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.

Emission of extensively-drug-resistant Acinetobacter baumannii from hospital settings to the natural environment.

BACKGROUND: Acinetobacter baumannii is a leading emerging pathogen that is frequently recovered from patients during hospital outbreaks. The role of environmental A. baumannii reservoirs is therefore of great concern worldwide. AIM: To investigate the connection between A. baumannii causing hospital outbreaks and environmental isolates from hospital wastewater, urban sewage and river water as the final natural recipient of wastewaters. METHODS: Clinical isolates from patients with hospital-acquired pneumonia and environmental isolates from water were collected during a two-month monitoring period. Recovery of A. baumannii was performed using CHROMagar Acinetobacter plates, incubated at 42°C for 48 h. Identification was performed by matrix-assisted laser desorption ionization-time of flight mass spectrometry and analyses of rpoB gene. The antibiotic resistance profiles were interpreted according to criteria given for clinical isolates of A. baumannii. The sequence types (ST) were retrieved by multi-locus sequence typing. RESULTS: Fourteen of 19 isolates recovered from patients, hospital wastewaters, urban sewage and river water belonged to ST-195. The remaining five isolates recovered from patients and river water were assigned to ST-1421. All isolates showed very strong relatedness and clustered into CC92, which corresponds to IC2. All isolates were non-susceptible to at least one agent in all but two or fewer antimicrobial categories, and thus were classified as 'extensively-drug-resistant' (XDR). Heteroresistance to colistin was found in two isolates from hospital wastewater. CONCLUSION: Close relatedness of clinical and environmental isolates suggests the emission of XDR A. baumannii via the untreated hospital wastewater in the natural environment.

Sarcoidosis and tuberculosis in South Croatia: are there epidemiological similarities or not?

OBJECTIVE: Tuberculosis and sarcoidosis are chronic granulomatous diseases. Clinical, pathologic and immunologic aspects are similar although different. The authors were interested to highlight possible epidemiological similarities of these two granulomatous diseases. The objective of this study was to evaluate incidence rate as well as age, sex and geographic distribution of sarcoidosis in South Croatia and to compare it with these epidemiological characteristics of tuberculosis. STUDY DESIGN: Retrospective. METHODS: The study was including ten years follow up period (1997-2006), and was performed in Split-Dalmatia County, Croatia. All data were collected retrospectively and analyzed using Statistica 7 programme. RESULTS: The mean annual incidence of sarcoidosis was 3.3/100,000 inhabitants with a mean of 15,6 cases per year. Woman accounted for 61% of all sarcoidosis cases. The mean sarcoidosis patient age was 44.94 ± 11.85 years. The peak age group was 40-49 years (31%). Significant difference according to incidence rate on the islands comparing to the rates on the coast and the mainland was observed (P = 0.003). The mean sarcoidosis mortality rate was 1.2/100,000. Statistically significant differences between sarcoidosis and tuberculosis were observed according the higher number of tuberculosis patients (P < 0.000), among males (P < 0.000), and females, too (P < 0.000) as well as in mortality rates (P = 0.401). Significantly more patients had tuberculosis on the mainland (P < 0.000) and on the coast (P < 0.000), but not in the islands (P = 0.260). CONCLUSIONS: The results from this study showed dissimilarities in classic epidemiological patterns between sarcoidosis and tuberculosis, incidence rates, as well as sex and geographic distribution. Our findings resulted from this study might be starting point for the future epidemiological, genetic, and immunological studies.

Antimicrobial susceptibility and beta-lactamase production of selected gram-negative bacilli from two Croatian hospitals: MYSTIC study results.

The meropenem yearly Susceptibility Test Information Collection (MYSTIC) programme is a global, longitudinal resistance surveillance network that monitors the activity of meropenem and compares its activity with other broadspectrum antimicrobial agents. We now report the antimicrobial efficacy of meropenem compared to other broad-spectrum agents within the selective Gram-negative pathogen groups from two Croatian Hospitals investigated between 2002-2007. A total of 1510 Gram-negative pathogens were tested and the minimum-inhibitory concentrations (MICs) were determined by broth microdilution method according to CLSI.There was no resistance to either imipenem or meropenem observed for Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis in both medical centers. High resistance rates of K. pneumoniae to ceftazidime (18%), cefepime (17%) and gentamicin (39%) are raising concern. Acinetobacter baumannii turned out to be the most resistant Gram-negative bacteria with 81% resistant to ceftazidime, 73% to cefepime, 69% to gentamicin and 71% to ciprofloxacin. Almost 20% of Pseudomonas aeruginosa strains were resistant to imipenem, 13% to meropenem, 69% to gentamicin and 38% to ciprofloxacin.The prevalence of extended-spectrum beta-lactamases (ESBLs) in E. coli was 10% and in K. pneumoniae 49%. PCR and sequencing of the amplicons revealed the presence of SHV-5 in nine E. coli strains and additional tem-1 beta-lactamase five strains. Five K. pneumoniae strains were positive for bla(SHV-5 )gene. Eight ESBL positive Enterobacter spp. strains were found to produce tem and CtX-m beta-lactamases. Plasmid-mediated AmpC beta-lactamases were not found among K. pneumoniae, E. coli and Enterobacter spp. Three A. baumannii strains from Zagreb University Center were identified by multiplex PCR as OXA-58 like producers. Six A. baumannii strains from Split University Center were found to possess an ISAba1 insertion sequence upstream of bla(OXA-51 )gene. According to our results meropenem remains an appropriate antibiotic for the treatment of severe infections caused by Gram-negative bacteria. These data indicate that despite continued use of meropenem, carbapenem resistance is not increasing among species tested, except for A. Baumannii, in the two study hospitals and suggest that clinicians can still administer carbapenems as a reliable and effective choice in managing serious nosocomial infections.