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Background: Severe sepsis and septic shock have a high 30-day mortality (10-50%), but the long-term mortality is not well described. The purpose of this study was to describe long-term mortality among patients with community-acquired severe sepsis or septic shock compared to a population-based reference cohort. Methods: Two hundred and twelve patients who, within the first 24 h after arrival at the hospital, presented with infection and had failure of at least 1 organ system were included. A population-based reference group of 79,857 patients was identified, and data on comorbidities were extracted from the National Danish Patient Registry. We analyzed the hazard ratio for mortality at predefined intervals. Results: Absolute mortality within the first 30 days was 69/211 (33%, 95% confidence interval (CI) 25-41%), with a cumulative mortality of 121/211 (57%, 95% CI 48-69%) for the entire follow-up. Among septic patients who survived the first 30 days, the mortality hazard ratio was 2.7 (95% CI 1.7-4.3) until day 365, and among septic patients who survived the first year, the 1-4 y mortality hazard ratio was 2.3 (95% CI 1.7-3.3), compared to the community-based reference persons (multivariate Cox regression controlling for age, sex, and Charlson comorbidity index). Conclusions: Patients with severe sepsis and septic shock who survived the first 30 days had a 2.7 times higher mortality hazard in the first year and a 2.3 times higher mortality hazard in the next 3 y, compared to persons of similar age, sex, and comorbidity level.

This study compared Neo-Sensitabs with Oxoid paper disks using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) disk diffusion antimicrobial susceptibility test on Mueller-Hinton agar. The EUCAST-recommended quality control strains (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212) (Part I) and clinical isolates (Part II) were investigated. In Part I of the study, 27 combinations of antimicrobial agents were tested on four quality control strains repeatedly up to 60 times and zone diameters of tablets and disks were compared. In Part II of the study, 351 clinical isolates were included to cover a broad range of species, as well as resistance mechanisms. In Part I, four major deviations (>1 mm outside quality control ranges) were observed with Neo-Sensitabs. In one case with P. aeruginosa ATCC 27853 (meropenem), there was a corresponding major deviation (2 mm) with the Oxoid disk. The three remaining major deviations with Neo-Sensitabs were observed with meropenem (2 mm) in E. coli ATCC 25922 and with ciprofloxacin (2 mm) and gentamicin (3 mm) in P. aeruginosa ATCC 27853. For Oxoid disks, there were only minor deviations (=1 mm outside quality control ranges) in these three cases. In Part II, there were six discrepancies, susceptible versus resistant, in 3,533 comparisons between the two methods with the clinical isolates. The Rosco Neo-Sensitabs appear to be a possible alternative to Oxoid paper disks for EUCAST disk diffusion antimicrobial susceptibility testing on Mueller-Hinton agar.

Background: In 2008, a set of 41 quality indicators for antibiotic treatment of respiratory tract infections (RTIs) in general practice were developed in an international setting as part of the European project HAPPY AUDIT. Objectives: To investigate Danish general practitioners' (GPs') assessment of a set of internationally developed quality indicators and to explore if there is an association between the GPs' assessment of the indicators and their practice characteristics as well as their antibiotic prescription pattern. Methods: A total of 102 Danish GPs were invited to assess the 41 quality indicators. The GPs were categorized into two groups according to their assessment of indicators. Data concerning practice characteristics and antibiotic treatment were obtained during a three-week registration of patients with RTIs and were linked to the GPs' assessments of the indicators. Results: A total of 62 (61%) responded. Quality indicators focusing on the frequency of prescribing of narrow-spectrum penicillin were rated as suitable by more than 80% of the Danish GPs, while quality indicators concerning cephalosporins or quinolones were rated suitable by less than half of the GPs. The antibiotic prescribing pattern differed significantly and the GPs who disagreed on most indicators prescribed more macrolides and less narrow-spectrum penicillin than the GPs who agreed on most indicators. Conclusion: Even though an international expert panel agreed on a set of quality indicators for antibiotic treatment of RTIs, only a few of them were rated suitable by the GPs, who are supposed to use them.

Accurate and fast yeast identification is important when treating patients with invasive fungal disease as susceptibility to antifungal agents is highly species related. Matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF-MS) provides a powerful tool with a clear potential to improve current diagnostic practice. Two MALDI-TOF-MS-systems (BioTyper/Bruker and Saramis/AXIMA) were evaluated using: (i) A collection of 102 archived, well characterised yeast isolates representing 14 different species and (ii) Prospectively collected isolates obtained from clinical samples at two participating laboratories. Of the 102 archived isolates, 81 (79%) and 92 (90%) were correctly identified by Saramis/AXIMA and BioTyper/Bruker respectively. Saramis/AXIMA was unable to separate Candida albicans, C. africana and C. dubliniensis in 13 of 32 isolates. After manual interpretation of the mass spectra output, all 13 isolates were correctly identified, resulting in an overall identification performance of 92%. No misidentifications occurred with the two systems. Of the routine isolates one laboratory identified 99/99 (100%) and 90/99 (91%) to species level by Saramis/Axima and conventional identification, respectively, whereas the other laboratory identified 83/98 (85%) to species level by both BioTyper/Bruker and conventional identification. Both MALDI-TOF-MS systems are fast, have built-in databases that cover the majority of clinically relevant Candida species, and have an accuracy that outperforms our conventional identification systems.

Recommendations for antibiotic treatment of acute otitis media (AOM) have changed over the years, and today many experts recommend initial observation. However, antibiotic prescribing should be considered in children aged2 years or if AOM is accompanied by discharging ear.To investigate the quality of treatment of AOM in general practice and to explore the influence of selected GP and patient characteristics on antibiotic prescribing.During the winter 2008, a prospective registration of patients diagnosed with AOM was conducted in general practice in Lithuania, Kaliningrad, Spain, Argentina, Sweden and Denmark. Some 1175 patients diagnosed with AOM were registered. Information about age and sex of the patient, duration of symptoms (days), temperature38.5°C, ear discharge and the antibiotic treatment given was recorded.Danish GPs had the lowest antibiotic prescription rate for AOM [72.7% (95% confidence interval (CI) = 67.0-77.8)] and GPs in Kaliningrad had the highest [97.1% (95% CI = 89.8-99.6)]. Narrow-spectrum penicillin was almost exclusively prescribed in the two Nordic countries, while broad-spectrum penicillins, often in combination with clavulanic acid, were prescribed in the other four countries. Macrolides comprised 5-10% of prescriptions. Antibiotic prescribing was associated with the following characteristics of the patients: symptoms for3 days, ear discharge and fever.The majority of patients with AOM were treated with antibiotics in all six countries, but considerable variations in both prescribing rate and choice of antibiotics were identified.

