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Purpose: Few data have been published on the efficacy of linezolid in enterococcal urinary tract infection (e-UTI). The aims of this study were to describe the characteristics of patients with enterococci UTI treated with linezolid, and to evaluate the efficacy and the tolerance of linezolid treatment. Methods: An observational multicentre retrospective study was conducted in 5 hospitals in France. Patients were included if they met the following criteria: ≥18 years, clinical and microbiological criteria for enterococcal UTI and linezolid treatment > 48 h. Primary outcome was clinical failure. Results: Eighty-one patients were included between January 2015 and December 2021. The median age was 73.0 [64; 83] years and 47 (58%) were men. The median Charlson comorbidity index was 3.00 [2; 6]. E. faecium was reported in 65 (80%) cases and E. faecalis in 26 cases (32%). Polymicrobial infections occurred in 41 (51%) cases. No enterococci was resistant to vancomycin. Before linezolid prescription an empiric antimicrobial treatment was started in 48 (59%) cases and was effective against enterococci in 19/48 (39.5%) patients for a median of 3.5 days [2.0; 4.0]. The median duration of linezolid antibiotic treatment was 13 days [10; 14]. Three adverse events were reported, none were serious but one led to discontinuation of treatment. Treatment failure was reported in 2 cases (2.5%). Conclusion: This study provides evidence for efficacy and safety of linezolid in enterococcal UTI. [ABSTRACT FROM AUTHOR]

Background: Infections caused by biofilm-forming bacteria have significantly linked to dental plaque and caries. The aim of this study is to assess efficacy of some natural compounds in inhibition and eradication of biofilm formed by bacterial isolates from dental infections. Results: Bacterial isolates were recovered from dental plaque/caries and identified using standard microbiological tests and 16S rDNA sequencing. The isolated bacterial strains include Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Klebsiella pneumoniae, and Escherichia coli. The antibiotic susceptibility was determined by disk diffusion method and revealed that the majority of isolates showed high antibiotic resistance, and 61% of isolates were found to be multidrug resistant. The biofilm formation capacity of isolates was investigated using microtiter plate assay. Among the 77 bacterial isolates, seventeen showed moderate biofilm formation capacity, twenty-two showed near-moderate, thirty-four had weak biofilm-forming capacity, and four were non-biofilm producers. The antibiofilm activity of tested compounds (rose and jasmine oils, propolis, vanillin, and vinegar) was evaluated against isolates with highest biofilm-forming capacity. The in vitro antibiofilm ability of tested substances were investigated alone or in combination with each other to evaluate their ability to prevent biofilm formation or destroy preformed single-/multispecies biofilms. Finally, antibiofilm ability of tested combination was evaluated ex vivo on natural teeth. Our results showed that vanillin in combination with rose or jasmine oils showed promising biofilm inhibition and biofilm eradication activities in both the in vitro and ex vivo models. Conclusions: Dental plaque and caries can be successfully prevented using combination of vanillin with rose or jasmine oils, and these compounds can be incorporated in new anticaries dental formulations. [ABSTRACT FROM AUTHOR]

Amongst all Enterococcus spp., E. faecalis and E. faecium are most known notorious pathogen and their biofilm formation has been associated with endocarditis, oral, urinary tract, and wound infections. Biofilm formation involves a pattern of initial adhesion, microcolony formation, and mature biofilms. The initial adhesion and microcolony formation involve numerous surface adhesins e.g. pili Ebp and polysaccharide Epa. The mature biofilms are maintained by eDNA, It’s worth noting that phage-mediated dispersal plays a prominent role. Further, the involvement of peptide pheromones in regulating biofilm maintenance sets it apart from other pathogens and facilitating the horizontal transfer of resistance genes. The role of fsr based regulation by regulating gelE expression is also discussed. Thus, we provide a concise overview of the significant determinants at each stage of Enterococcus spp. biofilm formation. These elements could serve as promising targets for antibiofilm strategies. [ABSTRACT FROM AUTHOR]

Purpose: To analyse recent epidemiological trends of bloodstream infections (BSI) caused by Enterococcus spp. In adult patients admitted to tertiary care centres in Germany. Methods: Epidemiological data from the multicentre R-NET study was analysed. Patients presenting with E. faecium or E. faecalis in blood cultures in six German tertiary care university hospitals between October 2016 and June 2020 were prospectively evaluated. In vancomycin-resistant enterococci (VRE), the presence of vanA/vanB was confirmed via molecular methods. Results: In the 4-year study period, 3001 patients with BSI due to Enterococcus spp. were identified. E. faecium was detected in 1830 patients (61%) and E. faecalis in 1229 patients (41%). Most BSI occurred in (sub-) specialties of internal medicine. The pooled incidence density of enterococcal BSI increased significantly (4.0–4.5 cases per 10,000 patient days), which was primarily driven by VRE BSI (0.5 to 1.0 cases per 10,000 patient days). In 2020, the proportion of VRE BSI was > 12% in all study sites (range, 12.8–32.2%). Molecular detection of resistance in 363 VRE isolates showed a predominance of the vanB gene (77.1%). Conclusion: This large multicentre study highlights an increase of BSI due to E. faecium, which was primarily driven by VRE. The high rates of hospital- and ICU-acquired VRE BSI point towards an important role of prior antibiotic exposure and invasive procedures as risk factors. Due to limited treatment options and high mortality rates of VRE BSI, the increasing incidence of VRE BSI is of major concern. [ABSTRACT FROM AUTHOR]

Purpose: Globally, colorectal cancer (CRC) is among the most prevalent cancers. One distinctive feature of colorectal cancer is its close relationship to the gut microbiota, which is a crucial component of the tumor microenvironment. Over the last ten years, research has demonstrated that colorectal cancer is accompanied with dysbiosis of gut bacteria, fungi, viruses, and Archaea, and that these alterations may be causal. Objectives: This study aimed to evaluate the disruption of the microorganism composition in the intestine, especially bacteria and to determine their relationship with colorectal cancer. Methods: An evaluation system for determining colorectal cancer (CRC) risk and prognosis can be established more easily with the help of accurate gut microbiota profiling. Stool samples from 14 CRC patients and 13 controls were collected and the flora relative abundance was measured using targeted quantitative PCR (qPCR) assays to evaluate diagnostic potential of selected biomarkers: Streptococcus gallolyticus and Enterococcus faecalis. Culture and MALDI-TOF mass spectrometry were coupled to identify the gut microbiota in both colorectal cancer and control groups. Results: Compared with controls, the gut microbiota of CRC patients showed an increase in the abundance of Enterococcus, Fusobacterium and Streptococcus. At the species level, the CRC enriched bacterium including Escherichia coli, Enterococcus faecalis, Fusobacterium nucleatum, Streptococcus gallolyticus, Flavoni fractorplautii and Eggerthella lenta acted as promising biomarkers for early detection of CRC. Conclusion: This study highlights the potential of gut microbiota biomarkers as a promising non-invasive tool for the accurate detection and distinction of individuals with CRC. [ABSTRACT FROM AUTHOR]

One of the most prevalent infectious diseases identified in both communities and hospitalized patients is urinary tract infection (UTI). Enterococcus is evolved into a clinically pertinent uropathogen due to its evolving resistance to multiple antimicrobial agents.This study, detects antimicrobial susceptibility patterns of Enterococcus species and molecular detection of Enterococcus faecalis from patients with urinary tract infections. In this cross-sectional observational study, 165 urine samples were obtained from clinically diagnosed patients with UTIs of different ages and gender. Enterococcus species were identified by standard microbiological procedure and PCR (by using species-specific primers for Enterococcus faecalis). A modified Kirby Bauer Disc diffusion method was used to identify the antimicrobial susceptibility pattern following Clinical and Laboratory Standards Institute (CLSI) guidelines. Out of 165 urine samples, 134 samples yielded positive cultures. Enterococcus species were isolated from 23 (17.1%) urine samples. Among all Enterococcus, 16 (69.6%) isolates were E. faecalis, detected by PCR assay. A higher (30.4%) proportion of Enterococcus-positive patients were from the age group 48–57 years and female patients (78.2%) had a higher prevalence. Enterococcal infection was found in 56.5% of non-catheterized patients and 43.5% of catheterized patients. Vancomycin and linezolid (78.3%) and meropenem (73.9%) sensitivity was prevalent among all Enterococcus species. They showed 100% resistance towards ceftriaxone, cefixime 95.7%, cefuroxime 91.3%, azithromycin 82.6%. This research indicated the occurrence of Enterococcus species and the advent of multidrug-resistant E. faecalis in patients with UTIs. Routine speciation and antimicrobial susceptibility testing of Enterococcus in various clinical samples is encouraged. [ABSTRACT FROM AUTHOR]

