David Fuglsang-Damgaard

Characterization of third-generation cephalosporin-resistant Escherichia coli from bloodstream infections in Denmark.

The aim of the study was to investigate the molecular epidemiology of 87 third-generation cephalosporin-resistant Escherichia coli (3GC-R Ec) from bloodstream infections in Denmark from 2009. Sixty-eight of the 87 isolates were extended-spectrum beta-lactamase (ESBL) producers, whereas 17 isolates featured AmpC mutations only (without a coexpressed ESBL enzyme) and 2 isolates were producing CMY-22. The majority (82%) of the ESBL-producing isolates in our study were CTX-M-15 producers and primarily belonged to phylogroup B2 (54.4%) or D (23.5%). Further, one of the two CMY-22-producing isolates belonged to B2, whereas only few of the other AmpCs isolates belonged to B2 and D. Pulsed-field gel electrophoresis revealed that both clonal and nonclonal spread of 3GC-R Ec occurred. ST131 was detected in 50% of ESBL-producing isolates. The remaining ESBL-producing isolates belonged to 17 other sequence types (STs), including several other internationally spreading STs (e.g., ST10, ST69, and ST405). The majority (93%) of the ESBL-producing isolates and one of the CMY-22-producing isolates were multiresistant. In conclusion, 3GC-R in bacteriaemic E. coli in Denmark was mostly due to ESBL production, overexpression of AmpC, and to a lesser extent to plasmid-mediated AmpC. The worldwide disseminated CTX-M-15-ST131 was strongly represented in this collection of Danish, bacteriaemic E. coli isolates.

An ST405 NDM-4-producing Escherichia coli isolated from a Danish patient previously hospitalized in Vietnam

This variant, designatedNDM-4, differs from NDM-1 by a single amino substitution and anincreased carbapenemase activity. NDM-4 was first detected inan Escherichia coli isolated from a urinary sample from a patientpreviously hospitalized in India. A second finding of the NDM-4variantwasreportedinanE.colirectalisolatefromapatienttrans-ferredfromCameroontoFrance.

The use of Gram stain and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry on positive blood culture: synergy between new and old technology

Fuglsang‐Damgaard D, Nielsen CH, Mandrup E, Fuursted K. The use of Gram stain and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry on positive blood culture: synergy between new and old technology. APMIS 2011; 119: 681–88.

Positive blood cultures and diagnosis of bacterial meningitis in cases with negative culture of cerebrospinal fluid

The diagnosis of bacterial meningitis may be established through positive blood cultures in cases with negative culture of cerebrospinal fluid (CSF). However, information is sparse regarding how often the aetiological diagnosis is determined by blood culture alone. Therefore, we undertook a retrospective study of patients with meningitis and a blood culture-based bacterial diagnosis in North Jutland County, Denmark in 1997–2005. Patients were included if they had 1) a microscopy- and culture-negative CSF specimen with a leukocyte count ≥100×106/l and an erythrocyte count ≤10,000×106/l and 2) a positive blood culture within an interval of 3 d. A total of 20 patients fulfilled these criteria: 7 with Streptococcus pneumoniae, 4 with Neisseria meningitidis, 4 with Staphylococcus aureus, and 5 with miscellaneous bacteria. Suitable antibiotic therapy was delayed up to 48 h after lumbar puncture, and in-hospital mortality was 25%. A discharge diagnosis of bacterial meningitis was missing in 6/20 cases, and 12/20 cases had not been reported to Danish health authorities. Thus, patients with CSF-culture negative bacterial meningitis are at risk of delayed therapy, a poor outcome, and incomplete notification to health authorities.

Use of whole-genome sequencing for detection of the spread of VIM-4-producing Escherichia coli between two patients in Denmark