A collection of 138 consecutive isolates from blood primarily identified as Gram-positive, cluster-forming, coagulase-negative cocci was examined by a conventional routine method for identification of clinical isolates of coagulase-negative Staphylococcus and Micrococcus species. The method was based on selected reactions from the Kloos & Schleifer scheme, utilizing the conventional media of Statens Seruminstitut. Double determinations for each isolate were performed by the conventional method. The results were compared with speciation by the commercial micromethods API-Staph and API-Staph-Ident. For control, 31 Staphylococcus and 13 Micrococcus reference strains were included. Of the 31 Staphylococcus spp. (reference strains), the conventional system, API-Staph, and API-Staph-Ident correctly identified 87%, 87% and 81%, respectively. Micrococcus spp. were only identified to genus level by the conventional method as well as by API-Staph. API-Staph-Ident is not designed for Micrococcus identification. Of 138 blood isolates, 121 belonged to the genus Staphylococcus while 17 were Micrococcus spp. S. epidermidis dominated with all three methods, constituting approx. 35% of the isolates tested. In only 57% of the isolates identification by all three methods agreed. The three methods were unable to put a name on 7.5% (conventional method), 10.7% (API-Staph) and 2.5% (API-Staph-Ident) of the isolates. Reproducibility was high with the conventional method (100% for the reference strains and 91% for blood culture isolates) as well as with API-Staph and API-Staph-Ident (88%/81% and 81%/81%, respectively). We concluded that our conventional system was able to identify most clinically significant staphylococcal species by means of relatively few tests with a high certainty and a high degree of reproducibility.

Population analyses of susceptibility to cefotaxime (CTX) and desacetyl-cefotaxime (DCTX) of strains of Staphylococcus and some genera of Enterobacteriaceae were carried out. DCTX, which is the main metabolite of CTX, has antimicrobial activity. The penicillinase-producing strains of S. aureus and S. epidermidis were homogeneous as regards susceptibility to both agents. CTX was about 4-8 times more active than DCTX. The methicillin-resistant strains contained a sub-population of resistant bacteria with both CTX and DCTX. The frequency of resistant bacteria was 10(-6) - 10(-5). The E. coli strain was homogeneous to both agents. The strains of Enterobacter cloacae and Citrobacter freundii had a sub-population of resistant bacteria with both agents. The frequency of resistant bacteria was 10(-7) - 10(-4.5). In Klebsiella pneumoniae no resistant sub-population was found. CTX was about four times more active than DCTX with the strains of the Enterobacteriaceae. DCTX had no advantage over CTX as regards homogeneity of susceptibility of the populations examined. CTX seems applicable for treatment of infections with E. coli, Klebsiella pneumoniae, and penicillinase-producing, methicillin-susceptible Staphylococcus, but should not be used alone in treatments of infections with Enterobacter cloacae or Citrobacter freundii.

Coagulase-negative Micrococcaceae from blood cultures were classified biochemically according to Baird-Parker and to Kloos & Schleifer and by means of antibiotic susceptibility testing, in an attempt to distinguish between bacterial growth due to contamination and growth due to bacteremia. S. epidermidis biotype 1 (according to Baird-Parker) accounted for c. 60% of the isolates and for c. 85% of the isolates considered to be of clinical importance. The more time- and resource-demanding classification of Kloos & Schleifer provided no further clinically useful information, as the predominant number of isolates turned out to be S. epidermidis. Isolates other than S. epidermidis (according to Kloos & Schleifer) accounted for c. 45% of the total number of isolates but for only 20% of the isolates considered of clinical importance. Resistance to multiple antibiotics was recorded more often in isolates from patients with positive clinical information than in isolates thought to be contaminators. The frequency of blood cultures contaminated with coagulase-negative Micrococcaceae was estimated at c. 1.5%. Records from 91 patients were reviewed. By correlating the clinical findings to biochemical classification, resistance types, and the massiveness of growth it was suggested that the quantitative rather than the qualitative findings are suitable for determining clinical significance of isolates of coagulase-negative Micrococcaceae from blood cultures.

A total of 965 strains of Micrococcaceae isolated from 200 patients, personnel, and air in a department of cardiac surgery were classified by means of Baird-Parker's scheme. The majority of strains were identified as Staphylococcus epidermidis (S. epidermidis) biotype 1, but S. epidermidis biotype 4 accounted for c. 25% of isolates from patients post-operatively. Pre-operative isolates were generally sensitive to most antibiotics tested while post-operative strains of coagulase-negative Micrococcaceae from patients and isolates from personnel and air were frequently multiply-resistant. Strains of Staphylococcus aureus (S. aureus) were sensitive or resistant only to penicillin. More patients were colonized with coagulase-negative Micrococcaceae after operation than at admission to the hospital (p less than 0.001), while the frequency of S. aureus carriers was the same before and after operation and equal to the frequency found earlier. The frequency of S. aureus carriers among the personnel, however, was lower than reported earlier (10%). Multiply-resistant strains of S. epidermidis seem to have replaced resistant strains of S. aureus as the predominant hospital saprophyte among Micrococcaceae.

