First evidence of KPC-producing ST258 Klebsiella pneumoniae in river water.

This paper reports the first case of Klebsiella pneumoniae carbapenemase (KPC)-2-producing K. pneumoniae in river water in Croatia. In total, four KPC-2-producing K. pneumoniae isolates were analysed. All isolates shared a similar genetic background, belonging to ST258. Isolates displayed uniform, multi-drug-resistant profiles susceptible to colistin. blaSHV-1, aac(3')-II, aac(6')-Ib and aph(3')-Ia genes were detected in all isolates. In all isolates, the blaKPC-2 gene was localized on a single non-conjugative IncFII plasmid that varied in size (∼140, ∼230, ∼225 and ∼220 kb). K. pneumoniae was viable in river water for up to 50 days, confirming its ability to survive and disseminate in the environment.

Spread of Enterobacter cloacae carrying blaNDM-1, blaCTX-M-15, blaSHV-12 and plasmid-mediated quinolone resistance genes in a surgical intensive care unit in Croatia.

The objective of this study was to describe a hospital cluster of NDM-1-producing Enterobacter cloacae infections observed in the surgical intensive care unit (ICU) of a tertiary-care hospital at Pula, Croatia. NDM-1-producing E. cloacae strains isolated from clinical samples were screened by PCR for the presence of carbapenemases. Genetic relatedness of NDM-1-producing E. cloacae strains was determined by multilocus sequence typing (MLST). During the period October 2013 to April 2014, four patients, with overlapping hospital stay in the surgical ICU, developed severe infections caused by E. cloacae demonstrated to produce carbapenemases. According to MLST, all strains belonged to ST133 and were positive by PCR for the blaNDM-1 carbapenemase gene, for blaCTX-M-15 and blaSHV-12 extended-spectrum ß-lactamase (ESBL) genes, and for blaTEM-1 and blaOXA-1 narrow-spectrum ß-lactamase genes. They were negative for other carbapenemases genes including blaOXA-48, blaVIM and blaKPC as well as for AmpC and the armA and rmtB aminoglycoside resistance genes. All strains were positive for the HI2 replicon, suggesting that an IncHI2 plasmid is likely the plasmid carrying the blaNDM-1 gene. Infection control measures were implemented after the first case although they were not effective in avoiding spread of this organism to other patients in the surgical ICU. In conclusion, the evolving epidemiology of NDM-producing micro-organisms and the interspecies diffusion of this resistance mechanism to emerging pathogens such as E. cloacae necessitate the setting up of strong and urgent joint measures to control the spread of NDM carbapenemase especially in the ICU setting.

Carbapenemase-producing Enterobacteriaceae in Europe: a survey among national experts from 39 countries, February 2013.

The spread of carbapenemase-producing Enterobacteriaceae (CPE) is a threat to healthcare delivery, although its extent differs substantially from country to country. In February 2013, national experts from 39 European countries were invited to self-assess the current epidemiological situation of CPE in their country. Information about national management of CPE was also reported. The results highlight the urgent need for a coordinated European effort on early diagnosis, active surveillance, and guidance on infection control measures.

A European survey of antibiotic management of methicillin-resistant Staphylococcus aureus infection: current clinical opinion and practice.

Although the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) varies across Europe, healthcare-associated MRSA infections are common in many countries. Despite several national guidelines, the approach to treatment of MRSA infections varies across the continent, and there are multiple areas of management uncertainty for which there is little clinical evidence to guide practice. A faculty, convened to explore some of these areas, devised a survey that was used to compare the perspectives of infection specialists from across Europe on the management of MRSA infections with those of the faculty specialists. The survey instrument, a web-based questionnaire, was sent to 3840 registered delegates of the 19th European Congress of Clinical Microbiology and Infectious Diseases, held in April 2009. Of the 501 (13%) respondents to the survey, 84% were infection/microbiology specialists and 80% were from Europe. This article reports the survey results from European respondents, and shows a broad range of opinion and practice on a variety of issues pertaining to the management of minor and serious MRSA infections, such as pneumonia, bacteraemia, and skin and soft tissue infections. The issues include changing epidemiology, when and when not to treat, choice of treatment, and duration and route of treatment. The survey identified areas where practice can be improved and where further research is needed, and also identified areas of pan-European consensus of opinion that could be applied to European guidelines for the management of MRSA infection.

Outpatient use of systemic antibiotics in Croatia.

According to the results of the European Surveillance of Antimicrobial Consumption (ESAC), outpatient antibiotic use in Croatia in 2001 (17.6 DDD per 1000 inhabitants per day) was comparable to the median use of 24 European countries, which were able to deliver valid data.

Transferable DHA-1 cephalosporinase in Escherichia coli.

Three Escherichia coli isolates resistant to third-generation cephalosporins but negative for extended-spectrum beta-lactamase production were isolated from hospitalised patients in Zagreb, Croatia, during June 2003 to February 2004. Resistance was due to the inducible production of a DHA-1 cephalosporinase. Each isolate contained an integron-associated bla(DHA-1)-ampR sequence carried by similar-sized plasmids, of which one was self-transferable. Serotyping and polymerase chain reaction typing using ERIC2 primer indicated that the isolates were distinct. This is the first description of DHA beta-lactamase production in E. coli.

Activity of telithromycin and seven other agents against 1034 pediatric Streptococcus pneumoniae isolates from ten central and eastern European centers.

