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Antibiotic resistant bacteria (ARB) in the aquatic environment are reported from all over the world and their presence in the environment has become quite common. The current most prominent example is the presence of beta-lactamases harboring Enterobacteriaceae. The aim of this study was to investigate the presence and diversity (on the genetic and phenotypic levels) of extended spectrum beta-lactamases (ESBL) and carbapenemases harboring Enterobacteriaceae from the River Mur in the center of Graz, Austria's second largest city. Thus over a period of four months water samples were taken, filtrated and screened for these bacteria. All samples revealed ESBL harboring Enterobacteriaceae, of which all with only one exception were Escherichia coli. Dominant ESBL gene family was CTX-M, represented by subgroups CTX-M-1 group, CTX-M-2 group and CTX-M-9 group. Surprisingly co-resistances to non-beta-lactam antibiotics were low, only resistance to trimethoprim was detected in 50% of all (70) isolates. One Klebsiella oxytoca with GES-1 was isolated. To date, GES ESBL has never been reported from Austria before and only rarely from other European countries. Screening for carbapenemase harboring Enterobacteriaceae revealed one Enterobacter cloacae with the gene for VIM-1. Members sharing the same multi-locus-sequence-type (MLST) as well as members of the same rep PCR clusters occurred at different sampling time points. ESBL-harboring Enterobacteriaceae are common in Austrian river water, is dominated by Escherichia coli and CTX-M enzymes. Furthermore, some of the isolates could be linked to different origins., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

We present an isolate of Klebsiella pneumoniae OXA-48 isolated in a 68-year-old man who underwent radical prostatectomy due to prostate cancer. The antibiotic susceptibility testing to a wide range of antibiotics was performed by disk diffusion method and determination of minimal inhibitory concentrations. The isolate was classified as multidrug-resistant. It showed intermediate susceptibility to imipenem and meropenem, resistance to ertapenem, and sensitivity only to colistin, amikacin, and trimethoprim-sulfamethoxazole. The isolate was positive for ESBLs, negative for AmpC. Polymerase chain reaction and sequencing revealed bla(OXA-48)', bla(CTX-M-15) and bla(SHV-11). The plasmid encoding OXA-48 ß-lactamase did not belong to any known PCR-based replicon typing. According to genotyping, the isolate belonged to ST37.

In a clinical setting it seems to be normal these days that a relevant proportion or even the majority of different bacterial species has already one or more acquired antibiotic resistances. Unfortunately, the overuse of antibiotics for livestock breeding and medicine has also altered the wild-type resistance profiles of many bacterial species in different environmental settings. As a matter of fact, getting in contact with resistant bacteria is no longer restricted to hospitals. Beside food and food production, the aquatic environment might also play an important role as reservoir and carrier. The aim of this study was the assessment of the resistance patterns of Escherichia coli and Klebsiella spp. out of surface water without prior enrichment and under non-selective culture conditions (for antibiotic resistance). In addition, the presence of clinically important extended spectrum beta lactamase (ESBL) and carbapenmase harboring Enterobacteriaceae should be investigated. During Joint Danube Survey 3 (2013), water samples were taken over the total course of the River Danube. Resistance testing was performed for 21 different antibiotics. Samples were additionally screened for ESBL or carbapenmase harboring Enterobacteriaceae. 39% of all isolated Escherichia coli and 15% of all Klebsiella spp. from the river Danube had at least one acquired resistance. Resistance was found against all tested antibiotics except tigecycline. Taking a look on the whole stretch of the River Danube the proportion of multiresistances did not differ significantly. In total, 35 ESBL harboring Enterobacteriaceae, 17 Escherichia coli, 13 Klebsiella pneumoniae and five Enterobacter spp. were isolated. One Klebsiella pneumoniae harboring NMD-1 carbapenmases and two Enterobacteriaceae with KPC-2 could be identified. Human generated antibiotic resistance is very common in E. coli and Klebsiella spp. in the River Danube. Even isolates with resistance patterns normally associated with intensive care units are present., Competing Interests: The authors have declared that no competing interests exist.

