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The aim of this study was to investigate antimicrobial susceptibilities and genomic characteristics of mupirocin-resistant MRSA isolates in Stockholm, Sweden. In total, 44 non-duplicate mupirocin-resistant MRSA isolates detected in Stockholm during 2010-2022 were investigated. Antimicrobial susceptibility testing was performed using broth microdilution method and further tested for high-level mupirocin-resistance (MuH) and rifampicin by Etest. All isolates were subjected to whole genome sequencing. 41 isolates presented MuH with MICs =1024 mg/L whilst three isolates displayed low-level mupirocin resistance (MuL). mupA-gene was detected in all MuH isolates. Point mutations in ileS gene leading to N213D and V588F were identified in the three MuL isolates. Mutation in rpoB (H481N) was detected in a rifampicin-resistant isolate. Among the isolates, 15 multi-locus sequence types (MLST) were identified, with the four most common sequence types (ST22, ST72, ST8, and ST125) accounting for 66% of the isolates. Mupirocin-resistant MRSA in Stockholm was uncommon, with a percentage of <0.5% among MRSA cases during 2010-2022. In the present study, most mupirocin-resistant isolates were MuH and mupA-positive, predominantly linked to ST22 or ST72 isolates. MuL-resistance was associated with a point mutation in the IleS protein. A multidrug-resistant ST1-MRSA-IV strain was resistant to both mupirocin and rifampicin. [ABSTRACT FROM AUTHOR]

Automated testing of antimicrobial susceptibility is common in clinical microbiology laboratories but their ability to detect low‐level resistance has been questioned. This Nordic multicentre study aimed to evaluate the performance of commercially available automated AST systems. A phenotypically well‐characterised collection of gram‐negative bacilli (Escherichia coli (n = 7), Klebsiella pneumoniae (n = 6) and Pseudomonas aeruginosa (n = 7)) with and without resistance mechanisms was examined by Danish (n = 1), Finnish (n = 6), Norwegian (n = 16) and Swedish (n = 5) laboratories. Minimum inhibitory concentrations (MICs) were determined for 12 antimicrobials with automated systems and compared with MICs obtained with gold standard broth microdilution. The automated systems used were VITEK 2 (n = 23), Phoenix (n = 4), MicroScan (n = 1), and ARIS (n = 1). Very major errors were identified for six antimicrobials; cefotaxime (6.9%), meropenem (0.4%), ciprofloxacin (0.7%), ertapenem (4.3%), amikacin (3.4%) and colistin (6.4%). Categorical agreement of MIC for the automated systems compared to broth microdilution ranged from 83% for imipenem to 100% for ampicillin and trimethoprim‐sulfamethoxazole. The analysis revealed several important antimicrobials where resistance was underestimated, potentially with significant consequences in patient treatment. The results cast doubt on the use of automated AST in the management of patients with serious infections and suggests that more work is needed to define their limitations. [ABSTRACT FROM AUTHOR]

Aim This study explored the incidence and aetiology of bloodstream infections after patients received the pneumococcal conjugate vaccination and a risk-based intrapartum antibiotic prophylaxis against early onset sepsis caused by group B streptococcus. We also monitored clinically relevant antimicrobial resistance. Method We studied 3986 positive blood cultures from children up to 17 years of age at a paediatric hospital in Stockholm, Sweden, using data from medical records before and after the initiatives, to reduce early onset sepsis, were introduced in 2007 and 2008. Results Bloodstream infections caused by Streptococcus pneumoniae declined by 42% overall (5.6 to 3.2/100 000) and by 62% in previously healthy children under 36 months of age (24.2 to 9.2/100 000). Early onset sepsis caused by group B streptococcus declined by 60% (0.5 to 0.2/1000 live born children). Bacterial meningitis caused by these bacteria decreased by 70%. Staphylococcus aureus and various Gram-negative bacteria became the dominant pathogens, in both previously healthy children and those with underlying disease. Overall, antimicrobial resistance remained low between the two 5-year study periods. Conclusion Pneumococcal conjugate vaccination and risk-based intrapartum antibiotic prophylaxis against group B streptococcus effectively decreased the incidence of bloodstream infections. Empirical antibiotic therapy should target Staphylococcus aureus in both community and hospital-acquired invasive bacterial infections. [ABSTRACT FROM AUTHOR]

