Whole-genome sequencing analysis reveals the spread of a vanB-carrying transposon among different vancomycin-resistant Enterococcus faecium clinical isolates in a non-endemic setting.

BACKGROUND: Vancomycin-resistant enterococci (VRE), particularly Enterococcus faecium (VREfm), can cause serious nosocomial infections, and have been responsible for healthcare-associated outbreaks. Spreading of VREfm can occur both clonally and by the dissemination of mobile genetic elements. AIM: To report prospective analysis of whole-genome sequencing (WGS) data, including both core-genome multi-locus sequence typing (cgMLST) and transposon analysis, during a vanB VREfm outbreak. METHODS: Screening for vanB-positive VREfm isolates was performed by real-time polymerase chain reaction (PCR) on an overnight enriched broth and, if positive, subculture was performed. vanB-positive VREfm isolates underwent WGS. Generated data were used for molecular typing that was performed by cgMLST using SeqSphere. For transposon characterization, sequence data were mapped against the reference sequence of transposon Tn1549 using CLC Genomics Workbench, or de-novo assemblies were used for BLASTN comparisons. RESULTS: In total, 1358 real-time PCRs were performed. Two hundred and fifty-one specimens from 207 patients tested positive on PCR for vanB, of which 13 specimens obtained from six patients were identified as vanB VREfm positive on culture. These six patients harboured seven unique isolates belonging to four cluster types: CT118 (N=2), CT2483 (N=3), CT2500 (N=1) and CT2501 (N=1). Transposon analysis revealed the presence of an identical vanB-carrying transposon in the isolates cultured from all six patients that could be linked based on epidemiological data. CONCLUSION: A vanB VREfm outbreak occurred in the study hospital, including six patients with isolates belonging to four cluster types. In-depth transposon analysis revealed that dissemination of transposon Tn1549 rather than clonal spread was the cause of the outbreak.

Short-course aminoglycosides as adjunctive empirical therapy in patients with Gram-negative bloodstream infection, a cohort study.

OBJECTIVE: Short-course aminoglycosides as adjunctive empirical therapy to ß-lactams in patients with a clinical suspicion of sepsis are used to broaden antibiotic susceptibility coverage and to enhance bacterial killing. We quantified the impact of this approach on 30-day mortality in a subset of sepsis patients with a Gram-negative bloodstream infection. METHODS: From a prospective cohort study conducted in seven hospitals in the Netherlands between June 2013 and November 2015, we selected all patients with Gram-negative bloodstream infection (GN-BSI). Short-course aminoglycoside therapy was defined as tobramycin, gentamicin or amikacin initiated within a 48-hour time window around blood-culture obtainment, and prescribed for a maximum of 2 days. The outcome of interest was 30-day all-cause mortality. Confounders were selected a priori for adjustment using a propensity score analysis with inverse probability weighting. RESULTS: A total of 626 individuals with GN-BSI who received ß-lactams were included; 156 (24.9%) also received aminoglycosides for a median of 1 day. Patients receiving aminoglycosides more often had septic shock (31/156, 19.9% versus 34/470, 7.2%) and had an eight-fold lower risk of inappropriate treatment (3/156, 1.9% versus 69/470, 14.7%). Thirty-day mortality was 17.3% (27/156) and 13.6% (64/470) for patients receiving and not receiving aminoglycosides, respectively; yielding crude and adjusted odds ratios for 30-day mortality for patients treated with aminoglycosides of 1.33 (95% CI 0.80-2.15) and 1.57 (0.84-2.93), respectively. CONCLUSIONS: Short-course adjunctive aminoglycoside treatment as part of empirical therapy with ß-lactam antibiotics in patients with GN-BSI did not result in improved outcomes, despite better antibiotic coverage of pathogens.

First report of a brain abscess caused by Nocardia veterana.

Among Nocardia species causing infections, Nocardia veterana is rarely isolated and is mostly described as causing pulmonary infections. This is the first presentation of a case of brain abscess attributable to an N. veterana infection in a patient with type 2 diabetes. Prolonged antibiotic therapy with trimethoprim-sulfamethoxazole led to successful clinical recovery.

[Relationship between rubella virus and Fuchs heterochromic uveitis; 2 patients]. / Samenhang tussen rubellavirus en fuchs-heterochromie-uveïtis; 2 patiënten.

Two otherwise healthy men, aged 26 and 29 years, were diagnosed with Fuchs heterochromic uveitis (FHU) on the basis of the presence of iris heterochromia or iris atrophy, stellate corneal precipitates, and/or cataract. Microbiological investigation of aqueous humour demonstrated intraocular antibody production against rubella virus, but not against Toxoplasma gondii, herpes simplex virus or varicella zoster virus. Microbial nucleic acid detection was negative for all pathogens. Some time later, both patients underwent cataract surgery, which improved their vision considerably. FHU is a chronic, generally unilateral iridocyclitis, accompanied by the above-mentioned ophthalmologic manifestations in the absence of systemic disease. Little is known about the pathogenesis ofFHU, but recent publications have provided evidence for the possible involvement of the rubella virus.

[Patients co-infected with HIV and hepatitis-B virus (HBV): the favourable effect of lamivudine, as part of combined antiretroviral therapy, on HBV may be dependent upon the number of CD4-cells]. / Patiënten met een coïnfectie van HIV en hepatitis-B-virus (HBV): gunstig effect van lamivudine, als onderdeel van antiretrovirale combinatiebehandeling, op HBV mogelijk afhankelijk van het CD4-celaantal.

OBJECTIVE: To determine the effect of lamivudine on HBV co-infection in HIV-infected patients. DESIGN: Retrospective METHOD: The HBsAg status and the use of lamivudine were determined retrospectively in a cohort of 800 HIV-infected patients under treatment at the Infectious Diseases outpatient clinic of the University Medical Centre in Utrecht, The Netherlands. In the group of HBsAg-positive patients using lamivudine 150 mg twice daily as part of highly active antiretroviral therapy (HAART), the HBV-DNA was measured quantitatively in the remaining plasma. In addition, the HBsAg, HBeAg, activity of alanineaminotransferase (ALAT) and CD4-count were obtained from the patient records. RESULTS: The study identified 29 (3.6%) HIV-infected patients to be HBsAg-positive. Plasma samples of 14 of these 29 patients were positive for HBV-DNA before the start of the therapy. Ten of these 14 patients had CD4 counts of at least 200 x 10(6) cells/l, while four patients had less than 200 x 10(6) cells/l. In contrast to the group with less than 200 x 10(6) cells/l, a significant decrease in HBV-DNA load was seen after six months of therapy in the patients with at least 200 x 10(6) CD4-cells/l (t-test for repeated measurements; p = o.oo1). The difference between the two groups in the effect of lamivudine was statistically significant (p = 0.021). At final evaluation after a mean follow-up of 32 and 13 months, respectively, HBV-DNA could no longer be detected in 7 patients; ALAT normalised in 9 patients (64%). CONCLUSION: In this retrospective study, lamivudine was effective in the therapy of HIV-infected patients with a HBV co-infection. The decrease in the amount of circulating HBV was associated with the number of CD4 cells.