As ethnic medicine, the whole grass of plants in Cirsium was used as antimicrobial. This review focuses on the antimicrobial activity of plants in Cirsium, including antimicrobial components, against different types of microbes and bacteriostatic mechanism. The results showed that the main antimicrobial activity components in Cirsium plants were flavonoids, triterpenoids and phenolic acids, and the antimicrobial ability varied according to the species and the content of chemicals. Among them, phenolic acids showed a strong antibacterial ability against Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterococcus faecium. The antibacterial mechanisms include: (1) damaging the cell membrane, cell walls, mitochondria and nucleus of bacteria; (2) inhibiting the synthesis of proteins and nucleic acids; (3) suppressing the synthesis of enzymes for tricarboxylic acid cycle pathways and glycolysis, and then killing the bacteria via inhibition of energy production. Totally, most research results on antimicrobial activity of Cirsium plants are reported based on in vitro assays. The evidence from clinical data and comprehensive evaluation are needed. [ABSTRACT FROM AUTHOR]

Urinary tract infections (UTIs) are among the most common bacterial infections, posing significant public health challenges due to increasing antimicrobial resistance (AMR). This study aims to assess the prevalence, demographic characteristics, microbial profile, and antimicrobial resistance patterns in Indian patients with UTIs admitted to intensive care unit. A total of 154 patients with positive UTIs were included in this cross-sectional study. The prevalence data including demographics, microbial isolates, and antimicrobial susceptibility patterns were collected. Additionally, risk factors for multidrug resistance uropathogens were assessed using multivariate analyses. The patient cohort had diverse demographic, with a slight male predominance of 52.6% (n = 81). The most common comorbidities were hypertension 59.1% (n = 91) and diabetes mellitus 54.5% (n = 84). The microbial profile was dominated by gram-negative bacteria, particularly Escherichia coli 26.62% (n = 41) and Klebsiella pneumoniae 17.53% (n = 27). The predominant gram-positive and fungal isolate was Enterococcus faecium 7.14% (n = 11) and Candida spp. 18.83% (n = 29), respectively. Substantial resistance was noted against common antimicrobials, with variations across different pathogens. Gram-negative bacteria, particularly Escherichia coli and Klebsiella pneumoniae, exhibited high MDR rates, emphasizing the challenge of antimicrobial resistance. Multivariate logistic regression identified age groups 50–65 and over 65, and prolonged catheterization as significant risk factors for MDR infections. A significantly high resistance rate among pathogens emphasizes the need for judicious antimicrobial use. Our findings emphasize the necessity of ongoing surveillance and tailored interventions based on local pathogen prevalence and antibiogram data to effectively address the threat of AMR threat for better management of UTI management in ICU settings. [ABSTRACT FROM AUTHOR]

The prevalence of multidrug resistant (MDR) bacterial infections continues to rise as the development of antibiotics needed to combat these infections remains stagnant. MDR enterococci are a major contributor to this crisis. A potential therapeutic approach for combating MDR enterococci is bacteriophage (phage) therapy, which uses lytic viruses to infect and kill pathogenic bacteria. While phages that lyse some strains of MDR enterococci have been identified, other strains display high levels of resistance and the mechanisms underlying this resistance are poorly defined. Here, we use a CRISPR interference (CRISPRi) screen to identify a genetic locus found on a mobilizable plasmid from Enterococcus faecalis involved in phage resistance. This locus encodes a putative serine recombinase followed by a Type IV restriction enzyme (TIV-RE) that we show restricts the replication of phage phi47 in vancomycin-resistant E. faecalis. We further find that phi47 evolves to overcome restriction by acquiring a missense mutation in a TIV-RE inhibitor protein. We show that this inhibitor, termed type IV restriction inhibiting factor A (tifA), binds and inactivates diverse TIV-REs. Overall, our findings advance our understanding of phage defense in drug-resistant E. faecalis and provide mechanistic insight into how phages evolve to overcome antiphage defense systems. Using a CRISPR interference genetic approach, Bullen, Johnson, and colleagues discover a type IV restriction (TIV-RE) enzyme encoded on a mobilizable plasmid of multidrug resistant Enterococcus faecalis. This TIV-RE is a potent inhibitor of bacteriophage infection. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the performance of a rapid multiplex microarray-based method (Unyvero BCU system, BCU) to identify microorganisms and detect antimicrobial resistance directly from positive blood culture (BC) bottles with polymicrobial growth, and to assess relevance of information provided for timely guidance of polymicrobial bloodstream infection treatment. Methods: Accuracy, time-to-actionable results and potential impact of BCU on antimicrobial treatment were compared with those of standard of care during a prospective study for the sample analysis (November 2017-November 2018) and a retrospective study for the clinical data analysis and the time-to-result analysis. The study was complemented with an experimental study, based on spiked blood cultures to assess the ability of the method to detect antimicrobial resistance genes. Results: Sixty-five clinical polymicrobial BC samples (163 total microorganisms) and 30 simulated polymicrobial BC samples (60 strains) were included. BCU reported 84.6% samples as polymicrobial, correctly identified all the bacteria of the mix for 72.3% samples (47/65) and detected bacteria that were missed by the conventional culture for 13.8% samples. All identifications and antimicrobial resistances were accurately detected for 61.5% (40/65) samples. Limitations concerned the detection of anaerobes, enterococci and enterobacterial susceptibility to third generation cephalosporins. BCU results would have guided antimicrobial treatment for 50.8% of the cases (33/65) in a timely and relevant manner, had no impact for 27.7% (18/65) and been misleading for 18.5% (12/65). Conclusions: Despite some limitations, the Unyvero BCU system is a rapid and reliable method for polymicrobial BC sample analysis. [ABSTRACT FROM AUTHOR]

E. innesii is a recently described Enterococcus species which may be difficult to differentiate from the more common E. casseliflavus. We present the first clinical report of invasive E. innesii infection, featuring two cases of biliary sepsis. Whole genome sequencing confirmed the taxonomic assignment and the presence of vanC-4. Analysis of public genomes identified 13 deposited E. innesii and 13 deposited E. casselifalvus/E.gallinarum genomes which could be reassigned as E. innesii. Improved laboratory diagnosis of E. innesii is expected to generate additional data concerning its clinical relevance and support the future diagnosis and treatment of this uncommon pathogen. [ABSTRACT FROM AUTHOR]

Bacteriocins are antimicrobial peptides that are naturally produced by many bacteria. They hold great potential in the fight against antibiotic resistant bacteria, including ESKAPE pathogens. Engineered live biotherapeutic products (eLBPs) that secrete bacteriocins can be created to deliver targeted bacteriocin production. Here we develop a modular bacteriocin secretion platform that can be used to express and secrete multiple bacteriocins from non-pathogenic Escherichia coli host strains. As a proof of concept we create Enterocin A (EntA) and Enterocin B (EntB) secreting strains that show strong antimicrobial activity against Enterococcus faecalis and Enterococcus faecium in vitro, and characterise this activity in both solid culture and liquid co-culture. We then develop a Lotka-Volterra model that can be used to capture the interactions of these competitor strains. We show that simultaneous exposure to EntA and EntB can delay Enterococcus growth. Our system has the potential to be used as an eLBP to secrete additional bacteriocins for the targeted killing of pathogenic bacteria. Bacteriocins are antimicrobial peptides that are naturally produced by many bacteria. In this work, authors develop a bacteriocin secretion platform, and test, in a proof-of-concept study, antimicrobial efficacy against Enterococcus faecalis and Enterococcus faecium. [ABSTRACT FROM AUTHOR]

Diabetic foot ulcer caused by vancomycin-resistant Enterococcus strains is a troublesome concern that threatens public health. Due to the few treatment options, it is essential to develop a new alternative antibacterial agent with high efficacy. In the current study, we evaluated the antibacterial and antibiofilm properties of usnic acid (UA), a secondary metabolite of lichens. Microdilution broth and crystal violet methods were applied to determine the antibacterial and antibiofilm activities, respectively. To further elucidate the biofilm inhibition, scanning electron microscopy imaging was performed. In addition, the antioxidant activity of UA was investigated by 2, 2-diphenyl-1-picrylhydrazyl assay. The bacteriostatic and bactericidal activity of UA was observed as an amount of 23.02 and 312.5 μg/mL, respectively. UA suppressed biofilm formation and motivated the reduction of free oxygen radicals. The present study may open up a new perspective to combat biofilm-forming enterococci. [ABSTRACT FROM AUTHOR]

In the face of the escalating global antibiotic resistance crisis, exploring alternative therapeutic avenues has become imperative. This study investigates the antibacterial potential of Punica granatum and Rhamnus sp. extracts from Dhahran, Saudi Arabia. Antibacterial activities were evaluated against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis using well diffusion, MIC, and MBC assays. The antibacterial tests of both plants revealed that ethanolic extracts demonstrate superior activity compared to chloroform extracts. Also, Enterococcus faecalis proves most susceptible (up to 33 mm inhibition), while Escherichia coli exhibits notable resistance. Although Escherichia coli was the most resistant organism using MIC, Pseudomonas aeruginosa was the most susceptible strain with MIC less than 12.5 mg/mL. The Punica granatum extracts and Rhamnus sp. chloroformed extract display primarily bactericidal effects, whereas the ethanolic extract of Rhamnus sp. has a mainly bacteriostatic effect. Phytochemical analysis reveals a rich composition, including different well-known antimicrobial chemical classes. These findings underscore the potent antibacterial activity of both plants against diverse bacterial strains. The study emphasizes the need for further research into their phytochemical characterization and mechanistic insights. [ABSTRACT FROM AUTHOR]