Aim This study evaluated risk factors for acute wheeze in preschool children and investigated whether subnormal levels of vitamin D were associated with increased risk for acute wheeze, atopy or viral/bacterial respiratory infections. Methods We recruited 130 children with acute wheeze, aged 6 months to 4 years, from paediatric emergency departments in Stockholm, Sweden, and 101 age-matched controls with no history of wheeze or sensitisation to airborne allergens. Parents answered standardised questionnaires, and blood samples were analysed for specific IgE to airborne and food allergens and levels of 25 hydroxyvitamin D (25( OH)D). Nasopharyngeal virus samples were collected during the emergency department visit in the group of children with wheeze, and a subset were also tested for bacteria. Results Vitamin D insufficiency (25(OH)D < 75 nmol/L (30 ng/mL)) was associated with an odds ratio of 2.7 (95% confidence interval 1.1-6.2) for acute wheeze. However, no association was found between vitamin D insufficiency and atopy, presence of virus or bacteria or recurrent infections. Children older than 24 months were particularly at risk of subnormal vitamin D levels, irrespective of wheezing history . Conclusion Our findings support the hypothesis that subnormal levels of vitamin D are associated with acute wheeze in young children. [ABSTRACT FROM AUTHOR]

Aim To study the aetiology of bloodstream infections ( BSI) in children 0-17 years, the influence of age and underlying co-morbidity on BSI rate, distribution of pathogens and outcome; and to provide data on antimicrobial susceptibility patterns. Methods A retrospective population-based study. Data on blood cultures were collected at yearly intervals during 1998-2008. Information about risk factors, focal infection and outcome was retrieved from the patient charts. Results We identified 1097 BSI. The incidence of BSI was 0.4/1000. The age-specific incidence was 2.3/1000 in neonates (0-28 days old) and 0.2/1000 in the age group 6-17 years. Staphylococcus aureus was the most common pathogen. The number of species causing BSI in previously healthy children was lower compared with children with co-morbidity. Most children requiring intensive care had a serious underlying illness. Antimicrobial resistance was rare and did not influence outcome. The case-fatality rate was 14.4% in neonates, 5.4% in children with co-morbidity and 1.7% in previously healthy children. Conclusion Mortality from BSI is low, and a limited spectrum of pathogens is isolated from previously healthy children compared with children with co-morbidity. When choosing empirical therapy for suspected BSI, age and presence of risk factors should be taken into account. [ABSTRACT FROM AUTHOR]

Herein, we describe the phenotypic and genotypic characterization of a multiresistant clone of Pseudomonas aeruginosa disseminating in a burn unit in Orumieh, Iran. A total of 58 isolates of P. aeruginosa were collected during August 2007 and June 2008. Minimum inhibitory concentrations (MICs) of P. aeruginosa isolates were determined against 11 antimicrobial agents by E test. Serotyping, pulsed-field gel electrophoresis ( PFGE), and multilocus sequence typing ( MLST) were used for studying the clonal relationship among the isolates. Antibiotic susceptibility testing revealed that most of the isolates were multidrug resistant and colistin was the antibiotic with the highest activity. Pseudomonas aeruginosa isolates fell into nine different serotypes, and O10 and O11 were the most common. PFGE analyses showed 12 different genotypes and 68.1% of isolates showed more than 80% similarity, indicating possible clonal relatedness. These isolates were found to belong to the same sequence type, ST773. This sequence type has earlier been reported from China, and a double locus variant of this ST has been found earlier in France in a PER-1 extended-spectrum β-lactamase-producing P. aeruginosa. [ABSTRACT FROM AUTHOR]

To determine the distribution of urinary tract pathogens with focus on Staphylococcus saprophyticus and analyse the seasonality, antibiotic susceptibility, and gender and age distributions in a large Swedish cohort. S. saprophyticus is considered an important causative agent of urinary tract infection ( UTI) in young women, and some earlier studies have reported up to approximately 40% of UTIs in this patient group being caused by S. saprophyticus. We hypothesized that this may be true only in very specific outpatient settings. During the year 2010, 113 720 urine samples were sent for culture to the Karolinska University Hospital, from both clinics in the hospital and from primary care units. Patient age, gender and month of sampling were analysed for S. saprophyticus, Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis. Species data were obtained for 42 633 (37%) of the urine samples. The most common pathogens were E. coli (57.0%), Enterococcus faecalis (6.5%), K. pneumoniae (5.9%), group B streptococci (5.7%), P. mirabilis (3.0%) and S. saprophyticus (1.8%). The majority of subjects with S. saprophyticus were women 15-29 years of age (63.8%). In this age group, S. saprophyticus constituted 12.5% of all urinary tract pathogens. S. saprophyticus is a common urinary tract pathogen in young women, but its relative importance is low compared with E. coli even in this patient group. For women in other ages and for men, growth of S. saprophyticus is a quite uncommon finding. [ABSTRACT FROM AUTHOR]

