Franz Allerberger

Clinical Course and the Role of Shiga Toxin–Producing Escherichia coli Infection in the Hemolytic-Uremic Syndrome in Pediatric Patients, 1997–2000, in Germany and Austria: A Prospective Study

Hemolytic-uremic syndrome (HUS) is mainly associated with foodborne infections by Shiga toxin-producing Escherichia coli (STEC). From January 1997 through December 2000, 394 children with HUS were evaluated in a prospective multicenter surveillance study in Germany and Austria (incidences, 0.7/100,000 and 0.4/100,000 children <15 years old, respectively). Blood leukocytosis was associated with increased detection of STEC in stool cultures (P<.01) and a more severe disease course. Risk of death was associated with cerebral involvement (P<.01). Most strikingly, non-O157:H7 STEC were detected in 43% of stool cultures of patients with HUS: O26 was detected in 15%, sorbitol-fermenting O157:H(-) in 10%, O145 in 9%, O103 in 3%, and O111 in 43%. Patients with O157:H7 serotypes required dialysis for a longer time and had bloody diarrhea detected more frequently, compared with patients with non-O157:H7 serotypes (P<.05). This large study in children with HUS underlines the rising importance of non-O157:H7 serotypes, and, despite increased public awareness, the number of patients remained unchanged.

Livestock-associated Methicillin-Resistant Staphylococcus aureus in Humans, Europe

To estimate the proportion of methicillin-resistant Staphylococcus aureus (MRSA) isolates from humans that were sequence type (ST) 398, we surveyed 24 laboratories in 17 countries in Europe in 2007. Livestock-associated MRSA ST398 accounted for only a small proportion of MRSA isolates from humans; most were from the Netherlands, Belgium, Denmark, and Austria.

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.

Changing bacterial ecology during a five year period of selective intestinal decontamination

The development of bacterial resistance during selective decontamination of the digestive tract (SDD) is controversial. We studied effects on bacterial resistance one year before and during a randomized, placebo-controlled trial of SDD in a surgical intensive care unit. We randomized patients within two different topical regimens (PTA, PCA) or placebo, administered four-times daily to both the oropharynx and gastrointestinal tract. All patients received intravenous ciprofloxacin (200 mg b.d.) for four days. Both SDD regimens successfully reduced aerobic Gram-negative intestinal colonization. There was no increase in resistance of Enterobacteriaceae or Pseudomonas aeruginosa. Acinetobacter calcoaceticus developed multi-resistance over one year, but differences between groups were not significant. We detected a shift towards Gram-positive organisms. Oxacillin-resistant Staphylococcus aureus increased in concert with ciprofloxacin resistance, from 17 to 80.7%, and frequencies of resistance were significantly higher in SDD patients (P < 0.001). Resistance of coagulase-negative staphylococci (CNS) to oxacillin increased initially (25 to 66.9%), but values returned to baseline in controls. Ciprofloxacin resistance in CNS remained higher (P < 0.001) in SDD-treated patients (52.5 vs. 23.3%). The incidence of late respiratory tract infections was unaltered by the prophylactic regimen (SDD 35.2%; Placebo 41.2%; n.s.). We cannot recommend SDD as a prophylactic tool in critically ill patients.

Strain-Specific Differences in the Amount of Shiga Toxin Released from Enterohemorrhagic Escherichia coli O157 following Exposure to Subinhibitory Concentrations of Antimicrobial Agents

Abstract There is no consensus regarding the benefit versus harm of antibiotic therapy for treatment of disease due to enterohemorrhagic Escherichia coli O157. The effects in vitro of subinhibitory concentrations of 13 antimicrobial agents on the release of Shiga toxin (Stx) by three different Escherichia coli O157 strains expressing Stx 1 or Stx 2 either alone or in combination were investigated. The Stx-induced cell death of Vero cells was determined using a colorimetric assay based on the measurement of lactate dehydrogenase (LDH) released into the supernatant from the cytosol of damaged cells. Growth of all O157 strains in broth cultures containing subinhibitory concentrations of cotrimoxazole, trimethoprim, azithromycin, or gentamicin was accompanied by a marked increase in the release of Stx. Exposure to cefixime, ceftriaxone, or erythromycin caused a marked increase in the release of Stx by the O157 strain producing Stx 2 alone, but decreased toxin production was observed with the Stx 1 producer and the strain producing Stx 1 and Stx 2. Exposure to ampicillin caused increased Stx release in the Stx 2-producing strain but had no effect on Stx production in the other two test isolates. Exposure to penicillin G, streptomycin, ciprofloxacin, fosfomycin, or sulfamethoxazole caused an increase in toxin production in two of the three test strains in each case, while decreases were observed for the other isolates. The response of Escherichia coli O157 isolates to subinhibitory concentrations of antibiotics seems to be highly dependent on the nature of the strain involved.

