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Analysis of molecular epidemiological characteristics and antimicrobial susceptibility of vancomycin-resistant and linezolid-resistant Enterococcus in China.

BACKGROUND: This study investigates the distribution and characteristics of linezolid and vancomycin susceptibilities among Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) and explores the underlying resistance mechanisms. METHODS: A total of 2842 Enterococcus clinical isolates from patients were retrospectively collected, and their clinical data were further analyzed. The minimum inhibitory concentrations (MICs) of vancomycin and linezolid were validated by broth dilution method. The resistance genes optrA, cfr, vanA, vanB and vanM were investigated using polymerase chain reaction (PCR). Housekeeping genes and resistance genes were obtianed through whole-genome sequencing (WGS). RESULTS: Of the 2842 Enterococcus isolates, 88.5% (2516) originated from urine, with E. faecium accounted for 60.1% of these. The vanA gene was identified in 27/28 vancomycin resistant Enterococcus (VRE) isolates, 4 of which carried both vanA and vanM genes. The remaining strain was vanM positive. The optrA gene was identified in all E. faecalis isolates among linezolid resistant Enterococcus (LRE). E. faecium showed a higher multiple antibiotic resistance index (MAR index) compared to E. faecalis. The multi-locus sequence typing (MLST) showed the sequence type of E. faecium mainly belongs to clonal complex (CC) 17, nearly E. faecalis isolates analyzed were differentiated into 7 characteristics of sequence types (STs), among which ST16 of CC16 were the major lineage. CONCLUSION: Urine was the primary source of VRE and LRE isolates in this study. E. faecium showed higher levels of resistance compared to E. faecalis. OptrA gene was detected in 91.6% of LRE, which could explain linezolid resistance, and van genes were detected in all vancomycin resistant Enterococcus strains, while vanA was a key resistance mechanism in VRE identified in this study.

Linezolid

Fingolimod as a potent anti-Staphylococcus aureus: pH-dependent cell envelope damage and eradication of biofilms/persisters.

BACKGROUND: The urgent need for new antibacterial drugs has driven interest in repurposing therapies to combat Gram-positive biofilms and persisters. Fingolimod, an Food and Drug Administration (FDA)-approved drug for multiple sclerosis, shows bactericidal activity, particularly against Methicillin-resistant Staphylococcus aureus (MRSA) and biofilm-related infections. With a well-documented safety profile and strong translational potential, it aligns with World Health Organization's goals for antimicrobial repurposing. However, the action mode and mechanism of Fingolimod against gram-positive bacteria remain elusive. METHODS: This study utilized clinical Staphylococcus aureus (S. aureus), Enterococcus faecalis (E. faecalis), Streptococcus agalactiae (S. agalactiae). And their susceptibility to Fingolimod and other antibiotics was tested via Minimum Inhibitory Concentration (MIC) assays. Biofilm inhibition and hemolytic activity were evaluated using crystal violet staining, Confocal Laser Scanning Microscopy (CLSM), and hemolysis assays, respectively, while the effect of phospholipids on Fingolimod efficacy was assessed with checkerboard assays. Membrane permeability and integrity were measured using SYTOX green staining and transmission electron microscopy. Whole-genome sequencing was performed on Fingolimod-resistant S. aureus isolates to identify Single Nucleotide Polymorphisms (SNPs) linked to resistance. RESULTS: Our data indicated that Fingolimod exerted bactericidal activity against a wide spectrum of gram-positive bacteria, including S. aureus, E. faecalis, S. agalactiae. Moreover, Fingolimod could significantly eliminate the persisters, inhibit biofilm formation and eradicate in-vitro mature biofilms of S. aureus. The mechanism by which Fingolimod rapidly eradicated S. aureus involved a pH-dependent disruption of bacterial cell permeability and envelope integrity. Concomitantly, exogenous supplementation of phospholipids in the culture medium resulted in a dose-dependent increase in the MIC of Fingolimod. Specifically, the addition of 64 μg/mL of cardiolipin (CL) and phosphatidylethanolamine (PE) completely nullified the bactericidal activity of Fingolimod at a concentration of 4 times the MIC. After four months of Fingolimod exposure, the MIC values of S. aureus showed a slight increase, indicating that it is not prone to developing drug resistance. CONCLUSION: Fingolimod exhibits bactericidal activity against diverse gram-positive bacteria, with remarkable effects on S. aureus (including MRSA), disrupting bacterial cell structural integrity in a pH-dependent way and eradicating biofilms and persisters of S. aureus.

