KPC-2 and OXA-48 carbapenemase-harbouring Enterobacteriaceae detected in an Austrian wastewater treatment plant.

Multiresistant Enterobacteriaceae, like carbapenemase-producing strains, have their primary reservoir in medical institutions. They can also be found with increasing tendency in other reservoirs. One possible way for entrance of multiresistant Enterobacteriaceae into the environment is via waste water. The aim of the study was to screen isolates from a wastewater treatment plant for the presence of carbapenemase-producing Enterobacteriaceae. Three isolates harboured carbapenemase genes, one Klebsiella pneumoniae harboured KPC-2 and one K. pneumoniae and one Escherichia coli harboured OXA-48. This is the first report of carbapenemase-harbouring Enterobacteriaceae isolated outside medical institutions in Austria and the first detection of KPC-harbouring K. pneumonia MLST ST 1245.

Water quality assessment of a Central European River - does the Directive 2000/60/EC cover all the needs for a comprehensive classification?

Over the last decades, European rivers have mostly shared the same fate. Until the mid 1980s many of them were in rather bad shape. Nobody would have thought about using the waters for recreation. However, continuous efforts over the last three decades have achieved success and today's water quality has changed to the better. This obvious change in water quality, which was proved by investigations done on the basis of the European Directive 2000/60/EC framework for Community action in the field of water policy, made people consider using the rivers for recreation or irrigation. In the present study a Central European River was investigated from hygienic perspectives (faecal indicators), and its toxicological status was examined - both of which are parameters which are not included in the Directive 2000/60/EC. The pivotal question was whether these parameters provide essential and additional information about the river's status and its quality. In general, the monthly values for all sampling points from the toxicological tests correlated with the determined physical and chemical statuses according to the national water management plan. Once, however, the toxicological values did increase significantly. This detection of a single toxicological burst within a year of observation underlines the applicability of the tests as screening methods for the detection of toxic substances. Moreover, our microbiological investigations show high numbers of faecal indicators and Salmonellae. These results are not consistent with the apparently good quality of the river. Therefore faecal indicators, which are directly related to the discharge of waste water, should be an integral part of the procedure of water classification.

Comparison of extended-spectrum-ß-lactamase (ESBL) carrying Escherichia coli from sewage sludge and human urinary tract infection.

For many years, extended-spectrum-beta-lactamase (ESBL) producing bacteria were a problem mainly located in medical facilities. Within the last decade however, ESBL-producing bacteria have started spreading into the community and the environment. In this study, ESBL-producing Escherichia coli from sewage sludge were collected, analysed and compared to ESBL-E. coli from human urinary tract infections (UTIs). The dominant ESBL-gene-family in both sample groups was bla(CTX-M), which is the most prevalent ESBL-gene-family in human infection. Still, the distribution of ESBL genes and the frequency of additional antibiotic resistances differed in the two sample sets. Nevertheless, phenotyping did not divide isolates of the two sources into separate groups, suggesting similar strains in both sample sets. We speculate that an exchange is taking place between the ESBL E. coli populations in infected humans and sewage sludge, most likely by the entry of ESBL E. coli from UTIs into the sewage system.

Concentrations of viable airborne fungal spores and trichloroanisole in wine cellars.

In wineries, unwanted microorganisms present not only hygienic problems but also have a negative influence on wine quality. An evaluation of Austrian/Styrian wine cellars with regard to the volume and the composition of the mycoflora is very important both for the process of wine production and for occupational safety. Thirty-six wine cellars of 20 vintners were investigated with regard to microorganisms in the air and on material surfaces. Moreover, the presence of trichloroanisole in the air was determined by means of solid-phase micro-extraction. Microorganisms were sampled using the six-stage Andersen-Cascade impactor. The results showed that the concentrations of xerophilic fungi in the air of cellars with large visible mold areas (> 80%) reached values up to 1.4 × 104 colony forming units per m³. In the wine cellars fourteen predominant fungal genera were found in the indoor air, the most frequent was Penicillium. Trichloroanisole was detected in the air of wine cellars with large visible moldy patches. The spore concentrations in the cellar air were two times higher in cellars with Zasmidium cellare growth than in cellars without Z. cellare. These results will serve as a database for further studies.

Lethal brain abscess due to the fungus Scedosporium apiospermum (teleomorph Pseudallescheria boydii) after a near-drowning incident: case report and review of the literature.

A 39-year-old healthy man developed a brain abscess weeks after a near-drowning incident. Scedosporium apiospermum, the anamorph of Pseudallescheria boydii, was isolated from the abscess. The patient died 153 days after the accident despite antifungal therapy. We discuss the role of antifungals and review the literature for comparable cases.

Transmission of Campylobacter spp. in a poultry slaughterhouse and genetic characterisation of the isolates by pulsed-field gel electrophoresis.

1. Contamination of retail products with Campylobacter spp. during the slaughter of poultry is a well-known problem of product hygiene. Mechanical evisceration often leads to intestinal rupture and discharge of gut contents, which can contain zoonotic and human pathogens. Processes along the slaughter line cause aerosols and airborne droplets, containing bacterial loads. 2. To estimate the possible transmission routes of intestinal Campylobacter, 36 measurements of the bioaerosol (Andersen sampler and SKC BioSampler), 30 cloacal (of three flocks), 10 equipment and 4 sedimentation samples were tested for the presence of Campylobacter species. 3. The results imply that, in addition to contaminated equipment, which was Campylobacter-positive in 80% of cases, aerosols with peak values of 4.0 x 10(4) (test series 1) and 1.4 x 10(4) (test series 2) CFU/m3 also provide a potential vector for horizontal transmission. 4. To explore the genetic similarities of isolates from different origins, 18 isolates recovered from air, 26 cloacal, 8 equipment and 4 sedimentation isolates were analysed by pulsed-field gel electrophoresis (PFGE), using the restriction enzymes Sma I and Sal I. The similarity of cloacal isolates with isolates from equipment, air and sediment, suggest that the contamination is of intestinal origin. 5. There were direct links between Campylobacter-positive flocks and the presence of the same strains in the aerosol of the slaughter hall. Air as a potential source for microbial transmission must be taken into account.