Introduction: This study aimed to determine the pathogen distribution and drug susceptibility of diabetic foot wound secretions in a tertiary hospital in a coastal area of southeastern China to guide clinical antibiotic selection. Methods: A retrospective analysis was conducted on 212 patients with diabetic foot hospitalized at Xiamen Third Hospital from 2018 to 2023, and foot wound secretions were collected for microbial culture and drug susceptibility testing. Results: Among 212 cases of patients with diabetic foot wound secretions, 163 cases (76.9%) were cultured with pathogenic bacteria, and a total of 207 strains of pathogenic bacteria were cultured, including 75 strains (36.23%) of Gram-positive (G+) bacteria, 118 strains of Gram-negative (G−) bacteria (57.00%), 14 strains of fungi (6.76%), 120 cases of single microorganism infection (73.62%), 43 cases of mixed infection (26.38%), and 15 strains of multidrug-resistant bacteria (7.25%). The top three pathogenic bacteria were Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa. G+ bacteria were dominated by S. aureus. Drug susceptibility results showed that G+ bacteria were highly susceptible to vancomycin, linezolid, tigecycline, quinupristin/dalfopristin, rifampicin, and furotoxin, and somewhat resistant to penicillin, erythromycin, clindamycin, and cefoxitin. Among G− bacterial infections, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Proteus were the major species. Drug susceptibility testing indicated that carbapenems such as imipenem and ertapenem were the most effective antibacterial drugs against G− strains, followed by amikacin, piperacillin, and tazabactams to which these bacteria were also relatively sensitive, while resistance to penicillins and first-generation cephalosporins increased significantly. We isolated one strain of pathogenic bacteria from a Wagner grade 1 ulcer, which was G+ bacteria. In Wagner grade 2 ulcers, the distribution of pathogenic bacteria was mainly G+ bacteria. In Wagner grade 3 and 4 ulcers, the distribution of pathogenic bacteria was mainly G− bacteria, and the increased rate of mixed infection was mainly due to mixed infection of G+ and G−. Two strains of pathogenic bacteria were isolated at Wagner grade 5, which were mixed infections of G+ and G−. Conclusions: Pathogenic bacteria in diabetic foot wounds are predominantly G− bacteria, followed by G+ bacteria. As the Wagner ulcer grade increases, the distribution of pathogenic bacteria changes from G+ bacteria to G− bacteria, and the mixed infection rate increases. G+ bacteria are highly susceptible to vancomycin, linezolid, tigecycline, quinupristin/dalfopristin, rifampicin, and furotoxin, and somewhat resistant to penicillin, erythromycin, clindamycin, and cefoxitin. G− bacteria are more sensitive to the antimicrobial drugs ertapenem, imipenem, amikacin, piperacillin tazobactam, and have high resistance to penicillin and first-generation cephalosporins. [ABSTRACT FROM AUTHOR]

To investigate the mechanism of Triton X-100 (TX-100) reducing the Ag+-resistance of Enterococcus faecalis (E. faecalis), and evaluate the antibacterial effect of TX-100 + Ag+ against the induced Ag+-resistant E. faecalis (AREf). The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of AgNO3 against E. faecalis with/without TX-100 were determined to verify the enhanced antibacterial activity. Transmission electron microscopy (TEM) was used to observe the morphological changes of E. faecalis after treatment. The intra- and extracellular concentration of Ag+ in treated E. faecalis was evaluated using inductively coupled plasma mass spectrometer (ICP-MS). The changes in cell membrane potential and integrity of treated E. faecalis were also observed using the flow cytometer. Moreover, AREf was induced through continuous exposure to sub-MIC of Ag+ and the antibacterial effect of TX-100 + Ag+ on AREf was further evaluated. The addition of 0.04% TX-100 showed maximal enhanced antibacterial effect of Ag+ against E. faecalis. The TEM and ICP-MS results demonstrated that TX-100 could facilitate Ag+ to enter E. faecalis through changing the membrane structure and integrity. Flow cytometry further showed the effect of TX-100 on membrane potential and permeability of E. faecalis. In addition, the enhanced antibacterial effect of TX-100 + Ag+ was also confirmed on induced AREf. TX-100 can facilitate Ag+ to enter E. faecalis through disrupting the membrane structure and changing the membrane potential and permeability, thus reducing the Ag+-resistance of E. faecalis and enhancing the antibacterial effect against either normal E. faecalis or induced AREf. [ABSTRACT FROM AUTHOR]

Nasally colonized staphylococci carry antibiotic resistance genes and may lead to serious opportunistic infections. We are investigating nasal carriage of Staphylococcus aureus and Staphylococci other than S. aureus (SOSA) among young volunteers in Egypt to determine their risk potential. Nasal swabs collected over 1 week in June 2019 from 196 volunteers were cultured for staphylococcus isolation. The participants were interviewed to assess sex, age, general health, hospitalization and personal hygiene habits. Identification was carried out using biochemical tests and VITEK 2 automated system. Disc diffusion and minimum inhibitory concentration tests were performed to determine antibiotic susceptibility. Screening for macrolide resistance genes (ermA, ermB, ermC, ermT and msrA) was performed using polymerase chain reaction. Thirty four S. aureus and 69 SOSA were obtained. Multi-drug resistance (MDR) was detected among most staphylococcal species, ranging from 30.77% among S. hominis to 50% among S. epidermidis. Phenotypic resistance to all tested antibiotics, except for linezolid, was observed. Susceptibility to rifampicin, vancomycin and teicoplanin was highest. ermB showed the highest prevalence among all species (79.41% and 94.2% among S. aureus and SOSA, respectively), and constitutive macrolide-lincosamide-streptogramin B (MLSB) resistance was equally observed in S. aureus and SOSA (11.11% and 16.22%, respectively), whereas inducible MLSB resistance was more often found in S. aureus (77.78% and 43.24%, respectively). The species or resistance level of the carried isolates were not significantly associated with previous hospitalization or underlying diseases. Although over all colonization and carriage of resistance genes are within normal ranges, the increased carriage of MDR S. aureus is alarming. Also, the fact that many macrolide resitance genes were detected should be a warning sign, particularly in case of MLSB inducible phenotype. More in depth analysis using whole genome sequencing would give a better insight into the MDR staphylococci in the community in Egypt. [ABSTRACT FROM AUTHOR]

Healthy cattle, sheep, and goats can be reservoirs for gastrointestinal pathogenic fecal enterococci, some of which could be multidrug-resistant to antimicrobials. The objective of this study was to determine the prevalence and diversity of Enterococcus species in healthy sheep, goat, and cattle carcasses, as well as to analyze the antimicrobial resistance phenotype/genotype and the virulence gene content. During 2019–2020, carcass surface samples were collected from 150 ruminants in a slaughterhouse. A total of 90 enterococci, comprising five species, were obtained. The overall prevalence of enterococci was found to be 60%, out of which 37.7% were identified as Enterococcus (E.) hirae, 33.3% as E. casseliflavus, 15.5% as E. faecium, 12.2% as E. faecalis, and 1.1% as E. gallinarum. Virulence-associated genes of efaA (12.2%) were commonly observed in the Enterococcus isolates, followed by gelE (3.3%), asaI (3.3%), and ace (2.2%). High resistance to quinupristin-dalfopristin (28.8%), tetracycline (21.1%), ampicillin (20%), and rifampin (15.5%) was found in two, four, four, and five of the Enterococcus species group, respectively. The resistance of Enterococcus isolates to 11 antibiotic groups was determined and multidrug resistant (MDR) strains were found in 18.8% of Enterococcus isolates. Characteristic resistance genes were identified by PCR with an incidence of 6.6%, 2.2%, 1.1%, 1.1%, 1.1%, and 1.1% for the tetM, ermB, ermA, aac(6ʹ)Ie-aph(2")-la, VanC1, and VanC2 genes in Enterococcus isolates, respectively. Efflux pump genes causing multidrug resistance were detected in Enterococcus isolates (34.4%). The results showed that there were enterococci in the slaughterhouse with a number of genes linked to virulence that could be harmful to human health. [ABSTRACT FROM AUTHOR]

The microbiota represents a crucial area of research in maintaining human health due to its potential for uncovering novel biomarkers, therapies, and molecular mechanisms relevant to population identification and experimental model characterization. Among these microorganisms, Enterococcus faecalis, a Gram-positive bacterium found in the gastrointestinal tract of humans and animals, holds particular significance. Strains of this bacterial species have sparked considerable debate in the literature due to their dual nature; they can either be utilized as probiotics in the food industry or demonstrate resistance to antibiotics, potentially leading to severe illness, disability, and death. Given the diverse characteristics of Enterococcus faecalis strains, this review aims to provide a comprehensive understanding of their impact on various systems within the host, including the immunological, cardiovascular, metabolic, and nervous systems. Furthermore, we summarize the bacterium-host interaction characteristics and molecular effects to highlight their targets, features, and overall impact on microbial communities and host health. [ABSTRACT FROM AUTHOR]

