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Objectives: Hospital-associated infections with vancomycin-resistant Enterococcus faecium (VREfm) have increased dramatically in Denmark. A cornerstone in infection control is effective cleaning and disinfection. This study investigated the survival and resuscitation/growth of clinical isolates of E. faecium exposed to the chlorine-releasing disinfectant, sodium dichloroisocyanurate plus detergent (NaDCC Plus). Methods: To assess biocide efficacy, we modified a method developed to characterise the dose-time-response of bacteria to antibiotics. E. faecium isolates (n = 59) were screened in 96-well plates containing 50–1400 ppm free available chlorine. Bacteria were exposed for 10 min, after which the biocide was inactivated with a neutralizer. Cells were collected by centrifugation, new broth added, and after 20–22 h, viability was recorded as growth/no growth. For a subset of strains the impact of shorter biocide exposure times were examined, as was the influence of longer incubation times. Results: E. faecium survived exposure to relatively high concentrations of NaDCC Plus, average 415 ppm of free available chlorine (SD ± 78 ppm), compared to recommended in-use concentration (1000 ppm). “Outbreak” clones did not prove more tolerant to NaDCC Plus compared to other VREfm clones, hospital-associated vancomycin-susceptible E. faecium (VSEfm) or community-associated VSEfm. Shorter exposure time and extended incubation time in broth both significantly increased the concentration needed to eradicate E. faecium, with some isolates surviving higher concentrations than the recommended in-use concentration. Conclusion: Our results indicate that if an exposure time of 10 min is not achieved, the efficacy of the disinfectant will not be sufficient.

From October 2022 through January 2023, nine patients with NDM-5/OXA-48-carbapenemase-producing Enterobacter hormaechei ST79 were detected in Denmark and subsequently one patient in Iceland. There were no nosocomial links between patients, but they had all been treated with dicloxacillin capsules. An NDM-5/OXA-48-carbapenemase-producing E. hormaechei ST79, identical to patient isolates, was cultured from the surface of dicloxacillin capsules in Denmark, strongly implicating them as the source of the outbreak. Special attention is required to detect the outbreak strain in the microbiology laboratory. From October 2022 through January 2023, nine patients with NDM-5/OXA-48-carbapenemase-producing Enterobacter hormaechei ST79 were detected in Denmark and subsequently one patient in Iceland. There were no nosocomial links between patients, but they had all been treated with dicloxacillin capsules. An NDM-5/OXA-48-carbapenemase-producing E. hormaechei ST79, identical to patient isolates, was cultured from the surface of dicloxacillin capsules in Denmark, strongly implicating them as the source of the outbreak. Special attention is required to detect the outbreak strain in the microbiology laboratory.

Summary Background Surveillance systems for surgical site infections (SSIs), as a measure of patient safety, help health institutions devise strategies to reduce or prevent them. No surveillance systems exist to monitor SSIs in Ghana. Aim To establish a system for monitoring trends and detecting outbreaks in order to create awareness of and control SSIs. Methods An active 30-day surveillance was undertaken at the general surgical unit of the Korle Bu Teaching Hospital, from July 1st, 2017 to December 31st, 2018 to identify SSI. It involved a daily inpatient surveillance of patients who had had a surgical procedure, followed by post-discharge surveillance by means of a healthcare personnel-based survey and a patient-based telephone survey. We supplied quarterly feedback of results to surgeons. Findings Among the 3267 patients included, 331 were identified with an SSI, a 10% incidence risk. Patients who acquired an SSI experienced increased morbidity including nine extra days in hospital and an adjusted relative mortality risk of 2.3 (95% confidence interval: 1.3 - 4.1; P=0.006) compared to patients without SSI. Forty-nine per cent (161/331) of SSIs were diagnosed post discharge using the healthcare personnel-based survey. The patient-based telephone survey contributed 12 additional cases. SSI incidence risk decreased from 12.8% to 7.5% during the study period. Conclusion Post-discharge surveillance is feasible using existing healthcare personnel, and the results highlight the high risk and burden of SSIs in Ghana. A surveillance system with feedback for monitoring SSIs may contribute to reducing SSIs; however, firm conclusions regarding the impact need longer observation time.