The increasing prevalence of ESBL-, AmpC- and carbapenemase-producing Enterobacteriaceae necessitates reliable phenotypic tests for detection and categorization. The main objective of this study was to evaluate ROSCO Neo-Sensitabs with different β-lactam-β-lactam inhibitor combinations for phenotypic detection and categorization of β-lactamases in Enterobacteriaceae. Using standard CLSI/EUCAST methodology, differences in zones of inhibitions between a β-lactam alone compared with the combination with a β-lactamase inhibitor as well as subjective synergy observations were determined for 172 well characterized Enterobacteriaceae strains with defined resistance mechanisms. The results showed that for all ESBL-positive strains (n = 66), combinations of clavulanic acid synergy with cefotaxime, ceftazidime or cefepime, were observed. All acquired AmpC β-lactamases (n = 17) were detected using cloxacillin combined with cefotaxime and/or ceftazidime (both combinations were required). Carbapenemase producers (n = 59) with the exception of one KPC-producer were correctly grouped using the combination of meropenem ± aminophenylboronic acid (APBA) or dipicolinic acid (DPA). Ethylene diamine tetraacetic acid (EDTA) also inhibited all metallo-β-lactamases, but as with DPA, one false positive result was observed. Based upon these data, we propose a tablet layout for 14 cm agar plates, which could be used as a whole or in a targeted approach for detection and categorizing of relevant acquired β-lactamases in Enterobacteriaceae. [ABSTRACT FROM AUTHOR]

Neonatal intensive care units ( NICUs) are vulnerable to nosocomial outbreaks caused by multiresistant Enterobacteriaceae, but no reports of NICU outbreaks of extended-spectrum β-lactamase ( ESBL) producing Klebsiella pneumoniae have previously been published from countries with a low level of antimicrobial resistance such as the Scandinavian countries. We describe a clonal outbreak of CTX- M-15 -producing Klebsiella pneumoniae affecting 58 infants in the neonatal intensive care unit at Stavanger University Hospital, Norway, during a period of 4 months, 2008-2009. The clone spread widely and rapidly in the NICU, and extensive interventions were required to terminate the outbreak. In contrast to previous outbreaks, only one infant acquired a systemic infection caused by the outbreak strain, probably due to a favourable epidemic strain lacking the most common virulence factors. A probable index case was identified, due to multiple positive breast milk samples collected from the infant's mother before and after the infant's transfer from another hospital. Breast milk samples from 3/18 (17%) mothers of colonized infants were positive for ESBL-producing K. pneumoniae. Vertical transmission of ESBL-producing bacteria has been shown previously,’but the possibility of transmission of ESBL-producing K. pneumoniae through expressed breast milk is reported here for the first time. The increasing occurrence of ESBL-producing’ Enterobacteriaceae should therefore encourage changes in diagnostic routines for bacterial screening of breast milk. [ABSTRACT FROM AUTHOR]

Background: Bacteremia is a major cause of morbidity and mortality in patients with hematological malignancies. Objectives: The aim of this study was to define temporal trends in species distribution, antimicrobial susceptibility, and all-cause mortality in bacteremic hospitalized patients receiving no antibacterial prophylaxis during chemotherapy-induced neutropenia. Methods: A total of 677 clinical episodes of bacteremia were identified in 463 patients during 2002-2008, and the results were compared with those published from the same institution during 1980-86 and 1988-2001. No major changes in patient selection were introduced during this period. Results: Between 2002 and 2008, the dominating pathogens were Escherichia coli (18%), coagulase-negative staphylococci (15%), viridans streptococci (14%), Klebsiella spp. (10%), and Enterococcus faecium (8%). The 7-d crude mortality rate was 5.2%. Polymicrobial bacteremia was seen in 25.7% of the patients who died within 7 d and in 13.1% of the survivors ( P = 0.04). Acquired resistance was rarely observed, but a statistically significant increase in ciprofloxacin resistance in E. coli was observed. Comparing 2002-2008 with historical data from the same institution, the proportion of Gram-positive isolates remained stable at 53-55% from 1988. Conclusions: The avoidance of fluoroquinolone prophylaxis may have contributed to a stable proportion of Gram-positive bacteremia. The crude mortality was low in an international perspective. Acquired resistance was uncommon, but ciprofloxacin resistance in E. coli increased significantly. We believe that an indiscriminate use of antibacterial prophylaxis could be avoided in neutropenic patients without a negative impact on mortality. [ABSTRACT FROM AUTHOR]

