Hydrogen Peroxide Vapor Decontamination in a Patient Room Using Feline Calicivirus and Murine Norovirus as Surrogate Markers for Human Norovirus.

OBJECTIVE: To determine whether hydrogen peroxide vapor (HPV) could be used to decontaminate caliciviruses from surfaces in a patient room. DESIGN: Feline calicivirus (FCV) and murine norovirus (MNV) were used as surrogate viability markers to mimic the noncultivable human norovirus. Cell culture supernatants of FCV and MNV were dried in triplicate 35-mm wells of 6-well plastic plates. These plates were placed in various positions in a nonoccupied patient room that was subsequently exposed to HPV. Control plates were positioned in a similar room but were never exposed to HPV. METHODS: Virucidal activity was measured in cell culture by reduction in 50% tissue culture infective dose titer for FCV and by both 50% tissue culture infective dose titer and plaque reduction for MNV. RESULTS: Neither viable FCV nor viable MNV could be detected in the test room after HPV treatment. At least 3.65 log reduction for FCV and at least 3.67 log reduction for MNV were found by 50% tissue culture infective dose. With plaque assay, measurable reduction for MNV was at least 2.85 log units. CONCLUSIONS: The successful inactivation of both surrogate viruses indicates that HPV could be a useful tool for surface decontamination of a patient room contaminated by norovirus. Hence nosocomial spread to subsequent patients can be avoided.

Risk factors for infections with extended-spectrum beta-lactamase-producing Escherichia coli in a county of Southern Sweden.

BACKGROUND: It is important to identify patients who are at risk for infections with extended-spectrum ß-lactamase (ESBL)-producing bacteria in order to reduce mortality, to avoid spread of resistant bacteria in hospitals, and to minimize the number of patients receiving unnecessary treatment with broad-spectrum antibiotics. A case-control survey among Swedish patients was performed at Skåne University Hospital to identify risk factors for developing an infection with ESBL-producing Escherichia coli in a low endemic country. METHODS: We used a computerized database to identify patients with growth of ESBL-producing E. coli (n = 109) in urine or blood cultures and an equal number of controls matched for age and gender with non ESBL-producing E. coli in urine and blood diagnosed between January and October 2008. We used unadjusted P-values. RESULTS: Patients with ESBL-producing E. coli had a significantly (P < 0.05) higher likelihood of having traveled to Asia including Turkey and the Middle East including Egypt (14/58) than the non-ESBL-positive group (4/53). Hospital stay during the previous year (P < 0.04), especially for more than one month, was another significant (P = 0.01) risk factor for infection with ESBL-producing E. coli (8/58). A stay in the surgical department was a further risk factor (P < 0.01). CONCLUSION: In this study, we identified 22 of 58 (38%) patients with ESBL-producing E. coli by considered significant risk factors before starting antibiotics.

Prevalence of extended-spectrum beta-lactamase-producing bacteria in food.

Extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae with Cefotaximase-München (CTX-M) enzymes are rapidly increasing worldwide and pose a threat to health care. ESBLs with CTX-M enzymes have been isolated from animals and different food products, but it is unknown if food imported from the Mediterranean area may be a possible reservoir of these bacteria. During 2007-2008, swab samples from food across different retail outlets (mostly food from the Mediterranean countries and Swedish chicken) were collected. Escherichia coli strains from Swedish meat and E. coli isolates from unspecified food from a Swedish food testing laboratory were also examined. In 349 of the 419 swab samples, growth of Enterobacteriaceae was found. In most of the samples, there was also growth of Gram-negative environmental bacteria. Air dry-cured products contained significantly less Enterobacteriaceae isolates compared to lettuces; however, none of the examined Enterobacteriaceae harbored ESBLs. This study did not support the theory that imported food from the Mediterranean area or Swedish domestic food might constitute an important vehicle for the dissemination of ESBL-producing Enterobacteriaceae; however, a spread from food to humans may have occurred after 2008.

Duration of colonization with extended-spectrum beta-lactamase-producing Escherichia coli in patients with travellers' diarrhoea.

BACKGROUND: Resistant Enterobacteriaceae have become a worldwide epidemic during the last decade and are a great threat to health care worldwide. International travel is a major risk factor for becoming colonized with extended-spectrum beta-lactamase (ESBL)-producing bacteria. Data on the persistence of colonization with ESBL-producing bacteria in the faecal flora are limited. METHODS: A prospective cohort study was performed between October 2007 and October 2010. Fifty-eight patients with faecal carriage of ESBL-producing Escherichia coli from a previous study of patients with travellers' diarrhoea were included. RESULTS: Forty-one of the patients had a complete follow-up. Ten of these patients (24%) carried ESBL-producing E. coli at the first follow-up point (3-8 months), of whom 4 had a new ESBL strain. At the 3-y follow-up, 4 patients carried ESBL (10%), of whom 1 had 2 new ESBL strains. CONCLUSIONS: The long duration of ESBL carriage is worrisome. These carriers may be an important source of the spread of ESBLs in the population and this has implications for the clinical management of patients.