Carbapenemases involved in acquired carbapenem resistance in Enterobacteriaceae belong to Ambler class A serin β-lactamases, class B metallo-β-lactamases (MBL) or class D OXA-48-like β-lactamases. The aim of the present study was to analyse the molecular epidemiology and the mechanisms and routes of spread of class B and class D carbapenemases in Croatia. In total 68 isolates were analyzed. Antibiotic susceptibility was determined by broth microdilution method. PCR was used to detect antibiotic-resistance genes. Genotyping was performed by rep-PCR and MLST. Sixty-five isolates were found to harbour VIM-1 carbapenemase, seven of which were positive also for NDM-1, while two strains harboured only NDM-1. OXA-48 was detected in three isolates, two of which coproduced VIM-1. Thirty-six strains possessed additional CTX-M-15 β-lactamase whereas 64 were positive for TEM-1. CMY was found in 18 Citrobacter freundii isolates and DHA-1 in one Enterobacter cloacae isolate. Four different plasmid-incompatibility groups were found: A/C, L/M, N and FIIAs. Unlike C. freundii and E. cloacae, Klebsiella pneumoniae showed high diversity of rep-PCR patterns. E. cloacae and C. freundii predominantly belonged to one large clone which was allocated to ST105 and ST24, respectively. Three different types of carbapenemases were identified showing the complexity of CRE in Croatia., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Spa-typing and microarray techniques were used to study epidemiological changes in methicillin-resistant Staphylococcus aureus (MRSA) in South-East Austria. The population structure of 327 MRSA isolated between 2002 and 2012 was investigated. MRSA was assigned to 58 different spa types and 14 different MLST CC (multilocus sequence type clonal complexes); in particular, between 2007 and 2012, an increasing diversity in MRSA clones could be observed. The most abundant clonal complex was CC5. On the respective SCCmec cassettes, the CC5 isolates differed clearly within this decade and CC5/SCCmecI, the South German MRSA, predominant in 2002, was replaced by CC5/SCCmecII, the Rhine-Hesse MRSA in 2012. Whereas in many European countries MLST CC22-MRSA (EMRSA 15, the Barnim epidemic MRSA) is predominant, this clone occurred in Austria nearly 10 years later than in neighbouring countries. CC45, the Berlin EMRSA, epidemic in Germany, was only sporadically found in South-East Austria. The Irish ST8-MRSA-II represented by spa-type t190 was frequently found in 2002 and 2007, but disappeared in 2012. Our results demonstrate clonal replacement of MRSA clones within the last years in Austria. Ongoing surveillance is warranted for detection of changes within the MRSA population., (© FEMS 2016.)

Background: An increased frequency of Proteus mirabilis isolates resistant to expanded-spectrum cephalosporins was observed recently in a long-term care facility in Zagreb (Godan). The aim of this study was the molecular characterization of resistance mechanisms to new cephalosporins in P. mirabilis isolates from this nursing home., Methods: Thirty-eight isolates collected from 2013-2015 showing reduced susceptibility to ceftazidime were investigated. Antibiotic susceptibilities were determined by broth microdilution method. Inhibitor-based tests were performed to detect extended-spectrum (ESBLs) and AmpC β-lactamases. AmpC β-lactamases were characterized by polymerase chain reaction (PCR) followed by sequencing of bla ampC genes. Quinolone resistance determinants (qnr genes) were characterized by PCR. Genotyping of the isolates was performed by repetitive element sequence (rep)-PCR and pulsed-field gel electrophoresis (PFGE)., Results: Presence of an AmpC β-lactamase was confirmed in all isolates by combined-disk test with phenylboronic acid. All isolates were resistant to amoxicillin alone and combined with clavulanate, cefotaxime, ceftriaxone, cefoxitin, and ciprofloxacin; but susceptible to cefepime, imipenem, and meropenem. PCR followed by sequencing using primers targeting bla ampc genes revealed CMY-16 β-lactamase in all but one strain. Bla cmy-16 was carried by a non-conjugative plasmid which did not belong to any known plasmid-based replicon typing (PBRT) group. Rep-PCR identified one large clone consisting of 15 isolates, three pairs or related isolates, one triplet, and four singletons. PFGE confirmed the clonality of the isolates., Conclusions: This is the first report of multidrug resistant P. mirabilis in a nursing home in Croatia. Cephalosporin resistance was due to plasmid-mediated AmpC β-lactamase CMY-16.