Recently, focus has been placed on renewable sources, as they can be provided in large quantities at the lowest possible cost, in order to create nanoparticles. One of these sources is Zygnema moss which used in the present investigation to create Copper oxide nanoparticles (CuONPs). Several phenols and flavonoids were identified the extract of Zygnema sp. via analysis of High performance liquid chromatography. These constituents served as reducing and stabilizing agents for CuONPs. Characterization of CuONPs was performed via UV-visible spectrum that demonstrated peak at 252 nm, Transmission electron microscopy that showed spherical CuONPs with mean diameter of 30.06 nm, Fourier transform infrared spectroscopy that confirm that presence of several functional groups aided to formation of CuONPs. The crystallographic pattern of CuONPs was recorded via X-ray diffraction analysis. Antimicrobial potential of CuONPs was compared to copper acetate and antibiotic/antifungal drug. CuONPs exhibited more inhibition zones against S. aureus (32 ± 0.1 mm), E. coli (36 ± 0.1 mm), S. typhi (27 ± 0.2 mm), E. faecalis (37 ± 0.1 mm), C. albicans (34 ± 0.3 mm) than copper acetate and antibiotic/antifungal drug. Promising MIC values of were recorded against S. aureus, E. coli, and S. typhi. CuONPs at 200 ppm inhibited the growth of C. lunata, F. oxysporium, A. flavus, and Mucor circinelloid with inhibtion of 76.92, 73.33, 63.63, and 53.84%, respectively regarded the control 100% growth. The photocatalytic role of CuONPs was recorded for degradation of reactive red (RR195) and reactive blue (RB) dyes with maximum degradation of 84.66% and 90.82%, respectively at 75 min. Moreover, the optimal dyes degradation was 84.66 and 90.82%, respectively at 40 °C. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to to compare the antimicrobial resistance rate and its relationship with the antibiotic consumption in two separate Intensive Care Units (ICUs) of the same hospital, one with and other without selective decontamination of the digestive tract (SDD). Methods: We performed a retrospective study in the two ICUs of the Araba University Hospital. Trauma and neurosurgical patients are admitted to the SDD-ICU, and general digestive surgery patients go to the no SDD-ICU. From 2014 to 2018 we analyzed the number of isolates, and the bacterial resistance trends of 47 antimicrobial-microorganism combinations. Additionally, antimicrobial consumption was estimated in both ICUs. Resistance rates were also compared with those reported in ENVIN-HELICS Spanish national registry. Results: In the ICU with SDD protocol, there was a significant decrease in the resistance of E. coli to amoxicillin/clavulanic acid and in the resistance of E. faecalis to high concentration of gentamycin and high concentration of streptomycin. A significant increase of resistance of Staphylococcus coagulasa negative (CoNS) to linezolid in the no SDD-ICU was also detected. Overall, the level of resistance in the SDD-ICU was lower or of the same order than in the ICU without SDD and that reported in the Spanish national registry. Conclusions: SDD had neither a clinically relevant impact on emergence and spread of resistance, nor in the overall systemic antimicrobial use. The patient type rather than the SDD protocol showed to condition the ecology and therefore, the resistance rate in the ICUs. [ABSTRACT FROM AUTHOR]

Background: Bacterial drug resistance represents a significant global concern, with vancomycin-resistant Enterococcus faecalis posing a particularly grave threat to contemporary healthcare systems. This study aims to reveal the reasons for the prevalence of VRE in China. Methods: This study collected data from the China Antimicrobial Resistance Detection System, China Statistical Yearbook, and China Meteorological Network. The collected data are meticulously organized and subjected to both single-factor and multi-factor analyses. An accurate multiple linear regression model was developed by utilizing this comprehensive dataset. Results: Single-factor analysis revealed significant regional variations in the resistance rate of vancomycin-resistant Enterococcus faecalis (P = 0.003). Specifically, there were noteworthy disparities observed between regions experiencing temperate and monsoon climates (P = 0.029; P = 0.005). Furthermore, multi-factor regression analysis demonstrated a negative correlation between the drug resistance rate and both rainfall and rGDP, while a positive correlation was observed with nPI. Conclusion: We successfully established a prediction model for the VRE and found that the resistance rate was low in areas with high rainfall and high per capita economic income, but high in areas with many specialized public health institutions. This is critical for public health strategies and helps policymakers and healthcare practitioners tailor antibiotic resistance approaches to local geography, meteorology, economic conditions. [ABSTRACT FROM AUTHOR]

The unique properties of biosurfactants obtained from microbes, including their activity at extreme temperatures, make them more attractive than synthetic alternatives. Henceforth, the principle objective is to isolate and detect the antibacterial and antifungal activities of the biosurfactants produced from bacteria of the economically competitive mangrove ecosystem. Using the serial dilution method, 53 bacterial strains were recovered from the Manakudy mangrove forest in Kanyakumari, India, for the investigation. Different biosurfactant screening methods and morphological and biochemical tests were opted to select the potential biosurfactant producer. After the initial screening, two strains were discovered by 16S rRNA gene sequencing followed by extraction using chloroform: methanol (2:1) by the precipitation method. The partially purified biosurfactants were then screened for antimicrobial properties against pathogens including Mucor sp., Trichoderma sp. Morphological, biochemical, and 16S rRNA gene sequencing identified the two strains to be gram-positive, rod-shaped bacteria namely Virgibacillus halodentrificans CMST-ZI (GenBank Accession No.: OL336402.1) and Pseudomonas pseudoalcaligenes CMST-ZI (GenBank Accession No (10 K): OL308085.1). The two extracted biosurfactants viz., 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester, as well as cycloheptane efficiently inhibited human pathogens, including Enterococcus faecalis, and fungi, including Mucor sp., Trichoderma sp., indicated by the formation of a zone of inhibition in pharmacological screening. Thus, there is a growing interest in the prospective application of these biosurfactants isolated from marine microbes, exhibiting antimicrobial properties which can be further studied as a potential candidate in biomedical studies and eco-friendly novel drug development. [ABSTRACT FROM AUTHOR]

The biodiversity of enterococci from pastırma (a traditional Turkish dry-cured meat product) by genotypic identification and the antagonistic activities of strains were investigated. Pastırma samples taken from 20 different small-scale factories were subjected to microbiological and physicochemical analysis. A hundred enterococci isolates were identified by 16S rRNA gene sequence analysis. To determine antagonistic activity of strains, Listeria monocytogenes and Staphylococcus aureus were used. The lactic acid bacteria and Micrococcus/Staphylococcus counts were ≥ 6 log cfu/g in 55% of the samples and 75% of the samples, respectively. Enterobacteriaceae was generally below the detectable level (< 2 log cfu/g). The enterococci count was higher than 6 log cfu/g in 30–35% of the samples, depending on the medium used. The enterococci isolates (100 isolates) were identified as E. faecium (80 strains), E. faecalis (19 strains) and E. hirae (1 strain) in genotypic identification. The nine E. faecium strains showed antagonistic activity against L. monocytogenes in the well diffusion test. In contrast, in the same antagonistic activity test, all of the strains had no antagonistic activity against S. aureus. Further studies could be planned to characterize E. faecium strains that show antagonistic activity against L. monocytogenes. [ABSTRACT FROM AUTHOR]

Bile microecology changes play an important role in the occurrence and development of choledocholithiasis. At present, there is no clear report on the difference of bile microecology between asymptomatic patients with gallbladder polyps and choledocholithiasis. This study compared bile microecology between gallbladder polyp patients and patients with choledocholithiasis to identify risk factors for primary choledocholithiasis. This study was conducted in 3 hospitals in different regions of China. Bile samples from 26 patients with gallbladder polyps and 31 patients with choledocholithiasis were collected by laparoscopic cholecystectomy and endoscopic retrograde choledocholithiasis cholangiography (ERCP), respectively. The collected samples were used for 16S ribosomal RNA sequencing and liquid chromatography mass spectrometry analysis. The α-diversity of bile microecological colonies was similar between gallbladder polyp and choledocholithiasis, but the β-diversity was different. Firmicutes, Proteobacteri, Bacteroidota and Actinobacteriota are the most common phyla in the gallbladder polyp group and choledocholithiasis group. However, compared with the gallbladder polyp patients, the abundance of Actinobacteriota has significantly lower in the choledocholithiasis group. At the genera level, the abundance of a variety of bacteria varies between the two groups, and Enterococcus was significantly elevated in choledocholithiasis group. In addition, bile biofilm formation—Pseudomonas aeruginosa was more metabolically active in the choledocholithiasis group, which was closely related to stone formation. The analysis of metabolites showed that a variety of metabolites decreased in the choledocholithiasis group, and the concentration of beta-muricholic acid decreased most significantly. For the first time, our study compared the bile of gallbladder polyp patients with patients with choledocholithiasis, and suggested that the change in the abundance of Actinobacteriota and Enterococcus were closely related to choledocholithiasis. The role of Pseudomonas aeruginosa biofilm in the formation of choledocholithiasis was discovered for the first time, and some prevention schemes for choledocholithiasis were discussed, which has important biological and medical significance. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the occurrence of vancomycin-variable enterococci (VVE) in a hospital in central Italy. Methods: vanA positive but vancomycin-susceptible Enterococcus faecium isolates (VVE-S) were characterized by antibiotic susceptibility tests, molecular typing (PFGE and MLST), and WGS approach. The reversion of VVE-S to a resistant phenotype was assessed by exposure to increasing vancomycin concentrations, and the revertant isolates were used in filter mating experiments. qPCR was used to analyze the plasmid copy number. Results: Eleven putative VVE-S were selected. WGS revealed two categories of vanA cluster plasmid located: the first type showed the lack of vanR, the deletion of vanS, and an intact vanH/vanA/vanX cluster; the second type was devoid of both vanR and vanS and showed a deletion of 544-bp at the 5′-end of the vanH. Strains (n = 7) carrying the first type of vanA cluster were considered VVE-S and were able to regain a resistance phenotype (VVE-R) in the presence of vancomycin, due to a 44-bp deletion in the promoter region of vanH/vanA/vanX, causing its constitutive expression. VVE-R strains were not able to transfer resistance by conjugation, and the resistance phenotype was unstable: after 11 days of growth without selective pressure, the revertants were still resistant but showed a lower vancomycin MIC. A higher plasmid copy number in the revertant strains was probably related to the resistance phenotype. Conclusion: We highlight the importance of VVE transition to VRE under vancomycin therapy resulting in a potential failure treatment. We also report the first-time identification of VVE-S isolates pstS-null belonging to ST1478. [ABSTRACT FROM AUTHOR]