Biofilms

Comparative genomic characterization and antimicrobial resistance of bacteremia-causing Enterococcus faecium and Enterococcus faecalis in a Chinese hospital.

Enterococci are common commensals of the human gut and important opportunistic pathogens, with Enterococcus faecium and Enterococcus faecalis being the most clinically prevalent species. A significant epidemiological shift has emerged with an increasing clinical burden of E. faecium. To compare genomic evolution of E. faecium and E. faecalis, we performed whole-genome sequencing on 93 E. faecium and 32 E. faecalis isolates causing bloodstream infections at a single hospital (2022-2024). Analysis of patient demographics revealed that E. faecium infections originated from fewer sources than E. faecalis, with a higher proportion deriving from intra-abdominal infections. Multilocus sequence typing identified ST78 and ST789 as the predominant sequence types for E. faecium, whereas ST16 and ST179 were most common for E. faecalis. E. faecium carried more antimicrobial resistance genes and putative virulence marker (PVM)-type virulence genes than E. faecalis, with vancomycin resistance predominantly mediated by vanHAX (33/93, 35.5%) and a single E. faecalis isolate also carrying vanHAX (1/32, 3.1%); the structurally incomplete vanHMX gene cluster was detected in 11 E. faecium isolates. Pan-genome analysis indicated a larger core genome in E. faecalis compared to E. faecium, consistent with greater plasmid replicon diversity in the latter. Intra-host comparisons showed that two E. faecalis pairs from the same patient were clonally related, with one isolate acquiring a vanHAX plasmid conferring vancomycin resistance. In contrast, E. faecium isolates exhibited marked genomic diversity even among clonally related pairs. These findings suggest that E. faecium possesses greater genomic plasticity and adaptive potential to the clinical environment.IMPORTANCEThis study provides a detailed comparison of clinical and genomic features between Enterococcus faecium and Enterococcus faecalis from the same hospital setting. We show that E. faecium isolates, mainly ST78/ST789, carry more antimicrobial resistance genes and a higher number of putative virulence marker (PVM) genes than E. faecalis, reflecting their hospital-adapted nature. E. faecium also exhibits a smaller core genome and greater diversity of plasmid replicon types, indicating higher genomic plasticity and capacity for horizontal gene transfer. By contrast, E. faecalis retains a larger core genome and a set of classical virulence factors, and its within-host isolates are clonally related. These distinct genomic profiles help to understand how the two species adapt to clinical environments and may inform more targeted infection control strategies and resistance surveillance.

Enterococcus faecium

Genome-wide analysis of Enterococcus faecalis genes that facilitate interspecies competition with Lactobacillus crispatus.

Enterococci are opportunistic pathogens notorious for causing a variety of infections. While both Enterococcus faecalis and Lactobacillus crispatus are commensal residents of the vaginal tract, the molecular mechanisms that enable E. faecalis to take advantage of a vaginal biome with lower counts of lactobacilli to colonize the vaginal tract and induce aerobic vaginitis remain unknown. Here, we show that L. crispatus eradicates E. faecalis in a contact-independent manner. Using transposon sequencing to identify E. faecalis OG1RF transposon (Tn) mutants that are either under-represented or over-represented when co-cultured with L. crispatus, we found that Tn mutants with disruption in the dltABCD operon, that encodes the proteins responsible for the D-alanylation of teichoic acids, and OG1RF_11697 encoding for an uncharacterized hypothetical protein are more susceptible to killing by L. crispatus. Inversely, Tn mutants with disruption in ldh1, which encodes for L-lactate dehydrogenase, are more resistant to L. crispatus killing. Using the Galleria mellonella infection model, we show that co-injection of L. crispatus with E. faecalis OG1RF enhances larvae survival while this L. crispatus-mediated protection was lost in larvae co-infected with either L. crispatus and E. faecalisΔldh1 or Δldh1Δldh2 strains. Last, using RNA sequencing to identify E. faecalis genes that are differently expressed in the presence of L. crispatus, we found major changes in the expression of genes associated with glycerophospholipid metabolism, central metabolism, and general stress responses. The findings in this study provide insights into how E. faecalis mitigate assaults by L. crispatus.IMPORTANCEEnterococcus faecalis is an opportunistic pathogen notorious for causing a multitude of infections. As vaginal commensals, E. faecalis must interact with Lactobacillus crispatus, but how E. faecalis overcomes or mitigate assaults by L. crispatus killing remains unknown. We show that L. crispatus eradicates E. faecalis temporally in a contact-independent manner. Using high-throughput molecular approaches, we identified genetic determinants that enable E. faecalis to compete with L. crispatus. This study represents an important first step for the identification of adaptive genetic traits required for enterococci to tolerate assaults by lactobacilli.