Spread and persistence of antibiotic resistance pose a severe threat to human health, yet there is still lack of knowledge about reservoirs of antibiotic resistant bacteria in the environment. We took the opportunity of the Joint Danube Survey 3 (JDS3), the world's biggest river research expedition of its kind in 2013, to analyse samples originating from different sampling points along the whole length of the river. Due to its high clinical relevance, we concentrated on the characterization of Pseudomonas spp. and evaluated the resistance profiles of Pseudomonas spp. which were isolated from eight sampling points. In total, 520 Pseudomonas isolates were found, 344 (66.0%) isolates were identified as Pseudomonas putida, and 141 (27.1%) as Pseudomonas fluorescens, all other Pseudomonas species were represented by less than five isolates, among those two P. aeruginosa isolates. Thirty seven percent (37%) of all isolated Pseudomonas species showed resistance to at least one out of 10 tested antibiotics. The most common resistance was against meropenem (30.4%/158 isolates) piperacillin/tazobactam (10.6%/55 isolates) and ceftazidime (4.2%/22 isolates). 16 isolates (3.1%/16 isolates) were multi-resistant. For each tested antibiotic at least one resistant isolate could be detected. Sampling points from the upper stretch of the River Danube showed more resistant isolates than downriver. Our results suggest that antibiotic resistance can be acquired by and persists even in Pseudomonas species that are normally not in direct contact with humans. A possible scenario is that these bacteria provide a reservoir of antibiotic resistance genes that can spread to related human pathogens by horizontal gene transfer.

Cystic fibrosis (CF) is the most common hereditary lung disease in the Caucasian population, characterized by viscous bronchial secretion, consecutive defective mucociliary clearance, and unavoidable colonization with microorganisms. Besides Pseudomonas aeruginosa, Staphylococcus aureus is the most common bacterial species colonizing the CF respiratory tract. Under antibiotic pressure S. aureus is able to switch to small colony variants (SCV). These small colony variants can invade epithelial cells, overcome antibiotic therapy inside the cells and can be the starting point for extracellular recolonization. The aim of the present study was the isolation and characterization of S. aureus small colony variants from Austrian cystic fibrosis patients. Samples collected from 147 patients were screened for the presence of S. aureus wild-type and small colony variants. Antibiotic susceptibility testing and determination of the small colony variants causing auxotrophism were performed. Wild-type isolates were assigned to corresponding small colony variants with spa typing. In total, 17 different small colony variant isolates and 12 corresponding wild-type isolates were obtained. 13 isolates were determined thymidine auxotroph, 2 isolates were auxotroph for hemin, and none of the tested isolates was auxotroph for both, respectively. The presence of SCVs is directly related to a poor clinical outcome, therefore a monitoring of SCV prevalence is recommended. This study revealed rather low SCV ratios in CF patients compared to other countries.

