Hanne Marie Holt

16S rRNA Gene Sequencing in Routine Identification of Anaerobic Bacteria Isolated from Blood Cultures

ABSTRACT A comparison between conventional identification and 16S rRNA gene sequencing of anaerobic bacteria isolated from blood cultures in a routine setting was performed (n = 127). With sequencing, 89% were identified to the species level, versus 52% with conventional identification. The times for identification were 1.5 days and 2.8 days, respectively.

Homogeneity of Danish Environmental and Clinical Isolates of Shewanella algae

ABSTRACT Danish isolates of Shewanella algae constituted by whole-cell protein profiling a very homogeneous group, and no clear distinction was seen between strains from the marine environment and strains of clinical origin. Although variation between all strains was observed by ribotyping and random amplified polymorphic DNA analysis, no clonal relationship between infective strains was found. From several patients, clonally identical strains of S. algaewere reisolated up to 8 months after the primary isolation, indicating that the same strain may be able to maintain the infection.

Trends in occurrence of antimicrobial resistance in Campylobacter jejuni isolates from broiler chickens, broiler chicken meat, and human domestically acquired cases and travel associated cases in Denmark.

Campylobacter jejuni is a frequent cause of bacterial gastroenteritis. Often it causes self-limiting disease but severe or prolonged cases may require antimicrobial treatment. The agricultural use of antimicrobial agents selects for resistance among C. jejuni which is transmitted to humans via food. In Denmark, the use of fluoroquinolones in animal husbandry has been restricted since 2003. The purpose of the present study was to look at trends in occurrence of resistance among C. jejuni from broiler chickens, broiler chicken meat and human domestically acquired or travel associated cases. From 1997 through 2007, C. jejuni isolates were obtained from The Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP) and susceptibility tested for ciprofloxacin, erythromycin, nalidixic acid, and tetracycline. Erythromycin resistance was at a low level in all the reservoirs during the study period. Resistance to ciprofloxacin, nalidixic acid and tetracycline was significantly higher in C. jejuni from imported broiler chicken meat compared to Danish broiler chicken meat. In domestically acquired human C. jejuni isolates, resistance to ciprofloxacin and nalidixic acid was for most years significantly higher compared to the level found in isolates from Danish broiler chicken meat, whereas the resistance level was similar to the level found in isolates from imported broiler chicken meat. Imported broiler chicken meat may therefore contribute to the high level of ciprofloxacin and nalidixic acid resistance in C. jejuni isolates from domestically acquired human infections. In 2006 and 2007, the occurrence of resistance to ciprofloxacin, nalidixic acid and tetracycline was significantly higher in travel associated C. jejuni isolates compared to isolates acquired domestically. Even though the use of fluoroquinolones is restricted for animal use in Denmark, Danes are still often infected by fluoroquinolone resistant C. jejuni from imported chicken meat or by travelling.

Risk factors for Clostridium difficile infection in the community: a case-control study in patients in general practice, Denmark, 2009-2011

To identify risk factors for Clostridium difficile infection (CDI) in Danish patients consulting general practice with gastrointestinal symptoms, a prospective matched case-control study was performed; cases (N = 259) had positive cultures for toxigenic C. difficile and controls (N = 455) negative cultures. Data were analysed by conditional logistic regression. In patients aged ⩾2 years (138 cases), hospitalization [odds ratio (OR) 8·4, 95% confidence interval (CI) 3·1-23], consumption of beef (OR 5·5, 95% CI 2·0-15), phenoxymethylpenicillin (OR 15, 95% CI 2·7-82), dicloxacillin (OR 27, 95% CI 3·6-211), and extended spectrum penicillins (OR 9·2, 95% CI 1·9-45) were associated with CDI. In patients aged <2 years none of these were associated with CDI, but in a subgroup analysis contact with animals was associated with CDI (OR 8·1, 95% CI 1·0-64). This study emphasizes narrow-spectrum penicillins, and suggests beef consumption, as risk factors for CDI in adults, and indicates a different epidemiology of CDI in infants.

Solobacterium moorei Bacteremia: Identification, Antimicrobial Susceptibility, and Clinical Characteristics

ABSTRACT We present five cases of Solobacterium moorei bacteremia. The isolates were identified with 16S rRNA gene sequencing and were susceptible to common antibiotics used for anaerobic infections. Bacteremia with S. moorei seems to be associated with debilitating conditions, but the prognosis of the infection appears to be good.

International travel and the risk of hospitalization with non-typhoidal Salmonella bacteremia. A Danish population-based cohort study, 1999-2008

