This national population-based study was conducted as part of the development of a national automated surveillance system for hospital-acquired bacteraemia and ascertains the utilization of blood cultures (BCs). A primary objective was to understand how local differences may affect interpretation of nationwide surveillance for bacteraemia. From the Danish Microbiology Database, we retrieved all BCs taken between 2010 and 2013 and linked these to admission data from the National Patient Registry. In total, 4 587 295 admissions were registered, and in 11%, at least one BC was taken. Almost 50% of BCs were taken at admission. The chance of having a BC taken declined over the next days but increased after 4 days of admission. Data linkage identified 876 290 days on which at least one BC was taken; 6.4% yielded positive results. Ten species, Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus pneumoniae, Enterococcus faecium, Enterococcus faecalis, Pseudomonas aeruginosa, Candida albicans, Enterobacter cloacae and Klebsiella oxytoca, accounted for 74.7% of agents for this purpose classified as pathogenic. An increase in BCs and positive BCs was observed over time, particularly among older patients. BCs showed a seasonal pattern overall and for S. pneumoniae particularly. A predominance of male patients was seen for bacteraemias due to S. aureus, E. faecium and K. pneumoniae. Minor differences in BCs and positive BCs between departments of clinical microbiology underpin the rationale of a future automated surveillance for bacteraemia. The study also provides important knowledge for interpretation of surveillance of invasive infections more generally.

The clinical significance of Dietzia papillomatosis is for the moment limited to the rare skin disease confluent and reticulated papillomatosis. We present a case of infection with D. papillomatosis in a 2-year-old boy with known syringomyelia. The microbiological diagnosis was done using 16S rRNA gene sequencing. This is the first report of bacteremia with D. papillomatosis. CASE REPORTA 2-year-old boy was approximately 5 months earlier diagnosed to have syringomyelia. As part of the syringomyelia elucidation, a cerebrospinal fluid (CSF) sample was obtained and tested for Borrelia IgG and IgM antibodies, and PCR was performed for herpes simplex and varicella-zoster; all were negative. In addition, a Gram stain was performed, but no bacteria were seen and the culture was negative. CSF leukocytes were at a level of 13 ϫ 10 6 /ml, but other parameters were reported normal. The patient had a ventriculoperitoneal shunt inserted for treatment of the syringomyelia. Due to malfunction of the shunt, the patient had an uncomplicated planned shunt revision. The day after the shunt revision, the boy was readmitted to the pediatric department due to high fever (39.2°C). He had a discrete truncal maculopapular exanthem but was not septic. He was suspected of having a viral infection. Chest X ray was normal, and there was no obvious focus of his infection. Two days later, the symptoms were unchanged, but the C-reactive protein rose from 9 to 108 mg/liter (normal value, Ͻ10 mg/liter), and his leukocyte count remained normal. A blood culture was obtained, and treatment with intravenous cefuroxime (100 mg/kg divided into three daily doses) was initiated. After another 2 days, the fever disappeared and the exanthem faded. Blood culture identified Dietzia papillomatosis by 16S rRNA gene PCR and sequencing. The boy was treated with cefuroxime for 4 days and then switched to oral amoxicillin, 36 mg/kg, for another 3 days and discharged. A blood culture, taken after antibiotic treatment was terminated, remained negative.The Dietzia species is an aerobic, Gram-positive coccus or short rod belonging to the group of actinomycetes. On the basis of its colony appearance and its microscopic resemblance, it has been classified as a Rhodococcus species. Gram staining of positive aerobic blood culture obtained from a peripheral vein revealed Gram-positive bacteria with an alternating morphology of cocci in clusters and coryneform rods. Blood was plated on blood agar and chocolate agar (Statens Serum Institute, Copenhagen, Denmark). The bacteria grew poorly on the first day under aerobic conditions and were later incubated on chocolate plates in a CO 2 atmosphere where growth was significantly improved. Repeated microscopy of colonies showed small Gram-positive rods. Fluorescence in situ hybridization using peptide nucleic acid probes (PNA-FISH; AdvanDx, Vedbaek, Denmark) using the probe set for identification of Staphylococcus aureus and coagulase-negative staphylococci was negative. Matrix-assisted laser desorption ionization-time of flight ...

In this study, RipSeq Mixed, a software resolving uninterpretable mixed DNA sequencing chromatograms, revealed the bacterial content of 15 polymicrobial samples. Direct sequencing combined with RipSeq Mixed constitutes a valuable supplement to cultivation, particularly when cultivation is negative and direct sequencing is inconclusive despite continued clinical indications of infection.

Here, we present a longitudinal shotgun sequencing metagenomics study of 16 healthy, Danish women in the reproductive age. The aim of the study was to investigate whether lactobacilli, orally consumed, had any impact on the vaginal microbiome and its functional potential. The 16 women aged 19–45 years were recruited from Copenhagen, Denmark. One baseline vaginal sample (Day 0) and two study samples (Days 25–30 and Days 55–60, respectively), were sampled. The vaginal samples were analyzed by shotgun metagenomics. We detected 26 species in the vaginal microbiota of the 16 women, of which six belonged to the Lactobacillus genus. We observed three vaginal microbiome clusters mainly dominated by Gardnerella vaginalis, Lactobacillus iners, or Lactobacillus crispatus. The oral probiotic had no detectable effect on either the composition or the functional potential of the vaginal microbiota. Most of the study subjects (11 out of 16 women) exhibited only minor changes in the vaginal microbiome during the treatment with probiotics. Any compositional changes could not be associated to the probiotic treatment. Future studies may benefit from an increased number of participants, and administration of the probiotics during conditions with bacterial imbalance (e.g., during/after antibiotic treatment) or the use of different Lactobacillus spp. known to colonize the vagina.