Objectives: An increase of severe infections caused by Streptococcus dysgalactiae subsp. equisimilis (SDSE) similar to infections caused by Streptococcus pyogenes has been reported over the last years. Little is known about infections with SDSE in Austria. Therefore, we investigated a collection of 113 SDSE invasive and non-invasive isolates from different infection sites and type of infections as well as patients' characteristics., Methods: The isolates were phenotypically identified and emm typed using the enlarged emm database from the Centers for Disease Control and Prevention. Additionally, 13 antimicrobial agents were tested using EUCAST guidelines and virulence genes were investigated., Results: Severe SDSE infections were most common in elderly men with underlying diseases especially diabetes mellitus. With VitekMS identification of SDSE isolates was successful to the species level only. Emm typing revealed 24 different emm types, one new type and one new subtype. StG485, stG6, stC74a, stG643, and stG480 were the predominant types in this study, stC74a and stG652 in invasive infections and stG643, stC74a and stG485 in non-invasive infections. Resistance was observed to tetracycline (62%), macrolides (13%) with one M phenotype, and clindamycin (12%) presenting 6 constitutive MLS(B) phenotypes and 8 inducible MLS(B) phenotypes. Levofloxacin resistance was detected only in one isolate. All isolates tested for virulence genes were positive for scpA, ska, saga and slo. Superantigenic genes were negative except speG(dys) (positive 17/34; 50%)., Conclusion: This paper presents the first report of SDSE infections in Austria. Severe SDSE infections were found mainly in elderly men with underlying diseases. SDSE isolates demonstrated substantial emm type diversity without association with infections site or invasiveness. Analysis of virulence genes showed no significant difference between invasive and non-invasive infections., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Acinetobacter baumannii is an increasingly common pathogen in healthcare settings globally. It is frequently resistant to multiple antimicrobial agents and there are recent reports on strains that are pandrug resistant. The aim of the study was to characterize the mechanisms of carbapenem resistance of A. baumannii strains from a nursing home in Zagreb and to genotype the strains. Antimicrobial susceptibility testing was performed by the broth microdilution method according to the Clinical and Laboratory Standards Institute (CLSI). PCR was used to detect genes encoding carbapenemases of groups A, B, and D and extended-spectrum β-lactamases. Genotyping of the strains was performed by rep-PCR. All strains were found to be resistant to ceftazidime, cefotaxime, ceftriaxone, cefepime, piperacillin/tazobactam, imipenem, meropenem, and ciprofloxacin. All, but one strain, were resistant to gentamicin. PCR revealed blaOXA-23 genes in 14, blaOXA-24 in 5, and blaVIM in 11 strains. All strains positive for blaVIM genes coharbored blaOXA-23 genes. The 14 strains with OXA-23 belonged to ICL II, whereas the 5 strains positive for blaOXA-24 belonged to ICL I. In contrast to hospitals where OXA-24/40-like β-lactamases and OXA-58 were the most prevalent, OXA-23-like β-lactamases are the dominant group in the nursing home. OXA-58-like β-lactamase, which is the most widespread group, was not found. Acquisition of blaMBL genes in A. baumannii strains was observed. Rep-PCR identified two clones. Two strains A10 and A13 were alocated to a novel sequence type ST 637. Nursing homes can act as a source of dissemination of blaOXA and blaMBL genes in the environment and the possible influx to the hospital environment.

Purpose: Implant-associated osteomyelitis still represents a demanding challenge due to unfavourable biological conditions, bacterial properties and incremental resistance to antibiotic treatment. Therefore different bactericide or bacteriostatic implant coatings have been developed recently to control local intramedullary infections. Controlled local release of gentamicin base from a highly lipophilic gentamicin palmitate compound achieves extended intramedullary retention times and thus may improve its bactericide effect., Methods: Forty male Sprague-Dawley rats were divided into two groups receiving an intramedullary femoral injection of 10(2) colony-forming units (CFU) of a common methicillin susceptible Staphylococcus aureus strain (MSSA Rosenbach) and either an uncoated femur nail (Group I) or a nail coated with gentamicin palmitate (Group II). Animals were observed for 28 and 42 days. Serum haptoglobin and relative weight gain were assessed as well as rollover cultures of explanted femur nails and histological scores of periprosthetic infection in dissected femurs., Results: Implants coated with gentamicin palmitate significantly reduced periprosthetic bacterial growth as well as signs of systemic inflammation compared with uncoated implants., Conclusions: Gentamicin palmitate appears to be a viable coating for the prevention of implant-associated infections. These findings will have to be confirmed in larger animal models as well as in clinical trials.