Population analyses of susceptibility to methicillin, vancomycin, cephalothin, cefamandole, and cefoxitin were carried out with two strains of Staphylococcus epidermidis (S. epidermidis) resistant to penicillin, streptomycin, and methicillin. Two strains of Staphylococcus aureus (S. aureus), one sensitive to penicillin the other resistant, were used as controls. All strains were clinical isolates. From the strains of S. epidermidis it was possible to select sub-populations highly resistant to methicillin, cephalothin, cefamandole, and cefoxitin. The mutation-rate varied between c. 10(-3) and 10(-7). The two strains were homogeneous as regards vancomycin susceptibility. Methicillin was shown to be more penicillinase-stable than the cephalosporines. Treatment in vivo, with cephalothin and vancomycin did not increase the frequency of highly resistant bacteria against any of the antibiotics, but during treatment with cephalothin the majority-population became eight times more resistant to vancomycin and may explain why the treatment in vivo was not successful.

Udgivelsesdato: May A semi-national laboratory-based surveillance programme for fungaemia was initiated in 2003 that now covers c. 3.5 million inhabitants (64%) of the Danish population. In total, 1089 episodes of fungaemia were recorded during 2004-2006, corresponding to an annual incidence of 10.4/100 000 inhabitants. The annual number of episodes increased by 17% during the study period. Candida spp. accounted for 98% of the fungal pathogens. Although Candida albicans remained predominant, the proportion of C. albicans decreased from 66.1% in 2004 to 53.8% in 2006 (p 1 mg/L, was detected in one (2.5%) C. glabrata isolate in 2004 and in 12 (14.0%) isolates in 2006 (p 0.03). Overall, the proportion of isolates with decreased susceptibility to fluconazole exceeded 30% in 2006. The incidence of fungaemia in Denmark was three-fold higher than that reported from other Nordic countries and is increasing. Decreased susceptibility to fluconazole is frequent, and a new trend towards C. glabrata isolates with elevated voriconazole MICs was observed.

Antecedentes En atencion primaria, el tratamiento de las infecciones del tracto respiratorio (ITR) es empirico. Los patrones de prescripcion de antibioticos son distintos entre paises y vienen dados por la prevalencia de resistencias antimicrobianas existentes. El objetivo de este estudio fue comparar que antibioticos prescriben los medicos de atencion primaria en las ITR en Espana y Dinamarca. Metodos Estudio observacional multicentico efectuado en atencion primaria de salud. Dos grupos de medicos de atencion primaria de Espana y de Dinamarca registraron todas las visitas con ITR durante un periodo de 3 semanas entre noviembre de 2001 y enero de 2002. Resultados Se registraron un total de 2.833 casos de ITR. Los antibioticos mas prescritos por los medicos espanoles fueron las penicilinas de amplio espectro, asociadas o no a inhibidores de betalactamasas (62,3%), seguido de los macrolidos (22,3%), mientras que en el caso de los daneses fueron las penicilinas de espectro reducido (58% de todos los antibioticos prescritos), seguido de los macrolidos (29%; p Conclusiones Las importantes diferencias existentes en el tratamiento antibiotico de las ITR entre los medicos participantes debe hacernos reflexionar sobre la utilizacion racional de los antibioticos. Esto podria ser producto de tradiciones, habitos, recomendaciones y presiones distintas entre ambos paises.

BACKGROUND: Hemovigilance has shown that bacteria cause more fatalities than other infections together. Surveillance for detection of bacteria in platelets (PLTs) was initiated. Concomitantly, the storage period for PLTs was extended from 5 to 7 days to reduce cost. STUDY DESIGN AND METHODS: Analysis was performed of all cases of a positive signal in a screening procedure for contaminated PLTs taking into account results of confirmative cultures and results of culture from blood components including bacteria strains. Records were assessed from patients transfused with blood components issued before the screening culture became positive. RESULTS: Samples were collected from 22,057 PLT units. An initial reaction was seen in 84 (0.38%). Growth was confirmed in 70 of these. Of the associated PLT units, 26 had been issued or outdated at the time when the culture was found to be reactive, in 27 bacteria were found, and in 17 cultures were negative. The bacteria found were mainly from normal skin flora. Sixty-six patients received 75 blood components issued before the screening system alarmed. None of these patients had a transfusion reaction reported. The outdating fell to less than 5 percent. CONCLUSION: A screening system for detection of bacterial contamination was implemented without increase in cost owing to extension of storage time to 7 days. Transfusion of several contaminated blood components was prevented.

Udgivelsesdato: 2010-Apr Abstract Carriage of pig-associated methicillin-resistant Staphylococcus aureus (MRSA) is known to occur in pig farmers. Zoonotic lineages of MRSA have been considered of low virulence and with limited capacity for inter-human spread. We present a case of family transmission of pig-associated MRSA ST398, which resulted in a severe infection in a newborn.

Our experience from attempts to identify bacteria isolated from boar bite/gore wounds is the background for a discussion of identification problems. Some organisms, although not very common or well-known, can be identified when using commercial kits or conventional methods, provided they are sufficiently characterized, as exemplified by Pasteurella aerogenes isolated from cases 1 and 2. Some organisms may be wrongly identified, or not identified, by both commercial kits and conventional methods, unless seen by experienced microbiologists with knowledge of the original literature. This is exemplified by case 3, in which the final identification result was Bisgaard's taxon 15. Sometimes isolates cannot be identified even in reference laboratories and by using available identification tables and databases. In such cases, the organism involved may turn out to belong to a previously undescribed taxon. This is illustrated by the strains isolated from cases 4 and 5.

Dysgonomonas capnocytophagoides, formerly known as CDC group DF-3, is an opportunistic pathogen associated with diarrhoea and very rarely bacteraemia. We report a case of D. capnocytophagoides found in blood cultures from a severely neutropenic patient treated for acute myeloid leukaemia. The isolate was found resistant to penicillin, cephalosporins, meropenem, aminoglycosides and ciprofloxacin, and susceptible to ampicillin, tetracycline, chloramphenicol, clindamycin and trimethoprim-sulphamethoxazole. It was identified using conventional phenotypic testing but remained unidentified by the automated identification system (Vitek-2) as this system did not contain DF-3 or D. capnocytophagoides in its database.