Development of a real-time SYBRGreen PCR assay for rapid detection of acquired AmpC in Enterobacteriaceae.

INTRODUCTION: Acquired AmpC enzymes, classified as miscellaneous extended-spectrum beta-lactamase (ESBL(M)) enzymes according to a recently proposed beta-lactamase classification, are increasing according to several publications. Simple and rapid methods for detection of ESBL(M) are needed for appropriate infection control. A gel-based multiplex PCR method for acquired bla(AmpC) detection and subtype classification has been available for several years. Here, we describe a modification of the protocol to suit real-time PCR platforms and to include novel genotypes. MATERIAL AND METHODS: Clinical isolates with clavulanic acid non-reversible non-susceptibility to extended-spectrum cephalosporins were subjected to combination disk testing with cefoxitin +/- cloxacillin at Malmö University Hospital. Phenotypical AmpC production was defined as cloxacillin reversible cefoxitin resistance. In this study 51 phenotypical AmpC-producing isolates, were subjected to the acquired bla(AmpC) real-time PCR assay. The acquired blaAmpC positive isolates were further characterized by DNA sequencing of the acquired AmpC encoding gene, Pulsed-Field Gel Electrophoresis (PFGE) and PCR-based replicon typing. RESULTS AND DISCUSSION: The real-time PCR assay was able to detect and sub-classify all acquired bla(AmpC) genes described to date. The assay can be performed in less than 3h, including pre-PCR preparations. Analysis of the isolate collection resulted in 18 of 51 phenotypical AmpC-producing isolates being positive in the acquired bla(AmpC) real-time multiplex PCR assay; 17 of subtype CIT and one DHA. Sequence analysis identified 16 isolates as blaCMY-2, one as blaCMY-16 and one as blaDHA-1. Detected plasmid replicon types were I1 and B/O. Two of the E. coli isolates were identical according to PFGE and the others were unrelated.

Bacterial aetiology in ventilator-associated pneumonia at a Swedish university hospital.

Ventilator-associated pneumonia (VAP) is a common complication of respiratory support and is associated with increased mortality, morbidity and costs, and a prolonged stay in the intensive care unit. Scandinavian data on the aetiology in VAP are lacking. We hereby present a retrospective study on the aetiology of VAP diagnosed by protective specimen brush culture at Malmö University Hospital in relation to early- and late-onset VAP, antibiotic treatment and the incidence of drug-resistant bacteria. Patients registered with a diagnosis of VAP between January 2004 and September 2007 were included in the study. Sixty-five of 109 patients diagnosed with VAP met the inclusion criteria, and 103 bacterial isolates were cultured from these patients. The most common findings among the 65 VAP episodes were Enterobacteriaceae (28), Pseudomonas aeruginosa (13), Haemophilus influenzae (12) and Staphylococcus aureus (8). Patients with no antibiotic treatment at the onset of VAP had significantly more H. influenzae (p = 0.035) and Gram-positive pathogenic bacteria (p = 0.019). There was no difference in incidence of P. aeruginosa between early- and late-onset VAP. Resistant bacteria were found in 18% of the patients.

Extended-spectrum beta-lactamase-producing Escherichia coli in patients with travellers' diarrhoea.

The identification of patients carrying extended-spectrum beta-lactamase (ESBL)-producing bacteria is important, since these patients are at risk of receiving inappropriate empirical therapy if they become infected. The purpose of this study was to investigate the occurrence of ESBL-producing bacteria in patients with travellers' diarrhoea. Patients with travellers' diarrhoea (N = 242) having delivered stool samples for the diagnosis of Salmonella, Shigella, Yersinia or Campylobacter, were also examined for ESBL-producing Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis. The overall prevalence of faecal carriage of ESBL-producing bacteria was 24% (58/242). Of the patients who had travelled in Europe, 3% (2/63) were found to be ESBL carriers in comparison to 36% (50/138) of those who had travelled outside Europe. ESBL-producing E. coli was especially common among patients returning from India (11/14), Egypt (19/38; 50%) and Thailand (8/38; 22%). In total, 90% of the genes of the ESBL-positive samples were of CTX-M type. The CTX-M-1 group dominated, followed by the CTX-M-9 group. The repetitive sequence-based PCR fingerprint pattern showed that there was no similarity between the ESBL strains found. Patients who have travelled outside Europe are at high risk of being colonized with ESBL-producing Enterobacteriaceae, and, if infected, are also at risk of receiving inappropriate empirical antibiotic therapy.

