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• Enterococci have become major human MDR nosocomial pathogens. • They are found ubiquitously in non-human habitats (animals and the environment). • The main threat for public health is the transfer of MDR enterococci and AMR genes. • Contamination of humans through the food chain seems to be minimal. • The environment (especially wastewaters) may play a critical role in AMR in humans. Enterococci are long-standing members of the gastrointestinal tract of humans and many animals and they are also ubiquitously distributed in natural environments. Classically as harmless bacteria, two main species (namely Enterococcus faecalis and Enterococcus faecium) have become a leading cause of human infections, especially in hospital settings, with the worldwide spread of multidrug-resistant isolates, especially vancomycin-resistant enterococci. In this review, it will be summarized what is known about genetic diversity and ecology of enterococci with a focus on E. faecalis and E. faecium from human and non-human habitats and related risks for public health. [ABSTRACT FROM AUTHOR]

Abstract Vancomycin-resistant enterococci (VRE) are important nosocomial pathogens. Invasive VRE infections are difficult to treat since common therapeutic options including ampicillin and glycopeptides often fail. In vitro , most VRE remain susceptible to last-resort antibiotics such as linezolid, tigecycline and daptomycin. However, neither tigecycline nor linezolid act in a bactericidal manner, and daptomycin has proven activity only at high dosages licensed for treating enterococcal endocarditis. Despite these pharmacological and therapeutic limitations, reports on resistance to these last-resort drugs in VRE, and enterococci in general, have increased in recent years. In this review, we briefly recapitulate the current knowledge on the mode of action as well as the known and novel mechanisms of resistance and describe surveillance data on resistance to linezolid, tigecycline and daptomycin in enterococci. In addition, we also suggest a common nomenclature for designating enterococci and VRE with resistances to these important last-resort antibiotics. [ABSTRACT FROM AUTHOR]

Objectives: The Enterobacter cloacae complex (Ecc), routinely referred to as "E. cloacae" in clinical microbiology, encompasses several species with 12 genetic clusters and one sequence crowd that can be identified based on hsp60 sequencing. Little is known about the pathogenicity and distribution of resistance to antibiotics among the Ecc.Methods and Results: In this prospective multicentre study, a total of 193 Ecc clinical isolates were collected from 10 academic hospitals distributed nationally across France and identified at the genetic cluster level on the basis of hsp60 sequencing. E. hormaechei isolates, which belong to clusters VI-VIII, were the largest group (53%), followed by cluster III that accounted for 28% of clinical isolates. All other Ecc clusters were present except cluster VII (E. hormaechei subsp. hormaechei). Cephalosporinase overproduction and ESBL were significantly more present in E. hormaechei (33% and 20%) than in other clusters (19% and 3%, respectively).Conclusions: These results suggest that rapid identification of "E. cloacae" at the genetic cluster level could improve adequacy of empirical antibiotic treatment and reduce the unnecessary use of broad spectrum antibiotics. [ABSTRACT FROM AUTHOR]

Summary: Objectives: Isolation rates of unusual non-fermentative Gram-negative bacilli (i.e. other than Pseudomonas aeruginosa and Acinetobacter baumannii) are increasing but studies are limited to few observations. We aimed at determining risk factors for infection and influence of antibiotic treatment on the outcome. Methods: A six-month (December 1, 2008–May 31, 2009) prospective multicenter cohort study was conducted in nine French teaching hospitals. Characteristics of patients colonized or infected by unusual NF-GNB, adequacy of antimicrobial therapies, and outcome were analyzed. Results: Analysis of 158 patients (median age, 62.7 years) was conducted. Stenotrophomonas maltophilia was the predominant bacterial species isolated (39%) followed by Achromobacter group (15%) and non-baumannii Acinetobacter species (13%). Compared to colonized patients, infected ones were more frequently immunocompromised [relative risk (RR) = 1.63, (95% confidence interval (CI) = 1.02–2.60, P = 0.05)], hospitalized within the last three months [RR 1.67 (95% CI 1.09–2.58, P = 0.02)], admitted in an intensive care unit with central venous catheter [RR 1.74 (95% CI 1.15–2.63, P = 0.01)]. The overall hospital mortality concerned 28 patients (18%) but no association with inadequate antimicrobial treatment was found except in the group of S. maltophilia infected cases [RR 2.81 (95% CI 1.01–7.83, P = 0.02)]. Conclusion: Naturally carbapenems-resistant S. maltophilia is the main unusual NF-GNB pathogen in hospitalized patients, leading to inappropriate empirical antibiotic treatment at the time of emerging extended-spectrum β-lactamase-producing bacteria. [ABSTRACT FROM AUTHOR]