Classifying spa Types in Complexes Improves Interpretation of Typing Results for Methicillin-Resistant Staphylococcus aureus

ABSTRACT A total of 382 isolates of methicillin-resistant Staphylococcus aureus originating from three Austrian regions and one adjacent Italian region (Vienna, Lower Austria, North Tyrol, and South Tyrol) were typed by DNA sequence analysis of the variable repeat region of the protein A gene (spa typing). The strain collection consisted of arbitrarily chosen isolates originating from clinical specimens taken in the years 2003 to 2005 at 17 hospitals. The most common spa types found were t001 (28.8% of all isolates), t190 (27.0%), t008 (14.1%), and t041 (11.3%). The 42 remaining spa types accounted for ≤2.4% each. The dominating spa types varied between the different regions. As short sequence DNA repeat units are unstable entities, the 46 spa types were classified into seven spa complexes with respect to short sequence repeat unit composition and organization. Such classification into complexes can provide additional information for the hospital epidemiologist, empowering one to differentiate the introduction of a new strain from mere variation of endemic spa types.

Clostridium difficile: a new zoonotic agent?

ZusammenfassungClostridium difficile wird meist als nosokomialer Erreger, der mit Diarrhö oder pseudomembranöser Colitis einhergeht, angesehen. Im Jahr 2007 wurden von Spitälern in Österreich 2.761 Clostridium-difficile-Infektionen (277 davon mit tödlichem Ausgang) gemeldet, im Vergleich dazu waren im Jahr 2003 nur 777 Fälle (53 davon mit tödlichem Ausgang) registriert worden. Neben diesem Anstieg der nosokomialen Erkrankungen wird auch über ein vermehrtes Auftreten von ambulant erworbenen C. difficile-Infektionen berichtet. Aktuelle Publikationen belegen das Vorkommen von C. difficile bei Tieren und in Lebensmitteln. Ziel der vorliegenden Studie war der Nachweis von C. difficile in Tieren und Lebensmitteln in Österreich. Zwischen März 2008 und Juli 2008 wurden Stuhl- und Kotproben von 67 Rindern, 61 Schweinen und 59 Broilern in österreichischen Schlachthöfen gesammelt. Zudem wurden Fleischproben (51 Rind [25 davon faschiert], 27 Schwein [17 faschiert] und 6 Hühnerfleischproben) zwischen Februar 2008 und April 2008 in Supermärkten gekauft und untersucht. Von den 187 getesteten Proben konnten in 8 C. difficile nachgewiesen werden: dabei waren 3/67 (4,5%) Kuhproben, 2/61 (3,3%) Schweineproben und 3/59 (5%) Hühnerproben positiv. Sechs der acht Isolate (2/67 [3%] Kuhproben, 2/61 [3,3%] Schweineproben und 2/59 [3,4%] Hühnerproben) waren toxinbildende C. difficile. In einem der zwei Toxin-positiven C. difficile Schweineproben war auch der Nachweis eines binären Toxin-Bildners des PCR-Ribotyps 126 möglich. In keiner der untersuchten Fleischproben konnte C. difficile nachgewiesen werden. Die Ergebnisse dieser Studie bestätigen, dass Tiere als Reservoir dienen können und eine Übertragung von C. difficile über Lebensmittel möglich ist.SummaryClostridium difficile is mainly considered a nosocomial pathogen associated with diarrhea and pseudomembranous colitis in hospitalized patients. Austrian hospitals reported 2761 cases of C. difficile infection (including 277 lethal outcomes) in 2007, compared with 777 cases (including 54 lethal outcomes) in 2003. The occurrence of community-acquired C. difficile infection is also increasingly reported. Recent studies have shown the occurrence of C. difficile in food and animals. The aim of the present study was to determine the occurrence of C. difficile in food and animals in Austria. Between March and July 2008, gut or fecal samples from 67 cows, 61 pigs and 59 broiler chickens were collected at Austrian abattoirs. Between February and April 2008 meat samples (51 beef [25 ground], 27 pork [17 ground] and 6 samples of chicken meat) were purchased at retail outlets. Of the 187 samples tested, eight yielded C. difficile: in cows 3/67 samples (4.5%) were positive, in pigs 2/61 (3.3%), in broiler chickens 3/59 (5%). Six of the eight isolates yielded toxigenic C. difficile (toxins A and B): 2/67 (3%) cow samples, 2/61 (3.3%) pig samples, 2/59 (3.4%) chicken samples. Genes for the binary toxin were detected in one of the two pig isolates, a PCR ribotype 126 strain. None of the 84 meat samples yielded C. difficile. The results of this Austrian study suggest that animal reservoirs are possible sources, via food, of human C. difficile infection.