Thirty-nine Danish cases of Capnocytophaga canimorsus septicemia were reviewed to determine the clinical course of this infection. The cases of septicemia were related to recent dog bites or other close contact with dogs. The period from the bite to the onset of symptoms ranged from 1 to 8 days. The mean age of the patients was 59.1 years (range, 28-83 years). Underlying conditions included previous splenectomy and alcoholism. Thirteen patients had previously been in good health. Common initial symptoms were fever, malaise, myalgia, vomiting, diarrhea, abdominal pain, dyspnea, confusion, headache and skin manifestations. Disseminated intravascular coagulation developed in 14 patients, meningitis in 5, and endocarditis in 1. Twelve of the patients died. All patients except two were treated with penicillin or ampicillin. Five patients had received antibiotics prior to admission. Attention should be drawn to C. canimorsus septicemia in cases of febrile illness following dog bites or contact with dogs, as well as those involving previously healthy persons. The incidence of this condition in Denmark is estimated to be 0.5 case per 1 million people per year.

Pasteurella aerogenes is rarely isolated from human specimens. The species is found in the digestive tract of pigs. From 1976 to 1994 7 strains were cultured in Denmark from wounds or ulcers. Five patients were bitten by pigs and 2 patients with ulcers were employed in pig farming. A mixture of bacterial species was often found. All 7 strains of P. aerogenes were susceptible to ampicillin, cephalosporins and ciprofloxacin. Ability to hydrolyse urea, to produce oxidase and catalase, to decarboxylate ornithine and to produce gas from glucose and inability to produce indole was characteristic for P. aerogenes. Most bite wounds were located on the lower lateral part of the thigh. Foul smelling pus and abscess formation was the rule. Incision, drainage and antibiotic treatment were usually necessary.

Background Influenza viruses such as swine-origin influenza A(H1N1) virus (A(H1N1)pdm09) generate genetic diversity due to the high error rate of their RNA polymerase, often resulting in mixed genotype populations (intra-host variants) within a single infection. This variation helps influenza to rapidly respond to selection pressures, such as those imposed by the immunological host response and antiviral therapy. We have applied deep sequencing to characterize influenza intra-host variation in a transmission chain consisting of three cases due to oseltamivir-sensitive viruses, and one derived oseltamivir-resistant case. Methods Following detection of the A(H1N1)pdm09 infections, we deep-sequenced the complete NA gene from two of the oseltamivir-sensitive virus-infected cases, and all eight gene segments of the viruses causing the remaining two cases. Results No evidence for the resistance-causing mutation (resulting in NA H275Y substitution) was observed in the oseltamivir-sensitive cases. Furthermore, deep sequencing revealed a subpopulation of oseltamivir-sensitive viruses in the case carrying resistant viruses. We detected higher levels of intra-host variation in the case carrying oseltamivir-resistant viruses than in those infected with oseltamivir-sensitive viruses. Conclusions Oseltamivir-resistance was only detected after prophylaxis with oseltamivir, suggesting that the mutation was selected for as a result of antiviral intervention. The persisting oseltamivir-sensitive virus population in the case carrying resistant viruses suggests either that a small proportion survive the treatment, or that the oseltamivir-sensitive virus rapidly re-establishes itself in the virus population after the bottleneck. Moreover, the increased intra-host variation in the oseltamivir-resistant case is consistent with the hypothesis that the population diversity of a RNA virus can increase rapidly following a population bottleneck.

Seventy-eight patients with culture-positive epidural catheters, were studied. Fifty-nine had symptoms of exit site infection and 11 patients had clinical meningitis, two of whom also had an epidural abscess. This corresponds to a local infection incidence of at least 4.3% and an incidence of central nervous system infection of at least 0.7% at Odense University Hospital. This degree of infection is of the same magnitude as that reported for intravascular devices. We found that the patients with generalized symptoms of infection had been catheterized for a longer time, and were older than patients with only local symptoms of infection. The microorganisms isolated from the tips of the epidural catheters were coagulase-negative staphylococci (41%), Staphylococcus aureus (35%), Gram-negative bacilli (14%) and others (10%). The Gram-negative bacilli and S. aureus caused serious infections more frequently than the others. We discuss the symptoms and diagnosis of spinal epidural abscess and suggest a proposal for prophylactic and diagnostic guidelines for epidural catheter-related infections.

Accurate and fast yeast identification is important when treating patients with invasive fungal disease as susceptibility to antifungal agents is highly species related. Matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF-MS) provides a powerful tool with a clear potential to improve current diagnostic practice. Two MALDI-TOF-MS-systems (BioTyper/Bruker and Saramis/AXIMA) were evaluated using: (i) A collection of 102 archived, well characterised yeast isolates representing 14 different species and (ii) Prospectively collected isolates obtained from clinical samples at two participating laboratories. Of the 102 archived isolates, 81 (79%) and 92 (90%) were correctly identified by Saramis/AXIMA and BioTyper/Bruker respectively. Saramis/AXIMA was unable to separate Candida albicans, C. africana and C. dubliniensis in 13 of 32 isolates. After manual interpretation of the mass spectra output, all 13 isolates were correctly identified, resulting in an overall identification performance of 92%. No misidentifications occurred with the two systems. Of the routine isolates one laboratory identified 99/99 (100%) and 90/99 (91%) to species level by Saramis/Axima and conventional identification, respectively, whereas the other laboratory identified 83/98 (85%) to species level by both BioTyper/Bruker and conventional identification. Both MALDI-TOF-MS systems are fast, have built-in databases that cover the majority of clinically relevant Candida species, and have an accuracy that outperforms our conventional identification systems.

Methods Patients with a first time discharge diagnosis of sepsis (IDC10 code A40.0 – A41.9) were identified from the Funen Patient Administrative System. Clinical details were evaluated manually and patients who within the first 24 hours after arrival to the hospital presented with infection and had failure of minimum one organ system were included. A population based reference group on 79,857 patients were identified. Data on co-morbidity were obtained from the National Danish Patient Registry.

