliva of patients that were divided into two groups: 10 patients with caries lesions and 10 caries-free patients. Material and methods: Saliva samples were collected with a sterile swab and inoculated in enterococcosel media for 48 hours. The positive samples were subcultured in broth agar - blood medium for storage and subsequent PCR analysis. The data were analyzed using the SPSS (χ2). Results: From the 20 cases included in the study, 3 were positive in the Enterococcosel medium, and both tests (culture and PCR) used confirmed that three of them belonged to the species E. faecalis. No samples were positive in Enterococcosel broth in the group of caries-free patients. From the three samples that were identified as Enterococcus in broth (positive bile esculin test), an amplified for E. faecalis PCR analysis (p > 0.005). Conclusion: Through the use of PCR, it was possible to identify the genus Enterococcus and the species E. faecalis in saliva of patients with carious lesions – the pathogen that may influence the prognosis of diseases of the oral cavity.

English

In order to investigate the reasons of lamb death, 11 bacteria were isolated from the brain, liver and other tissues of natural infectious lambs presenting neurological symptoms and septicaemia as the main clinical feature from different farm of two regions in Xinjiang, China in recent years. They were identified as Enterococcus faecalis according to morphological, cultural and biochemical characteri-zation. The infecting strains were identified as belonging to serotype D, G or to an unidentified serotype with Lancefild Group A to G Latex Diagnostic Kit. All 11 E. faecalis pathogens were resistant to norfloxacin, penicillin, tetracycline, streptomycin, gentamicin and erythromycin at varying degrees. PCR screening for nine common virulence factor genes associated with Enterococcusspecies revealed the presence of eight virulence factor genes (esp, cylA, asa1, ace, efa, gelE, ef0591 and ef3314) in 11 of the isolates, five genes were present in all strains examined and gelE was present in 1 pathogenic strain examined. Nine virulence factor genes were not present in two strains examined. Virulence gene analy-sis revealed a 96% homology to Enterococcal genes from pathogenic E. faecalis sequences deposited in GenBank. Some genus from Enterococcus can also lead to the infection and death of lambs.   Key words: Lamb, encephalitis, identification of Enterococcus faecalis, virulence factor gene

A novel method for simultaneous Enterococcus species identification/typing and van genotyping by high resolution melt analysis

In order to develop a typing and identification method for van gene containing Enterococcus faecium, two multiplex PCR reactions were developed for use in HRM-PCR (High Resolution Melt-PCR): (i) vanA, vanB, vanC, vanC23 to detect van genes from different Enterococcus species; (ii) ISR (intergenic spacer region between the 16S and 23S rRNA genes) to detect all Enterococcus species and obtain species and isolate specific HRM curves. To test and validate the method three groups of isolates were tested: (i) 1672 Enterococcus species isolates from January 2009 to December 2009; (ii) 71 isolates previously identified and typed by PFGE (pulsed-field gel electrophoresis) and MLST (multi-locus sequence typing); and (iii) 18 of the isolates from (i) for which ISR sequencing was done. As well as successfully identifying 2 common genotypes by HRM from the Austin Hospital clinical isolates, this study analysed the sequences of all the vanB genes deposited in GenBank and developed a numerical classification scheme for the standardised naming of these vanB genotypes. The identification of Enterococcus faecalis from E. faecium was reliable and stable using ISR PCR. The typing of E. faecium by ISR PCR: (i) detected two variable peaks corresponding to different copy numbers of insertion sequences I and II corresponding to peak I and II respectively; (ii) produced 7 melt profiles for E. faecium with variable copy numbers of sequences I and II; (iii) demonstrated stability and instability of peak heights with equal frequency within the patient sample (36.4±4.5days and 38.6±5.8days respectively for 192 patients); (iv) detected ISR-HRM types with as much discrimination as PFGE and more than MLST; and (v) detected ISR-HRM types that differentiated some isolates that were identical by PFGE and MLST. In conjunction with the rapid and accurate van genotyping method described here, this ISR-HRM typing and identification method can be used as a stable identification and typing method with predictable instability based on recombination and concerted evolution of the rrn operon that will complement existing typing methods.

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

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).

Identification of Enterococcus faecalis in Root-filled Teeth With or Without Periradicular Lesions by Culture-dependent and—Independent Approaches

Enterococcus faecalis is the most commonly found species in root-filled teeth evincing recalcitrant periradicular lesions and as a consequence, a role in causation of endodontic treatment failure has been suggested. The purpose of this study was to evaluate the prevalence of this bacterial species in root-filled teeth with or without periradicular lesions. Identification of E. faecalis was carried out by polymerase chain reaction (PCR) or conventional culture procedures. Overall, E. faecalis was detected by species-specific 16S rRNA gene-based PCR in 40/50 teeth (80%), while culture revealed the occurrence of this species in 8/50 teeth (16%). PCR was significantly more effective than culture in detecting this bacterial species (p < 0.001). Of 27 root-filled teeth with no periradicular lesions, E. faecalis was found in 22 cases (81.5%) by PCR and in five cases (18.5%) by culture. Of 23 root-filled teeth with periradicular lesions, E. faecalis was identified in 18 cases (78%) by PCR and in three cases (13%) by culture. Regardless of the identification technique used, no significant difference was observed when comparing the occurrence of E. faecalis in root-filled teeth with and without periradicular lesions (p > 0.05). Although these findings apparently put into question the status of E. faecalis as the main species causing endodontic treatment failure, other related factors still need to be clarified before this assumption turns into certainty.