Summary: Actinobaculum schaalii is a facultative anaerobic, Gram-positive rod-shaped species phylogenetically related to Actinomyces that is likely part of the commensal flora of the human genitourinary tract. Because of its fastidious growth under aerobic conditions and its resemblance to bacteria of the resident flora, A. schaalii is frequently overlooked or considered as a contaminant. It is also difficult to identify phenotypically, still requiring molecular identification. Note that the recent technology of matrix-assisted laser desorption/ionisation time-of-flight-mass spectrometry could be a promising tool for its identification. Recent studies using sensitive PCR assays showed that its clinical significance was largely underestimated. Since its first description in 1997, A. schaalii has been responsible for numerous urinary tract infections (UTIs), mainly in elderly (usually >60 years) and patients with underlying urological conditions. Infected urines usually show many Gram-positive rods with significant leukocyturia and a negative test for nitrites. Numerous cases of severe infections have also been described, such as urosepsis, bacteremia, cellulitis, spondylodiscitis, and endocarditis. In vitro, A. schaalii is highly susceptible to β-lactams but it is resistant to ciprofloxacin and cotrimoxazole, first-choice antimicrobials for the oral treatment of UTIs. A penicillin (e.g. amoxicillin) or a cephalosporin (e.g. cefuroxime, ceftriaxone) should be the preferred treatment. [ABSTRACT FROM AUTHOR]

Abstract: Helicobacter pylori eradication by standard therapy is decreasing due to clarithromycin and metronidazole resistance. Fluoroquinolones are valuable drugs for alternative therapy, but their activity needs to be updated. We determined minimum inhibitory concentrations (MICs) of the newly marketed fluoroquinolones (levofloxacin, moxifloxacin and gatifloxacin) and assessed the prevalence of resistance in 128 H. pylori strains isolated in 2004–2005. The quinolone resistance-determining region (QRDR) of gyrA was sequenced for all strains. Gatifloxacin MICs (MIC50 =0.25mg/L) were two- to four-fold lower than those of the other fluoroquinolones. The prevalence of resistance (ciprofloxacin MIC>1mg/L) was 17.2% (22 strains). All resistant strains harboured one gyrA mutation at codons 86, 87 or 91, including three new mutations (Asp86Asn, Thr87Ile and Asn87Tyr). Ciprofloxacin-susceptible strains were devoid of such gyrA mutations, but harboured a polymorphism at codon 87 that distinguished 18 isolates (17%) with a Thr87 like the reference strain J99 from 88 strains with Asn87 like the reference strain 26695. Strains with Thr87 were four-fold more susceptible to nalidixic acid, pefloxacin, ciprofloxacin and levofloxacin and were equally susceptible to moxifloxacin and gatifloxacin. The high rate of quinolone resistance in H. pylori requires the use/implication of a ‘test and treat’ strategy that can confidently rely on QRDR gyrA sequencing. [Copyright &y& Elsevier]

