Multidrug-resistant K. pneumoniae producing OXA-48-like carbapenemase are emerging as important pathogens in Spain due to intra- and inter-hospital, clonal and non-clonal dissemination.

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The results for indicators related with sample collection indicate that the process is stable and well controlled. However, based on the results for Hemolyzed serum sample, the group saw the need for installing centrifuges in all phlebotomy centers and established a recommendation to unify the system for measuring hemolysis. The indicator External control exceeds acceptance limit clearly highlighted the need to rigorously monitor the analytic phase of the clinical laboratory. The values obtained for the indicator Reports from referred tests exceed delivery time show that there is considerable deviation regarding the expected report delivery time, whereas for in-house laboratory reports, delivery time is satisfactory.

The prevalence of Neisseria meningitidis isolates that are moderately susceptible to penicillin and ampicillin has increased very quickly in Spain; the current prevalences in our hospital are 48.4 and 55.6%, respectively. We studied the reliability of the disk diffusion method for discriminating between fully susceptible (MIC, .0.06 ,ug/ml) and moderately susceptible (MICs, 0.12 to 1.0 pg/ml) isolates. Thirty-eight isolates (12 Pens AmpS, 18 Penr Ampr, and 8 Pen' Ampr) were assayed by the disk diffusion and plate dilution methods by use of disks of 1, 2, 5, and 10 U of penicillin, 2 and 10 pg of ampicillin, and 1 pg of oxacillin on Mueller-Hinton agar with or without 5% sheep blood. Breakpoints were generated, and their discriminative values were determined with a collection of 195 meningococcal strains isolated consecutively from cerebrospinal fluid or blood. None of the strains produced 1-lactamase. Penicillin-and ampicillin-susceptible strains (MIC, <0.06 ,ug/ml) had oxacillin inhibition zones of >11 mm on blood agar. Moderate susceptibility to ampicillin and penicillin was suggested by an inhibition zone of < 10 mm around the 1-,ug oxacillin disk and an inhibition zone of .26 mm around the 2-U penicillin disk. We conclude that discrimination between fuly susceptible and moderately susceptible populations ofN. meningitidis can be accomplished by the disk diffusion method by use of Mueller-Hinton agar plus 5% sheep blood with the 1-,ug oxacillin disk (which is especially suited for screening methods) and the 2-U penicillin disk. The lack of an oxacillin inhibition zone indicates moderate susceptibility to either penicillin or ampicillin or both.Neisseria meningitidis isolates that are moderately susceptible to penicillin (MIC, 20.12 jg/ml) have been described in the last few years in Spain, England, and South Africa (1, 3, 11-13, 15, 16). Most of these isolates do not produce ,-lactamase, and modifications of the penicillinbinding proteins are believed to be a major mechanism of resistance (4, 5, 13). Current penicillin MICs for moderately susceptible isolates (0.12 to 1.0 ug/rml) are only 2-to 20-fold higher than that for the fully susceptible ones (.0.06 ,ug/ml) (8,(13)(14)(15).With regard to penicillin and ampicillin susceptibilities, three phenotypes have been described in N. meningitidis: susceptible to penicillin and ampicillin (Pens AmpS), moderately susceptible to both penicillin and ampicillin (Penr Ampr), and susceptible to penicillin and moderately susceptible to ampicillin (Pen' Ampr) (11, 14). The existence of Pen' Ampr strains (penicillin MIC, <0.06 ,ug/ml; ampicillin MIC, >0.12 ,ug/ml) is well documented (11,(13)(14)(15) moderately susceptible populations are of clinical and epidemiological interest. In the present study, we addressed the question of whether the three phenotypes mentioned above can be reliably discriminated by the disk susceptibility testing method. MATERIALS AND METHODSStrains. Identification of strains as N. meningitidis was carried out by standard bacteriological methods. ...

With the spread of new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), there is a need to assess the protection conferred by both previous infections and current vaccination. Here we tested the neutralizing activity of infected and/or vaccinated individuals against pseudoviruses expressing the spike of the original SARS-CoV-2 isolate Wuhan-Hu-1 (WH1), the D614G mutant and the B.1.1.7 variant. Our data show that parameters of natural infection (time from infection and nature of the infecting variant) determined cross-neutralization. Uninfected vaccinees showed a small reduction in neutralization against the B.1.1.7 variant compared to both the WH1 strain and the D614G mutant. Interestingly, upon vaccination, previously infected individuals developed more robust neutralizing responses against B.1.1.7, suggesting that vaccines can boost the neutralization breadth conferred by natural infection.

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The genetic relatedness of 42 penicillin-resistant Neisseria meningitidis isolates obtained during a 2-year period from a single hospital was studied by multilocus enzyme electrophoresis and by restriction fragment length polymorphism (RFLP) analysis of penicillin-binding protein (PBP) 2 genes. The PBP 2 genes of 7 susceptible strains gave identical RFLP profiles. Sixteen different PBP 2 RFLP profiles were found among the 42 resistant strains, but 4 were found in greater than 1 resistant isolate. Multilocus enzyme electrophoresis revealed a high level of genetic diversity. Four clusters of resistant strains could be distinguished at a genetic distance of 0.75. Resistant strains with the most common PBP 2 RFLP profile were restricted to one of these clusters and may be derived from a common ancestral strain. However, resistant strains with the 3 other common RFLP profiles were distributed in two or more of the clusters.

Objectives: The study aimed to characterize the clonal spread of resistant bacteria and dissemination of resistance plasmids among carbapenem-resistant Enterobacterales at a tertiary hospital in Catalonia, Spain.Methods: Isolates were recovered from surveillance rectal swabs and diagnostic samples. Species identification was by matrix-assisted laser desorption ionization-time time of flight mass spectrometry (MALDI-TOF MS). Molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Antimicrobial susceptibility was assessed by gradient-diffusion and carriage of bla genes was detected by PCR. Plasmid typing, conjugation assays, S1-PFGE studies and long-read sequencing were used to characterize resistance plasmids.Results: From July 2018 to February 2019, 125 Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacterales were recovered from 101 inpatients from surveillance (74.4%) or clinical samples (25.6%), in a tertiary hospital in Barcelona. Clonality studies identified a major clone of Klebsiella pneumoniae belonging to sequence type ST15 and additional isolates of K. pneumoniae, Escherichia coli and Enterobacter sp. from different STs. All isolates but one carried the blaKPC–2 allelic variant. The blaKPC–2 gene was located in an IncFIIk plasmid of circa 106 Kb in a non-classical Tn4401 element designated NTEKPC-pMC-2-1. Whole-genome sequencing revealed different rearrangements of the 106 Kb plasmid while the NTEKPC-pMC-2-1 module was highly conserved.Conclusion: We report a hospital outbreak caused by the clonal dissemination of KPC-producing ST15 K. pneumoniae but also the intra- and inter-species transmission of the blaKPC–2 gene associated with plasmid conjugation and/or transposon dissemination. To our knowledge, this is the first report of an outbreak caused by KPC-producing Enterobacterales isolated from human patients in Catalonia and highlights the relevance of surveillance studies in the early detection and control of antibiotic resistant high-risk clones.