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We described a case of a 65-year-old man with a brain abscess caused by Porphyromonas gingivalis and Eubacterium nodatum. The patient presented right central facial nerve palsy, mutism and right hemiparesis at the examination. The patient underwent a left frontal craniotomy with evacuation of the brain abscess. Specimens were collected for microbiological analysis and intravenous treatment was started with levetiracetam, dexamethasone, meropenem (1 g/8 h) and linezolid (600 mg/12 h). After identification of anaerobic bacteria the antibiotic treatment was changed to piperacillin/tazobactam (4 g/0,5 g/8 h), fulfilling 8 weeks of antibiotic with good clinical and radiological evolution., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Aim: This study aimed to assess the ultra-fast method using MinION™ sequencing for rapid identification of β-lactamase-producing Klebsiella pneumoniae clinical isolates from positive blood cultures. Methods: Spiked-blood positive blood cultures were extracted using the ultra-fast method and automated DNA extraction for MinION sequencing. Raw reads were analyzed for β-lactamase resistance genes. Multilocus sequence typing and β-lactamase variant characterization were performed after assembly. Results: The ultra-fast method identified clinically relevant β-lactamase resistance genes in less than 1 h. Multilocus sequence typing and β-lactamase variant characterization required 3-6 h. Sequencing quality showed no direct correlation with pore number or DNA concentration. Conclusion: Nanopore sequencing, specifically the ultra-fast method, is promising for the rapid diagnosis of bloodstream infections, facilitating timely identification of multidrug-resistant bacteria in clinical samples.

Background: Fast diagnosis of bloodstream infections remains the most important challenge for clinical microbiologists. The introduction of the mass-spectrometry represents a breakthrough, although several methods are already commonly used for the direct identification from positive blood cultures we present a faster method (ultra fast) for Lytic anaerobic flasks. Methods: We compare the ultra-fast (UF) method with the extensively employed differential centrifugation method (DC) and both to routine identification after 18-24 h of incubation. UF and DC method correlation rates to the gold standard were calculated, and statistical significance was proved with the Z test. Results: UF performed better overall than DC, with this difference being statistically significant. This tendency was observed in every subanalysis.

The objective of this study was the phenotypic and genotypic characterization of a carbapenem resistant Acinetobacter baumannii (CRAB) isolate. The isolate, recovered in Northern Spain in 2019, was identified by MALDI-TOF to the species level. Antimicrobial susceptibility testing was performed using the Phoenix BD NMIC-502 Panel, E-test, and broth microdilution methods. The presence of a metallo- β -lactamase (MBL) was verified by PCR and immunochromatographic assays. The genetic location of the MBL was confirmed using S1-pulsed-field gel electrophoresis (S1-PFGE) followed by Southern blot hybridization. Whole genome sequencing (WGS) was completed using the Miseq and MinION platforms, followed by core-genome MLST (cgMLST) and seven-locus MLST analysis. The CRAB was assigned ST85 (Pasteur scheme) and ST957 (Oxford scheme) representing international clone (IC) 9 and harbored the intrinsic β -lactamase OXA-94 with IS Aba1 upstream of it, and the MBL bla NDM-6 . Hybridization experiments revealed that the bla NDM-6 was encoded on the chromosome. Using WGS the bla NDM-6 environment could be identified arranged in the following order: IS Aba14 , aphA6 , IS Aba125 , bla NDM-6 , ble MBL , trpF , dsbC , cutA , and IS Aba14 . Downstream, a 10,462 bp duplication was identified, including a second copy of bla NDM-6 in the following genetic composition: IS Aba125 , bla NDM-6 , ble MBL , trpF , dsbC , cutA , and IS Aba14 . To our knowledge, this is the first description of bla NDM-6 in A. baumannii . The MBL was present in two copies in the chromosome in a new genetic environment associated with IS elements highlighting the contribution of mobile genetic elements in the dissemination of this gene., (Copyright © 2020 Xanthopoulou, Urrutikoetxea-Gutiérrez, Vidal-Garcia, Diaz de Tuesta del Arco, Sánchez-Urtaza, Wille, Seifert, Higgins and Gallego.)

Objective: Nowadays, the use of flow-cytometry for the screening of urine samples is extended, but appropriate cut-off points for each population group are yet to be established. The objective of this study was to evaluate the Sysmex UF-1000i® cytometer as a screening method for detection of asymptomatic bacteriuria (AB) and Group B Streptococcus (GBS) in pregnant women., Methods: Urine samples obtained during pregnancy between January-July 2019 were both processed with the Sysmex UF-1000i® and also cultured. Demographic data, flow-cytometry parameters and the result of the urine culture were collected. To assess the performance of the flow-cytometer for detection of AB and GBS, receiver operating characteristic (ROC) curves for the BACT/μL variable were applied., Results: A total of 33,687 urine samples were received, among which 1,443 (4.3%) belonged to pregnant women. Urine culture was positive in 82 (5.7%) samples, 1,295 (89.7%) were negative and 66 (4.6%) were considered contaminated. GBS was isolated in 69 (4.8%) patients and 46 (66.7%) with a microbial count below 10E4 CFU/mL. For AB detection, the cut-off point of 550 BACT/μL yielded a sensitivity of 91.5%, a negative predictive value of 99.3% and could avoid culturing 74.1% of the samples. No cut-off value could be established for GBS detection., Conclusions: Although the Sysmex UF-1000i® system is a valid screening method for the AB detection in pregnant population, it is not useful for the identification of GBS bacteriuria in our area. Therefore, the conventional urine culture is still required during pregnancy., (©The Author 2020. Published by Sociedad Española de Quimioterapia. This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)(https://creativecommons.org/licenses/by-nc/4.0/).)