Abstract: The genus Haemophilus constitutes a heterogeneous group of Pasteurellaceae species, and conventional identification of isolates other than Haemophilus influenzae and Haemophilus parainfluenzae is often challenging. Here, simple colony-PCR and sequencing assays with the same pair of degenerate primers were used to characterize a 449- to 458-bp fragment (sodA int) internal to the sodA gene encoding the manganese-dependent superoxide dismutase in type strains of all 15 Haemophilus species and Actinobacillus actinomycetemcomitans. The topology of a sodA int-based phylogenetic tree was in general agreement with that inferred from the analysis of 16S rRNA and other housekeeping gene sequences, but allowed more confident delineation of the main clusters of species. The sodA int sequences showed a markedly higher divergence than those of the corresponding 16S rRNA genes, and 38 independent human clinical isolates were identified by comparing their sodA int sequence to those of the type species. Except for one Haemophilus aphrophilus strain, all isolates were unambiguously characterized in spite of a high intraspecific sodA int sequence diversity. This study provides a comprehensive sequence-based phylogenetic analysis of the entire genus Haemophilus, and confirms that sodA is a potent target for the identification of clinical isolates of Pasteurellaceae. This approach might contribute to the taxonomic reappraisal of this family, and to the development of diagnostic tools. [Copyright &y& Elsevier]

Résumé: Les bactéries peuvent résister aux antibiotiques par exportation active grâce à des transporteurs membranaires appelés pompes d’efflux. Ces protéines sont spécifiques d’une classe d’antibiotiques ou au contraire responsables de MDR (multidrug resistance). Pour fonctionner, les pompes d’efflux utilisent l’énergie fournie par dissipation d’un gradient de protons (familles MFS, RND et SMR) ou d’ions sodium (famille MATE) ou encore par hydrolyse d’ATP (famille ABC). Plusieurs fonctions physiologiques ont été décrites chez les procaryotes, comme la protection vis-à-vis de toxiques environnementaux et le maintien de l’homéostasie cellulaire, qui contribuent indirectement à la virulence bactérienne. Chez les bactéries à Gram négatif, les systèmes d’efflux sont souvent des complexes protéiques ternaires avec une pompe transmembranaire, une protéine périplasmique de jonction et une porine de la membrane externe. Les pompes les plus fréquemment rencontrées sont de type RND comme AcrB chez Escherichia coli ou MexB chez Pseudomonas aeruginosa. Chez les bactéries à Gram positif, les systèmes d’efflux ne sont constitués que de la pompe. Les plus étudiés sont les pompes MFS comme NorA ou QacA chez Staphylococcus aureus et PmrA chez Streptococcus pneumoniae. Quelques transporteurs ont aussi été décrits chez les mycobactéries. Malgré la caractérisation d’un nombre croissant de pompes, les conséquences cliniques de la résistance par efflux sont mal connues du fait de niveaux variables d’expression et du manque de marqueurs spécifiques pour les détecter en routine. Enfin, l’utilisation d’inhibiteurs spécifiques des pompes bactériennes semble constituer une approche thérapeutique intéressante, cependant difficile à mettre en pratique, en raison de leur toxicité pour les cellules eucaryotes. [Copyright &y& Elsevier]

Polyphasic taxonomic analysis was performed on a novel bacterium, designated UR159T, isolated in 2016 from human blood of a septic patient hospitalized in France. Preliminary 16S rRNA gene sequence-based phylogenetic analysis indicated that strain UR159T belonged to the family Flavobacteriaceae , forming a distinct phyletic line distantly related (<94% sequence similarity) to known species of the family. Further phenotypic, chemotaxonomic and genomic analyses were performed. Cells were non-motile, oxidase-negative, catalase-positive Gram-negative rods. It was strictly aerobic yielding yellow-pigmented colonies, and was metabolically rather inert. Major fatty acids were iso-branched fatty acids, predominantly iso-C 15:0 (55.5%) and iso-C 17:1 ω 9 c (8.8%). Whole genome sequencing revealed a 2.3-Mbp genome encoding a total of 2262 putative genes with a genomic DNA G + C content at 37.6 mol%. The average nucleotide identity (ANI) and in silico DNA-DNA hybridization (isDDH) values between strain UR159T and the most closely related members of the Flavobacteriaceae family were <75% and <39%, respectively, much below the established cut-offs for ANI (<95–96%) and isDDH (<70%) for species and genus delineation. Average Amino Acid Identity (AAI) percentages were also estimated and were lower than 65% (cut-off proposed for genus delineation for uncultivated prokaryotes) in all cases, except for F. marinum that was just at the limit (65.1%). Based on these findings, we propose it as a new genus and species, Avrilella dinanensis gen. nov., sp. nov. (type strain UR159T = CIP 111616T = DSM 105483T). [ABSTRACT FROM AUTHOR]