Enterococcus faecalis

The adjacent ATP-binding protein-encoding genes of the Enterococcus faecalis phosphate-specific transport (pst) locus have non-overlapping cellular functions.

UNLABELLED: The widely conserved pst-phoU operon encodes a low-velocity, high-affinity, ATP-dependent importer for inorganic phosphate (Pi). The pstB gene encodes the ATPase that powers the import of Pi into the cell. In some Firmicutes, including the gastrointestinal commensal and opportunistic pathogen Enterococcus faecalis, the pst-phoU locus contains adjacent pstB genes. In this work, we compared the functionality of E. faecalis pstB1 and pstB2. E. faecalis pstB1 and pstB2 share sequence similarities with verified PstB ATPases from Escherichia coli and Streptococcus pneumoniae and only share ~60% amino acid identity with each other. Deletion of pstB1 was associated with a growth defect in low Pi-containing chemically defined medium (CDM), reduced Pi uptake, and a moderate increase in alkaline phosphatase (AP) activity. Deletion of pstB2 fully inhibited growth in CDM regardless of inorganic phosphorus source but did not hinder growth in rich, undefined medium. The ΔpstB2 mutant also exhibited a significant increase in AP activity that was associated with extracellular Pi accumulation. Overexpression of pstB2 in the pstB1 mutant was sufficient to restore growth in low-Pi CDM, Pi uptake, and AP activity, but this was not recapitulated with overexpression of pstB1 in the ΔpstB2 mutant. Deletion of either pstB paralog increased expression of the tandem paralog, and overexpression of pstB2 in ΔpstB2 reduced pstB1 expression. These results suggest that the E. faecalis pstB2-encoded ATPase is required for Pi import, while the pstB1-encoded ATPase has an accessory role in Pi import that can be duplicated by the presence of excess PstB2. IMPORTANCE: Phosphate is critical for all microbial life. In many bacteria, inorganic phosphate (Pi) is imported by the high-affinity, low-velocity Pst-PhoU system. The pstB gene encodes the ATPase that powers Pi import. The pst-phoU operon in many Firmicutes, including the human commensal and opportunistic pathogen Enterococcus faecalis, contains adjacent pstB genes, pstB1 and pstB2. No studies on the relative biological contributions of tandem pstB paralogs in any microbe have been published. This genetic study indicates that E. faecalis pstB1 and pstB2 do not have equivalent functions. The pstB2 gene encodes an ATPase that is required for Pi import, while the ATPase encoded by pstB1 has an accessory role in Pi import that can be duplicated by the presence of excess PstB2.

Enterococcus faecalis

Hidden diversity in Enterococcus faecalis revealed by CRISPR2 screening: eco-evolutionary insights into a novel subspecies.