Background Carbapenemase-producing Escherichia coli are increasing worldwide. In recent years, an increase in OXA-244-producing E. coli isolates has been seen in the national surveillance of carbapenemase-producing organisms in Denmark. Aim Molecular characterisation and epidemiological investigation of OXA-244-producing E. coli isolates from January 2016 to August 2019. Methods For the epidemiological investigation, data from the Danish National Patient Registry and the Danish register of civil registration were used together with data from phone interviews with patients. Isolates were characterised by analysing whole genome sequences for resistance genes, MLST and core genome MLST (cgMLST). Results In total, 24 OXA-244-producing E. coli isolates were obtained from 23 patients. Among the 23 patients, 13 reported travelling before detection of the E. coli isolates, with seven having visited countries in Northern Africa. Fifteen isolates also carried an extended-spectrum beta-lactamase gene and one had a plasmid-encoded AmpC gene. The most common detected sequence type (ST) was ST38, followed by ST69, ST167, ST10, ST361 and ST3268. Three clonal clusters were detected by cgMLST, but none of these clusters seemed to reflect nosocomial transmission in Denmark. Conclusion Import of OXA-244 E. coli isolates from travelling abroad seems likely for the majority of cases. Community sources were also possible, as many of the patients had no history of hospitalisation and many of the E. coli isolates belonged to STs that are present in the community. It was not possible to point at a single country or a community source as risk factor for acquiring OXA-244-producing E. coli.

Background Carbapenemase-producing Escherichia coli are increasing worldwide. In recent years, an increase in OXA-244-producing E. coli isolates has been seen in the national surveillance of carbapenemase-producing organisms in Denmark. Aim Molecular characterisation and epidemiological investigation of OXA-244-producing E. coli isolates from January 2016 to August 2019. Methods For the epidemiological investigation, data from the Danish National Patient Registry and the Danish register of civil registration were used together with data from phone interviews with patients. Isolates were characterised by analysing whole genome sequences for resistance genes, MLST and core genome MLST (cgMLST). Results In total, 24 OXA-244-producing E. coli isolates were obtained from 23 patients. Among the 23 patients, 13 reported travelling before detection of the E. coli isolates, with seven having visited countries in Northern Africa. Fifteen isolates also carried an extended-spectrum beta-lactamase gene and one had a plasmid-encoded AmpC gene. The most common detected sequence type (ST) was ST38, followed by ST69, ST167, ST10, ST361 and ST3268. Three clonal clusters were detected by cgMLST, but none of these clusters seemed to reflect nosocomial transmission in Denmark. Conclusion Import of OXA-244 E. coli isolates from travelling abroad seems likely for the majority of cases. Community sources were also possible, as many of the patients had no history of hospitalisation and many of the E. coli isolates belonged to STs that are present in the community. It was not possible to point at a single country or a community source as risk factor for acquiring OXA-244-producing E. coli.

Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) is mostly known as an opportunistic pathogen found in horses and as a rare human zoonosis. An 82-year-old male, who had daily contact with horses, was admitted in a septic condition. The patient presented with dyspnea, hemoptysis, impaired general condition, and severe pain in a swollen left shoulder. Synovial fluid from the affected joint and blood cultures showed growth of S. equi subsp. zooepidemicus. Transesophageal echocardiography showed a vegetation on the aortic valve consistent with endocarditis. Arthroscopic revision revealed synovitis and erosion of the rotator cuff. Technetium-99m scintigraphy showed intense increased activity in the left shoulder, suspicious of osteitis. The infection was treated with intravenous antibiotics over a period of five weeks, followed by oral antibiotics for another two months. The patient recovered without permanent sequelae.