OBJECTIVE: To test the activity of telithromycin against 1034 Streptococcus pneumoniae isolates from pediatric patients in ten centers from ten central and eastern European countries during 2000-2001, and to compare it with the activities of erythromycin A, azithromycin, clarithromycin, clindamycin, and quinupristin-dalfopristin. METHODS: The minimum inhibitory concentrations (MICs) of telithromycin, erythromycin A, azithromycin, clarithromycin, clindamycin, levofloxacin, quinupristin-dalfopristin and penicillin G were tested by the agar dilution method with incubation in air, and mechanisms of resistance to macrolides and quinolones were investigated. RESULTS: Strains were isolated from sputum, tracheal aspirates, ear, eye, blood, and cerebrospinal fluid. Among S. pneumoniae strains tested, 36% had raised penicillin G MICs (>/= 0.12 mg/L). Susceptibilities were as follows: telithromycin, quinupristin-dalfopristin and levofloxacin, >/= 99%; clindamycin, 83%; and erythromycin A, azithromycin and clarithromycin, 78%. Of 230 (22.3%) erythromycin A-resistant S. pneumoniae strains, 176 (79.6%) had erm(B), 38 (16.1%) had mef(A), and 10 (4.3%) had mutations in 23S ribosomal RNA or in ribosomal protein L4. The rates of drug-resistant S. pneumoniae are high in all centers except Kaunas, Riga, and Prague. CONCLUSION: Telithromycin had low MICs against all strains, irrespective of macrolide, azalide or clindamycin resistance. Ribosomal methylation was the most prevalent resistance mechanism among all resistant strains, except in Sofia, where the prevalence of the efflux mechanism was higher.

Activity of telithromycin compared with seven other agents against 1039 Streptococcus pyogenes pediatric isolates from ten centers in central and eastern Europe.

In total, 1039 pediatric Streptococcus pyogenes isolates from Bulgaria, Croatia, the Czech Republic, Hungary, Latvia, Lithuania, Poland, Romania, Slovakia and Slovenia were studied. All strains were susceptible to penicillin G, levofloxacin, and quinupristin-dalfopristin, 91-100% to telithromycin, and 82-100% to erythromycin, azithromycin, and clarithromycin, and 90-100% to clindamycin. Macrolide resistance occurred mainly in Slovakia (25%), the Czech Republic (17.3%), and Croatia (15.8%). Overall, 9.7% of S. pyogenes isolates were erythromycin resistant due to erm(B)- or erm(A)-encoded methylases (72.3%) or to a mef(A)-encoded efflux pump (25.7%). One strain had alterations of both 23S rRNA (A2058G Escherichia coli numbering) and ribosomal protein L22 (G95D).

Bacterial and fungal microleakage of AH26 and AH Plus root canal sealers.

AIM: To evaluate the penetration of Candida albicans alone and a combination of bacteria through root canals filled with gutta-percha and one or other root canal sealers, AH26 and AH Plus. METHODOLOGY: Eighty teeth were randomly divided into two groups of 40 teeth each and obturated with gutta-percha using either AH26 or AH Plus sealer. A further 10 teeth served as negative controls and 10 as positive controls. The external surface of each root, except the apical 2 mm, was covered with two layers of nail varnish. The teeth were inserted into Eppendorf plastic tubes and suspended in glass bottles containing sterile Schaedler broth. Streptococcus mutans, Streptococcus mitis, Prevotella melaninogenica and Lactobacillus acidophilus were placed in the access cavities of 20 teeth filled with AH26 and 20 with AH Plus. Candida albicans was placed in the access cavities of the other teeth. The culture medium with microorganisms was changed every 7 days. Every 72 h bacterial or fungal growth in the broth was tested up to a period of 90 days. RESULTS: Leakage in the experimental teeth occurred between 14 and 87 days. Leakage was present in 47% of all samples. From the samples with AH26, 45% leaked bacteria and 60% leaked fungi; whilst from the samples with AH Plus, 50% leaked bacteria and 55% fungi. There was no statistically significant difference in penetration of bacteria and fungi between the sealers. CONCLUSION: In this in vitro study, gutta-percha and the sealers AH26 and AH Plus allowed leakage of bacteria and fungi.

Failure of bacteriophage typing to detect an inter-hospital outbreak of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb subsequently identified by random amplification of polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE).

OBJECTIVE: To establish the extent of inter-hospital spread of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb and to determine the most suitable method for typing local strains. METHODS: We analyzed a collection of 33 MRSA isolates from three Zagreb hospitals together with five unrelated British MRSA isolates by antibiogram typing, bacteriophage typing, randomly amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis (PFGE) after digestion with Smal restriction endonuclease. Bacteriophage typing was done with the international set of S. aureus typing phages. RAPD and PFGE profiles were analyzed visually and by using the 'GelCompar' computer program. RESULTS: Antibiogram typing provided eight profiles. Thirty (91%) of the 33 Croatian strains of MRSA were non-typable by phage typing. Visual analysis of RAPD products identified six, and visual analysis of PFGE fragments nine, distinct profiles. Computer analysis of RAPD data separated British isolates from the Croatian ones, but did not cluster the visually determined RAPD types. PFGE computer analysis separated British isolates and clustered isolates in concordance with visual interpretation. Thirty-one of the 38 isolates (82%) were visually grouped in the same clusters by both molecular methods. The dominant strain was present in each of the three hospitals. CONCLUSIONS: Bacteriophage typing was unhelpful for the analysis of Croatian MRSA, since most strains were untypable with the international set of bacteriophages. RAPD and PFGE were more successful in typing the organisms and showed evidence of inter-hospital spread of one predominant MRSA strain in all three Zagreb hospitals. Thus RAPD and PFGE proved to be a useful aid in elucidating the epidemiology of MRSA infection in Zagreb hospitals and should be established in Croatia for typing MRSA.