A prerequisite for rational antibiotic treatment is the suspicion that the disease is caused by bacteria and that treatment will reduce symptoms and complications. In primary health care most infections are caused by bacteria. The European project HAPPY AUDIT found that an intervention targeting doctors and patients in primary care led to a considerable decrease in antibiotic prescribing. Teaching of hospital doctors, combined with a restriction in the choice of antibiotics, resulted in a decrease in multidrug-resistant strains. A concerted effort is needed to curb the growing resistance problem.

A three-month laboratory-based prospective survey was conducted at four major university hospitals covering one-third of the Danish population in order to determine the prevalence, significance, and susceptibility pattern of aspergilli in airway samples. Samples received in January-March 2007 for routine microbiologic investigation were examined for Aspergillus following routine procedures and with extended incubation (5 days). Identification was done by morphologic criteria and susceptibility testing using EUCAST method for azoles and amphotericin B E-test. Invasive aspergillosis (IA) was evaluated using modified EORTC/MSG criteria. A total of 11,368 airway samples were received. Growth of Aspergillus spp. was found in 129 and 151 patients using routine and extended incubation, respectively. Three patients had proven IA (2%), 11 probable (7%), four had allergic bronchopulmonary aspergillosis (ABPA) (3%), but the majority was colonised (88%). Underlying conditions were cystic fibrosis in 82 patients (55%), chronic obstructive pulmonary disease in 19 (13%) and haematological disorder in 11 (7%). Twenty-six patients (18%) were at intensive care unit and 69 (47%) received steroid treatment. Azole MICs were elevated for five isolates as follows (itraconazole, posaconazole, voriconazole MICs [mg/L]): two A. fumigatus isolates (>4; >4; 2 and >4; 0.125; 1), one A. lentulus isolate (2; 2; 0.5) and two A. terreus isolates (2; 2; 2 and 2; 0.125; 1). For four isolates the amphotericin B MIC was >1 μg/ml (3/112 A. fumigatus, 1/2 A. terreus). In conclusion, Aspergillus appears to be an important pathogen in Denmark. Elevated itraconazole MICs were detected in 4% of the isolates including a multi-azole resistant isolate.

Different molecular methods for the discrimination of Candida glabrata, C. bracarensis and C. nivariensis were evaluated and the prevalence of these species among Danish blood isolates investigated. Control strains were used to determine fragment length polymorphism in the ITS1, ITS2, ITS1-5.8S-ITS2 regions and in the D1/D2 domain of 26S rDNA using primers designed for this study. A total of 133 blood isolates previously identified as C. glabrata were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and the peptide nucleic acid-fluorescent in situ hybridization (PNA-FISH) method. The size of ITS1 allowed differentiation between C. glabrata (483), C. nivariensis (361) and C. bracarensis (385), whereas the ITS2 region was of similar size in C. nivariensis (417) and C. glabrata (418). Sequence analysis of the ITS region suggested that many restriction enzymes were suitable for RFLP differentiation of the species. Enzymatic digestion of the D1/D2 domain with TatI produced unique band sizes for each of the three species. PCR-RFLP and PNA-FISH were in agreement for all of the isolates tested. None of the 133 Danish blood isolates were C. nivariensis or C. bracarensis. Fragment size polymorphism of ITS1 and RFLP of the D1/D2 domain or the ITS region are useful methods for the differentiation of the species within the C. glabrata group. C. bracarensis and C. nivariensis are rare among Danish C. glabrata blood isolates.

Background Excessive use of antibiotics is worldwide the most important reason for development of antimicrobial resistance. As antibiotic resistance may spread across borders, high prevalence countries may serve as a source of bacterial resistance for countries with a low prevalence. Therefore, bacterial resistance is an important issue with a potential serious impact on all countries. Initiatives have been taken to improve the quality of antibiotic prescribing in primary care, but only few studies have been designed to determine the effectiveness of multifaceted strategies across countries with different practice setting. The aim of this study was to evaluate the impact of a multifaceted intervention targeting general practitioners (GPs) and patients in six countries with different health organization and different prevalence of antibiotic resistance. Methods GPs from two Nordic countries, two Baltic Countries and two Hispano-American countries registered patients with respiratory tract infections (RTIs) in 2008 and 2009. After first registration they received individual prescriber feedback and they were offered an intervention programme that included training courses, clinical guidelines, posters for waiting rooms, patient brochures and access to point of care tests (Strep A and C-Reactive Protein). Antibiotic prescribing rates were compared before and after the intervention. Results A total of 440 GPs registered 47011 consultations; 24436 before the intervention (2008) and 22575 after the intervention (2009). After the intervention, the GPs significantly reduced the percentage of consultations resulting in an antibiotic prescription. In patients with lower RTI the GPs in Lithuania reduced the prescribing rate by 42%, in Russia by 25%, in Spain by 25%, and in Argentina by 9%. In patients with upper RTIs, the corresponding reductions in the antibiotic prescribing rates were in Lithania 20%, in Russia 15%, in Spain 9%, and in Argentina 5%. Conclusion A multifaceted intervention programme targeting GPs and patients and focusing on improving diagnostic procedures in patients with RTIs may lead to a marked reduction in antibiotic prescribing. The pragmatic before-after design used may suffer from some limitations and the reduction in antibiotic prescribing could be influenced by factors not related to the intervention.

We compared two matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) systems (Shimadzu/SARAMIS and Bruker) on a collection of consecutive clinically important anaerobic bacteria ( n = 290). The Bruker system had more correct identifications to the species level (67.2% versus 49.0%), but also more incorrect identifications (7.9% versus 1.4%). The system databases need to be optimized to increase identification levels. However, MALDI-TOF MS in its present version seems to be a fast and inexpensive method for identification of most clinically important anaerobic bacteria.

Summary On the occasion of five Danish human Pasteurella aerogenes from pig bite lesions, a comparison was made between 6 isolates from man and 15 animal isolates, mainly from pigs. The strains originated from 6 different countries (USA, Canada, Czechoslovakia, France, Belgium and Denmark). The 21 isolates were characterized by conventional biochemical tests, antibiogram and the API 20 NE kit; finally ribotyping was carried out by hybridizing EcoRI -digested chromosomal DNA with a probe derived from E. coli ribosomal RNA. By ribotyping, 19 of the 21 strains clustered at a similarity level of 81% or more; both phenotypical tests and ribotyping indicated that the remaining two strains did not belong to the species P. aerogenes . In conclusion, despite minor differences our P. aerogenes isolates constituted a well-defined group and they could not be subdivided on basis of animal or geographical origin.