Titelman E, Iversen A, Kahlmeter G, Giske CG. Antimicrobial susceptibility to parenteral and oral agents in a largely polyclonal collection of CTX-M-14 and CTX-M-15-producing Escherichia coli and Klebsiella pneumoniae. APMIS 2011; 119: 853-63. Activity of oral and parenteral antimicrobials against consecutively isolated extended-spectrum β-lactamase (ESBL)-producing Escherichia coli (n = 149) and Klebsiella pneumoniae (n = 20) was determined, and susceptibility test methods were compared for parenteral β-lactams. Polymerase chain reaction (PCR) targeting blaCTX-M, blaSHV and blaTEM, and DNA sequencing and epidemiological typing with pulsed-field gel electrophoresis were performed. PCR targeting pabB was screened for E. coli O25b-ST131. Minimum inhibitory concentrations (MICs) were determined using Etest and broth microdilution. Disc diffusion was performed according to European Committee on Antimicrobial Susceptibility Testing (EUCAST). Dominating genotypes were blaCTX-M-15 (75%) and blaCTX-M-14 (23%). Four E. coli clusters (7-18 isolates) were found. Forty-two per cent of E. coli belonged to O25b-ST131. Ciprofloxacin resistance was 72%, trimethoprim resistance was 70%. Among E. coli, resistance to mecillinam (13%), nitrofurantoin (7%) and fosfomycin (3%) was low, although resistance was high in K. pneumoniae (25%, 60%, 85%). Susceptibility to ertapenem was 99%, piperacillin-tazobactam 91%, tigecycline 96% and temocillin 76%. Susceptibility rates obtained with broth microdilution and Etest were in agreement for cefotaxime (2 vs 1%) and ceftazidime (9 vs 11%), but not for piperacillin-tazobactam (59 vs 91%). With disc diffusion major errors occurred with piperacillin-tazobactam (18/169). Several therapeutic alternatives exist for ESBL-producing E. coli, but few exist for K. pneumoniae. Disc diffusion and Etest can accurately predict susceptibility to cefotaxime and ceftazidime, but not to piperacillin-tazobactam with the present breakpoints. [ABSTRACT FROM AUTHOR]

Dumpis U, Iversen A, Balode A, Saule M, Miklaševičs E, Giske CG. Outbreak of CTX-M-15-producing Klebsiella pneumoniae of sequence type 199 in a Latvian teaching hospital. APMIS 2010; 118: 713–6. Previous studies on the epidemiology of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in Latvia are lacking. ESBL-producing Klebsiella pneumoniae (n = 32) were subjected to pulsed-field gel electrophoresis (PFGE) and selected isolates to multi-locus sequence typing (MLST). Species identification and susceptibility testing were performed using VITEK2, and sequencing of blaCTX-M was performed in selected isolates. PFGE revealed one major clone (n = 23), with most of the isolates derived from the ICU. The clone harboured blaCTX-M-15, was sequence type 199 and comprised two ertapenem non-susceptible isolates. This is the first report of an ESBL outbreak in Latvia, and calls for increased epidemiological typing of ESBL-producing Enterobacteriaceae, as well as improved infection control routines. [ABSTRACT FROM AUTHOR]

El Amin N, Giske CG, Jalal S, Keijser B, Kronvall G, Wretlind B. Carbapenem resistance inPseudomonas aeruginosa: alterations of porin OprD and efflux proteins do not fully explain resistance patterns observed in clinical isolates. APMIS 2005;113:187–96.Imipenem resistance inPseudomonas aeruginosais considered to be associated with loss of the porin OprD combined with activity of chromosomalβ-lactamase (AmpC), while overexpression of multidrug efflux pumps is considered to confer meropenem resistance. Carbapenem resistance can also result from production of metallo-β-lactamases. Transcription ofoprDand efflux pump genesmexB,mexYandmexFwas analysed in 23 clinical isolates ofP. aeruginosaby quantitative RT-PCR.oprDwas sequenced in all, and mexR, regulator of efflux pump MexAB-OprM, in selected isolates. Four isolates that were imipenem susceptible had significant reduction ofoprDmRNA and presence ofoprDmutations causing frameshift or translational stop. In strains only resistant to imipenem no significant difference in transcription ofoprDwas observed between low-level and high-level resistant isolates. The differences could not be explained by either pattern ofoprDmutations. Increased transcription ofmexBgenerally correlated well with meropenem resistance. One high-level meropenem-resistant isolate showed no signicant change in mexB mRNA, but sequencing confirmed presence of analBmutation. Furthermore, one meropenem-susceptible isolate showed significant increase inmexBtranscription, but nomexRmutations. In summary, our findings indicate that the resistance patterns observed cannot be fully explained by the currently described carbapenem resistance mechanisms. [ABSTRACT FROM AUTHOR]