Enterococcus faecalis is a commensal bacterium that colonizes the gut of humans and animals and is a major opportunistic pathogen, known for causing multidrug-resistant healthcare-associated infections (HAIs). Its ability to thrive in diverse environments and disseminate antimicrobial resistance genes (ARGs) across ecological niches highlights the importance of understanding its ecological, evolutionary, and epidemiological dynamics. The CRISPR2 locus has been used as a valuable marker for assessing clonality and phylogenetic relationships in E. faecalis. In this study, we identified a group of E. faecalis strains lacking CRISPR2, forming a distinct, well-supported clade. We demonstrate that this clade meets the genomic criteria for classification as a novel subspecies, here referred to as "subspecies B." Through a comprehensive pangenome analysis and comparative genomics, we explored the adaptive ecological traits underlying this diversification process, identifying clade-specific features and their predicted functional roles. Our findings suggest that the frequent isolation of subspecies B from meat products and processing facilities may reflect dissemination routes involving environmental contamination (e.g., water, plants, soil) from avian species. The absence of key virulence traits required for pathogenicity in mammals, particularly humans, and the lack of clinically relevant resistance determinants indicate that subspecies B currently poses minimal threat to public health compared with the broadly disseminated "subspecies A." Nevertheless, the unclear potential for genetic exchange between these subspecies and the frequent association of subspecies B with food sources calls for continued genomic surveillance of E. faecalis from a One Health perspective to detect and mitigate the emergence of high-risk variants in advance.IMPORTANCEExploring intraspecific genetic variability in generalist bacteria with pathogenic potential, such as Enterococcus faecalis, is a key to uncovering stable evolutionary trends. By screening the CRISPR2 locus across a representative set of genomes from diverse sources, this study reveals a previously unrecognized lineage within the population structure of E. faecalis, associated with underexplored nonhuman and nonhospital reservoirs. These findings broaden our knowledge of the species' genetic landscape and shed light on its adaptive strategies and patterns of ecological dissemination. By bridging phylogenetic patterns with variation in genetic defense systems and accessory traits, the study generates testable hypotheses about the genomic determinants and corresponding selective pressures that shape the species' behavior and long-term dissemination. This work offers new perspectives on the eco-evolutionary dynamics of E. faecalis and highlights the value of genomic surveillance beyond clinical settings, in alignment with One Health principles.

Enterococcus faecalis

Emergence of an optrA-positive Enterococcus faecalis ST699 lineage in animal-derived foods in Beijing, China.

Enterococci from animal-derived foods are key reservoirs for antimicrobial resistance (AMR) in the food chain. However, comparative genomic studies investigating the distribution of the oxazolidinone resistance gene optrA among food- and human-derived Enterococci remain limited. This study assessed linezolid-resistant Enterococci from retail meat and healthy humans in Beijing, China (2023-2024). Among 87 isolates, E. faecalis and E. faecium predominated. Food-derived isolates showed broader resistance profiles than human isolates. Fourteen optrA-positive strains were identified, accounting for 92.9% of food isolates. optrA frequently co-localized with erm(A), ant(9)-Ia, and fexA on Tn554-family transposons, suggesting a potentially transferable multidrug resistance module. Notably, an optrA-positive E. faecalis ST699 clone was identified for the first time in Chinese retail meat. This clone formed a distinct lineage and carried a complete Tn554-optrA island. A representative ST699 isolate exhibited enhanced fitness and virulence potential in the Galleria mellonella model. These findings highlight animal-derived foods as important reservoirs of linezolid-resistant Enterococci and provide genomic evidence consistent with their role as potential sources of optrA-mediated resistance. The emergence of a multidrug-resistant E. faecalis ST699 clone with enhanced fitness characteristics underscores the need for continued surveillance of foodborne antimicrobial resistance within the One Health framework.

Enterococcus faecalis

Novelty, diversity, and genetic dark matter in enterococci of invertebrates.

Enterococci appear to have originated in the guts of early terrestrializing arthropods and invertebrates over 425 million years ago-hosts that are now highly diverse and widespread in nature today. Yet most knowledge of the genus comes from human infection-associated lineages with genomes swollen by the recent accretion of foreign DNA conveyed by mobile elements. Because invertebrates dominate terrestrial animal diversity and biomass, they would be predicted to constitute a major but little-explored reservoir of enterococcal diversity. We therefore systematically examined Enterococcus association and species diversification in invertebrate hosts of the comparatively natural, isolated, but well-characterized environment of the Azorean island of Terceira. Over 100 invertebrate specimens were examined for associated enterococci, which were taxonomically classified by whole-genome sequencing. Supporting the existence of a large pool of uncharacterized enterococci and Enterococcus-adapted genes, 40% (eight of 20) of the Enterococcus species identified were either undescribed, including four candidate new species described here, or very recently discovered. In contrast, control isolates from vertebrates were exclusively of known species typical of sampling elsewhere, discounting geographic isolation as a main driver of the novelty observed. Further, because of the abundance of E. casseliflavus and E. flavescens in this collection, we obtained the resolution necessary to quantify the divergence and decipher the drivers of speciation in the controversial division between these naturally vancomycin-resistant species. These findings provide robust support for the existence of a large pool of new species and unexplored adaptive traits in invertebrate-associated enterococci-diverse environmental survival traits optimized for expression in an enterococcal background, and well positioned for transmission into human-associated enterococcal strains.IMPORTANCEEnterococci are auxotrophic gut-associated bacteria that co-evolved with their terrestrial hosts over many eons. In the last 75 years-the "antibiotic era"-E. faecalis and E. faecium gained genes for antibiotic resistance and enhanced virulence, emerging as leading causes of multidrug-resistant infection. Little is known about the source of those genes or the pathway by which they entered human-associated strains. A recent global survey suggested a potentially large repository of uncharacterized genetic diversity in the enterococci of invertebrates. We directly tested this prospect by examining enterococci of invertebrate hosts in a largely natural and pastoral environment. Our findings provide clear evidence that invertebrates naturally harbor vast unexplored enterococcal diversity. Moreover, associations are likely driven by intrinsic host selection factors rather than geographic isolation. This expands our knowledge of Enterococcus biodiversity, including the identification of four novel species, identifying a vast reservoir of enterococcal genes available to species that colonize and infect humans.

