Showing papers by "Guido Werner published in 2018"

The importance of adjusting for enterococcus species when assessing the burden of vancomycin resistance: a cohort study including over 1000 cases of enterococcal bloodstream infections

Abstract: Infections caused by vancomycin-resistant enterococci (VRE) are on the rise worldwide. Few studies have tried to estimate the mortality burden as well as the financial burden of those infections and found that VRE are associated with increased mortality and higher hospital costs. However, it is unclear whether these worse outcomes are attributable to vancomycin resistance only or whether the enterococcal species (Enterococcus faecium or Enterococcus faecalis) play an important role. We therefore aimed to determine the burden of enterococci infections attributable to vancomycin resistance and pathogen species (E. faecium and E. faecalis) in cases of bloodstream infection (BSI). We conducted a retrospective cohort study on patients with BSI caused by Enterococcus faecium or Enterococcus faecalis between 2008 and 2015 in three tertiary care hospitals. Data was collected on true hospital costs (in €), length of stay (LOS), basic demographic parameters, and underlying diseases including the results of the Charlson comorbidity index (CCI). We used univariate and multivariable regression analyses to compare risk factors for in-hospital mortality and length of stay (i) between vancomycin-susceptible E. faecium- (VSEm) and vancomycin-susceptible E. faecalis- (VSEf) cases and (ii) between vancomycin-susceptible E. faecium- (VSEm) and vancomycin-resistant E. faecium-cases (VREm). We calculated total hospital costs for VSEm, VSEf and VREm. Overall, we identified 1160 consecutive cases of BSI caused by enterococci: 596 (51.4%) cases of E. faecium BSI and 564 (48.6%) cases of E. faecalis BSI. 103 cases of E. faecium BSI (17.3%) and 1 case of E. faecalis BSI (0.2%) were infected by vancomycin-resistant isolates. Multivariable analyses revealed (i) that in addition to different underlying diseases E. faecium was an independent risk factor for in-hospital mortality and prolonged hospital stay and (ii) that vancomycin-resistance did not further increase the risk for the described outcomes among E. faecium-isolates. However, the overall hospital costs were significantly higher in VREm-BSI cases as compared to VSEm- and VSEf-BSI cases (80,465€ vs. 51,365€ vs. 31,122€ p < 0.001). Our data indicates that in-hospital mortality and infection-attributed hospital stay in enterococci BSI might rather be influenced by Enterococcus species and underlying diseases than by vancomycin resistance. Therefore, future studies should consider adjusting for Enterococcus species in addition to vancomycin resistance in order to provide a conservative estimate for the burden of VRE infections.

Whole-Genome Sequencing of Recent Listeria monocytogenes Isolates from Germany Reveals Population Structure and Disease Clusters.

Abstract: Listeria monocytogenes causes foodborne outbreaks with high mortality. For improvement of outbreak cluster detection, the German consiliary laboratory for listeriosis implemented whole-genome sequencing (WGS) in 2015. A total of 424 human L. monocytogenes isolates collected in 2007 to 2017 were subjected to WGS and core-genome multilocus sequence typing (cgMLST). cgMLST grouped the isolates into 38 complexes, reflecting 4 known and 34 unknown disease clusters. Most of these complexes were confirmed by single nucleotide polymorphism (SNP) calling, but some were further differentiated. Interestingly, several cgMLST cluster types were further subtyped by pulsed-field gel electrophoresis, partly due to phage insertions in the accessory genome. Our results highlight the usefulness of cgMLST for routine cluster detection but also show that cgMLST complexes require validation by methods providing higher typing resolution. Twelve cgMLST clusters included recent cases, suggesting activity of the source. Therefore, the cgMLST nomenclature data presented here may support future public health actions.

Emergence and control of linezolid-resistant Staphylococcus epidermidis in an ICU of a German hospital.