We investigated sinks as possible sources of a prolonged Klebsiella pneumonia carbapenemase (KPC)-producing Klebsiella oxytoca outbreak. Seven carbapenem-resistant K. oxytoca isolates were identified in sink drains in 4 patient rooms and in the medication room. Investigations for resistance genes and genetic relatedness of patient and environmental isolates revealed that all the isolates harbored the blaKPC-2 and blaTEM-1 genes and were genetically indistinguishable. We describe here a clonal outbreak caused by KPC-2-producing K. oxytoca, and handwashing sinks were a possible reservoir., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Background: Cystic fibrosis (CF) is the most common life-limiting inherited disease in Caucasian populations. While pathological changes can be seen in various organs, morbidity and mortality are mainly related to the respiratory tract, with patients suffering from chronic bronchopulmonary infections with characteristic pathogens including Staphylococcus aureus., Objectives: To date, there is only very limited data on the genetic and phenotypic characteristics of S. aureus in CF patients. Therefore, in our study, we characterized 58 S. aureus isolates collected from CF patients in Austria by spa typing, DNA microarray profiling, as well as antimicrobial susceptibility testing in order to determine common genomic and antimicrobial resistance features. The tested strain collection exhibited high genomic diversity., Results: The 58 isolates were assigned to 16 clonal complexes and 48 spa types and differed greatly regarding their virulence and resistance gene profiles. The predominant clonal complexes were MLST CC30 (22%), CC15 (16%), CC45 (14%), and CC5 (12%), complexes that are highly prevalent worldwide among S. aureus strains isolated from humans colonized or infected with S. aureus. DNA microarray profiles showed a wide variety of genes encoding antimicrobial resistance and virulence factors such as various leukocidins, haemolysins, enterotoxins, exfoliative toxins, toxic shock syndrome toxin, as well as genes involved in adhesion and immune evasion., Conclusions: While a large number of strains exhibited resistance to one or several antimicrobial agents, methicillin-resistant S. aureus was found at a low prevalence of 3% (n = 2) only. The two methicillin-resistant S. aureus isolates were assigned to CC152/t355 (SCCmecV) and CC5/t001 (SCCmecI). This is the first study to genetically characterize S. aureus isolates in CF patients in Austria., (© 2015 S. Karger AG, Basel.)

Multidrug resistant bacteria (MDR bacteria), such as extended spectrum beta-lactamase (ESBL) Enterobacteriaceae, methicillin resistant Staphylococcus aureus (MRSA), and vancomycin-resistant Enterococci (VRE), pose a challenge to the human health care system. In recent years, these MDR bacteria have been detected increasingly outside the hospital environment. Also the contamination of food with MDR bacteria, particularly of meat and meat products, is a concern. The aim of the study was to evaluate the occurrence of MDR bacteria in chicken meat on the Austrian market. For this study, 50 chicken meat samples were analysed. All samples originated from chickens slaughtered in Austrian slaughterhouses and were marked as produced in Austria. Samples were analysed for the presence of ESBL Enterobacteriaceae, methicillin resistant Staphylococci and VRE. Resistance genes of the isolated bacteria were characterised by PCR and sequencing. In the present study 26 ESBL producing E. coli, five mecA gene harbouring Staphylococci (but no MRSA), and four VRE were detected in chicken meat samples of Austrian origin. In 24 (48%) of the samples no ESBL Enterobacteriaceae, MRSA, methicillin resistant coagulase negative Staphylococcus (MRCNS) or VRE could be detected. None of the samples contained all three types of investigated multiresistant bacteria. In concordance to previous studies, CTX-M-1 and SHV-12 were the dominant ESBL genes.