Bacteriophage therapy is a promising approach to address antimicrobial infections though questions remain regarding the impact of the immune response on clinical effectiveness. Here, we develop a mouse model to assess phage treatment using a cocktail of five phages from the Myoviridae and Siphoviridae families that target Vancomycin-Resistant Enterococcus gut colonization. Phage treatment significantly reduces fecal bacterial loads of Vancomycin-Resistant Enterococcus. We also characterize immune responses elicited following administration of the phage cocktail. While minimal innate responses are observed after phage administration, two rounds of treatment induces phage-specific neutralizing antibodies and accelerate phage clearance from tissues. Interestingly, the myophages in our cocktail induce a more robust neutralizing antibody response than the siphophages. This anti-phage immunity reduces the effectiveness of the phage cocktail in our murine model. Collectively, this study shows phage-specific immune responses may be an important consideration in the development of phage cocktails for therapeutic use. Bacteriophage can be used to target bacterial infection and used as a therapeutic approach for antibiotic resistant bacteria. Here the authors show that the use of bacteriophage to target antibiotic resistant bacteria in a mouse model of infection can induce phage specific immune responses and may impair their therapeutic efficacy. [ABSTRACT FROM AUTHOR]

Self-disinfecting materials that are both safe and scalable for production are increasingly in demand, particularly in healthcare settings where they can be used to combat hospital-acquired infections (HAIs). Here, we employed the natural food colorant chlorophyllin (E140ii) as a photosensitizer to prepare photodynamic antimicrobial materials through both chemical conjugation and electrospinning, resulting in chlorophyllin-grafted cotton fabric (Chl-fabric) and chlorophyllin-embedded polyacrylonitrile nanofibers (Chl-NF), respectively. The materials were characterized by a number of physical methods, as was their ability to generate singlet oxygen upon visible light illumination. The best results with Chl-fabric yielded 99.998% inactivation of vancomycin-resistant E. faecium and 99.994% of methicillin-resistant S. aureus after 60 min visible light illumination (400–700 nm, 80 ± 5 mW/cm2), whereas Chl-NF inactivated both bacteria by 99.9999%. Feline calicivirus was also photodynamically susceptible, with 99.8% inactivation by both materials. Gram-negative Klebsiella pneumoniae was not initially susceptible to photodynamic inactivation by Chl-NF, however addition of the photothermal agent MoS2 fully inactivated (99.9999%) this pathogen under NIR illumination, indicative of synergistic photothermal and photodynamic activities. These findings suggest that chlorophyllin can be used in photodynamic antimicrobial materials against drug-resistant Gram-positive bacteria, and that its efficacy can be synergistically amplified in the presence of a photothermal agent against Gram-negative pathogens. [ABSTRACT FROM AUTHOR]

Zinc and copper have been used as growth promotors in alternative to antibiotics in pig's diet. The aim was the ascertainment of the Zn and Cu concentrations in piglets' liver and kidney and their impact in the reduced susceptibility to Zn, Cu, and antibiotics in enterococci, used as microbiota biomarker. Zn and Cu were determined in the livers and kidneys of 43 piglets slaughtered in Portugal, by flame atomic absorption spectrometry. Enterococci were isolated from feces for determining the identification of species (E. faecalis, E. faecium, and Enterococcus spp.); susceptibility to vancomycin, ciprofloxacin, linezolid, tigecycline, ampicillin, imipenem, and metals; and Cu tolerance genes. In piglets with Zn and Cu high or toxic levels, enterococci had reduced susceptibility to ions, reinforced by the presence of Cu tolerance genes and by resistance to antibiotics. The study relevance is to show the relationship between these metals' levels and decreased susceptibility to Cu, Zn, and antibiotics by enterococci. From the results, it could be supposed that the piglets were being fed with high doses of Zn and Cu which could select more resistant bacteria to both antibiotics and metals that could spread to environment and humans. [ABSTRACT FROM AUTHOR]

Background: Enterococci, known for their disturbing involvement in nosocomial infections, possess a diverse set of virulence factors, regulated by multiple genes. A key virulence regulator is the fecal Streptococcus regulator (Fsr) quorum sensing system. Multiple reports describe the involvement of fsr genes in several virulence mechanisms, notably gelatinase production and biofilm formation; however, the presence of fsr genes does not necessarily predict those virulence phenotypes. This study investigates the factors affecting the relation between molecular detection of fsr genes and accurate prediction of gelatinase activity and biofilm formation intensity. Methods: One hundred enterococcal samples were collected from patients suffering from urinary tract infections. The isolates were identified through the use of a polymerase chain reaction (PCR) technique targeting the ddl gene. Biofilm formation was quantified by the crystal violet assay, while gelatinase activity was evaluated on gelatin agar plates. PCR was used to detect the fsrA and fsrB genes, as well as the gelatinase enzyme-encoding gene (gelE). Results: Out of the collected 100 isolates, 93% were identified as Enterococcus faecalis. The isolates formed biofilm with different intensities: 47% were strong biofilm producers, 28% moderate, and 21% weak, while only four isolates (4%) did not form biofilm. Only 14% of all isolates had detectable gelatinase activity. The fsrA and fsrB genes were detected in 26% and 28% of the tested isolates, respectively, while gelE was detected in 57% of the isolates. Whereas no association was found between biofilm formation intensity and fsr locus genes or gelatinase activity, a strong positive correlation (r = 1) was found between the detection of both fsrA and fsrB genes and the gelatinase activity. Conclusion: fsrA and fsrB have a diagnostic value and may be used as biomarkers for gelatinase activity in E. faecalis. [ABSTRACT FROM AUTHOR]

Background: The incidence of comorbidity between type 2 diabetes mellitus (T2DM) and metabolic-associated fatty liver disease (MAFLD) is high, and patients tend to be younger. When people develop metabolic diseases such as T2DM and MAFLD, the original homeostasis of the gut microbiota in the body is disrupted, and gut flora drift occurs. This study investigated the relationship between the number of gut flora and MAFLD in young-onset T2DM. Methods: This retrospective study analyzed 44 adolescent T2DM patients who were divided into a non-MAFLD group and a MAFLD group. Anthropometric measurements, clinical and biochemical markers, inflammatory markers, thyroid function assessments, and stool specimens were collected. Real-time PCR was performed to quantify several important gut flora constituents at the genus level. Student's t-test and the chi-square test were applied for group comparisons, and binary regression models were used to explore the relationship between gut flora and MAFLD in young-onset T2DM. Results: Among the 44 subjects, 26 (59.1%) were diagnosed with MAFLD, and 18 (40.9%) were not. Compared with the non-MAFLD group, body mass index (BMI), abdominal circumference, and levels of blood uric acid and thyroid stimulating hormone (TSH) in the MAFLD group were significantly increased, and age level and high-density lipoprotein cholesterol (HDL-C) were significantly decreased (p < 0.05). Compared with the non-MAFLD group, the abundance of Faecalibacterium prausnitzii and Bifidobacterium in the MAFLD group was significantly reduced, and the abundance of Enterococcus and Lactobacillus was significantly increased (p < 0.05). In the multivariate regression analysis, Faecalibacterium prausnitzii and Bifidobacterium were independent protective factors for MAFLD in young-onset T2DM, after excluding confounding factors. Conclusion: In young-onset T2DM, there was a difference in gut flora between patients with MAFLD and those without MAFLD. Faecalibacterium prausnitzii and Bifidobacterium were independent protective factors for MAFLD in young-onset T2DM. [ABSTRACT FROM AUTHOR]