Abstract: The qnr genes are transferable genes that confer low-level quinolone resistance by protection of topoisomerase. The occurrence of mutations in DNA gyrase (gyrA, gyrB) and topoisomerase IV (parC, parE) genes in strains harbouring qnr was investigated in 28 qnrA-positive clinical isolates, among which 7 strains also harboured qnrS. Topoisomerase mutations were found in 25 (89%) of the 28 strains, with at least two mutations (gyrA and parC) in 13 strains and one mutation in 12 strains. Isolates of the Enterobacter cloacae complex were compared with reference strains of the new Enterobacter species. gyrA mutations were found at position 83 (Ser or Thr for Ile, Tyr, Leu or Phe depending on the species), and new gyrB mutations were described (S463A, S464F). qnrA had an additive effect of a 10-fold increase in the minimum inhibitory concentration (MIC) whatever the number of topoisomerase mutations, and qnrS was additive to qnrA with a further 2- to 10-fold increase in the MIC. Comparison of MICs with susceptibility breakpoints showed that strains combining qnrA and topoisomerase mutations were resistant to fluoroquinolones, but the three strains lacking a topoisomerase mutation were susceptible using ciprofloxacin and levofloxacin but not using nalidixic acid or moxifloxacin testing. [Copyright &y& Elsevier]

• Cephalosporinase-overproducer Enterobacterales are significantly more resistant to temocillin than ESBL-producing Enterobacterales (37.7% vs 23.5%; P < 0.01); • The rate of temocillin resistance is correlated to the number of inactive beta-lactams, ranging from 3.7% to 60.0% in strain susceptible to all beta-lactams and those resistant to 3 beta-lactams; • Among third-generation cephalosporin-resistant Enterobacterales, temocillin was active against 64.9% of the strains nonsusceptible to piperacillin-tazobactam, ofloxacin, and cotrimoxazole. • In ESBL-producing strains, the rate of resistance to temocillin trend to increase from 13.9% in 2014 to 23.9% in 2017 (P < 0.01). Temocillin is used for several years in some European countries but, only since 2015 in France. We assessed the susceptibility of Enterobacterales strains isolated from blood culture 1 year before (2014) and 2 years after (2017) its use in France. 1,387 strains were included by 17 clinical laboratories located throughout France: 363 in 2014 and 1,024 in 2017. The rate of resistance to temocillin was 4.6% and 26.7% in 3rd generation cephalosporin (3GC) susceptible and resistant strains respectively. Cephalosporinase-overproducer (COPE) strains were significantly more resistant to temocillin (37.7%) than ESBL-producer (ESBL-PE) (23.5%) (P < 0.01). The rate of temocillin resistance was correlated to the number of inactive beta-lactams. The rate of resistance to temocillin trend to increase from 13.9% in 2014 to 23.9% in 2017 (P < 0.01). Temocillin remains highly active against Enterobacterales but the trend in resistance should be assessed over time. [ABSTRACT FROM AUTHOR]

To evaluate performances of the rapid multiplex PCR assay BioFire FilmArray Pneumonia Panel (FA-PP) for detection of bacterial pathogens and antibiotic resistance genes in sputum, endotracheal aspirate (ETA) and bronchoalveolar lavage (BAL) specimens. This prospective observational study was conducted in 11 French university hospitals (July to December 2018) and assessed performance of FA-PP by comparison with routine conventional methods. A total of 515 respiratory specimens were studied, including 58 sputa, 217 ETA and 240 BAL. The FA-PP detected at least one pathogen in 384 specimens, yielding an overall positivity rate of 74.6% (384/515). Of them, 353 (68.5%) specimens were positive for typical bacteria while eight atypical bacteria and 42 resistance genes were found. While identifying most bacterial pathogens isolated by culture (374/396, 94.4%), the FA-PP detected 294 additional species in 37.7% (194/515) of specimens. The FA-PP demonstrated positive percentage agreement and negative percentage agreement values of 94.4% (95% CI 91.7%–96.5%) and 96.0% (95% CI 95.5%–96.4%), respectively, when compared with culture. Of FA-PP false-negative results, 67.6% (46/68) corresponded to bacterial species not included in the panel. At the same semi-quantification level (in DNA copies/mL for FA-PP versus in CFU/mL for culture), the concordance rate was 43.4% (142/327) for culture-positive specimens with FA-PP reporting higher semi-quantification of ≥1 log 10 in 48.6% (159/327) of cases. Interestingly, 90.1% of detected bacteria with ≥106 DNA copies/mL grew significantly in culture. FA-PP is a simple and rapid molecular test that could complement routine conventional methods for improvement of diagnosis accuracy of pneumonia. [ABSTRACT FROM AUTHOR]

