Clostridium difficile ribotypes in Austria: a multicenter, hospital-based survey

A prospective, noninterventional survey was conducted among Clostridium difficile positive patients identified in the time period of July until October 2012 in 18 hospitals distributed across all nine Austrian provinces. Participating hospitals were asked to send stool samples or isolates from ten successive patients with C.difficile infection to the National Clostridium difficile Reference Laboratory at the Austrian Agency for Health and Food Safety for PCR-ribotyping and in vitro susceptibility testing. A total of 171 eligible patients were identified, including 73 patients with toxin-positive stool specimens and 98 patients from which C. difficile isolates were provided. Of the 159 patients with known age, 127 (74.3 %) were 65 years or older, the median age was 76 years (range: 9–97 years), and the male to female ratio 2.2. Among these patients, 73 % had health care-associated and 20 % community-acquired C. difficile infection (indeterminable 7 %). The all-cause, 30-day mortality was 8.8 % (15/171). Stool samples yielded 46 different PCR-ribotypes, of which ribotypes 027 (20 %), 014 (15.8 %), 053 (10.5 %), 078 (5.3 %), and 002 (4.7 %) were the five most prevalent. Ribotype 027 was found only in the provinces Vienna, Burgenland, and Lower Austria. Severe outcome of C. difficile infection was found to be associated with ribotype 053 (prevalence ratio: 3.04; 95 % CI: 1.24, 7.44), not with the so-called hypervirulent ribotypes 027 and 078. All 027 and 053 isolates exhibited in vitro resistance against moxifloxacin. Fluoroquinolone use in the health care setting must be considered as a factor favoring the spread of these fluoroquinolone resistant C. difficile clones

High genetic diversity ofVibrio choleraein the European lake Neusiedler See is associated with intensive recombination in the reed habitat and the long-distance transfer of strains

Coastal marine Vibrio cholerae populations usually exhibit high genetic diversity. To assess the genetic diversity of abundant V. cholerae non-O1/non-O139 populations in the Central European lake Neusiedler See, we performed a phylogenetic analysis based on recA, toxR, gyrB and pyrH loci sequenced for 472 strains. The strains were isolated from three ecologically different habitats in a lake that is a hot-spot of migrating birds and an important bathing water. We also analyzed 76 environmental and human V. cholerae non-O1/non-O139 isolates from Austria and other European countries and added sequences of seven genome-sequenced strains. Phylogenetic analysis showed that the lake supports a unique endemic diversity of V. cholerae that is particularly rich in the reed stand. Phylogenetic trees revealed that many V. cholerae isolates from European countries were genetically related to the strains present in the lake belonging to statistically supported monophyletic clades. We hypothesize that the observed phenomena can be explained by the high degree of genetic recombination that is particularly intensive in the reed stand, acting along with the long distance transfer of strains most probably via birds and/or humans. Thus, the Neusiedler See may serve as a bioreactor for the appearance of new strains with new (pathogenic) properties.The study was financed by the Austrian Science Fund FWF, project nr P21625-B20. In addition, CP and ISD were partially supported by the Austrian Science Fund FWF, P25745-B20. CA thanks to the Ministerio de Educacion, Cultura y Deporte and FEDER funds for the grant AGL2014-58933-P. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.Peer Reviewe

Characterization of Clostridium difficile isolates using capillary gel electrophoresis-based PCR ribotyping

We have developed a Clostridium difficile PCR ribotyping method based on capillary gel electrophoresis and have compared it with conventional PCR ribotyping. A total of 146 C. difficile isolates were studied: five isolates were reference strains (PCR ribotypes 001, 014, 017, 027 and 053); 141 were clinical isolates comprising 39 Austrian PCR ribotypes collected in the period 2006–2007 at 25 Austrian healthcare facilities. Capillary gel electrophoresis yielded up to 11 fragments per isolate and 47 ribotype patterns. All but one of the five PCR ribotypes of reference strains were clearly reflected in the chromatograms of capillary-based typing. Capillary gel electrophoresis divided 24 isolates belonging to PCR ribotype type 014 into seven subgroups, whereas subtyping the same isolates using multiple-locus variable-number tandem-repeat analysis yielded three unrelated subgroups, without obvious correlation to sr subgroups. Using a web-based software program (http://webribo.ages.at), we were able to correctly identify these 014 isolates by simply allocating the seven subgroup patterns to one ribotype, i.e. to PCR ribotype 014. We consider capillary gel electrophoresis-based PCR ribotyping to be a way of overcoming the problems associated with inter-laboratory comparisons of typing results, while at the same time substantially diminishing the hands-on time for PCR ribotyping