Molecular epidemiology of metallo-beta-lactamase-producing Pseudomonas aeruginosa isolates from Norway and Sweden shows import of international clones and local clonal expansion.

Scandinavia is considered a region with a low prevalence of antimicrobial resistance. However, the number of multidrug-resistant (MDR) Gram-negative bacteria is increasing, including metallo-beta-lactamase (MBL)-producing Pseudomonas aeruginosa. In this study MBL-producing P. aeruginosa isolates identified in Norway (n = 4) and Sweden (n = 9) from 1999 to 2007 were characterized. Two international clonal complexes (CC), CC111 (n = 8) and CC235 (n = 2), previously associated with MBL-producing isolates, were dominant. CC111 isolates (ST111/229; serotype O12; bla(VIM-2)) included clonally related isolates identified in Skåne County, Sweden (n = 6), and two isolates associated with importation from Greece and Denmark. In all CC111 isolates, bla(VIM-2) was located in integron In59.2 or In59 variants. The two CC235 isolates (ST235/ST230; serotype O11; bla(VIM-4)) were imported from Greece and Cyprus, were possibly clonally related, and carried bla(VIM-4) in two different integron structures. Three isolates imported from Ghana (ST233; serotype O6; bla(VIM-2)), Tunisia (ST654; serotype O11; bla(VIM-2)), and Thailand (ST260; serotype O6; bla(IMP-14)) were clonally unrelated. ST233 was part of a new CC (CC233) that included other MBL-producing isolates, while ST654 could also be part of a new CC associated with MBL producers. In the isolates imported from Ghana and Tunisia, bla(VIM-2) was part of unusual integron structures lacking the 3' conserved segment and associated with transposons. The bla(VIM) gene was found to be located on the chromosome in all isolates. Known risk factors for acquisition of MBL were reported for all patients except one. The findings suggest that both import of successful international clones and local clonal expansion contribute to the emergence of MBL-producing P. aeruginosa in Scandinavia.

Antimicrobial activity of tigecycline and comparative agents against clinical isolates of staphylococci and enterococci from ICUs and general hospital wards at three Swedish university hospitals.

The activities of tigecycline and comparative agents on staphylococci and enterococci isolated from patients at general hospital wards (GHWs) and intensive care units (ICUs) at 3 university hospitals in Sweden were investigated. Oxacillin disc diffusion and minimal inhibitory concentration with E-test were used. The presence of mecA, vanA or vanB genes was determined with PCR. Statistically significant higher incidence of clindamycin, fusidic acid, rifampicin and multidrug-resistant CoNS was found at ICUs compared to GHWs. Resistance rates were low among S. aureus. Tigecycline, linezolid and vancomycin were the only agents with high activity against methicillin-resistant S. aureus and multidrug-resistant CoNS. Resistance rates were low among E. faecalis, except for high-level gentamicin-resistant (HLGR) E. faecalis. E. faecium showed high resistance rates to ampicillin, piperacillin/tazobactam and imipenem. The HLGR rates among E. faecium were lower than the rates for E. faecalis. Tigecycline and linezolid were the only drugs with high activity against all enterococci including vancomycin-resistant enterococci. No statistically significant differences in susceptibility rates were found between the ward levels for S. aureus and enterococcal isolates and no statistically significant differences were found between the hospitals.

Pharmacodynamic studies of vancomycin, metronidazole and fusidic acid against Clostridium difficile.

BACKGROUND: Pharmacodynamic studies of antibiotics have attracted great interest in recent years. However, studies on the pharmacodynamics of different antibiotics against Clostridium difficile are scarce. METHODS: The postantibiotic effects (PAE) and the postantibiotic sub-minimum inhibitory concentration (MIC) effects (PA SME) of vancomycin, metronidazole and fusidic acid were investigated by viable counts against three different strains of C. difficile. The killing rate and extent of the three antibiotics against the same strains were also studied by adding 2, 4, 8, 16 and 32x MIC of the three antibiotics, respectively. RESULTS: Metronidazole exerted a very rapid bactericidal effect at concentrations of 8x MIC and above against all three strains investigated. Vancomycin gave overall less kill in comparison to metronidazole and was bacteriostatic against two of the three strains. Fusidic acid exerted a concentration-dependent killing against two of the strains. Vancomycin exerted short PAEs and PA SMEs against all three strains. Significantly longer PAEs and PA SMEs were noted for fusidic acid. Metronidazole gave similar short PAEs like vancomycin but longer PA SMEs were noted against two of the investigated strains. CONCLUSION: Metronidazole exerted the most prominent bactericidal effect greater than fusidic acid and greater than vancomycin. Fusidic acid gave overall the longest PAEs and PA SMEs greater than metronidazole and greater than vancomycin.

The throat: an important site for MRSA colonization.