Abstract: Objectives To investigate an outbreak of linezolid-resistant Staphylococcus epidermidis (LRSE) in an interdisciplinary ICU, linezolid consumption and infection control measures taken. Methods Routine surveillance of nosocomial infections revealed colonization and infection with LRSE affecting 14 patients during a 15 month period. LRSE isolates were analysed with respect to their clonal relatedness, antimicrobial susceptibility, the presence of cfr and/or mutations in the 23S rRNA, rplC, rplD and rplV genes. cfr plasmids were characterized by Illumina sequencing. Medical records were reviewed and antibiotic consumption was determined. Results Molecular typing identified the presence of three different LRSE clusters: PFGE type I/ST168 (n = 5), PFGE type II/ST5 (n = 10) and PFGE type III/ST2 (n = 1). Ten strains harboured the cfr gene; we also detected mutations in the respective ribosomal protein genes. WGS revealed an almost identical 39 kb cfr plasmid obtained from strains of different genetic background (ST2, ST5, ST168) that shows high similarity to the recently published LRSE plasmid p12-02300. Due to an increase in the number of patients treated for infections with MRSA, a significant increase in linezolid usage was noted from January to July 2014 (from 5.55 to 20.41 DDDs/100 patient-days). Conclusions Here, we report the molecular epidemiology of LRSE in an ICU. Our results suggest the selection of resistant mutants under linezolid treatment as well as the spread of cfr-carrying plasmids. The reduction of linezolid usage and the strengthening of contact precautions proved to be effective infection control measures.

Dissemination of linezolid-dependent, linezolid-resistant Staphylococcus epidermidis clinical isolates belonging to CC5 in German hospitals.

Abstract: Objectives Linezolid-resistant Staphylococcus epidermidis (LRSE) and linezolid-dependent ST22 strains have been shown to predominate in tertiary care facilities all over Greece. We report herein the dissemination of ST22 but also ST2, ST5 and ST168 linezolid-dependent LRSE clones in four unrelated German hospitals. Methods Fourteen LRSE clinical isolates recovered during 2012-14 from five distantly located German hospitals were tested by for MIC determination broth microdilution and Etest, PCR/sequencing for cfr and for mutations in 23S rRNA, rplC, rplD and rplV genes, MLST, PFGE and growth curves without and with linezolid at 16 and 32 mg/L. Results Most (11, 78.6%) isolates had linezolid MICs >256 mg/L. Five isolates carried the cfr gene. Eight isolates belonged to ST22, two isolates each to ST168 and ST2 and one isolate each to ST5 and ST23. Ten isolates [seven belonging to ST22 and one to each of ST2, ST5 and ST168; all these STs belong to clonal complex (CC) 5] exhibited linezolid-dependent growth, growing significantly faster in linezolid-containing broth. Four isolates were non-dependent (one belonging to each of ST22, ST2, ST23 and ST168). Four isolates came from three different hospitals, whereas four and six isolates were recovered during outbreaks of LRSE in two distinct hospitals. Conclusions The multi-clonal dissemination of CC5 linezolid-dependent LRSE throughout German hospitals along with the clonal expansion of ST22 linezolid-dependent LRSE in Greek hospitals is of particular concern. It is plausible that this characteristic is inherent and provides a selective advantage to CC5 LRSE under linezolid pressure, contributing to their dissemination throughout hospitals in these countries.

Identification of a Novel Genomic Island Associated with vanD -Type Vancomycin Resistance in Six Dutch Vancomycin-Resistant Enterococcus faecium Isolates.

Abstract: Genomic comparison of the first six Dutch vanD-type vancomycin-resistant Enterococcus faecium (VRE) isolates with four vanD gene clusters from other enterococcal species and anaerobic gut commensals revealed that the vanD gene cluster was located on a genomic island of variable size. Phylogenetic inferences revealed that the Dutch VRE isolates were genetically not closely related and that genetic variation of the vanD-containing genomic island was not species specific, suggesting that this island is transferred horizontally between enterococci and anaerobic gut commensals.

Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus in Workers in the Food Industry

Abstract: Staphylococcus aureus is part of the common flora on the skin and mucous membranes of mammals and approximately 20–30% of humans are persistently colonized, mainly by mostly susceptible human-adapted isolates. In contrast, colonization with methicillin-resistant S. aureus is rare (approximately 1%), predominantly transient and associated with prior contact to the health care system. Additionally, in recent years livestock-associated S. aureus clones contributed to colonization in humans, especially in those working in close contact to farm animals. A considerable percentage of colonizing S. aureus isolates is equipped with enterotoxin genes. Humans carrying enterotoxigenic isolates represent a contamination source when handling food, thus generating a continuous risk of S. aureus food intoxication. Molecular characterization of isolates colonizing humans and obtained from food, respectively, enables the tracing of food-related outbreaks back to the source of food intoxication. We will summarize current knowledge about the S. aureus population colonizing humans, including those in close contact to animals and food, respectively. Additionally, we will review data on the molecular characterization of S. aureus isolates related to staphylococcal foodborne disease and the elucidation of staphylococcal foodborne outbreaks. Staphylococcal food poisoning is a common foodborne disease, mediated by the ingestion of enterotoxins produced by enterotoxigenic strains of S. aureus. For several outbreaks of foodborne S. aureus disease, colonized personnel could be identified as the source of food contamination. However, because of the widespread occurrence of enterotoxigenic strains as human colonizers and the often transient nature of colonization, the source of contamination cannot always be identified unambiguously. Therefore, compliance with hygiene measures is the most important requirement to prevent food contamination by both human colonization and environmental S. aureus reservoirs.