Botulism commonly occurs when the anaerobic, gram-positive bacterium Clostridium botulinum, under suitable conditions, produces botulinum neurotoxins. Named A-F, these toxins are the immediate causative agent of the clinical symptoms of symmetrical, descending neurological deficits, including respiratory muscle paralysis. We present five cases of foodborne botulism occurring in Greenland, two with fatal outcome, caused by ingestion of tradionally preserved eider fowl. In the cases of the survivors, antitoxin and supportive care, including mechanical ventilation, were administered. In these cases recovery was complete. Microbiological assays, including toxin neutralization bioassay, demonstrated the presence of neurotoxin E in two survivors. The third survivor was shown by PCR to have the BoNT type E gene in faeces. This is the first report of cases of fatal botulism in Greenland. It underscores the importance of prompt coordinated case management effort in a geographically isolated area such as Greenland.

Purpose: To investigate 1) the development in the incidence of ESBL-producing bacteria in hospitals and primary health care, 2) the contribution of primary health care to the incidence of ESBL-producing bacteria, and 3) the development in resistance patterns for all Escherichia coli and Klebsiella pneumoniae isolates in relation to antimicrobial consumption in hospitals and primary health care. Methods: ESBL-data were retrospectively collected from bacterial isolates from all specimens received at the Department of Clinical Microbiology from 2003 to 2011 together with the corresponding patient data. ESBL-production was detected in isolates from 1067 of 59,373 patients (1.8%) with an E. coli infection and in 263 of 8660 patients (3.0%) with a K. pneumoniae infection. Results: From 2003 to 2009, an increase in patients with an ESBL-producing isolate occurred in both hospitals and primary health care at the same time as an increased consumption of broad-spectrum antimicrobial agents was seen. Interventions to reduce prescription of cephalosporins and ciprofloxacin at the hospitals from 2010 resulted in a remarkable decrease in patients with ESBL-producing K. pneumoniae whereas a continuing increase was seen in patients with ESBL-producing E. coli both at hospitals and in primary health care. The proportion of patients with community-acquired ESBL-producing E. coli was stable with an increase of only 1.4% from 2007 to 2011. Conclusions: Reduction in prescription of broad-spectrum antimicrobial agents at the hospital level had an important impact on the incidence of ESBL-producing K. pneumoniae, but not on ESBL-producing E. coli.

Carbapenemase-producing Enterobacteriaceae (CPE) are extremely multiresistant bacteria with few or no treatment options. Infections with CPE are associated with a mortality of 40-50%. In Denmark, CPE were first detected in 2008. Prior to 2013 the CPE incidence was low, but since then the incidence has increased significantly. Seven outbreaks have occurred, and at least 25 patients have been infected or colonized. The rise in CPE incidence emphasizes the need for a national intervention to reduce the spread. This can be obtained through systematic surveillance, infection control and reduction of antibiotic consumption.

SUMMARYIncidence of various diseases can increase following a flood. We aimed to identify professionals in Copenhagen who became ill after contact with 2 July 2011 floodwater/sediment and determine risks and protective factors associated with illness. We conducted a cohort study of employees engaged in post-flood management activities. Participants completed a questionnaire collecting information about demographics, floodwater/sediment exposure, compliance with standard precautions, and symptoms of illness. Overall, 257 professionals participated, with 56 (22%) cases. Risk of illness was associated with not washing hands after floodwater/sediment contact [relative risk (RR) 2·45], exposure to floodwater at work and home (RR 2·35), smoking (RR 1·92), direct contact with floodwater (RR 1·86), and eating/drinking when in contact with floodwater (RR 1·77). Professionals need to follow standard precautions when in contact with floodwater/sediment, especially proper hand hygiene after personal protective equipment use and before eating/drinking and smoking.