Introduction: Resistant bacteria are a well-known public health problem. This study was conducted to investigate the prevalence and genetic characteristics of extended spectrum β-lactamase (ESBL) producing enterobacteria, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) in mixed minced meat from pork and beef., Methods: One hundred samples of mixed minced meat were collected from supermarkets (n = 70) and local butcher shops (n = 30) in the city of Graz (Austria). After enrichment and inoculation on selective media, bacteria were identified with MALDI-TOF MS or Vitek2 systems, tested for antibiotic resistance and further characterized with PCR and sequencing., Results: In 20 of the 100 meat samples 24 ESBL positive Escherichia coli isolates were found. The most common ESBL among the isolates was CTX-M-1. Other detected bla genes contained CTX-M-14, CTX-M-32, SHV-12 and TEM-52 types. Nine samples were tested positive for MRSA and spa-typed. Detected spa-types were hospital-acquired t3928, as well as livestock-associated t011, t034 and t2241. No VRE were found., Conclusion: A contamination of meat with ESBL-producing E. coli and MRSA was confirmed in this study. The large diversity of ESBL producing E. coli could indicate a growing dissemination of ESBL genes in E. coli found in meat products from porcine and bovine origin., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The aim was to determine the prevalence of E. coli and coliform bacteria in playground sand of all public children's sandpits in Graz (n = 45), Austria, and to assess the frequency of antimicrobial resistance in E. coli. Molecular characterization included the discrimination of O-serotypes and H-antigens and the determination of virulence and resistance genes, using a microarray technology. E. coli isolates were tested for susceptibility to a set of antibiotics by VITEK2 system and disk diffusion method. In total, 22 (49%) and 44 (98%) sandpits were positive for E. coli and coliform bacteria. Median concentrations of E. coli and coliform bacteria in the sand samples were: 2.6 × 10(4) CFU/100 g and 3.0 × 10(5) CFU/100 g. Resistance rates were: ampicillin, 12.5%; piperacillin, 10.4%; amoxicillin/clavulanic acid, 9.4%; cotrimoxazole, 6.3%; tetracycline, 6.3%; piperacillin/tazobactam, 5.2%. No ESBL- or carbapenemase-producing isolates were found. The most prevalent serogroups were O15, O6 and O4. Isolates harbored 0 up to 16 different virulence genes., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A total of 112 Staphylococcus aureus bloodstream isolates were genetically characterized. Spa typing and DNA microanalysis exhibited high diversity, resulting in 64 different spa types and 9 different SplitsTree clusters. Methicillin-resistant S. aureus (MRSA) were found in 6 cases only, including the first case of life-stock-associated MRSA (MRSA ST398) in bloodstream infection in Austria., (© 2013.)

For some time now, antibiotic-resistant bacterial strains have been found in the human population, in foods, in livestock and wild animals, as well as in surface waters. The entry of antibiotics and resistant bacterial strains into the environment plays an important role in the spread of antibiotic resistance. The goal of the present study was to monitor the entry of antibiotic resistances into the environment through the contamination of wastewater. To assess the extent of transmission of antibiotic resistances from human sources into the environment, the resistance patterns of Escherichia coli strains isolated from human patients have been compared to those found in strains isolated from sewage sludge. Our results may indicate if resistances to particular antibiotics are more prone than others to spread into the environment. To monitor the increase of specific resistances over time, samples taken in the years 2000 and 2009 were analysed. Our study shows that for some antibiotics a parallel development of resistance patterns has taken place in both patient and environmental samples over time. For other sets of antibiotics, independent developments have occurred in the samples. A clear increase of multi-resistant E. coli strains over time was observed in samples from both sources.

Objective: Fecal carriage of extended-spectrum β-lactamase-producing enterobacteriaceae may contribute to the spread of extended-spectrum β-lactamase-producing enterobacteriaceae into the community. The objective of this study was to assess the duration of fecal carriage after discharge and the occurrence of intrafamilial transmission., Design: Case series., Setting: Quaternary care children's hospital., Patients: Patients colonized with extended-spectrum β-lactamase-producing enterobacteriaceae at the neonatal ICU and the respective household members., Interventions: Screening for intestinal extended-spectrum β-lactamase-producing enterobacteriaceae colonization was done at 1, 2, 4, 6, 9, and 12 months after discharge. Genetic relatedness of isolated extended-spectrum β-lactamase-producing enterobacteriaceae strains was determined using automated rep-PCR., Results: Twenty-five neonates (case-patients) colonized with extended-spectrum β-lactamase-producing enterobacteriaceae (one extended-spectrum β-lactamase-Escherichia coli; six extended-spectrum β-lactamase-Klebsiella pneumoniae; 11 extended-spectrum β-lactamase-Klebsiella oxytoca; and seven extended-spectrum β-lactamase-Serratia marcescens) were included. Duration of fecal carriage was longer (up to 1 yr) in case-patients colonized with Klebsiella species than in case-patients colonized with Serratia marcescens (<4 months). During follow-up, strains and species of extended-spectrum β-lactamase-producing enterobacteriaceae different from the primary strain were found in four and three case-patients, respectively. In nine of 49 (18.4%) included household members, extended-spectrum β-lactamase-producing enterobacteriaceae were found during the follow-up period. In two of nine colonized household members, the isolated extended-spectrum β-lactamase-producing enterobacteriaceae was identical to the primary strains of the respective case-patients., Conclusions: After intestinal colonization with extended-spectrum β-lactamase-producing enterobacteriaceae at the neonatal ICU, infants potentially remain carriers during the first year after discharge. Intrafamilial spread has been proven.