Animals

Bacteriocin-mediated intraspecies competition driven by acquired Bac41 operon in epidemic Enterococcus faecalis ST179.

Enterococcus faecalis is a common gut commensal and an opportunistic pathogen causing hospital-acquired infections. Despite its clinical importance, comprehensive global genomic and epidemiological data remain limited. Here, we analyzed 5,895 E. faecalis genomes collected between 2000 and 2020 and identified ST179, a human-derived single-operon variant of the high-risk CC16 clonal complex, as an emerging epidemic clone in China. Spot-killing assays revealed that ST179 strongly inhibited other clinical E. faecalis sequence types. Biochemical purification and proteomic analyses identified BacL1 as a key effector associated with this species-specific antibacterial activity. Functional assays confirmed its inhibitory phenotype, providing ST179 with a lineage-specific, bacteriocin-mediated competitive advantage. The high prevalence of the Bac41 operon likely contributed to the epidemiological success and ecological fitness of ST179. These findings highlight the role of bacteriocin-mediated intraspecies competition in shaping E. faecalis population dynamics and suggest that ST179 might become an emerging high-risk lineage in China.IMPORTANCEEnterococcus faecalis is a common gut bacterium and an opportunistic pathogen. We identify ST179 as an emerging epidemic clone in China and show that it outcompetes other strains via the bacteriocin Bac41. This competitive advantage helps explain its rapid spread. Our findings highlight how bacterial competition shapes population dynamics and provide insights into the emergence of high-risk E. faecalis lineages, informing strategies for monitoring and infection control.

Enterococcus faecalis

The NagY antiterminator in Enterococcus faecalis: a novel regulatory mechanism and its impact on cell metabolism.

The Enterococcus genus is the most controversial group of lactic acid bacteria. While some strains are used as probiotics, other species, including Enterococcus faecalis, are responsible for health-related pathologies. Under conditions of infection, the N-acetylglucosamine metabolism of E. faecalis undergoes significant changes in expression, even more important than those of virulence factors. This metabolism is mediated by the nagY-nagE operon, which is regulated by the transcriptional antiterminator NagY. In this report, we focus on the regulatory mechanism of NagY and its impact on bacterial metabolism. We showed that NagY requires the interaction with the RNase III to achieve optimal induction of its own expression by cleaving the 5' untranslated region of the nagY mRNA. The NagY regulon was identified and the central role of the antiterminator in the E. faecalis metabolism was demonstrated, highlighting its importance in the opportunistic nature of the bacterium. This study provides a valuable advance in the understanding of regulation and the importance of post-transcriptional actors in E. faecalis adaptation.IMPORTANCEAs a commensal, Enterococcus faecalis colonizes the gastrointestinal tract of 31 to 80% of the intestinal microbiota in adults and is considered ubiquitous, due to its strong environmental stress resistance capabilities. However, in immunocompromised patients, the poorly understood transition from commensal to opportunistic pathogen occurs, and many studies suggest that the metabolism plays a central role in this process. In this study, we focus on the regulator NagY, which is involved in the metabolism of N-acetylglucosamine, an important carbon source for bacterial pathogens in the human host. We characterized a novel regulatory mechanism involving the NagY antiterminator and the ribonuclease RNase III. In addition, we identified the target genes of the regulator, through which we were able to demonstrate that NagY has a strong impact on the metabolism of β-glucosides. Overall, this work highlights the importance of regulation of the bacterial metabolic adaptation in the host.