BackgroundInformation is sparse regarding the association between international travel and hospitalization with non-typhoidal Salmonella bacteremia. The aim of this study was to determine the proportion, risk factors and outcomes of travel-related non-typhoidal Salmonella bacteremia.MethodsWe conducted a 10-year population-based cohort study of all patients hospitalized with non-typhoidal Salmonella bacteremia in three Danish counties (population 1.6 million). We used denominator data on Danish travellers to assess the risk per 100,000 travellers according to age and travel destination. We used patients contemporaneously diagnosed with travel-related Salmonella gastroenteritis as reference patients to estimate the relative risk of presenting with travel-related bacteremia as compared with gastroenteritis. To evaluate clinical outcomes, we compared patients with travel-related bacteremia and patients with domestically acquired bacteremia in terms of length of hospital stay, number of extraintestinal focal infections and mortality after 30 and 90 days.ResultsWe identified 311 patients hospitalized with non-typhoidal Salmonella bacteremia of whom 76 (24.4%) had a history of international travel. The risk of travel-related bacteremia per traveller was highest in the age groups 15-24 years (0.8/100,000 travellers) and 65 years and above (1.2/100,000 travellers). The sex- and age-adjusted relative risk of presenting with bacteremia was associated with travel to Sub-Saharan Africa (odds ratio 18.4; 95% confidence interval [6.9-49.5]), the Middle East (10.6; [2.1-53.2]) and South East Asia (4.0; [2.2-7.5]). We found high-risk countries in the same three regions when estimating the risk per traveller according to travel destination. Patients hospitalized with travel-related bacteremia had better clinical outcomes than patients with domestically acquired bacteremia, they had a shorter length of hospital stay (8 vs. 11 days), less extraintestinal focal infections (5 vs. 31 patients) and a lower risk of death within both 30 days (relative risk 0.2; [0.1-0.7]) and 90 days (0.3; [0.1-0.7]). A healthy traveller effect was a plausible explanation for the observed differences in outcomes.ConclusionsInternational travel is a notable risk factor for being hospitalized with non-typhoidal Salmonella bacteremia and the risk differs between age groups and travel destinations. Healthy travellers hospitalized with bacteremia are less likely to have poor outcomes than patients with domestically acquired bacteremia.

Report of the First Human Case of Caulobacter sp. Infection

ABSTRACT A Caulobacter sp. isolate was recovered from the dialysis fluid of a patient undergoing peritoneal dialysis. Bacterial identification included electron microscopy and 16S rDNA sequencing. To our knowledge, this is the first report of human Caulobacter infection. Special growth requirements suggest that Caulobacter spp. may be overlooked in the clinical microbiology laboratory.

Detection of Legionella bozemanae, a New Cause of Septic Arthritis, by PCR Followed by Specific Culture

ABSTRACT Legionella bozemanae is a rare isolate in clinical specimens. We describe a case of joint infection due to L. bozemanae in an immunocompromised patient with dermatomyositis. Without the use of PCR screening or culture on specialized medium, the organism would not have been detected.

Need for species-specific detection for the diagnosis of amoebiasis in a non-endemic setting.

Abstract The diagnosis of amoebiasis caused by Entamoeba histolytica is traditionally based on microscopy. However, the specificity of this method may be questioned, especially in areas where infections by E. histolytica are rare. In the present study, a species-specific real-time PCR was used for the identification of the morphologically similar species E. histolytica and Entamoeba dispar. Out of 15 microscopy-positive stool samples, all were negative for E. histolytica and positive for E. dispar. In 2 cases, a suspicion of amoebic liver abscesses was confirmed by detection of E. histolytica DNA in stored sample material. Microscopy alone is clearly insufficient for the detection of E. histolytica in a setting where this parasite is rare. Microscopy-positive stool samples should be further tested by species-specific tests to distinguish E. histolytica from the non-pathogenic parasite E. dispar. On specific suspicion of amoebiasis, such as the suspicion of amoebic liver abscesses, species-specific tests can be applied even after storage.

Efficacy and safety of faecal microbiota transplantation in patients with psoriatic arthritis: protocol for a 6-month, double-blind, randomised, placebo-controlled trial

Introduction An unbalanced intestinal microbiota may mediate activation of the inflammatory pathways seen in psoriatic arthritis (PsA). A randomised, placebo-controlled trial of faecal microbiota transplantation (FMT) infused into the small intestine of patients with PsA with active peripheral disease who are non-responsive to methotrexate (MTX) treatment will be conducted. The objective is to explore clinical aspects associated with FMT performed in patients with PsA. Methods and analysis This trial is a randomised, two-centre stratified, double-blind (patient, care provider and outcome assessor), placebo-controlled, parallel-group study. Eighty patients will be included and randomised (1:1) to either placebo (saline) or FMT provided from an anonymous healthy donor. Throughout the study, both groups will continue the weekly self-administered subcutaneous MTX treatment, remaining on the preinclusion dosage (15–25 mg/week). The clinical measures of psoriasis and PsA disease activity used include the Short (2-page) Health Assessment Questionnaire, the Dermatology Quality of Life Index, the Spondyloarthritis Research Consortium of Canada Enthesitis Index, the Psoriasis Area Severity Index, a dactylitis digit count, a swollen/tender joint count (66/68), plasma C reactive protein as well as visual analogue scales for pain, fatigue and patient and physician global assessments. The primary end point is the proportion of patients who experience treatment failure during the 6-month trial period. The number of adverse events will be registered throughout the study. Ethics and dissemination This is a proof-of-concept clinical trial and will be performed in agreement with Good Clinical Practice standards. Approvals have been obtained from the local Ethics Committee (DK-S-20150080) and the Danish Data Protection Agency (15/41684). The study has commenced in May 2017. Dissemination will be through presentations at national and international conferences and through publications in international peer-reviewed journal(s). Trial registration number NCT03058900; Pre-results.