Mucus is an integral polydisperse molecule around the epithelium layer throughout the gastrointestinal tract and acts as a nutrition source for gut microbiota. In the present study, three Enterococcus strains, i.e., Enterococcus faecium KS2, Enterococcus faecium KS3, and Enterococcus gallinarum KS4, having mucin degrading ability, were isolated from human feces and identified by metagenomic technique. The cultures exhibited halos when grown in a mucin-containing medium, whereas quantitative data indicated a decrease in oligosaccharide concentration ranging between 28.74 and 73.05%, further confirming its degradation behavior. The physiological properties revealed that E. gallinarum KS4 lacks virulence, whereas E. faecium KS2 and E. faecium KS3 had the virulence factors. All identified isolates could tolerate pH 2.0 for up to three hours and exhibited survivability ranging between 75 and 111%. They also exhibited more than 70% survivability after eight hours against 0.5% (w/v) bile concentration. Antibiotic susceptibility data revealed that the KS2 strain showed susceptibility against 12 out of 26 antibiotics, including tetracycline, chloramphenicol, gentamicin, and kanamycin. At the same time, KS3 did not show susceptibility against ceftazidime, cefixime, and nalidixic, while E. gallinarum KS4 strain was susceptible to all tested antibiotics except cefixime and nalidixic. The study highlights mucin degrading behavior of human-derived Enterococcus strains and physiological traits specifically depicted E. faecium KS2 and KS3 as opportunistic pathogens. However, E. gallinarum KS4 showed promising functional attributes and required to be corroborated using a suitable in-vivo model. [ABSTRACT FROM AUTHOR]

Increasing antimicrobial resistance in Enterococcus faecium necessitates the search for novel treatment agents, such as bacteriocins. In this study, we conducted an in vivo assessment of five bacteriocins, namely Lacticin Z, Lacticin Q, Garvicin KS (ABC), Aureocin A53 and Microbisporicin (NAI-107), against vanB-resistant Enterococcus faecium using a Galleria mellonella model. Our in vitro experiments demonstrated the efficacy of all five bacteriocins against vanB-resistant E. faecium with only NAI-107 demonstrating in vivo efficacy. Notably, NAI-107 exhibited efficacy across a range of tested doses, with the highest efficacy observed at a concentration of 16 µg/mL. Mortality rates in the group treated with 16 µg/mL NAI-107 were lower than those observed in the linezolid-treated group. These findings strongly suggest that NAI-107 holds promise as a potential alternative therapeutic agent for treating infections caused by resistant E. faecium and warrants further investigation. [ABSTRACT FROM AUTHOR]

In hospital environments, droplets generated by urination within shared toilets may represent a route of dissemination for bacteria such as vancomycin-resistant Enterococcus faecium (VREfm), which contributes significantly to the burden of hospital-acquired infections. We investigated the potential activity of a foam in preventing the generation of droplets containing Enterococcus spp. during urination. A uniform layer of foam was deposited in the inner walls and at the bottom of an experimental toilet contaminated with suspensions of Enterococcus strains (including a VREfm strain). Human urination was simulated, and colonies of Enterococcus were recovered through a toilet lid where agar plates had been placed. Results showed that the foam was able to suppress production of droplets containing Enterococcus spp. generated by a liquid hitting inner toilet walls. Conversely, Enterococcus colonies were recovered in absence of foam. Moreover, the foam did not show antibacterial activity. We propose a new non-antimicrobial approach aimed at limiting transmission of multidrug-resistant bacteria, particularly in healthcare settings. [ABSTRACT FROM AUTHOR]

Background: Enterococcus faecalis is a common cause of healthcare-associated infections. Its resistance to linezolid, the antibiotic of last resort for vancomycin-resistant enterococci, has become a growing threat in healthcare settings. Methods: We analyzed the data of E. faecalis isolates from 26 medical institutions between 2018 and 2020 and performed univariate and multivariate logistic regression analyses to determine the independent predictors for linezolid-resistant E. faecalis (LREFs). Then, we used the artificial neural network (ANN) and logistic regression (LR) to build a prediction model for linezolid resistance and performed a performance evaluation and comparison. Results: Of 12,089 E. faecalis strains, 755 (6.25%) were resistant to linezolid. Among vancomycin-resistant E. faecalis, the linezolid-resistant rate was 24.44%, higher than that of vancomycin-susceptible E. faecalis (p < 0.0001). Univariate and multivariate regression analyses showed that gender, age, specimen type, length of stay before culture, season, region, GDP (gross domestic product), number of beds, and hospital level were predictors of linezolid resistance. Both the ANN and LR models constructed in the study performed well in predicting linezolid resistance in E. faecalis, with AUCs of 0.754 and 0.741 in the validation set, respectively. However, synthetic minority oversampling technique (SMOTE) did not improve the prediction ability of the models. Conclusion: E. faecalis linezolid-resistant rates varied by specimen site, geographic region, GDP level, facility level, and the number of beds. At the same time, community-acquired E. faecalis with linezolid resistance should be monitored closely. We can use the prediction model to guide clinical medication and take timely prevention and control measures. [ABSTRACT FROM AUTHOR]

Lactic acid bacteria (LAB) coexist with Clostridium spp. in hydrogen production processes from complex substrates; however, the role of LAB is still unclear. This study analyzed the fermentation products in a wide range of initial pH (pHi, 5.5–6.9) and total solids (TS%, 8–22%) to determine the activity of these two microbial groups over time (from 24 to 120 h). Agave bagasse served as the feedstock for hydrogen production via consolidated bioprocess (CBP), while the inoculum source was the indigenous mature microbiota. In the early stage of the CBP, hydrogen production from lactic acid occurred only at pHi ≥ 6.0 (ρ = 0.0004) with no effect of TS%; lactic acid accumulated below this pHi value. In this stage, lactic acid production positively correlated with a first cluster of LAB represented by Paucilactobacillus (r = 0.64) and Bacillus (r = 0.81). After 72 h, hydrogen production positively correlated with a second group of LAB led by Enterococcus (r = 0.71) together with the hydrogen producer Clostridium sensu stricto 1 (r = 0.8) and the acetogen Syntrophococcus (r = 0.52) with the influence of TS% (ρ < 0.0001). A further experiment showed that buffering the pH to 6.5 increased and lengthened the lactic acid production, doubling the hydrogen production from 20 to 41 mL H2/gTSadded. This study confirmed the prevalence of distinct groups of LAB over time, whose microbial activity promoted different routes of hydrogen production. [ABSTRACT FROM AUTHOR]

An alarming global public health and economic peril has been the emergence of antibiotic resistance resulting from clinically relevant bacteria pathogens, including Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumonia, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species constantly exhibiting intrinsic and extrinsic resistance mechanisms against last-resort antibiotics like gentamycin, ciprofloxacin, tetracycline, colistin, and standard ampicillin prescription in clinical practices. The discovery and applications of antimicrobial peptides (AMPs) with antibacterial properties have been considered and proven as alternative antimicrobial agents to antibiotics. In this study, we have designed, produced, and purified a recombinant novel multifunctional hybrid antimicrobial peptide LL-37_Renalexin for the first time via the application of newly designed flexible GS peptide linker coupled with the use of our previously characterized small metal-binding proteins SmbP and CusF3H+ as carrier proteins that allow for an enhanced bacterial expression, using BL21(DE3) and SHuffle T7(DE3) Escherichia coli strains, and purification of the hybrid peptide via immobilized metal affinity chromatography. The purified tag-free LL-37_Renalexin hybrid peptide exhibited above 85% reduction in bacteria colony-forming units and broad-spectrum antimicrobial effects against Staphylococcus aureus, Escherichia coli, Methicillin-resistant Staphylococcus aureus (MRSA), and Klebsiella pneumoniae bacteria clinical isolates at a lower minimum inhibition concentration level (10–33 μM) as compared to its counterpart single-AMPs LL-37 and Renalexin (50–100 μM). Key points: • The hybrid antimicrobial peptide LL-37_Renalexin has been designed using a GS linker. • The peptide was expressed with the carrier proteins SmbP and CusF3H+. • The hybrid peptide shows antibacterial potency against clinical bacterial isolates. [ABSTRACT FROM AUTHOR]

Purpose: The extensive use of vancomycin has led to the development of Staphylococcus aureus strains with varying degrees of resistance to vancomycin. The present study aimed to explore the molecular causes of vancomycin resistance by conducting a proteomics analysis of subcellular fractions isolated from vancomycin-intermediate resistant S. aureus (VISA) and vancomycin-sensitive S. aureus (VSSA) strains. Methods: We conducted proteomics analysis of subcellular fractions isolated from 2 isogenic S. aureus strains: strain 11 (VSSA) and strain 11Y (VISA). We used an integrated quantitative proteomics approach assisted by bioinformatics analysis, and comprehensively investigated the proteome profile. Intensive bioinformatics analysis, including protein annotation, functional classification, functional enrichment, and functional enrichment-based cluster analysis, was used to annotate quantifiable targets. Results: We identified 128 upregulated proteins and 21 downregulated proteins in strain 11Y as compared to strain 11. The largest group of differentially expressed proteins was composed of enzymatic proteins associated with metabolic and catalytic activity, which accounted for 32.1% and 50% of the total proteins, respectively. Some proteins were indispensable parts of the regulatory networks of S. aureus that were altered with vancomycin treatment, and these proteins were related to cell wall metabolism, cell adhesion, proteolysis, and pressure response. Conclusion: Our proteomics study revealed regulatory proteins associated with vancomycin resistance in S. aureus. Some of these proteins were involved in the regulation of cell metabolism and function, which provides potential targets for the development of strategies to manage vancomycin resistance in S. aureus. [ABSTRACT FROM AUTHOR]

