Alice Friis-Møller

Elevation of Francisella philomiragia subsp. noatunensis Mikalsen et al. (2007) to Francisella noatunensis comb. nov. [syn. Francisella piscicida Ottem et al. (2008) syn. nov.] and characterization of Francisella noatunensis subsp. orientalis subsp. nov., two important fish pathogens.

Aims:  This study was conducted to clarify the taxonomic status of Francisella sp. strain Ehime‐1, a fish pathogen, in relation to the fish pathogens F. piscicida and F. philomiragia subsp. noatunensis and to F. philomiragia subsp. philomiragia.

Elevation of Francisella philomiragia subsp. noatunensis to Francisella noatunensis comb. nov. [syn. Francisella piscicida Ottem et al. (2008) syn. nov.] and characterization of Francisella noatunensis subsp. orientalis subsp. nov., two important fish pathogensMikalsen et al. (2007)

Aims:  This study was conducted to clarify the taxonomic status of Francisella sp. strain Ehime‐1, a fish pathogen, in relation to the fish pathogens F. piscicida and F. philomiragia subsp. noatunensis and to F. philomiragia subsp. philomiragia.

A randomized, double-blind, placebo-controlled multicenter trial evaluating topical zinc oxide for acute open wounds following pilonidal disease excision.

The purpose of this randomized, double‐blind, placebo‐controlled multicenter trial was to compare topical zinc oxide with placebo mesh on secondary healing pilonidal wounds. Sixty‐four (53 men) consecutive patients, aged 17–60 years, were centrally randomized to either treatment with 3% zinc oxide (n=33) or placebo (n=31) by concealed allocation. Patients were followed with strict recording of beneficial and harmful effects including masked assessment of time to complete wound closure. Analysis was carried out on an intention‐to‐treat basis. Median healing times were 54 days (interquartile range 42–71 days) for the zinc and 62 days (55–82 days) for the placebo group (p=0.32). Topical zinc oxide increased (p<0.001) wound fluid zinc levels to 1,540 (1,035–2,265) μM and decreased (p<0.05) the occurrence of Staphylococcus aureus in wounds. Fewer zinc oxide (n=3) than placebo‐treated patients (n=12) were prescribed postoperative antibiotics (p=0.005). Serum‐zinc levels increased (p<0.001) postoperatively in both groups but did not differ significantly between the two groups on day 7. Zinc oxide was not associated with increased pain by the visual analog scale, cellular abnormalities by histopathological examination of wound biopsies, or other harmful effects. Larger clinical trials will be required to show definitive effects of topical zinc oxide on wound healing and infection.

Prevalence and molecular characterization of clinical isolates of Escherichia coli expressing an AmpC phenotype

OBJECTIVES To establish the prevalence of the AmpC beta-lactamase phenotype in clinical isolates of Escherichia coli and characterize the genetic resistance mechanisms causing the observed phenotype. METHODS Clinical E. coli (n = 74) with reduced susceptibility to third-generation cephalosporins and resistance to cefoxitin were collected from the Department of Clinical Microbiology at Hvidovre Hospital, Denmark, in 2006. The AmpC disc test was used to confirm expression of AmpC, and test-positive strains were selected for further antimicrobial susceptibility testing and molecular characterization. Hyperproduction of AmpC beta-lactamase was confirmed by isoelectric focusing (IEF). The presence of a plasmid-mediated ampC gene (pAmpC) was detected by multiplex PCR. The promoter and the entire reading frame of the chromosomal ampC gene were sequenced to identify promoter mutations associated with hyperproduction and gene mutations associated with extended-spectrum AmpC (ESAC) beta-lactamase activity. RESULTS Twenty-four isolates exhibited a positive AmpC disc test. IEF confirmed AmpC expression in all isolates except one. Four isolates contained a bla(CMY-2) gene. These were not clonally related by multilocus sequence typing (MLST). The remaining isolates all had mutations or insertions in the promoter region, which could explain increased expression of the chromosomal AmpC enzyme. Mutations in the ampC gene associated with extended activity were rare and did not cause resistance to cefepime. Sequencing of ampC showed that most isolates were not clonally related. CONCLUSIONS E. coli expressing an AmpC phenotype occur sporadically and cause significant resistance to cephalosporins. The majority of these are hyperproducing chromosomal ampC although some isolates have acquired pAmpC.

Active ulcerative colitis associated with low prevalence of Blastocystis and Dientamoeba fragilis infection

Department of Microbiology and Infection Control, Statens Serum Institut, Copenhagen, Denmark, Department of Gastroenterology, Hvidovre University Hospital, Copenhagen, Denmark, Department of Clinical Microbiology, Slagelse Hospital, Slagelse, Denmark, Department of Clinical Microbiology, Hvidovre University Hospital, Copenhagen, Denmark, and National Food Institute, Technical University of Denmark, Copenhagen, Denmark

The prevalence and clinical significance of intestinal parasites in HIV-infected patients in Denmark