Enterococcus faecalis

Antimicrobial resistance among Gram-positive agents of bacteraemia in the UK and Ireland: trends from 2001 to 2019.

OBJECTIVES: The BSAC Bacteraemia Resistance Surveillance collected isolates from UK and Irish hospitals for central testing. Concurrent UKHSA surveillance collated English hospitals' own susceptibility data. Results were collated and compared. METHODS: BSAC Surveillance collected quotas of isolates per site annually from 2001 to 2019. MIC testing was by BSAC agar dilution, with resistance mechanisms identified by synergy tests, interpretive reading and PCR. The UKHSA sought hospitals' data on all bacteraemia isolates. RESULTS: Both surveillance systems recorded dramatic falls in MRSA, from c. 40% of bloodstream Staphylococcus aureus in 2001 to <10% by 2019. Both noted rises in the proportion of MRSA (especially) and MSSA resistant to fusidic acid, along with declines of ciprofloxacin and macrolide resistance amongst MRSA. Methicillin resistance also fell among coagulase-negative staphylococci, albeit only modestly; fusidic acid resistance rose. Shifts for pneumococci were complex, reflecting vaccine-contingent serotype displacements; resistance rates remained low, with high-dose penicillin almost universally active. Enterococcus faecium became more prevalent relative to Enterococcus faecalis; vancomycin resistance averaged 29% among E. faecium versus 2% in E. faecalis, without trend. Erythromycin resistance rose among groups B, C and G (but not group A) streptococci. Oxazolidinones, tigecycline, daptomycin and anti-PBP2' cephalosporins retained near-universal activity against target species, except that tigecycline has been compromised by breakpoint reductions for streptococci. CONCLUSIONS: Gram-positive pathogens were the dominant historical pathogens of bacteraemia. The trends seen here-with many near-universally active antibiotics-indicate little hazard of this situation returning. Nevertheless, few treatments exist in some settings, notably multi-resistant E. faecium endocarditis.

Humans

[Examinations on the behaviour of grampositive and gramnegative bacteria in aqua bidest and tap-water at different initial colony counts and different temperatures (author's transl)].

Experiments were done about the behaviour of 5 different strains of Staph. aureus, Strept. faecalis, E. coli, Ser. marcescens and P. aeruginosa in aqua bidest and sterile tap water at 20 degrees C, 30 degrees C and 40 degrees C. The behaviour of 3 different initial colony counts in each case was examined. Colony forming units (CFU) were evaluated at the beginning and after 1, 2, 4, 6 and 24 h. CFU were reduced at Staph. aureus and E. coli in aqua bidest and tap water not earlier than after 4 h, but mostly after 24 h. In Strept. faecalis CFU were reduced in aqua bidest at 40 degrees C after 24 h, in tap water at 40 degrees C after 4 h, but there were no changes of CFU in both media at 20 degrees C or 30 degrees C. Ser. marcescens strains were growing in aqua bidest and tap water after 24 h at 20 degrees C. P. aeruginosa strains were reduced in aqua bidest at all temperatures but they were growing in tap water after 24 h. Reduction or growth of all strains did not depend on initial colony count during examination time, changing of CFU turned out to be quicker at 40 degrees C than at 20 degrees C. It is the conclusion that in humidifiers and nebulizers CFU will be different from initial colony count not earlier than after 6 h. Growth will be seen after 6 until 24 h if there are no particular nutritive organic compounds.

Bacteria

Genomic insights into an optrA-carrying plasmid associated with linezolid resistance in clinical Enterococcus faecalis isolates, Argentina.

The spread of the transferable optrA gene poses an increasing threat to the clinical efficacy of oxazolidinones. Here, we characterized a novel optrA-carrying plasmid, pEfa-optrA-Arg, from a linezolid-resistant Enterococcus faecalis clinical isolate from Argentina. The 68,653-bp conjugative plasmid harbored optrA together with multiple antimicrobial resistance genes and showed high similarity to a plasmid previously identified in a bovine isolate from Switzerland. pEfa-optrA-Arg, or a closely related variant, was also detected in E. faecalis isolates from several Argentinian hospitals, highlighting the role of horizontal gene transfer in the spread of antimicrobial resistance across human and animal reservoirs within the One Health continuum.