To date, no outbreak of carbapenemase-producing bacteria has been reported for Austria. While outbreaks of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae have been increasingly reported, no outbreak caused by KPC-producing Klebsiella oxytoca has been described yet, to the best of our knowledge. We report an outbreak of KPC-producing K. oxytoca. In 5 months, 31 KPC-producing Klebsiella oxytoca strains were isolated from five patients. All patients were admitted to the same medical intensive care unit in Austria.

Pseudomonas aeruginosa is the major pathogen in the cystic fibrosis (CF) lung, the predominant source of its acquisition, however, is under discussion. In order to study the molecular epidemiology, we evaluated 86 P. aeruginosa isolates from 43 CF patients from southeast Austria. The DiversiLab system was used to identify genetic relationships among the isolates. Antibiotic susceptibilities were tested with a broth microdilution method (Micronaut Merlin). A total of 39 unrelated P. aeruginosa genotypes were found of which 34 were unique to a single patient and one was unique to a sibling pair. We found low rates of resistance for β-lactams with resistance to piperacillin/tazobactam and ceftazidime ranging from 4 to 6%. Resistance rates for meropenem and ciprofloxacin were 11% and 15%, respectively. The prevalence of multidrug-resistant isolates was 2%. We conclude that the majority of P. aeruginosa isolates from CF patients originate from environmental sources and patient-to-patient spread is very uncommon in our centre.

The reduction of implant related infections plays a pivotal role in orthopaedic surgery as an increasing number of people require implants (up to 200,000 per year in the United States (source: Joint Implant Surgery & Research Foundation 2010)). The aim of the current study is to prevent and thus decrease the number of bacterial infections. Both pre and post operative systemic antibiotic treatment and gentamicin containing bone cements (polymethylmethacrylate, PMMA) are commonly used strategies to overcome infections. In this study, the antimicrobial efficacy of gentamicin sulfate loaded bone cement was compared with titan discs coated with a new form of gentamicin, gentamicin palmitate. Adherence prevention, killing rates and killing kinetics were compared in an in vitro model, using Staphylococcus aureus (S. aureus), which together with Staphylococcus epidermidis (S. epidermidis) represents 60% of bacteria found responsible for hip implant infections (An and Friedman, 1996, J Hosp Infect 33(2):93-108). In our experiments gentamicin, which was applied as gentamicin palmitate on the surface of the implants, showed a high efficacy in eliminating bacteria. In contrast to gentamicin sulfate containing bone cements, gentamicin palmitate is released over a shorter period of time thus not inducing antibiotic resistance. Another benefit for clinical application is that it achieves high local levels of active ingredient which fight early infections and minimize toxic side effects. Furthermore, the short term hydrophobic effect of gentamicin palmitate can successfully impede biofilm formation. Thus, the use of self-adhesive antibiotic fatty acid complexes like gentamicin palmitate represents a new option for the anti-infective coating of cementless titan implants.