Background The first case of methicillin-resistant Staphylococcus aureus (MRSA) in Greenland was diagnosed in 2000 and led to the first guideline on screening and treatment for MRSA. Up to 2015 there were only 13 patients with MRSA but since then a nearly 4-fold increase in incidence has been seen. The objectives of this study were to analyze the reasons for this increase. Methods MRSA data were collected from the laboratory surveillance database at Dronning Ingrids Hospital, typing results from the Reference Laboratory for Antimicrobial Resistance and Staphylococci at SSI, and the patient records. Results From 2000 to 2017, 48 patients (15 children and 33 adults) have been diagnosed with MRSA. Thirty patients were colonized with MRSA, predominantly in the nose and throat. Eighteen patients had infections: conjunctivitis, middle ear infections, wounds, skin abscesses, mastitis, surgical site infections, for example. The increase since 2015 was mainly due to three large outbreaks in three different cities: Aasiaat in 2014/2015 (seven persons with MRSA; three children and four adults), the capital Nuuk in 2016 (six persons with MRSA; two children and four adults) and Tasiilaq in 2017 (13 persons with MRSA; three children and ten adults). The first two outbreaks were community-acquired with transmission in families and the last one was community-acquired or community-onset hospital acquired. Each outbreak was caused by a specific MRSA-type: t902 CC22 in Aasiaat (unknown epidemiology), t3979 CC5 in Nuuk (probably from Australia), and t304 CC6 in Tasiilaq (probably from Denmark). MRSA was mainly imported from Denmark or abroad due to admission to hospital or due to traveling to high-endemic countries like Australia, but in some cases the epidemiology was unknown. Transmission occurred mainly in families with close contact. Conclusion The increasing number of patients with MRSA in Greenland can be explained by factors such as import from Denmark or abroad due to admission to hospital or traveling, and transmission in Greenland. An ongoing surveillance, compliance to screening procedures (especially patients admitted to hospitals abroad) and guidelines for infection prevention and control are necessary in order to combat MRSA in Greenland in the future. Disclosures All authors: No reported disclosures.

Consecutively collected synovial fluids were examined for presence of bacterial DNA (a 700-bp fragment of the bacterial 23S rRNA gene) followed by DNA sequencing of amplicons, and by conventional bacteriological methods. One or more microorganisms were identified in 22 of the 227 synovial fluids (9,7%) originating from 17 patients. Sixteen of the patients had clinical signs of arthritis. For 11 patients molecular and conventional bacterial examinations were in agreement. Staphylococcus aureus, Streptococcus dysgalactiae subspecies equisimilis and Streptococcus pneumoniae, were detected in synovial fluids from 6, 2 and 2 patients, respectively. In 3 patients only 23S rRNA analysis was positive; 2 synovial fluids contained S. dysgalactiae subspecies equisimilis and 1 S. pneumoniae). The present study indicates a significant contribution by PCR with subsequent DNA sequencing of the 23S rRNA gene analysis in recognizing and identification of microorganisms from synovial fluids.

A modified rat model of endocarditis with catheterization for 2 days was established in female Lewis rats using different inocula of Enterococcus faecalis (strain no. EF 19) in order to measure IgG antibodies in serum during the course of infection. Increasing the inocula intravenously resulted in an increase in the CFU/g vegetation and the CFU/g spleen, the ID50 being about 10 CFU/ml and the ID90 about 1x10(2) CFU/ml. The lowest bacterial inoculum infecting 100% of the rats was 3x10(3) CFU/ml, and for further investigations we used this inoculum size. Rats were sacrificed on day 2, 5, 7, 9, 11 and 28 after infection. The CFU/g vegetation and the CFU/g spleen increased until day 7 and then decreased. Serum samples were collected from 129 rats at different times after challenge. Three different ELISA systems were established to measure the IgG antibody responses: E. faecalis sonicate ELISA (a pool of four sonicates of strain no. EF 10, EF 11, EF 19 and EF 48), E. faecalis whole cell ELISA (strain no. EF 19) and E. faecalis purified cell wall ELISA (strain no. EF 19). An IgG antibody response was detected already on day 2, and except for a minor decrease on day 6/7 the antibody response continued to increase until day 14 (whole cell ELISA and sonicate ELISA) and day 21 (purified cell wall ELISA) when a plateau was reached. Significant increases in IgG antibody responses (p

