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Background Effective antimicrobial treatment is key for survival in bloodstream infection (BSI), but the impact of timing of treatment remains unclear. Our aim was to assess the association between time to appropriate antimicrobial treatment and 30-day mortality in BSI patients. Methods This was a retrospective cohort study using electronic health record data from a large academic center in Sweden. Adult patients admitted between the years 2012 and 2019, with onset of BSI at the emergency department or general wards, were included. Pathogen-antimicrobial drug combinations were classified as appropriate or inappropriate based on reported in vitro susceptibilities. To avoid immortal time bias, the association between appropriate therapy and mortality was assessed with multivariable logistic regression analysis at pre-specified landmark times. Results We included 10 628 BSI-episodes, occurring in 9192 unique patients. The overall 30-day mortality was 11.8%. No association in favor of a protective effect between appropriate therapy and mortality was found at the 1, 3 and 6 hours landmark after blood culture collection. At 12 hours, the risk of death increased with inappropriate treatment (adjusted odds ratio 1.17 [95% confidence interval {CI}, 1.01–1.37]) and continued to increase gradually at 24, 48, and 72 hours. Stratifying by high or low SOFA score generated similar odds ratios, with wider confidence intervals. Conclusions Delays in appropriate antimicrobial treatment were associated with increased 30-day mortality after 12 hours from blood culture collection, but not at 1, 3, and 6 hours, in BSI. These results indicate a benchmark for providing rapid microbiological diagnostics of blood cultures.

Background Infantile central nervous system infections (CNSIs) can be life-threatening and severe sequelae can be observed in encephalitis and bacterial meningitis. The causative microorganism is unknown in > 40% of patients with aseptic infections. This study aimed to analyze metagenome for detection of pathogen, and transcriptome for host reaction of infection in a single cerebrospinal fluid (CSF) sample using two different next-generation sequencing (NGS) platforms, Illumina and Nanopore. Methods Twenty-eight CNSIs patients (< 12 months), treated between June 2012 and April 2020, were enrolled. A total of 49 clinical samples (28 CSF and 21 blood) from 28 patients were collected. Extracted RNA, which was obtained from 23 CSF in sufficient quantities, was sequenced using both Nanopore and Illumina platforms to compare their performances in the detection of pathogens. Human-derived reads subtracted during pathogen detection were used for host transcriptomic analysis from both Nanopore and Illumina sequencing. All the extracted DNA from 28 CSF and 21 blood, was sequenced using Illumina sequencer for the detection of pathogens. Data analysis was performed on the in-house PATHDET pipeline. A flowchart for pathogen detection and transcriptome analysis Pathogen detection was performed on the in-house PATHDET pipeline and transcriptome analysis was performed by using the secondary outputs. Results RNA sequencing of CSF samples in 23 cases revealed potential viral pathogens in 10 cases: coxsackievirus B5 (4 cases), coxsackievirus B4 (3 cases), coxsackievirus B2 (1 case), echovirus E7 (1 case), and human parechovirus 3 (1 case). DNA sequencing was performed in 28 cases using Illumina sequencing, and Proteus mirabilis (1 case, consistent with the culture test) and human parvovirus B19 (1 case) were detected. The results of Nanopore sequencing and Illumina sequencing were consistent. However, the mapping coverage and depth to the detected pathogen genome of Nanopore sequencing was superior to that of Illumina sequencing. MX1, ISG15, and OAS1 were differentially expressed genes in patients with identified pathogens via metagenomic NGS, both the Nanopore and Illumina sequencing, and were associated with antiviral roles in innate immunity. A pie chart for patients with central nervous system infections (CNSIs) Results of next-generation sequencing (NGS) and the detected pathogen candidates using DNA/RNA workflows. Performance of Nanopore and Illumina sequencing for pathogen genome mapping The sequencing output, which is 100,000 reads randomly extracted from original sequencing reads, was mapped to the pathogen reference genome to determine (A) mapping coverage and (B) mapping depth. Volcano plot of all genes analyzed in Nanopore and Illumina sequencing Volcano plots showing differentially expressed genes in the cerebrospinal fluid between identified-pathogen and unidentified-pathogen samples from (A) Nanopore sequencing and (B) Illumina RNA sequencing. Conclusion The use of Nanopore sequencing for metagenomic diagnostics of CSF samples should help to understand both pathogens and host immune responses of CNSI and could shed light on the pathogenesis of these infections. Disclosures All Authors: No reported disclosures.