A lincosamide-resistant and macrolide-susceptible phenotype has not been described to date in Streptococcus pyogenes [group A streptococcus (GAS)]. The aim of this study was to characterize a GAS isolate susceptible to macrolides but resistant to lincosamide, streptogramin A and pleuromutilin antibiotics. Antimicrobial susceptibility was tested using the microdilution broth method and the resistance phenotype was tested by D-test. The GAS2887HUB isolate was subjected to whole-genome sequencing. The isolate showed a positive Gots' test (clindamycin inactivation). Whole-genome sequencing revealed that the strain was ST10 and emm 93, and had five resistance genes [ lnu (B) , ant (6)-Ia , aph (3′)-III, tet (M) and dfrG ]. The tet (M) gene was located in a Tn 916 -like transposon. The lsa (E)− lnu (B) - containing sequence (inserted downstream of the rumA gene) was formed by a 39.6-kb prophage, followed by a gene cluster encoding aminoglycoside–streptothricin resistance [ ant (6)Ia− sat4 − aph (3′)III] and lsa (E)− lnu (B) genes. This structure was not transferred by conjugation. This study identified a new genetic element carrying a determinant of lincosamide resistance in a GAS. Further molecular epidemiological surveys are needed to determine the prevalence of this mechanism of resistance in GAS. [ABSTRACT FROM AUTHOR]

Abstract The commercial Rapid Polymyxin NP test was evaluated to detect colistin-resistant Enterobacteriaceae. A total of 223 enterobacterial isolates corresponding to 136 resistant (including 38 MCR-like producers), 19 heteroresistant, and 78 colistin-susceptible isolates were tested. The test was performed according to the manufacturer's instruction, and the color of the wells was read after 2 and 3 hours of incubation. The results were compared with those of the homemade Rapid Polymyxin NP test, and manual broth microdilution according to EUCAST guidelines was used as the reference method to determine the performance of the test. Excellent performance of the commercial Rapid Polymyxin NP test was found with a very major error rate, a major error rate, a sensitivity, and a specificity of 1.9%, 5.1%, 98.1%, and 94.9%, respectively. The performance of the homemade Polymyxin NP test was similar, with a slightly better value for the very major error (1.2 %). [ABSTRACT FROM AUTHOR]

Dual resistance to linezolid and glycopeptides is a milestone reached by certain extensively drug-resistant (XDR) enterococci. This paper describes the molecular and epidemiological investigations of a linezolid-resistant and vancomycin-resistant Enterococcus faecium (E. faecium) (LVR Ef) outbreak in the French overseas territory of Reunion Island (Indian Ocean). All vancomycin-resistant Enterococcus (VRE) isolates detected on Reunion Island between 2015 and 2019 were included in the study. The VRE isolates were phenotypically characterised and genetically explored by whole-genome sequencing (WGS). Sixteen vancomycin-resistant E. faecium (VR Ef) isolates were retrieved between 2015 and 2019. Seven isolates obtained in 2019 were involved in the outbreak. These seven LVR Ef isolates from the 2019 outbreak at the University Hospital of Reunion Island (UHRI) were suspected to be related to a linezolid-susceptible VR Ef strain imported from India. An epidemiological link was highlighted for six of the seven outbreak cases. All the LVR Ef outbreak isolates were obtained from rectal swabs (colonisation) and resistant to vancomycin (MIC > 128 mg/L) and linezolid (MIC 8–32 mg/L); one isolate was also resistant to daptomycin (MIC 8 mg/L). The seven outbreak isolates were positive for the vanA and optrA genes and belonged to ST761. These results argue for the strict application of control and prevention measures for VRE clones at high risk of spread, particularly in areas such as Reunion Island where the risk of importation from the Indian subcontinent is high. The regional spread of optrA linezolid-resistance genes in VRE isolates is a matter of concern, due to possibility of treatment failure. [ABSTRACT FROM AUTHOR]