Fatal Pseudomonas aeruginosa pneumonia in a previously healthy woman was most likely associated with a contaminated hot tub

Community-acquired pneumonia due to Pseudomonas aeruginosa in previously healthy individuals is a rare disease that is associated with high fatality. On 14 February 2010 a previously healthy 49-year-old woman presented to an emergency room with signs and symptoms of pneumonia, 2 days after returning from a spa holiday in a wellness hotel. Blood cultures and respiratory specimens grew P. aeruginosa. Despite adequate antimicrobial therapy, the patient died of septic multiorgan failure on day nine of hospitalization. On February 26, nine water samples were taken from the hotel facilities used by the patient: In the hot tub sample 37,000 colony-forming units of P. aeruginosa/100 ml were detected. Two of five individual colonies from the primary plate used for this hot tub water sample were found to be genetically closely related to the patients’ isolates. Results from PFGE, AFLP and MLST analysis allowed the two lung isolates gained at autopsy and the whirlpool bathtub isolates to be allocated into one cluster. The patient most likely acquired P. aeruginosa from the contaminated water in the hotel’s hot tub. The detection of P. aeruginosa in high numbers in a hot tub indicates massive biofilm formation in the bath circulation and severe deficiencies in hygienic maintenance. The increasing popularity of hot tubs in hotels and private homes demands increased awareness about potential health risks associated with deficient hygienic maintenance

Application of three different methods to determine the prevalence, the abundance and the environmental drivers of culturable Vibrio cholerae in fresh and brackish bathing waters

Aims Three cultivation methods were used to study the prevalence and abundance of Vibrio cholerae in Eastern Austrian bathing waters and to elucidate the main factors controlling their distribution. Methods and Results Vibrio cholerae abundance was monitored at 36 inland bathing sites with membrane filtration (MF), a standard most probable number (MPN) approach and direct plating (DP). Membrane filtration yielded the most reliable and sensitive results and allowed V. cholerae detection at 22 sites with concentrations up to 39 000 CFU per 100 ml, all belonging to serogroups other than O1 and O139 and not coding for cholera toxin and toxin coregulated pilus. Direct plating turned out as an easy method for environments with high V. cholerae abundances, conductivity was the only significant predictor of V. cholerae abundance in the bathing waters at warm water temperatures. Conclusions Vibrio cholerae nonO1/nonO139 are widely prevalent in Eastern Austrian bathing waters. Instead of the standard MPN approach, MF and DP are recommended for V. cholerae monitoring. Conductivity can be used as a first easytomeasure parameter to identify potential bathing waters at risk. Significance and Impact of the Study Vibrio cholerae nonO1/nonO139 infections associated with bathing activities are an increasing public health issue in many countries of the northern hemisphere. However, there are only limited data available on the prevalence and abundance of V. cholerae in coastal and inland bathing waters. For monitoring V. cholerae prevalence and abundance, reliable and simple quantification methods are needed. Moreover, prediction of V. cholerae abundance from environmental parameters would be a helpful tool for risk assessment. This study identified the best culturebased quantification methods and a first quick surrogate parameter to attain these aims.(VLID)340099

Listeriosis outbreak caused by acid curd cheese ‘Quargel’, Austria and Germany 2009

We report an outbreak of listeriosis in Austria and Germany due to the consumption of ‘Quargel’ cheese produced by an Austrian manufacturer. At the time of writing this report, the outbreak was known to account for 14 outbreak cases in 2009, including four cases with lethal outcome. On 23 January 2010, the cheese product was voluntarily withdrawn from the market