During a period of 3.5 years, endocarditis was suspected in 151 patients admitted to Rigshospitalet. 140 were available for this study. In 59 of the 140 patients, the diagnosis was confirmed, and 36 had positive blood cultures. Echocardiographic findings compatible with the diagnosis were present in 92% of the 59 cases. The most common causes of endocarditis were Staphylococcus aureus and viridans streptococci. Patients with endocarditis caused by S. aureus had significantly (p = 0.002) more embolic episodes compared to patients having endocarditis caused by the viridans streptococci. The diagnosis was established at a mean of 3–4 weeks after the onset of symptoms and 2 weeks after admission to hospital. In order to minimize diagnostic delay, the following aspects may be important: (1) earlier detection of endocarditis among physicians examining patients at risk; (2) educating patients with cardiac disease and cardiac valve prosthesis; (3) earlier antibiotic therapy, and (4) developing further diagnostics for endocarditis.

Studies on the value of culture-independent molecular identification of bacteria in cardiac valves are mostly restricted to comparing agreement of identification to what is obtained by culture to the number of identified bacteria in culture-negative cases. However, evaluation of the usefulness of direct molecular identification should also address weaknesses, their relevance in the given setting, and possible improvements. In this study cardiac valves from 56 Danish patients referred for surgery for infective endocarditis were analysed by microscopy and culture as well as by PCR targeting part of the bacterial 16S rRNA gene followed by DNA sequencing of the PCR product. PCR and DNA sequencing identified significant bacteria in 49 samples from 43 patients, including five out of 13 culture-negative cases. No rare, exotic, or intracellular bacteria were identified. There was a general agreement between bacterial identity obtained by ribosomal PCR and DNA sequencing from the valves and bacterial isolates from blood culture. However, DNA sequencing of the 16S rRNA gene did not discriminate well among non-haemolytic streptococci, especially within the Streptococcus mitis group. Ribosomal PCR with subsequent DNA sequencing is an efficient and reliable method of identifying the cause of IE, but exact species identification of some of the most common causes, i.e. non-haemolytic streptococci, may be improved with other molecular methods.

By means of enzyme-linked immunosorbent assays (ELISAs) the antibody responses were detected in sera from normal persons (n = 275), patients with various types of bacteremia (n = 137), and patients with endocarditis (n = 28). Four ELISAs were evaluated. As antigens ultrasonic extracts of Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, and nonhemolytic streptococci were used. The antibody levels against S. aureus increased with age, while antibodies against the other bacteria showed no age variation. The different ELISA systems showed many serological cross-reactions. The ELISA in this limited material was 72% successful in identifying cases of bacterial endocarditis caused by S. aureus (diagnostic sensitivity 84%; diagnostic specificity 89%), while in other cases of bacterial endocarditis it was less successful.

Sera from 151 patients suspected of having endocarditis were obtained during a period of 3 1/2 years at Rigshospitalet, Copenhagen. The sera were examined by crossed immunoelectrophoresis for antibodies to bacteria causing endocarditis. The patients were divided into four groups: 1. Patients with definite endocarditis, 2. Patients with culture-negative endocarditis, 3. Patients with uncertain endocarditis, and 4. Patients without endocarditis. In sera from patients suffering from endocarditis caused by viridans streptococci, precipitating antibodies were demonstrated by crossed immunoelectrophoresis (diagnostic specificity = 86%; diagnostic sensitivity = 100%) while other bacterial etiologies of endocarditis were less reliably demonstrated by this method.