We investigated the role of a novel small RNA expressed in Enterococcus faecium (named Ern0030). We revealed that ern0030 was encoded within the 5′untranslated region of tet (M), a gene conferring tetracycline resistance through ribosomal protection. By RACE mapping, we accurately determined the boundaries of ern0030 , which corresponded to P tet. This upstream sequence of tet (M), P tet , was previously described within transcriptional attenuation mechanism. Here, Northern blot analyses revealed three transcripts of different lengths (ca. 230, 150 and 100 nucleotides) expressed from P tet. Phenotypically, the total deletion of ern0030 conferred a decrease in tetracycline MICs that was consistent with gene expression data showing no significant tet (M) induction under tetracycline SIC in ern0030 -deleted mutant as opposed to a 10-fold increase of tet (M) expression in the wild-type strain. We investigated the transcriptional attenuation mechanism by toeprint assay. Whereas the expected tet (M) ribosome-binding site (RBS) was detected, the RBS of the putative leader peptide was not highlighted by toeprint assay, suggesting the transcriptional attenuation was unlikely. Here, we demonstrate that Ern0030 has a role in regulation of tet (M) expression and propose a novel model of tet (M) regulation alternative or complementary to transcriptional attenuation. [ABSTRACT FROM AUTHOR]

Abstract: The purpose of this retrospective study was to evaluate the pathogenic role of Corynebacterium species in lower respiratory tract infections as well as their routine laboratory investigation. From April 2007 to August 2009, 27 clinical isolates were significantly recovered from respiratory specimens of 27 different patients clinically suspected of having lower respiratory tract infections. The average age of patients was 65 years, while 22 (81%) patients presented at least 1 predisposing condition. Of the 27 patients, 15 (56%) were classified as infected according to Centers for Disease Control and Prevention/National Healthcare Safety Network criteria, with 93% of infections being hospital acquired. All isolates were accurately identified to the species level using molecular methods (i.e., 17 Corynebacterium pseudodiphtheriticum, 7 Corynebacterium striatum, and 3 Corynebacterium accolens), whereas phenotypic methods remained frequently unreliable for identifying C. striatum and C. accolens strains. All tested isolates were susceptible to amoxicillin, imipenem, vancomycin, linezolid, and tigecycline, whereas most of them were resistant to erythromycin. [Copyright &y& Elsevier]

Abstract: We have assessed the performance of semi-automated rep-PCR (Diversilab®) and multilocus sequence typing (MLST) in comparison to pulsed-field gel electrophoresis (PFGE) for typing a collection of 29 epidemiologically characterized vancomycin-resistant Enterococcus faecium (VRE). Sixteen strains that harbored the Tn1546 element were typed by PCR mapping. The discriminative power of the typing methods was calculated by the Simpson''s index of diversity, and the concordance between methods was evaluated by the Kendall''s coefficient of concordance. Semi-automated rep-PCR appeared as discriminative as PFGE and was further compared with PFGE for typing 67 VRE isolated during a hospital outbreak. Rep-PCR appeared to be more discriminative than PFGE for this second set of strains. Reproducibility of DiversiLab® was also tested against 35 selected isolates. Only three showed less than 97% similarity, indicating high reproducibility at this level of discrimination. In conclusion, semi-automated rep-PCR is a useful tool for rapid screening of VRE isolates during an outbreak, although cost of the system may be limiting for routine implementation. PFGE, which remains the reference method, should be used for confirmation and evaluation of the genetic relatedness of epidemic isolates. [Copyright &y& Elsevier]