Candida orthopsilosis and Candida metapsilosis are recentlydescribed species phenotypically indistinguishable from Candida parapsilosis. We evaluated phenotyping and molecular methods for the detection ofthese species among 79 unique blood culture isolates of the C. parapsilosis group obtained during the years 2004–2008. The isolates were screenedby PCR amplification of the secondary alcohol dehydrogenase-encoding gene (SADH) followed by digestion with the restriction enzyme BanI, using C. parapsilosis ATCC 22019, C. orthopsilosisATCC 96139 and C. metapsilosis ATCC 96144 as controls. Isolates withRFLP patterns distinct from C. parapsilosis were characterized bysequence analysis of the ITS1–ITS2, 26S rRNA (D1/D2) and SADH regions. Restriction patterns for the 3 species with each of 610restriction enzymes were predicted in silico using 12 available sequences.By PCR-RFLP of the SADH gene alone, four isolates (5.1 %)had a pattern identical to the C. orthopsilosis reference strain.Sequence analysis of SADH and ITS (internal transcribed spacer)regions identified two of these isolates as C. metapsilosis. Theseresults were confirmed by creating a phylogenetic tree based on concatenatedsequences of SADH, ITS and 26S rRNA gene sequence regions. Optimaldifferentiation between C. parapsilosis, C. metapsilosisand C. orthopsilosis was predicted using digestion with NlaIII,producing discriminatory band sizes of: 131 and 505 bp; 74, 288 and 348 bp;and 131, 217 and 288 bp, respectively. This was confirmed using the referencestrains and 79 clinical isolates. In conclusion, reliable discrimination wasobtained by PCR-RFLP profile analysis of the SADH gene after digestionwith NlaIII but not with BanI. C. metapsilosisand C. orthopsilosis are involved in a small but significant numberof invasive infections in Denmark.

Background Excessive and inappropriate use of antibiotics is considered to be the most important reason for development of bacterial resistance to antibiotics. As antibiotic resistance may spread across borders, high prevalence countries may serve as a source of bacterial resistance for countries with a low prevalence. Therefore, bacterial resistance is an important issue with a potential serious impact on all countries. The majority of respiratory tract infections (RTIs) are treated in general practice. Most infections are caused by virus and antibiotics are therefore unlikely to have any clinical benefit. Several intervention initiatives have been taken to reduce the inappropriate use of antibiotics in primary health care, but the effectiveness of these interventions is only modest. Only few studies have been designed to determine the effectiveness of multifaceted strategies in countries with different practice setting. The aim of this study is to evaluate the impact of a multifaceted intervention targeting general practitioners (GPs) and patients in six countries with different prevalence of antibiotic resistance: Two Nordic countries (Denmark and Sweden), two Baltic Countries (Lithuania and Kaliningrad-Russia) and two Hispano-American countries (Spain and Argentina). Methods/Design HAPPY AUDIT was initiated in 2008 and the project is still ongoing. The project includes 15 partners from 9 countries. GPs participating in HAPPY AUDIT will be audited by the Audit Project Odense (APO) method. The APO method will be used at a multinational level involving GPs from six countries with different cultural background and different organisation of primary health care. Research on the effect of the intervention will be performed by analysing audit registrations carried out before and after the intervention. The intervention includes training courses on management of RTIs, dissemination of clinical guidelines with recommendations for diagnosis and treatment, posters for the waiting room, brochures to patients and implementation of point of care tests (Strep A and CRP) to be used in the GPs'surgeries. To ensure public awareness of the risk of resistant bacteria, media campaigns targeting both professionals and the public will be developed and the results will be published and widely disseminated at a Working Conference hosted by the World Association of Family Doctors (WONCA-Europe) at the end of the project period. Discussion HAPPY AUDIT is an EU-financed project with the aim of contributing to the battle against antibiotic resistance through quality improvement of GPs' diagnosis and treatment of RTIs through development of intervention programmes targeting GPs, parents of young children and healthy adults. It is hypothesized that the use of multifaceted strategies combining active intervention by GPs will be effective in reducing prescribing of unnecessary antibiotics for RTIs and improving the use of appropriate antibiotics in suspected bacterial infections.

Udgivelsesdato: Mar OBJECTIVE: To develop a set of quality indicators focusing on the diagnosis and treatment of respiratory tract infections in general practice. DESIGN: A modified 2-round Delphi study. SETTING: General practice. SUBJECTS: A panel of 27 experts (13 countries) comprising mainly general practitioners, clinical microbiologists, and clinical pharmacologists were asked to rate the relevance of 59 quality indicators for diagnosis and treatment of respiratory tract infections with regard to reducing antimicrobial resistance and improving patient health. A thorough literature review was carried out to ensure that all potential quality indicators were considered. OUTCOME: Consensus for a quality indicator was reached if > or =75% of experts scored the item > or =5 on a 7-point Likert scale, ranging from 1 (=completely disagree) through 4 (=uncertain) to 7 (=completely agree). RESULTS: A 96% response rate was achieved in both Delphi rounds. A total of 41 of the proposed 59 quality indicators attained consensus. None of the quality indicators focusing on the diagnostic process achieved consensus. Consensus was attained for 14 quality indicators focusing on the decision regarding antibiotic treatment and for 27 quality indicators focusing on the choice of antibiotics. CONCLUSION: This study resulted in a final set of 41 quality indicators concerning respiratory tract infections in general practice. These quality indicators may be used to strengthen general practitioners' focus on their management of patients with respiratory tract infections and to identify where it is possible to make improvements.