Infection of Red Deer, Cattle, and Humans with Mycobacterium bovis subsp. caprae in Western Austria

ABSTRACT Twelve cases of Mycobacterium bovis subsp. caprae infection have occurred in four humans, three cattle, and five red deer in western Austria since 1994. DNA-fingerprinting of the isolates suggested transmission in and between these species over several years. Contact with cattle, but not with goats, was found to be associated with three of four human cases.

Incidence of Fecal Carriage of Listeria monocytogenes in Three Healthy Volunteers: A One-Year Prospective Stool Survey

The aim of this study was to establish the incidence of fecal carriage of Listeria monocytogenes in healthy adults. A total of 868 stool specimens from three healthy volunteers (1 male and 2 females; ages 44, 39, and 60 years) were collected between 1 October 2000 and 30 September 2001. Culture was performed using Fraser broth and Palcam selective agar plates. Polymerase chain reaction (PCR) was performed using Probelia Listeria monocytogenes (BioRad, France). Overall, Listeria monocytogenes was detected in 31 of the 868 (3.57%) stool specimens using PCR. Sixteen of the 31 positive results were single events, i.e., samples collected from the same patient the day before and the day after the positive result were both negative. Positive results on two consecutive days were found four times, on three consecutive days one time, and on four consecutive days one time. Listeria monocytogenes was cultured from 10 of 868 (1.15%) stool specimens. These culture-positive samples, all positive by PCR as well, accounted for five independent episodes. Using automated ribotyping on up to 40 single colonies per stool specimen, 9 of 10 culture-positive samples yielded more than one strain. There was no obvious seasonal clustering of positive results. None of the documented episodes of Listeria monocytogenes carriage, all of which involved serotypes 1/2a and 1/2b, coincided with overt illness. The results of PCR indicate an incidence of five to nine exposures to Listeria monocytogenes per person per year. On average, the incidence of culture-confirmed fecal carriage in healthy adults is two episodes of Listeria monocytogenes carriage per person per year. Fecal shedding was of short duration (maximum 4 days), which argues against the appropriateness of routine stool screening in dairy workers as a tool for prevention of listeriosis.

Spectrum of enteropathogens detected by the FilmArray GI Panel in a multicentre study of community-acquired gastroenteritis

The European, multicentre, quarterly point-prevalence study of community-acquired diarrhoea (EUCODI) analysed stool samples received at ten participating clinical microbiology laboratories (Austria, Finland, France, Germany, Greece, Ireland, Italy, Portugal, Romania, and the UK) in 2014. On four specified days, each local laboratory submitted samples from ≤20 consecutive patients to the Austrian Study Centre for further testing with the FilmArray GI Panel (BioFire Diagnostics, Salt Lake City, UT, USA). Of the 709 samples from as many patients received, 325 (45.8%) tested negative, 268 (37.8%) yielded only one organism, and 116 (16.4%) yielded multiple organisms. Positivity rates ranged from 41% (30 of 73 samples) in France to 74% (59 of 80 samples) in Romania. With the exception of Entamoeba histolytica and Vibrio cholerae, all of the 22 targeted pathogens were detected at least once. Enteropathogenic Escherichia coli, Campylobacter species, toxigenic Clostridium difficile, enteroaggregative E. coli, norovirus and enterotoxigenic E. coli were the six most commonly detected pathogens. When tested according to local protocols, seven of 128 positive samples (5.5%) yielded multiple organisms. Overall, the FilmArray GI Panel detected at least one organism in 54.2% (384/709) of the samples, as compared with 18.1% (128/709) when testing was performed with conventional techniques locally. This underlines the considerable potential of multiplex PCR to improve routine stool diagnostics in community-acquired diarrhoea. Classic culture methods directed at the isolation of specific pathogens are increasingly becoming second-line tools, being deployed when rapid molecular tests give positive results. This optimizes the yield from stool examinations and dramatically improves the timeliness of diagnosis.