The metallo-lactamase (MBL) blaVIM-4, a one amino acid varint of blaVIM-1, was first described in a Pseudomonas aeruginosa solate recovered from a Greek patient. It has since been reported n various Enterobacteriaceae from the Mediterranean region or uggesting import from there [1,2]. Here we describe the first detecion, to our knowledge, of blaVIM-4 in Denmark as well as the first eport of transmission of MBL-producing Escherichia coli between wo Danish patients. The first E. coli isolate (strain 180298) was recovered in October 012 from a urine sample of a 73-year-old patient with chronic bstructive pulmonary disease. In December 2012, a positive urine ample from an 80-year-old patient yielded an E. coli isolate (strain 21994). The latter patient had been hospitalised on several occaions during the previous 6 months due to chronic disease. A eview of the patient histories revealed an epidemiological link s they had concurrently been hospitalised in the same ward in ctober 2012. Due to the suspicion of MBL production, both isoates were sent to Statens Serum Institut (Copenhagen, Denmark) or further analysis. The two isolates were tested for antimicroial susceptibility to 26 antimicrobial agents using Trek Sensititre® anels (Thermo Scientific, Waltham, MA) following Clinical and aboratory Standards Institute (CLSI) guidelines. Susceptibility to igecycline and ertapenem was investigated using a Liofilchem MIC est (Liofilchem s.r.l., Roseto degli Abruzzi, Italy) (Table 1). E. coli TCC 25922 was used for quality control. Minimum inhibitory conentration (MIC) results were interpreted using CLSI guidelines. he isolates shared phenotypic susceptibility profiles and were ultidrug-resistant. Antimicrobial agents active in vitro included iprofloxacin, tigecycline and colistin (Table 1). PCR screening for MBL genes and sequencing identified the laVIM-4 gene in both isolates. For whole-genome sequencing, DNA as extracted from the isolates using a DNeasy Blood and Tissue it (QIAGEN, Hilden, Germany) and fragment libraries were contructed using a NexteraTM XT Kit (Illumina, San Diego, CA) followed y 250-bp paired-end sequencing (MiSeqTM; Illumina). By analysis f the sequences for antimicrobial resistance genes using Resinder v.2.1 (http://cge.cbs.dtu.dk/services/ResFinder/; accessed 8.09.14), three additional extended-spectrum -lactamase genes blaCTX-M-9, blaCTX-M-15 and blaOXA-1) were identified. Furtherore, resistance genes to aminoglycosides [aac(3)-IIa, aadA24],

Rare Elizabethkingia anophelis meningitis case in a Danish male

Introduction. Elizabethkingia anophelis is a Gram-negative, aerobic, non-motile rod belonging to the family Flavobacteriaceae. Over the last 5 years, it has emerged as an opportunistic human pathogen involved in neonatal meningitis and sepsis, as well as nosocomial outbreaks. It has been isolated from the midgut of the Anopheles gambiae mosquito, but there is no evidence for a role of the mosquito in human infections, and very little is known regarding the routes of transmission to humans. Recent studies, primarily from South-East Asia, suggest that E. anophelis, and not Elizabethkingia meningoseptica, is the predominant human pathogen of this genus. However, identification to the species level has been difficult due to the limitations of the current MALDI-TOF MS (matrix-associated laser desorption ionization-time of flight MS) systems for correct species identification. Case presentation. Here, we present a rare case of E. anophelis meningitis in a Danish male, who had a travel exposure to Malaysia 7 weeks before hospitalization. A multidrug-resistant Elizabethkingia species was isolated from blood and cerebrospinal fluid, and genomic sequencing was used to characterize the phylogenetic position of the isolate, which was determined as associated with previously described sublineage 11. The patient was successfully treated with intravenous moxifloxacin and rifampicin for 2 weeks with no major sequelae, but we did not find the source of transmission. Conclusion. All clinical microbiologists should be aware of the present limitations of the MALDI-TOF MS systems for correct species identification, and therefore we recommend the use of genome sequencing for the correct identification at the species and sublineage level.

Clostridium hastiforme bacteraemia secondary to pyometra in a 64-year-old woman

The Gram-negative, rod-shaped, anaerobe bacteria Tissierella praeacuta was first described in 1908 by Tissier. However, during the past decade, Clostridium hastiforme has been recognised as a later synonym of T. praeacuta. C. hastiforme/T. praeacuta has only rarely been described in previous literature as a cause of human infection. We present here a case report of C. hastiforme/T. praeacuta bacteraemia secondary to pyometra in a 64-year-old woman with a history of multiple sclerosis and an intrauterine device inserted three decades earlier. C. hastiforme/T. praeacuta was isolated from blood as well as pus from the site of infection. The patient was cured with a combination of drainage and antibiotic therapy.

First report of metronidazole resistant, nimD-positive, Bacteroides stercoris isolated from an abdominal abscess in a 70-year-old woman

We here present the first case of a metronidazole resistant nimD positive Bacteroides stercoris. The isolate originated from a polymicrobial intra-abdominal abscess in a 70-year-old woman. The nimD gene was detected by use of whole-genome shotgun sequencing and the subsequent use of the ResFinder 2.1 web service.