Enterococcus faecalis

Whole-Genome Sequencing of Feline Uropathogens Reveals Multidrug Resistance and Zoonotic Potential in Domestic Cats in Tunisia.

BACKGROUND: Urinary tract infections (UTIs) in cats are increasingly recognized as clinically relevant conditions frequently associated with multidrug-resistant (MDR) bacteria of potential zoonotic origin, yet genomic data on feline uropathogens remain scarce in Tunisia. METHODS: We used whole-genome sequencing to characterize seven bacterial isolates recovered from six cats with clinical signs of UTI: Mammaliicoccus lentus (n = 2), Staphylococcus schleiferi (n = 1), Mammaliicoccus sciuri (n = 1), Enterococcus faecalis (n = 1), Enterococcus casseliflavus (n = 1), and Klebsiella aerogenes (n = 1). RESULTS: Resistome analysis revealed determinants conferring resistance to &#x3b2;-lactams (blaZ, blaCMY-132), methicillin (mecC-type), macrolides (erm(43), ermB), tetracyclines (tet(M), tet(45), tetB), fosfomycins (fosI, fosB, fosA5), and aminoglycosides (aac(6'), aph(3')-IIIa, aph(6)-Id), alongside efflux pump genes (efrA, sepA, sdrM, oqxA, KpnE/F/G), vancomycin-operon genes (vanT, vanY, vanC, vanG), and biofilm/biocide-tolerance genes (salB, qacG). Notably, M. lentus S104 carried mecC-type elements, the first such report in Tunisia, while K. aerogenes displayed an extensive MDR profile, including blaCMY-132 and fosA5. Multilocus sequence typing/ribosomal multilocus sequence typing (MLST/rMLST) identified diverse lineages, including the internationally distributed E. faecalis ST19 and the rarely reported K. aerogenes ST242. Plasmids were absent in all isolates; a Tn916/1545-type transposon occurred in E. casseliflavus, and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems were unevenly distributed. CONCLUSIONS: These findings highlight companion animals as reservoirs of clinically important resistance genes, reinforcing the need for One Health AMR surveillance.

Animals

[Examinations on the behaviour of grampositive and gramnegative bacteria on aluminium foil (author's transl)].

The survival times of Staph. aureus, Strept. faecalis, E. coli, Klebs. pneumoniae, Ser. marcescens and P. aeruginosa were found out on aluminium foil. It was examined whether there exists a dependence on the initial colony count and in mixed cultures a mutual influence of the several species of bacteria. The longest survival times were found among the grampositive cocci. Ser. marcescens and Klebs. pneumoniae could no more be indicated after 19 days in case of initial colony count of log 7.0/cm2; E. coli could no more be shown after 14 days. P. aeruginosa had the shortest survival time. The dependence on the initial colony count was ascertained in 3 different concentrations of bacteria. The highest concentrations showed the smallest reduction of cocci and the longest survival times of Enterobacteriaceae respectively. The middle and lowest initial colony counts showed quicker reduction or shorter survival times. Ser. marcescens was more resistant to drying than Klebs. pneumoniae. The quickest dying rates showed P. aeruginosa without any dependence on initial colony count. In mixed cultures with Staph. aureus or Strept. faecalis survival times of Enterobacteriaceae were longer; the same result gave Klebs. pneumoniae in culture together with Ser. marcescens. The following explanation might be possible: because of the drying resistance of grampositive cocci the total bacteria number in these mixed cultures after several days is higher than in monocultures of E. coli, Klebs. pneumoniae or Ser. marcescens. This protects the Enterobacteriaceae against external influences for a longer time. Therefore it is understandable that the effect increases with the length of duration. The same interpretation could apply to the mixed cultures of Klebs. pneumoniae/Ser. marcescens.

Aluminum

[Development of a new testmethod for surface disinfection procedures. III. The impression method: influence of the test surface material and the types of microorganisms on the recovery rate (author's transl)].