Colonisation and infection with extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) is an emerging problem. The aim of this study was to investigate whether colistin, which is reported to be effective against multiresistant enterobacteria, prevents ESBL-E colonisation in neonates. For prophylaxis of necrotising enterocolitis, oral gentamicin (15 mg/kg/day) is routinely used in all neonates hospitalised at the Neonatal Intensive Care Unit of University Hospital Graz (Austria). During the study period from May 2005 to September 2007, three ESBL-E outbreaks (total duration 18 months) occurred. During these outbreaks, gentamicin was immediately replaced by oral colistin (8 mg/kg/day) in all hospitalised neonates. All neonates colonised with ESBL-E during the study period were retrospectively analysed with regard to the influence of colistin on ESBL-E colonisation. Genetic relatedness of isolates was assessed by repetitive sequence-based polymerase chain reaction (rep-PCR). During the study period, 30 (4.5%) of 667 neonates were colonised with ESBL-E. Twelve of twenty-one patients colonised with Klebsiella pneumoniae (ESBL-Kp) and one of nine patients colonised with Klebsiella oxytoca (ESBL-Ko) had received oral colistin at time of colonisation with ESBL-E. Amongst ESBL-Kp, the rate of colistin resistance was significantly higher in the colistin group (P=0.0075). Four different clones of ESBL-Kp and three different clones of ESBL-Ko were isolated, indicating the occurrence of patient-to-patient transmission. Colistin-resistant as well as colistin-susceptible isolates were detected within the same clones, indicating induction of resistance. At the dosage used, oral colistin did not prevent colonisation with ESBL-E and appeared to select colistin-resistant strains or to induce colistin resistance., (Copyright © 2010 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.)

Carnobacterium species have been isolated from the environment and are not regarded as human pathogens, although they are known to cause disease in fish. Only two reports describing isolation of Carnobacterium species from human pus were found in the literature. We report what we believe to be the first isolation of Carnobacterium sp. from a human blood culture.

The aim of this study was to investigate the degree of contamination of sewage sludge with ESBL-producing Escherichia coli strains and the effectiveness of different sewage sludge treatment methods. Monthly sewage sludge samples were collected between January and September 2009 in 5 different sewage treatment plants and tested for the presence of ESBL E. coli. In addition, the number of colony forming units (CFU) of E. coli and coliform bacteria before and after the different sludge treatment methods (aerobic/anaerobic digestion, lime stabilization, and thermal treatment) was investigated. Of the 72 sewage sludge samples investigated, ESBL-positive E. coli were found in 44 (61.1%) sewage sludge samples. The classification of beta-lactamase groups was carried out in 15 strains resulting in the detection of 2 different groups (CTX-M and TEM) of bla genes. All 15 of them had a CTX-M gene and 4 of these strains furthermore carried a TEM gene. With regard to the CFU of E. coli and coliform bacteria, thermal treatment and lime stabilization following dehydration sufficiently reduced pathogen concentrations. The plants using merely stabilization and dehydration showed an increase of E. coli and coliform bacteria and thus also an increase in ESBL-producing E. coli., (Copyright 2009 Elsevier Ltd. All rights reserved.)

In this retrospective study, the occurrence and genetic relatedness of methicillin-resistant Staphylococcus aureus (MRSA) ST398 in Austrian MRSA patients was investigated. From 2002 to 2008, 14 MRSA ST398 were detected. First occurrence of MRSA ST398 was already found in 2004. Spa ribotyping assigned 12 isolates to spa type t011 and 1 each to spa type t034 and spa type t1451. Isolated MRSA ST398 was nontypeable by pulsed-field gel electrophoresis (NT-MRSA) using restriction enzyme SmaI; therefore, genotyping was performed using automated repetitive-sequence-based PCR (rep-PCR) on the DiversiLab system. Rep-PCR results assigned 10 (71%) of the 14 MRSA ST398 into 1 cluster with a similarity >95%; there was 1 cluster consisting of 2 isolates with a similarity >99% and 2 unique MRSA ST398 isolates. In conclusion, MRSA ST398 was continuously detected in Southeast Austria; first in 2004 with up to 5 MRSA ST398 isolates in 2008. Automated rep-PCR proved as a reliable technique in determining genetic relatedness of NT-MRSA ST398 and demonstrates clonal spread of MRSA ST398 in the investigated region., (2010 Elsevier Inc. All rights reserved.)