The use of high-throughput sequencing to investigate an outbreak of glycopeptide-resistant Enterococcus faecium with a novel quinupristin-dalfopristin resistance mechanism

Abstract: High-throughput sequencing (HTS) has successfully identified novel resistance genes in enterococci and determined clonal relatedness in outbreak analysis. We report the use of HTS to investigate two concurrent outbreaks of glycopeptide-resistant Enterococcus faecium (GRE) with an uncharacterised resistance mechanism to quinupristin-dalfopristin (QD). Seven QD-resistant and five QD-susceptible GRE isolates from a two-centre outbreak were studied. HTS was performed to identify genes or predicted proteins that were associated with the QD-resistant phenotype. MLST and SNP typing on HTS data was used to determine clonal relatedness. Comparative genomic analysis confirmed this GRE outbreak involved two distinct clones (ST80 and ST192). HTS confirmed the absence of known QD resistance genes, suggesting a novel mechanism was conferring resistance. Genomic analysis identified two significant genetic determinants with explanatory power for the high level of QD resistance in the ST80 QD-resistant clone: an additional 56aa leader sequence at the N-terminus of the lsaE gene and a transposon containing seven genes encoding proteins with possible drug or drug-target modification activities. However, HTS was unable to conclusively determine the QD resistance mechanism and did not reveal any genetic basis for QD resistance in the ST192 clone. This study highlights the usefulness of HTS in deciphering the degree of relatedness in two concurrent GRE outbreaks. Although HTS was able to reveal some genetic candidates for uncharacterised QD resistance, this study demonstrates the limitations of HTS as a tool for identifying putative determinants of resistance to QD.

Effektives Management eines Ausbruchs mit multiresistenten Klebsiella pneumoniae in der Neurorehabilitation

Abstract: Neben Akutkliniken sind auch Rehabilitationskliniken zunehmend von der Problematik der multiresistenten Erreger betroffen. Lange Verweildauern und hohe Behandlungsintensitaten stellen dabei besondere Herausforderungen an die Hygiene dar. Systematische Untersuchung eines Ausbruchs mit „Extended Spectrum Beta-Lactamasen“-Klebsiella-pneumoniae (ESBL-K. pneumoniae) in einer Neurorehabilitationsklinik. Gesicherter Fall: Patient mit einem ESBL-K.-pneumoniae-Nachweis im Ausbruchszeitraum und Zugehorigkeit zum Ausbruchsklon, wahrscheinlicher Fall: kein typisiertes Isolat, aber epidemiologischer Link zum Ausbruchsgeschehen. Insgesamt 53 ESBL-K.-pneumoniae-Isolate wurden mittels Next Generation Sequencing (NGS) typisiert. Umgebungsuntersuchungen wurden durchgefuhrt und ein systematisches Screening wurde implementiert. Neben einer Starkung der Standardhygiene wurde eine Kohortierung von Patienten mit ESBL-K.-pneumoniae veranlasst. Insgesamt wurden 30 gesicherte und 6 wahrscheinliche Falle dem Ausbruchsgeschehen zugeordnet (Cluster 1, ST15). In der Typisierung konnten 2 weitere Cluster identifiziert werden: Cluster 2 (ST405) bei 7 Patienten und Cluster 3 (ST414) bei 8 Patienten. Bei 2 Patienten entwickelte der Ausbruchsstamm ST15 weitere Antibiotikaresistenzen (Colistin bzw. Carbapenem). ESBL-K.-pneumoniae wurde in Umgebungsuntersuchungen nicht identifiziert. Der Ausbruch konnte nach strikter Kohortierung beendet werden. Die Erregerausbreitung erfolgte vermutlich von Mensch zu Mensch. Die Ubertragungswahrscheinlichkeit stieg mit dem Grad der Interventionsintensitat, wahrend die Infektionsrate durch die Morbiditat der Patienten bedingt war. Die Identifikation eines Ausbruchsklons mit Entwicklung zusatzlicher Antibiotikaresistenzen und die Entdeckung weiterer Cluster demonstrieren die Relevanz einer Erregertypisierung und eines effektiven Ausbruchsmanagements.