Abstract To investigate the prevalence and clinical significance of intestinal parasites in human immunodeficiency virus (HIV)-infected patients, faecal specimens from 96 HIV-infected patients were submitted to microbiological analyses, including microscopy and polymerase chain reaction for protozoa and enteropathogenic bacteria. Results of microbiological analyses were compared with self-reported gastrointestinal complaints collected using a validated questionnaire. Thirty-two (33%) patients were positive for parasites. However, opportunistic parasites (Isospora and Cryptosporidium) were detected in only 2 instances. Entamoeba dispar was detected in 10 cases, 9 of which represented men who have sex with men (MSM). Despite generally low HIV RNA loads and high CD4+ T-cell counts, 42% of the 76 patients reporting symptoms complained of diarrhoea, 31% of whom were parasite-positive. The presence of diarrhoea was not associated with the presence or absence of parasites; neither was it associated with receiving highly active anti-retroviral therapy (HAART) in general, or protease inhibitors (PI) in particular. A CD4+ T-cell count <200 cells/mm3 was not associated with parasitic infection or with diarrhoea. The data show that diarrhoea is a common symptom among HIV-infected patients in Denmark, but do not indicate that the diarrhoea is due to intestinal parasites.

Bedside diagnosis of imported malaria using the Binax Now malaria antigen detection test

Malaria may be misdiagnosed in non-endemic countries when the necessary experience for rapid expert microscopy is lacking. Rapid diagnostic tests may improve the diagnosis and may play a role as a bedside diagnostic tool. In a multicentre study we recruited patients suspected of malaria over a period of 14 months. The Binax Now Malaria rapid test was used at the bedside and in the clinical microbiology laboratory. The training of clinical staff was monitored and their experience with the use of the test was recorded. 542 patients were included, 80 of whom had malaria diagnosed by microscopy. The rapid test used at the bedside had a sensitivity of 88% for the detection of P. falciparum compared to 95% when the test was performed in the microbiology laboratory. The risk of technical problems and invalid tests was highest when the test was used at the bedside. The rapid diagnostic test may be useful for the diagnosis of P. falciparum malaria when used by routine laboratory staff, but could lead to misdiagnoses when used at the bedside. Microscopy is still essential in order to identify the few missed diagnoses, to determine the degree of parasitaemia, and to ensure species diagnosis, including mixed infections.

Bacteriological aetiology and antimicrobial treatment of pleural empyema.

Abstract Background: Our aims were to describe the aetiologies of culture-positive pleural infections and to evaluate the choice of empiric antimicrobial treatment regimens according to antimicrobial sensitivity, and to evaluate the possible influence of this on outcome. Methods: All cases over a 9-y period were identified from 3 hospitals using the laboratory databases of the clinical microbiology departments, and were verified by evaluating the medical records. Results: We identified 291 isolates in pleural fluid cultures from 158 patients. These included viridans streptococci (25%), Staphylococcus aureus (18%), anaerobic bacteria (17%), Enterobacteriaceae (12%), Staphylococcus epidermidis (10%), and Streptococcus pneumoniae (7%), with differences between nosocomial and community-acquired infections. The mortality (overall 27%) was highest among the patients with Enterobacteriaceae (50%) and S. aureus (36%) infections, and in patients with mixed infections (34%). The actual empiric treatment or the recommended penicillin plus metronidazole had low antimicrobial coverage (49%) compared to the proposed cefuroxime plus metronidazole (78%). Thoracentesis was often delayed (median 2 days). The adequacy of empiric antimicrobial therapy was independently correlated with mortality (odds ratio 0.43, 95% confidence interval 0.30–0.62). Conclusions: The early diagnosis of pleural infection could be optimized. In this North-European patient population, we suggest that the recommended empiric antimicrobial treatment be changed to cefuroxime plus metronidazole for community-acquired and nosocomial infections.

Bacteriology in acute exacerbation of chronic obstructive pulmonary disease in patients admitted to hospital

We investigated the bacterial flora and antimicrobial sensitivity in sputum from patients admitted to hospital with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) in order to recommend the best empirical treatment for these patients. The survey was a retrospective study of all patients admitted to Hvidovre hospital with the diagnosis AECOPD during 2004. A total of 118 patients were included. Microscopy, culture and sensitivity testing investigated their sputums. Clinical and paraclinical features were collected from the patients’ files. Among the 118 patients, 59 (50%) had a positive sputum culture. The group with positive cultures had a higher blood neutrophil count (p=0.03). There were no other clinical differences between the groups with and without positive sputum culture. The majority of patients with positive sputum cultures were infected with bacteria other than S. pneumonia, mostly H. influenzae and Moraxella catarrhalis. Patients with low FEV1<1.0 litre were more likely to be infected with Pseudomonas aeruginosa (p=0.03). The sensitivity patterns of the bacteria showed that the majority were resistant to penicillin. If antibiotics are initiated empirically for AECOPD we recommend either cefuroxime for intravenous treatment or amoxicillin-clavulanate for oral treatment.

Problems in Identification of Francisella philomiragia Associated with Fatal Bacteremia in a Patient with Chronic Granulomatous Disease

ABSTRACT Francisella philomiragia is a rare gram-negative, halophilic coccobacillus with bizarre spherical forms on primary isolation. A case of F. philomiragia bacteremia in a 24-year-old patient with chronic granulomatous disease is reported. Identification of F. philomiragia was problematic with conventional tests but was done correctly and rapidly by kit 16S ribosomal DNA sequencing.