In order to evaluate the value of bacterial cultures taken from the throat, 266 patients with MRSA were retrospectively assessed. At the time when MRSA was first detected in the patient, the most frequent sites positive for MRSA were a skin lesion (110 patients, 41%), the anterior nares (109 patients, 41%), and the throat (102 patients, 38%). In 26%, 17%, and 17% of the patients, a skin lesion, the anterior nares, and the throat, respectively, were the only site where MRSA was seen. In 123 patients cultured for MRSA because of a close contact with an already known MRSA patient, 65 patients (53%) were positive for MRSA in their throat and in 40 patients (33%), throat was the only sample site with MRSA at the time when the patient was found to be MRSA positive. 146 of the 266 patients (55%) were colonized with MRSA in the throat any time throughout the period they were MRSA positive. We conclude that throat is an important reservoir for MRSA and that samples taken from the throat should be included in screening patients for MRSA.

Penicillin-resistant pneumococci in Sweden 1997-2003: increased multiresistance despite stable prevalence and decreased antibiotic use.

Antimicrobial resistance patterns and capsular groups of penicillin-resistant Streptococcus pneumoniae (PRP; MIC penicillin G > or = 0.5 mg/ml) in Sweden between 1997 and 2003 were described, and trends in resistance and antibiotic sales during the same period were compared. The most common serogroups were in descending order 9, 19, 14, 23, and 6. Despite a low and stable annual PRP rate (proportion of PRP out of all pneumococci) of around 2% during the study period, the proportion of PRP resistant to other antibiotics increased. Of all tested PRP isolates, 82% were also resistant to trimethoprim/sulfamethoxazole, 32% had additional resistance to tetracycline, and 26% to erythromycin. Antibiotic sales figures for all studied antibiotic subgroups decreased during the same period. Little correlation was found between antibiotic sales and PRP resistance rates, indicating that there are still other poorly defined factors contributing to the reported resistance levels in the population. However, although PRP strains in Sweden are becoming more commonly resistant to antibiotics other than beta-lactams, the low and further reduced antibiotic sales still might have delayed the development and rapid spread of PRP in the population.

Epidemiological MIC cut-off values for tigecycline calculated from Etest MIC values using normalized resistance interpretation.

OBJECTIVES: To apply the normalized resistance interpretation (NRI) method to Etest MIC results which have higher precision than conventional log2 dilution MIC tests due to the inclusion of intermediate values. If successful, NRI might provide an objective tool for the definition of epidemiological MIC cut-off values. METHODS: MICs of tigecycline and other antimicrobial agents were determined for 4771 clinical isolates comprising five Gram-positive and 13 Gram-negative species or species groups using the Etest. Histograms of MIC values were constructed for each species and NRI calculations were applied to them. An upper MIC limit of 2.5 SD above the theoretical mean of the normalized distribution was used for setting the epidemiological cut-off values. RESULTS: Calculated cut-off values for wild-type strains were between 0.11 and 0.96 mg/L for Gram-positive species, and between 0.44 and 8.3 mg/L for Gram-negative species, except for Pseudomonas aeruginosa, which had a cut-off value of 450 mg/L, consistent with earlier reports on the lack of activity of tigecycline against this species. CONCLUSIONS: NRI offers an objective method for the analysis of MICs produced using Etests and the determination of epidemiological MIC cut-off values.

Activity of three disinfectants and acidified nitrite against Clostridium difficile spores.

OBJECTIVE: To identify environmentally safe, rapidly acting agents for killing spores of Clostridium difficile in the hospital environment. DESIGN: Three classic disinfectants (2% glutaraldehyde, 1.6% peracetyl ions, and 70% isopropanol) and acidified nitrite were compared for activity against C. difficile spores. Four strains of C. difficile belonging to different serogroups were tested using a dilution-neutralization method according to preliminary European Standard prEN 14347. For peracetyl ions and acidified nitrite, the subjective cleaning effect and the sporicidal activity was also tested in the presence of organic load. RESULTS: Peracetyl ions were highly sporicidal and yielded a minimum 4 log10 reduction of germinating spores already at short exposure times, independent of organic load conditions. Isopropanol 70% showed low or no inactivation at all exposure times, whereas glutaraldehyde and acidified nitrite each resulted in an increasing inactivation factor (IF) over time, from an IF greater than 1.4 at 5 minutes of exposure time to greater than 4.1 at 30 minutes. Soiling conditions did not influence the effect of acidified nitrite. There was no difference in the IF among the 4 strains tested for any of the investigated agents. Acidified nitrite demonstrated a good subjective cleaning effect and peracetyl ions demonstrated a satisfactory effect. CONCLUSIONS: Cidal activity was shown against C. difficile spores by glutaraldehyde, peracetyl ions, and acidified nitrite. As acidified nitrite and peracetyl ions are considered to be environmentally safe chemicals, these agents seem well suited for the disinfection of C. difficile spores in the hospital environment.