The main purpose of the study was to investigate the frequency of ESBL-producing E. coli and Klebsiella strains in the Greater Copenhagen area. Four collections of strains were investigated: A) 380 consecutive E. coli and Klebsiella isolates primarily from urine, B) 200 gentamicin-resistant E. coli and Klebsiella isolates primarily from urine, C) 210 consecutive E. coli isolates from blood cultures, and D) 68 cefuroxime-resistant E. coli and Klebsiella isolates primarily from urine. Only one strain per patient was included. Strains with a zone diameter for cefpodoxime

Summary Background Staphylococcus epidermidis develops resistance to ciprofloxacin rapidly. That this antibiotic is excreted in apocrine and eccrine sweat of healthy individuals might be the reason for the development of such resistance. We assessed whether S epidermidis isolated from the axilla and nasal flora of healthy people could develop resistance to ciprofloxacin after a 1-week course of this antibiotic. Methods The concentration of ciprofloxacin in sweat was measured in seven volunteers after oral administration of 750 mg ciprofloxacin twice daily for 7 days, and the development of resistance in S epidermidis from axilla and nostrils was monitored during and 2 months after the treatment. Genotyping of S epidermidis was done by restriction fragment length polymorphism. Findings The mean concentration of ciprofloxacin in sweat increased during the 7 days of treatment—from 2·2 μg/mL 2·5 h after the first tablet to 2·5 μg/mL after the fifth tablet, and 5·5 μg/mL after the 13th tablet. All persons harboured susceptible S epidermidis (minimal inhibitory concentration [MIC] 0·25 μg/mL) in axilla and nostrils before treatment. Four resistant strains were detected, two intermediate-level (MIC 4–12 μg/mL) and two high-level (MIC >32 μg/mL). Three of these strains were found in all the participants, and a ciprofloxacin-sensitive variant of one of the high-level resistant strains was also found before the start of the treatment. The high-level resistant strains were also resistant to methicillin, erythromycin, gentamicin, sulphonamide, and trimethoprim. A mean of 2·7 days after the start of the treatment, development of ciprofloxacin resistance was detected in S epidermidis from the axilla of all persons, compared with 11 days for the appearance of resistant S epidermidis in nostrils. The resistant strains persisted for an average of 37 and 39 days in axilla and nostrils, respectively, after the end of the treatment. Interpretation The rapid development of resistance to ciprofloxacin due to excretion of this drug into the sweat might be involved in the development of multiresistant S epidermidis and possibly other skin bacteria in hospitals and in communities with high use of ciprofloxacin or related drugs.

Enterococci are part of the normal human fecal flora and also part of the fecal flora of many animals. Vancomycin- and ampicillin-resistant enterococci give rise to infections that may virtually be untreatable with antibiotics. Antibiotic use in humans is a risk factor for development or selection of vancomycin-resistant enterococci. In animals the related glycopeptide avoparcin is used, especially in poultry, as a food additive to promote growth. Selective pressures for vancomycin-resistant enterococci are high in Denmark with the production of 105 million poultry and the use of 24,000 kg avoparcin per year. The possible impact on vancomycin resistance among human isolates of enterococci remains to be defined. Furthermore, there has been a 3-fold increase in vancomycin usage during the last 5 years at our hospital, from 2.4 to 7 kg/year. We examined 91 stool specimens from 67 patients in risk units at our hospital. Using a selective medium (KAA agar), 17 strains of Enterococcus faecium grew on the selective medium and 3 (18%) were vancomycin-resistant (MIC > 256 mg/l). Using PCR and an internal probe, vanA was found in the vancomycin-resistant enterococci. No association between vancomycin therapy and carriage of VRE was demonstrated in these patients.

Det kraftige skybrud over København den 2. juli 2011 skabte en lang række uvante situationer, der kunne føre til både sygdomme og ulykker. Ny undersøgelse afdækker nu de mulige sundhedsmæssige påvirkninger, hos de mennesker, der har været direkte påvirket af oversvømmelserne