Background The outpatient setting is a key scenario for the implementation of antimicrobial stewardship (AMS) activities, considering that overconsumption of antibiotics occurs mainly outside hospitals. This publication is the result of a joint initiative by the JPIAMR ARCH and COMBACTE-MAGNET EPI-Net networks, which is aimed at formulating a set of target actions for linking surveillance data with AMS activities in the outpatient setting. Methods A scoping review of the literature was carried out in three research areas: AMS leadership and accountability; antimicrobial usage and AMS; antimicrobial resistance and AMS. Consensus on the actions was reached through a RAND-modified Delphi process involving over 40 experts in infectious diseases, clinical microbiology, AMS, veterinary medicine or public health, from 18 low-, middle- and high-income countries. Results Evidence was retrieved from 38 documents, and an initial 25 target actions were proposed, differentiating between essential or desirable targets according to clinical relevance, feasibility and applicability to settings and resources. In the first consultation round, preliminary agreement was reached for all targets. Further to a second review, 6 statements were re-considered and 3 were deleted, leading to a final list of 22 target actions in the form of a practical checklist. Conclusions This White Paper is a pragmatic and flexible tool to guide the development of calibrated surveillance-based AMS interventions specific to the outpatient setting, which is characterized by substantial inter- and intra-country variability in the organization of healthcare structures, maintaining a global perspective and taking into account the feasibility of the target actions in low-resource settings.

Pseudomonas aeruginosa is an environmental bacterium and a nosocomial pathogen with clone C one of the most prevalent clonal groups. The P. aeruginosa clone C specific genomic island PACGI-1 harbors a xenolog of ftsH encoding a functionally diverse membrane-spanning ATP-dependent metalloprotease on the core genome. In the aquatic isolate P. aeruginosa SG17M, the core genome copy ftsH1 significantly affects growth and dominantly mediates a broad range of phenotypes, such as secretion of secondary metabolites, swimming and twitching motility and resistance to aminoglycosides, while the PACGI-1 xenolog ftsH2 backs up the phenotypes in the ftsH1 mutant background. The two proteins, with conserved motifs for disaggregase and protease activity present in FtsH1 and FtsH2, have the ability to form homo- and hetero-oligomers with ftsH2 distinctively expressed in the late stationary phase of growth. However, mainly FtsH1 degrades a major substrate, the heat shock transcription factor RpoH. Pull-down experiments with substrate trap-variants inactive in proteolytic activity indicate both FtsH1 and FtsH2 to interact with the inhibitory protein HflC, while the phenazine biosynthesis protein PhzC was identified as a substrate of FtsH1. In summary, as an exception in P. aeruginosa, clone C harbors two copies of the ftsH metallo-protease, which cumulatively are required for the expression of a diversity of phenotypes.

The SIR system for antimicrobial susceptibility testing (AST) has not changed over several decades. The European Committee on Antimicrobial Susceptibility Testing will from 1 January 2019 implement a new SIR definition, which will also apply in Sweden. The main change is that the I-group (previously »intermediate«) will be renamed as »susceptible, increased exposure«. If »I« is included in the AST report it does not mean that the antimicrobial agent should be avoided, but rather that the dosing regimen needs to compensate for the decreased susceptibility.

The members of the European Survey on Carbapenemase-Producing Enterobacteriaceae, (EuSCAPE) Working Group are: Portugal—Manuela Caniça and Vera Manageiro BACKGROUND: Gaps in the diagnostic capacity and heterogeneity of national surveillance and reporting standards in Europe make it difficult to contain carbapenemase-producing Enterobacteriaceae. We report the development of a consistent sampling framework and the results of the first structured survey on the occurrence of carbapenemase-producing Klebsiella pneumoniae and Escherichia coli in European hospitals. METHODS: National expert laboratories recruited hospitals with diagnostic capacities, who collected the first ten carbapenem non-susceptible clinical isolates of K pneumoniae or E coli and ten susceptible same-species comparator isolates and pertinent patient and hospital information. Isolates and data were relayed back to national expert laboratories, which made laboratory-substantiated information available for central analysis. FINDINGS: Between Nov 1, 2013, and April 30, 2014, 455 sentinel hospitals in 36 countries submitted 2703 clinical isolates (2301 [85%] K pneumoniae and 402 (15%) E coli). 850 (37%) of 2301 K pneumoniae samples and 77 (19%) of 402 E coli samples were carbapenemase (KPC, NDM, OXA-48-like, or VIM) producers. The ratio of K pneumoniae to E coli was 11:1. 1·3 patients per 10 000 hospital admissions had positive clinical specimens. Prevalence differed greatly, with the highest rates in Mediterranean and Balkan countries. Carbapenemase-producing K pneumoniae isolates showed high resistance to last-line antibiotics. INTERPRETATION: This initiative shows an encouraging commitment by all participants, and suggests that challenges in the establishment of a continent-wide enhanced sentinel surveillance for carbapenemase-producing Enterobacteriaeceae can be overcome. Strengthening infection control efforts in hospitals is crucial for controlling spread through local and national health care networks. European Centre for Disease Prevention and Control info:eu-repo/semantics/publishedVersion

Patients with severe sepsis/septic shock have a high mortality. Beta-lactam antibiotics are normally first line treatment. This antimicrobial class has been associated with time-dependent efficacy. It is therefore plausible that administration as prolonged infusion will increase the therapeutic effect, as compared to short term bolus injections, which is the most common practice today. We have reviewed 14 randomized controlled studies to investigate whether prolonged infusion provides lower mortality and/or increased clinical cure. In summary, convincing advantages with prolonged infusion could not be found, however randomized studies are heterogeneous, and it cannot be excluded that some subgroups of critically ill patients could benefit from such treatment.