Purpose: Vancomycin-resistant enterococci (VRE) are a leading cause of hospital-acquired infections with limited therapeutic options. Combination of at least two antimicrobials is a possible strategy to obtain rapid and sustained bactericidal effects and overcome the emergence of resistance. We revised the literature on linezolid synergistic properties from in vitro studies to assess its activity in combination with molecules belonging to other antibiotic classes against Enterococcus spp. Methods: We performed a systematic review of the literature from three peer-reviewed databases including papers evaluating linezolid synergistic properties in vitro against Enterococcus spp. isolates. Results: We included 206 Enterococcus spp. isolates (92 E. faecalis, 90 E. faecium, 2 E. gallinarum, 3 E. casseliflavus, 19 Enterococcus spp.) from 24 studies. When an isolate was tested with different combinations, each combination was considered independently for further analysis. The most frequent interaction was indifferent effect (247/343, 72% of total interactions). The highest synergism rates were observed when linezolid was tested in combination with rifampin (10/49, 20.4% of interactions) and fosfomycin (16/84, 19.0%, of interactions). Antagonistic effect accounted for 7/343 (2.0%) of total interactions. Conclusion: Our study reported overall limited synergistic in vitro properties of linezolid with other antibiotics when tested against Enterococcus spp. The clinical choice of linezolid in combination with other antibiotics should be guided by reasoned empiric therapy in the suspicion of a polymicrobial infection or targeted therapy on microbiological results, rather than on an intended synergistic effect of the linezolid-based combination. [ABSTRACT FROM AUTHOR]

Background/aim: Enterococcus faecalis and Enterococcus faecium cause human infections including bacteraemia and infective endocarditis (IE). Only few studies describing non-faecalis and non-faecium Enterococcus (NFE) infections have been conducted. We aimed to describe the incidence, prognosis, and focus of infection of bacteraemia with NFE. Methods: This retrospective population-based study included all episodes of patients having a blood culture with growth of NFE between 2012 and 2019 in Region Skåne, Sweden. Information was collected from medical records. Episodes of bacteraemia caused by NFE were compared to episodes of bacteraemia caused by E. faecalis and E. faecium. Results: During the study period, 136 episodes with NFE bacteraemia were identified corresponding to an incidence of NFE bacteraemia of 16 cases per 1,000,000 person-years among adults. Enterococcus casseliflavus (n=45), Enterococcus gallinarum (n=34), and Enterococcus avium (n=29) were the most common species. The most common foci of infection were biliary tract infections (n=17) followed by gastrointestinal infections (n=7). Urinary tract infections were not commonly caused by NFE (n=1), and no episodes of IE were caused by NFE. Polymicrobial bacteraemia was more common with NFE (73%) than with E. faecalis (35%) and E. faecium (42%). Community acquired infections were more common in bacteraemia with NFE compared to E. faecium. 30- and 90-day survival rates were 76% and 68%, respectively, and recurrent NFE bacteraemia was seen after 3% of the episodes. Conclusion: Bacteraemia caused by NFE is rare and is often polymicrobial. Biliary tract focus is common in NFE bacteraemia whereas IE and urinary tract focus are uncommon. [ABSTRACT FROM AUTHOR]

Surveillance has revealed an increase of multidrug-resistant organisms (MDROs), even in low-prevalent settings such as Norway. MDROs pose a particular threat to at-risk populations, including persons with cancer. It is necessary to include such populations in future infection surveillance. By combining existing data sources, we aimed to describe the epidemiology of MDROs in persons diagnosed with cancer in Norway from 2008 to 2018. A cohort was established using data from the Cancer Registry of Norway, which was then linked to notifications of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin- and/or linezolid-resistant enterococci (V/LRE), and carbapenemase-producing Gram-negative bacilli (CP-GNB) from the Norwegian Surveillance System for Communicable Diseases, and laboratory data on third-generation cephalosporin-resistant Enterobacterales (3GCR-E) from Oslo University Hospital (OUH). We described the incidence of MDROs and resistance proportion in Enterobacterales from 6 months prior to the person's first cancer diagnosis and up to 3 years after. The cohort included 322,005 persons, of which 0.3% (878) were diagnosed with notifiable MDROs. Peak incidence rates per 100,000 person-years were 60.9 for MRSA, 97.2 for V/LRE, and 6.8 for CP-GNB. The proportion of 3GCR-E in Enterobacterales in blood or urine cultures at OUH was 6% (746/12,534). Despite overall low MDRO incidence, there was an unfavourable trend in the incidence and resistance proportion of Gram-negative bacteria. To address this, there is a need for effective infection control and surveillance. Our study demonstrated the feasibility of expanding the surveillance of MDROs and at-risk populations through the linkage of existing laboratory and register data. [ABSTRACT FROM AUTHOR]

Analyzing the influence of the bed allocation and utilization efficiency in healthcare institutions on the isolation proportion of Multidrug-resistant organisms (MDROs) to provide data to support prevention and control of MDROs. In this study, the provincial panel data from 2014 to 2020 in China on health resource indicators, including the number of beds per 1,000 population, hospital bed utilization rate, and average hospital stay from 2014 to 2020 in China were used to analyze the relationship between bed allocation or utilization efficiency and MDROs by the panel data quantile regression model. It was shown that the number of beds per 1,000 population had a negative effect on the isolation proportion of methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, vancomycin-resistant Enterococcus faecium, penicillin-resistant Streptococcus pneumoniae, methicillin-resistant coagulase-negative Staphylococcus, and cefotaxime or ceftriaxone resistant Escherichia coli (regression coefficient < 0, P < 0.05). The utilization rate of hospital bed had a positive effect on the isolation proportion of methicillin-resistant Staphylococcus aureus, methicillin-resistant coagulase-negative Staphylococcus, vancomycin-resistant Enterococcus faecium, penicillin-resistant Streptococcus pneumoniae, cefotaxime or ceftriaxone resistant Escherichia coli, carbapenem-resistant Escherichia coli, cefotaxime or ceftriaxone resistant Klebsiella pneumoniae, carbapenem-resistant Klebsiella pneumoniae, carbapenem-resistant Pseudomonas aeruginosa, and carbapenem-resistant Acinetobacter baumannii (regression coefficient > 0, P < 0.05). The average hospital stay had a positive effect on the isolation proportion for several antibiotic-resistant organisms, including methicillin-resistant Staphylococcus aureus, methicillin-resistant coagulase-negative Staphylococcus, vancomycin-resistant Enterococcus faecalis, vancomycin-resistant Enterococcus faecium, penicillin-resistant Streptococcus pneumoniae, cefotaxime or ceftriaxone resistant Escherichia coli, carbapenem-resistant Escherichia coli, quinolone-resistant Escherichia coli, cefotaxime or ceftriaxone resistant Klebsiella pneumoniae, carbapenem-resistant Pseudomonas aeruginosa, and carbapenem-resistant Acinetobacter baumannii (regression coefficient > 0, P < 0.05). Bed allocation and utilization efficiency in healthcare institutions may affect the isolation proportion of MDROs in varying degrees. [ABSTRACT FROM AUTHOR]

Introduction and hypothesis: Studies within the past decade have suggested associations among composition of the urinary microbiota, local immune responses, and urinary incontinence symptoms. To investigate these relationships, we evaluated the structure of the urinary microbiome, local inflammatory markers, and patient responses prior to and at 6-weeks after treatment with anticholinergic medication for urgency urinary incontinence (UUI). Methods: Using a prospective pilot study, we enrolled women who presented with UUI symptoms and were prescribed treatment with anticholinergics. Catheterized urine samples were collected from participants at their baseline and 6-week follow-up visits for microbiological (standard and 16S rRNA gene phylotyping analyses) and cytokine analysis along with the UDI-6 questionnaire and 2-day bladder diary. Results: Patients were Caucasian, post- menopausal, with a median age of 64 and median BMI of 30.1 kg/m2. Among the patients, 75% had UUI symptoms for less than 2 years, but with a frequency of at least a few times a week or every day. Most women were prescribed 10 mg oxybutynin ER daily at enrollment. Patients had varied urinary microbiota by culture and 16S phylotyping, with species of Lactobacillus being the most common, in six samples, in addition to taxa associated with Enterococcus, Staphylococcus, and mixed flora. Cytokine levels showed no differences before and after treatment with anticholinergics, nor correlation with urinary bacteria or microbiome composition. Conclusions: Our pilot study suggests factors in addition to the urinary microbiome and local immune responses may be involved in patients' response to anticholinergics for UUI. [ABSTRACT FROM AUTHOR]

The genomes of Enterococcus faecalis and Enterococcus faecium bacteriophages were analysed for gene shuffling in the lytic cassettes of bacteriophages infecting. It was found that enterococcal bacteriophages could be classified into well-defined groups based on the size of their genomes and each size group had its own conserved gene composition of lytic cassettes. Enterococcal bacteriophages use a relatively broad spectrum of holins and endolysins with variable cell-wall binding (CWB) and catalytic domains, and most of them utilise a lytic cassette with more than two genes. Enterococcal bacteriophages most commonly use endolysins with amidase catalytic domains and the CWB domain SH3_5. Some bacteriophages possess in their lytic cassette a holin-like gene with the XhlA domain protein, characteristic of hemolysin. Regardless of the shuffling of genes encoding holins and endolysins in lytic modules, a novel example of CWB domain shuffling within enterococcal endolysins was identified. [ABSTRACT FROM AUTHOR]