Actinomyces pyogenes (Corynebacterium pyogenes), a well-known pathogen in many animals, was isolated from 11 Danish patients since 1968. Bacteriologic characteristics and clinical pictures of the patients are described. Ability to hydrolyze gelatine, to produce beta-glucuronidase, to reach with antisera against group-G streptococci, and to produce acid from xylose differentiates A. pyogenes from Arcanobacterium haemolyticum, with which it has at times been confused. Actinomyces pyogenes is an established, but often misrecognized, human pathogen that should be better known to clinical microbiologists.

Udgivelsesdato: 2009 Objective. To investigate whether short-term treatment with pivmecillinam was more effective than sulfamethizole in patients with acute uncomplicated urinary tract infection (UTI). Design. Randomized controlled trial. Setting. General practice, Denmark. Subjects. Patients (n =167) with uncomplicated UTI confirmed by positive urine phase-contrast microscopy. Main outcome measures. Drug efficacy based on clinical and bacteriological cure. Results. Urinary symptoms disappeared first in patients treated with pivmecillinam, but after five days there was no significant difference in clinical cure rate between the two antibiotics. At the follow-up visit 7-10 days after initiation of treatment, 95.4% of patients treated with pivmecillinam and 92.6% of patients treated with sulfamethizole had no persistent cystitis symptoms (difference 2.8%, CI -4.5%; 10.0%). Bacteriological cure was observed in 68.8% of patients randomized to pivmecillinam and in 77.9% randomized to sulfamethizole (difference -9.2%, CI -24.7%; 6.3%). Some 26.8% of patients randomized to pivmecillinam experienced a new UTI within 6 months after treatment compared with 18.4% of patients randomized to sulfamethizole (difference 8.4%, CI -4.5%;21.4%). No patients developed septicaemia with urinary pathogens within one year after initial treatment. Conclusion. Patients treated with a three-day regime of pivmecillinam experienced faster relief of symptoms compared with patients treated with a three-day regime of sulfamethizole. Five days after initiation of treatment there was no significant difference in clinical and bacteriological cure between the two antibiotic regimes.

The adherence of radiolabeled staphylococci to silicone catheters was investigated in vitro. Staphylococcus aureus and Staphylococcus epidermidis strains bound to the same extent to the catheters. Also, S. epidermidis strains isolated from patients with plastic-related infections showed binding similar to that of other S. epidermidis strains. By preincubation of catheters the influence of purified staphylococcal cell surface components on the binding was evaluated. The most potent inhibitors of the binding of S. aureus were the two surface proteins, clumping factor and protein A, and the cytoplasmic membrane. Surface proteins and the cell membrane of S. epidermidis also blocked the binding. Only protein-containing surface proteins inhibited the binding. The production of slime correlated with the degree of S. epidermidis binding. Human plasma and serum, as well as purified albumin and IgG, inhibited the binding of both staphylococcal species. Fibrinogen, and to a certain extent fibronectin, inhibited the binding of S. epidermidis, while both these purified plasma proteins enhanced the binding of S. aureus.

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.

Treatment for respiratory tract infections (RTI) in the primary care setting is empirical. Antibiotic prescribing patterns differ among countries and are based on the prevalence of antibiotic resistance in the geographical area. The aim of this study was to compare the antibiotics prescribed by general practitioners (GPs) for RTIs between Spain and Denmark.Observational multicenter survey carried out in the primary healthcare setting. Two groups of GPs in Spain and Denmark recorded all contacts with RTI patients during a 3-week period between November 2001 and January 2002.A total of 2833 RTI cases were registered. Broad-spectrum penicillins and combinations of these drugs plus beta-lactamase inhibitors were the antibiotics most frequently prescribed by Spanish GPs (62.3%), followed by macrolides (22.3%). In contrast, narrow-spectrum penicillins were most commonly prescribed by Danish GPs (58% of all prescriptions), followed by macrolides (29%) (P.001). Antibiotics most frequently prescribed for ear, tonsillar, sinus and bronchopulmonary infections were broad-spectrum penicillins among Spanish GPs and narrow-spectrum penicillins in Denmark. Spanish GPs prescribed penicillin V only for tonsillitis, accounting for 5.1% of the antibiotics used for this condition, whereas this drug accounted for 91.7% of the prescriptions by their Danish colleagues for the same indication.The substantial differences in RTI management between the participating GPs should make us reflect on the rational use of antibiotics. The discrepancies disclosed may indicate dissimilarities in recommendations, traditions, habits, or antibiotic pressures between the countries studied.

The aim of this study was to present the first set of comprehensive data on fungemia in Denmark including the distribution of species and range of susceptibility to major antifungal compounds based on a seminational surveillance study initiated in 2003. The catchment area of the participating hospitals had a population of 2.8 million, or 53% of the Danish population. A total of 303 episodes of fungemia were registered (annual rate, 11 of 100,000 people or 0.49 of 1,000 hospital discharges). Candida species accounted for 97.4% of the fungal pathogens. C. albicans was the predominant species (63%), but the proportion varied from 57% to 72% among participating departments of clinical microbiology. C. glabrata was the second most frequent species (20%; range, 8% to 32%). C. krusei was a rare isolate (3%) and occurred only at two of the participating hospitals. Retrospective data retrieved from the Danish laboratory systems documented a continuous increase of candidemia cases since the early 1990s. For the 272 susceptibility-tested isolates, MICs of amphotericin B and caspofungin were within the limits expected for the species or genus. However, decreased azole susceptibility, defined as a fluconazole MIC of >8 μg/ml and/or itraconazole MIC of >0.125 μg/ml, was detected for 11 Candida isolates that were neither C. glabrata nor C. krusei . Including intrinsically resistant fungi, we detected decreased susceptibility to fluconazole and/or itraconazole in 87 (32%) current Danish bloodstream fungal isolates. We showed a continuous increase of fungemia in Denmark and an annual rate in 2003 to 2004 higher than in most other countries. The proportion of bloodstream fungal isolates with reduced susceptibility to fluconazole and/or itraconazole was also notably high.