The preceding test results appear to favour the impression method used for the testing of surface disinfection procedures. Further investigations were concerned with the selection of a suitable germ carrier. The high recovery rate of the bacteria and the invariably uniform quality of the germ carrier are the decisive criteria. Since a dependence on the type of germ was also to be expected, we checked Staph. aureus, Str. faecalis, E. coli, Kl. pneumoniae and P. aeruginosa for their behaviour on PVC, varnished wooden surfaces, V 2A, rough and smooth glass, resopal, ceramic tiles with dull and glazing finish as well as on plywood. The germ carriers were contaminated quantitatively, dried, sprayed with distilled water to stimulate disinfection and examined after they were allowed to react for a period of 6 hours. A set of 10 germ carriers was examined simultaneously with 10 Rodac impressions. The data were checked by variance analytical methods and evaluated on the mean (or median) values. It was found that while the initial germ number was kept equal OP tiles and V 2A steel produced the highest germ yield. Plywood proved largely unsuitable. Among the test germs, E. coli and Kl. pneumoniae had the lowest adhesion; it was inferior to that of staphylococci and pseudomonads. However, for these 4 germ types 3 impressions per germ carrier suffice for a reliable assessment based on computations. The yield amounts to more than 90 per cent. 94 per cent of the Streptococci were also recovered with 3 impressions, but the absolute germ quantity was low. The use of this type of germ can be recommended only with reservation.

Bacteriological Techniques

Antimicrobial Resistance in Nontyphoidal Salmonella and Clinically Relevant Enterococcus From Faecal Samples of Conservation-Priority Captive Ungulates in a United Arab Emirates Urban Zoo: A Cross-Sectional Baseline Study.

Antimicrobial resistance (AMR) is a One Health challenge driven by microbial exchange among humans, animals and the environment. Zoological institutions offer useful settings for environmental AMR surveillance. This single-zoo cross-sectional study examined the occurrence, antimicrobial susceptibility and genomic characteristics of nontyphoidal Salmonella enterica (NTS) and clinically relevant Enterococcus spp. in faecal samples from 101 clinically healthy captive ungulates representing seven conservation-priority species at a major urban zoo in the United Arab Emirates. NTS was detected in 4/101 samples (3.9%), including serovars Schwarzengrund (n&#x2009;=&#x2009;2), Kentucky (n&#x2009;=&#x2009;1) and Chester (n&#x2009;=&#x2009;1). Among the four recovered NTS isolates, all met the study MDR definition within the tested panel, including a Salmonella Kentucky ST198 isolate carrying multiple resistance genes and quinolone-associated mutations. Enterococcus spp. were detected in 77/101 samples (76.2%), dominated by Enterococcus faecium and Enterococcus casseliflavus (each 41.5%). Among 33 E. faecium/Enterococcus faecalis isolates tested phenotypically, resistance was generally low, with erythromycin and ciprofloxacin resistance each observed in 9.1%. One clinically important E. faecium isolate showed glycopeptide resistance and genetic markers associated with reduced daptomycin susceptibility. These single-institution cross-sectional data provide an initial regional baseline for AMR-relevant enteric bacteria in conservation-managed ungulates and identify priorities for broader longitudinal and interface-based surveillance.

Animals

[Studies on the antimicrobial effect of natural and synthetic humic acids (author's transl)].

Preparations of humic acids extracted from different soils by various methods and model humus substances obtained synthetically by oxidation of hydroquinone and pyrocatechin are tested for growth inhibition of representative strains of human pathogenic microorganisms using a micro serial dilution technique. Within the concentration range of less than or equal to 2500 micrograms/ml 57 of 81 natural and also the two synthetic humic acids show antimicrobial activity with differing spectra. These substances inhibit St. epidermidis, St. aureus, Str. pyogenes, S. typhimurium, Prot. vulgaris, Ent. cloacae, Ps. aeruginosa and C. albicans, but not Str. faecalis and E. coli. The degree of activity or the sensitivity of test organisms, respectively, amounts to 2500--1250 micrograms/ml predominantly, partially 625--312 micrograms/ml and can reach values of up to 39 micrograms/ml with synthetic hydroquinone humic acid. The spectrum and degree of activity vary according to the origin and extraction mode of the natural humic acids. The in vitro evidence of efficiacy against human pathogenic microorganisms gives a rational basis of therapeutic use of substances of humic acid type in infectious conditions.

Anti-Bacterial Agents