The demand for thorough disinfection within ambulances is essential, given the in-vehicle medical procedures and the potential high risk of infections due to patients' open wounds. One solution that can address this hygiene challenge involves the application of reactive products generated from atmospheric (air) oxygen and water vapor, activated through the use of cold plasma. Cold plasma's charged particles perforate the cell membranes of microorganisms. This process does not work in human cells, as proteins in the form of enzymes within the body break down the cold plasma and protect the cells. The study was done on an ambulance that was contaminated in eight places. Samples were taken from each site, and two surfaces measuring approximately 8 × 8 cm were carefully sealed and marked. These surfaces were deliberately contaminated by applying an Enterococcus faecium suspension of 8.5 × 107 CFU/mL using a sterile cotton swab. It was followed by the disinfection procedure, that was initiated with the PLASMOCAR device. It was positioned on the front workspace and operated for a duration of 30 min, utilizing the vehicle's onboard voltage. Throughout the operation, all doors and windows were closed and the vehicle's air conditioning system remained active. After the completion of the disinfection process, samples were collected from the surfaces for bacterial counts. A reduction of 3.73 log levels in initial bacteria was accomplished within the rescue vehicle for Enterococcus faecium, equivalent to a 10–fourfold reduction in bacteria, eliminating up to 99.99% of the initial microorganisms. This success makes the process well-suited and convenient as an ongoing "background" procedure to enhance the established disinfection procedures. The established disinfection procedures outlined in the hygiene plan must be promptly implemented whenever mechanical surface cleaning is required. The use of PLASMOCAR offers an extra layer of protection and security, significantly decreasing the risk of microorganism transmission through cross-contamination and aerosols. This is a significant benefit for the well-being of both staff and patients. [ABSTRACT FROM AUTHOR]

Colorectal cancer (CRC) is a significant global health burden with a rising incidence worldwide. Distinct bacterial populations are associated with CRC development and progression, and it is thought that the relationship between CRC and associated gut bacteria changes during the progression from normal epithelium to benign adenoma and eventually malignant carcinoma and metastasis. This study compared the interaction of CRC-associated species Enterotoxigenic Bacteroides fragilis, Enterococcus faecalis and Fusobacterium nucleatum and one probiotic species, Escherichia coli Nissle 1917 with a colorectal adenoma (S/RG/C2) and a colorectal adenocarcinoma (HCT116) derived cell line. Gentamicin protection assays showed that all species displayed higher attachment to benign tumour monolayers when compared to malignant monolayers. However, invasion of 3/4 species was higher in the HCT116 cells than in the adenoma cells. All species were found to persist within tumour cell monolayers for a minimum of 48 h under standard aerobic cell culture conditions, with persistence significantly higher in HCT116 cells. Downstream assays were performed to analyse the behaviour of S/RG/C2 and HCT116 cells post-infection and revealed that all species increased the tumour cell yield of both cell lines. The migratory and invasive potential of HCT116 cells was increased after infection with F. nucleatum; however, no species significantly altered these characteristics in S/RG/C2 cells. These results add to the growing evidence for the involvement of microorganisms in CRC progression and suggest that these interactions may be dependent on tumour cell-specific characteristics. [ABSTRACT FROM AUTHOR]

Cyanogenic glycosides in forage species and the possibility of cyanide (CN) poisoning can have undesirable effects on ruminants. The literature estimates that unknown rumen bacteria with rhodanese activity are key factors in the animal detoxification of cyanogenic glycosides, as they are capable of transforming CN into the less toxic thiocyanate. Therefore, identifying these bacteria will enhance our understanding of how to improve animal health with this natural CN detoxification process. In this study, a rhodanese activity screening assay revealed 6 of 44 candidate rumen bacterial strains isolated from domestic buffalo, dairy cattle, and beef cattle, each with a different colony morphology. These strains were identified as belonging to the species Enterococcus faecium and E. gallinarum by 16S ribosomal DNA sequence analysis. A CN-thiocyanate transformation assay showed that the thiocyanate formation capacity of the strains after a 12 h incubation ranged from 4.42 to 25.49 mg hydrogen CN equivalent/L. In addition, thiocyanate degradation resulted in the production of ammonia nitrogen and acetic acid in different strains. This study showed that certain strains of enterococci substantially contribute to CN metabolism in ruminants. Our results may serve as a starting point for research aimed at improving ruminant production systems in relation to CN metabolism. [ABSTRACT FROM AUTHOR]

Quorum sensing inhibitor (QSI) has been attracting attention as anti-virulence agent which disarms pathogens of their virulence rather than killing them. QSI marking cyclic peptide-mediated QS in Gram-positive bacteria is an effective tool to overcome the crisis of antibiotic-dependent chemotherapy due to the emergence of drug resistance strain, e.g., methicillin resistant Staphylococcus aureus (MRSA) and Vancomycin resistant Enterococci (VRE). From a semi-large-scale screening thus far carried out, two Epoxide compounds, Ambuic acid and Synerazol, have been found to efficiently block agr and fsr QS systems, suggesting that the Epoxide group is involved in the mode of action of these QSIs. To address this notion, known natural Epoxide compounds, Cerulenin and Fosfomycin were examined for QSI activity for the agr and fsr systems in addition to in silico and SAR studies. As a result, most of investigated Epoxide containing antibiotics correlatively interfere with QSI activity for the agr and fsr systems under sublethal concentrations. [ABSTRACT FROM AUTHOR]

Enterococcus species have been described as core members of the microbial community of Spodoptera frugiperda (Lepidoptera: Noctuidae) and have been previously reported as insecticide degrading agents. This study aimed to investigate the molecular composition of these microbial symbionts of S. frugiperda to better understand their association with the host and their potential for insecticide metabolization. Through phenotypic assays and comparative genomic analyses of several pesticide-degrading Enterococcus isolated from the gut of S. frugiperda larvae, we identified two new species: Enterococcus entomosocium n. sp. and Enterococcus spodopteracolus n. sp. Their identities as new species were confirmed by whole genome alignment, utilizing cut-offs of 95–96% for the average nucleotide identity (ANI) and 70% for the digital DNA: DNA hybridization (dDDH) values. The systematic positioning of these new species within the genus Enterococcus was resolved using genome-based analysis, revealing Enterococcus casseliflavus as a sister group of E. entomosocium n. sp., and Enterococcus mundtii as a sister group of E. spodopteracolus n. sp. Comparative genomic analyses of several isolates of E. entomosocium n. sp. and E. spodopteracolus n. sp. provided a better assessment of the interactions established in the symbiotic association with S. frugiperda and led to the discovery of misidentified new species of Enterococcus associated with insects. Our analyses indicated that the potential of E. entomosocium n. sp. and E. spodopteracolus n. sp. to metabolize different pesticides arises from molecular mechanisms that result in rapid evolution of new phenotypes in response to environmental stressors, in this case, the pesticides their host insect is exposed to. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the rate of bacterial contamination of semitendinosus (ST) tendons during graft harvest in anterior cruciate ligament reconstruction (ACLR), in order to precisely specify the underlying pathogens and obtain data on their susceptibility to potential antibiotics. Methods: In a prospective study, a total of 59 consecutive patients undergoing primary ACLR were recruited from one centre. No patient had history of previous surgery to the knee or showed clinical signs of infection. Four tissue samples of harvested ST tendons for anterior cruciate ligament (ACL) autografts (case group; ST) were examined for evidence of bacterial colonisation and compared to four tissue samples of the native ACL as negative controls (control group; ACL). Three of the respective samples were subjected to cultural microbiological examination and one to 16S rRNA-PCR. Antibiotic susceptibility testing was performed for each pathogen that was identified. Results: A total of 342 samples were analysed by culture. Significantly more patients showed a positive culture of the ST (33.9%; n = 20/59) compared to 3.4% of patients (n = 2/59) with positive culturing of the ACL (p < 0.0001). Including 16S rRNA-PCR, in a total of 42.4% (25/59) of patients, bacteria were detected in at least one ST sample either by PCR and/or culture. All species found (n = 33) belong to the typical skin flora with Staphylococcus epidermidis (39.4%; n = 13/33) being the most common species, followed by Staphylococcus capitis (24.2%; n = 8/33). All tested isolates (n = 29) were susceptible to vancomycin (29/29, 100%), 69% (n = 20/29) to oxacillin and 65.5% (n = 19/29) to clindamycin. Conclusion: ST autografts for ACLR were commonly contaminated with skin commensal bacteria during harvest. One-third of the isolates showed resistance to typical perioperative intravenous antibiotics, whereas all isolates were sensitive to vancomycin. Therefore, routine prophylactic decontamination of all hamstring autografts before implantation should be recommended, preferably with topical vancomycin. Level of evidence: Level III. [ABSTRACT FROM AUTHOR]