Summary Meningitis due to Mycoplasma hominis in adults is rarely described, with only three cases having been reported to date. A case of fatal meningitis in a 39-year-old patient after a neurosurgical procedure for a subarachnoid haemorrhage is reported herein. Identification and treatment were significantly delayed because of the rarity of the aetiology and difficulty identifying this organism with the routinely used conventional methods, such as Gram staining and agar growth on standard agar plates. Clinical procedures and the treatment of ‘culture-negative’ central nervous system infections is a real challenge for clinical microbiologists and clinicians, and M. hominis has to be considered as a potential, although very uncommon, pathogen. [ABSTRACT FROM AUTHOR]

All the 31 isolates of Alloscardovia omnicolens exhibited low MICs for β-lactams, glycopeptides, linezolid, tetracyclines, and cotrimoxazole. One strain showed MICs ≥256 μg/mL for both erythromycin and clindamycin with a single point mutation in 23S rRNA. One strain likely had acquired fluoroquinolone resistance associated with a unique mutation in ParC. [ABSTRACT FROM AUTHOR]

Abstract: We report a case of a recurrent peritonitis due to Microbacterium resistens in a 71-year-old male patient undergoing peritoneal dialysis (PD). Importantly, this Gram-positive rod was intrinsically resistant to cephalosporins and vancomycin, classically used in PD-related peritonitis treatment. His infection resolved after several weeks of appropriate therapy (amoxicillin plus gentamicin) and PD catheter removal. [Copyright &y& Elsevier]

• Excellent activity of eravacycline (ERC) against enterococci, including vancomycin-resistant isolates. • First description of enterococcal clinical isolates resistant to ERC. • Resistance to ERC is associated with resistance to all tetracyclines. • ERC-resistant isolates have at least two tet genes and one rpsJ mutation. Eravacycline (ERC), the first fluorocycline, is a new tetracycline with superior activity to tigecycline (TGC) against many bacterial species. This work aimed to determine the in vitro activity of ERC compared with other tetracyclines against enterococcal clinical isolates and to analyse corresponding resistance mechanisms. A collection of 60 enterococcal strains was studied: 54 epidemiologically unrelated clinical isolates (46 Enterococcus faecium and 8 Enterococcus faecalis) including 42 vancomycin-resistant enterococci (VRE) (33 vanA and 9 vanB), 3 in vitro TGC-resistant mutants (E. faecium AusTig, HMtig1 and HMtig2) and 3 reference wild-type strains (E. faecium Aus0004 and HM1070, E. faecalis ATCC 29212). In vitro susceptibility was determined using Etest strips (for ERC) or by broth microdilution (for TGC, doxycycline, minocycline and tetracycline). Resistance genes tet (M), tet (L), tet (O) and tet (S)] were screened by PCR for TGC- and/or ERC-resistant strains as well as sequencing of the rpsJ gene (encoding ribosomal protein S10). MIC 50/90 values were 0.016/0.08, ≤0.03/0.5, 4/32, 8/16 and 32/>32 mg/L for ERC, TGC, doxycycline, minocycline and tetracycline, respectively. According to EUCAST guidelines, nine strains were categorised as resistant to TGC (MIC, 0.5–8 mg/L), including four E. faecium vanA (+) strains also resistant to ERC (MIC, 0.19–1.5 mg/L). These four strains all possessed at least one mutation in rpsJ and two tet determinants: tet (M) + tet (L) (n = 2); and tet (M) + tet (S) (n = 2). Although ERC has excellent in vitro activity against enterococci (including VRE), emergence of resistance is possible due to combined mechanisms (rpsJ mutations + tet genes). [ABSTRACT FROM AUTHOR]

Abstract: The detection of vancomycin-resistant enterococci using a novel commercial multiplex real-time polymerase chain reaction assay (Xpert™ vanA/vanB, Cepheid, Sunnyvale, CA) was evaluated on 804 rectal swab specimens. Compared to enriched culture, sensitivity and negative predictive value of this method were 100%. Many false-positive results were recovered (sensitivity, 85.4%; positive predictive value, 8.7%), especially for the vanB gene. [Copyright &y& Elsevier]