Sinusitis is associated with overuse of antibiotics. The aim of this study was to determine whether GPs who use the CRP rapid test (CRP) have a lower antibiotic prescribing rate for sinusitis. During a three-week period, a group of GPs registered all patients with respiratory tract infections (n = 17,792). GPs using CRP prescribed antibiotics for 59% and GPs not using CRP prescribed antibiotics for 78% of the patients with sinusitis. CRP was the factor exerting the greatest influence on the prescribing of antibiotics. Implementing CRP in general practice may lead to a reduction in antibiotic prescribing to patients with sinusitis.

Dysgonomonas capnocytophagoides, formerly known as CDC group DF-3, is an opportunistic pathogen associated with diarrhoea and very rarely bacteraemia. We report a case of D. capnocytophagoides found in blood cultures from a severely neutropenic patient treated for acute myeloid leukaemia. The isolate was found resistant to penicillin, cephalosporins, meropenem, aminoglycosides and ciprofloxacin, and susceptible to ampicillin, tetracycline, chloramphenicol, clindamycin and trimethoprim-sulphamethoxazole. It was identified using conventional phenotypic testing but remained unidentified by the automated identification system (Vitek-2) as this system did not contain DF-3 or D. capnocytophagoides in its database.

The occurrence of the two Shewanella species found in clinical specimens, Shewanella algae and Shewanella putrefaciens, correlates with the temperature and salinity of seawater. This means that Shewanella infections occur in warm climates or during especially warm summers in temperate climates. The infections described most commonly involve ears, skin and soft tissue, with or without bacteraemia. Primary bacteraemia with a fulminant course is also seen in immunocompromised patients. Important differential characteristics between the two species include the ability of S. algae to produce mucoid colonies with beta-haemolysis on sheep blood agar, to grow at 42 degrees C and in NaCl 6% w/v, and to reduce nitrite, and an inability to produce acid from maltose, all of which are in contrast to the characteristics of S. putrefaciens. Automated identification systems fail to differentiate between S. algae and S. putrefaciens, as S. algae is not included in the databases of these systems. Presumably for this reason, most Shewanella infections reported during recent years have been attributed to S. putrefaciens. However, when extensive phenotypic characterisation is performed, most human infections are seen to be caused by S. algae. As the two species seem to have different pathogenic potential for humans, correct identification is important, and this is possible in routine clinical microbiology laboratories.

Symptoms of bacterial sinusitis overlap with viral sinusitis, and it is difficult to distinguish between the two conditions based only on a clinical examination. Uncertain diagnosis results in the significant overuse of antibiotics, which is considered to be one of the most important reasons for development of bacterial resistance to antibiotics. A raised C-reactive protein (CRP) level is an indicator of bacterial infection and the CRP rapid test has been shown to be useful for the diagnosis of bacterial sinusitis in general practice.To examine whether general practitioners (GPs) who use the CRP rapid test in their practice have a lower antibiotic prescribing rate for sinusitis than GPs who do not use the test.Observational design.General practice in Denmark.A group of GPs registered all contacts (n = 17 792) with patients who had respiratory tract infections during a 3-week period between 1 November 2001 and 31 January 2002. GPs who used a CRP rapid test were compared with GPs who did not, and the treatment of their patients (n = 1444) with suspected sinusitis was compared.A CRP rapid test was used by 77% (n = 281) of the GPs. In the group of GPs using a CRP rapid test, the rate of antibiotic prescribing was 59% (95% confidence interval [CI] = 56 to 62) compared with 78% (95% CI = 73 to 82) in the group of GPs who did not use a CRP test. Performing a CRP rapid test was the factor that exerted the greatest influence on whether the patients were prescribed antibiotics, and the level of CRP had a strong influence on the prescribing rate.The CRP rapid test has a substantial influence on the treatment of sinusitis, and implementing the test in general practice may lead to a reduction in antibiotic prescribing to patients with sinusitis.

Objectives. To elucidate and improve quality of diagnosis and treatment of respiratory tract infections in Greenland. Study design. All district medical officers and nursing staff in the Greenlandic coastal health services were invited to participate in the study. Twenty-five district medical officers and the nursing staff from nine districts completed the project and registered in a 3-week period 1,163 contacts involving respiratory tract infections. Methods. Self-registration according to the Audit Project Odense (APO) method on a simple APO registration chart. All contacts involving respiratory tract infections were registered with regard to sex, type of contact, contact form, infection focus, diagnosis and treatment, origin of infection, antibiotic treatment, choice of antibiotics and possible sick-leave. Results. Of the 1,163 registered contacts lung infections represented 26%, throat infections 22% and otitis media 16%. Paraclinical tests were performed in 32% of the cases, 47% of the cases were treated with antibiotics, in 2/3 of the cases with penicillin V. The use of paraclinical tests was somewhat lower and the use of antibiotics was higher than in Denmark. Conclusion. The study showed that it is possible to carry out an APO audit in Greenland, and that there was a moderate difference in the diagnosis and treatment between Greenland and Denmark. An increased use of paraclinical tests may result in quality improvement. Keywords: Greenland, respiratory tract infections, audit, general practice, antibiotics

Hemovigilance has shown that bacteria cause more fatalities than other infections together. Surveillance for detection of bacteria in platelets (PLTs) was initiated. Concomitantly, the storage period for PLTs was extended from 5 to 7 days to reduce cost.Analysis was performed of all cases of a positive signal in a screening procedure for contaminated PLTs taking into account results of confirmative cultures and results of culture from blood components including bacteria strains. Records were assessed from patients transfused with blood components issued before the screening culture became positive.Samples were collected from 22,057 PLT units. An initial reaction was seen in 84 (0.38%). Growth was confirmed in 70 of these. Of the associated PLT units, 26 had been issued or outdated at the time when the culture was found to be reactive, in 27 bacteria were found, and in 17 cultures were negative. The bacteria found were mainly from normal skin flora. Sixty-six patients received 75 blood components issued before the screening system alarmed. None of these patients had a transfusion reaction reported. The outdating fell to less than 5 percent.A screening system for detection of bacterial contamination was implemented without increase in cost owing to extension of storage time to 7 days. Transfusion of several contaminated blood components was prevented.