Antibiotic resistance of E. coli in sewage and sludge.

The aim of the study is the evaluation of resistance patterns of E. coli in wastewater treatment plants without an evaluation of basic antibiotic resistance mechanisms. Investigations have been done in sewage, sludge and receiving waters from three different sewage treatment plants in southern Austria. A total of 767 E. coli isolates were tested regarding their resistance to 24 different antibiotics. The highest resistance rates were found in E. coli strains of a sewage treatment plant which treats not only municipal sewage but also sewage from a hospital. Among the antimicrobial agents tested, the highest resistance rates in the penicillin group were found for Ampicillin (AM) (up to 18%) and Piperacillin (PIP) (up to 12%); in the cephalosporin group for Cefalothin (CF) (up to 35%) and Cefuroxime-Axetil (CXMAX) (up to 11%); in the group of quinolones for Nalidixic acid (NA) (up to 15%); and for Trimethoprime/Sulfamethoxazole (SXT) (up to 13%) and for Tetracycline (TE) (57%). Median values for E. coli in the inflow (crude sewage) of the plants were between 2.0 x 10(4) and 6.1 x 10(4)CFU/ml (Coli ID-agar, BioMerieux 42017) but showed a 200-fold reduction in all three plants in the effluent. Nevertheless, more than 10(2)CFU E. coli/ml reached the receiving water and thus sewage treatment processes contribute to the dissemination of resistant bacteria in the environment.

Comparative investigation of airborne culturable microorganisms in sewage treatment plants.

The present study investigated emissions and emmissions of airborne microorganisms (mesophilic bacteria, Escherichia coli, molds, Aspergillus fumigatus, thermophilic actinomycetes/bacilli) in sewage treatment plants. For the aerobiological investigations three sewage treatment facilities with an activated-sludge process, capacities between 2000 and 28,000 PE and different cleaning steps were selected. The measurements of microorganism emission were conducted in the area of the intake (screen), in the area of biological treatment (activated sludge tank) and at a distance of 10 m from the activated sludge tanks. In order to determine the emmission, additional measurements were conducted leeward of the plant at a distance of 200 m. Samples were taken using four parallel six-stage Andersen 1 AFCM volumetric samplers. In the area of the intake counts for bacteria were 7.4 x 10(2) CFU/m3 (median), for thermophilic actinomycetes 1.8 x 10(1) CFU/m3, for thermophilic bacilli 7.1 x 10(1) CFU/m3, for molds 2.4 x 10(3) CFU/m3 and for Aspergillus fumigatus 1.8 x 10(1) CFU/m3. Only isolated airborne coliform recoveries, i.e. E. coli, were detected. In the area of the activated sludge tank, in the adjoining area (10 m) and in the vicinity of the plants (200 m), the counts for all microorganism groups investigated corresponded to natural conditions. The results show that the counts of culturable aerogenic microorganisms in and in the immediate surrounding of the sewage plants investigated are low. Although the possibility of an infection through inhalation cannot be ruled out, the direct contact with sewage is much more critical.

Comparative investigations of airborne culturable microorganisms in selected waste treatment facilities and in neighbouring residential areas.

The evaluation of airborne microorganisms in waste treatment facilities is complicated by different measuring systems, a lack of measuring standards and large variations between individual counts. In the present study, different sectors of the waste management industry were compared by determining median values of airborne culturable microorganisms from numerous parallel counts over a prolonged time period. The samples were taken during the warm season using the six-stage Andersen volumetric sampler in a large composting plant and its immediate vicinity, in an agricultural composting plant, a waste disposal site, and a sorting facility for recyclable materials. Control samples were taken at a site not influenced by the waste management industry in an open and largely uninhabited area. The highest median values for culturable bacteria (37 degrees C) found were 1.1 x 10(5) CFU/m3, for moulds (25 degrees C) 1.4 x 10(5) CFU/m3, and for A. fumigatus (37 degrees C) 1.7 x 10(4) CFU/m3 in the sorting cabins of the sorting facility (p < 0.001). The highest median values for thermophilic bacteria (actinomycetes and bacillaceae, 50 degrees C) were 7.3 x 10(3) CFU/m3 in the large composting facility. In all other facilities as well as in the neighbouring residential areas of all facilities investigated, all median values were significantly lower and corresponded to the naturally occurring levels: approx. 10(2) CFU/m3 for bacteria, approx. 10(3) CFU/m3 for moulds and approx. 10(1) CFU/m3 for A. fumigatus and thermophilic bacteria. Only in the neighbouring residential area of the large composting plant, the median values for thermophilic bacteria were approx. 10(2) CFU/m3, but an additional impact from farms cannot be excluded in this case. These results show permanent increased loads of the investigated microorganisms inside large composting facilities and especially in the sorting cabins for recyclable materials. The increasing number of reports on potential health hazards in these areas require adequate measures on the part of occupational medicine in order to limit the health risk to a minimum. The most important task is the automatization of the sorting process for recyclable materials.

[Waste management--health: a longitudinal study]. / Abfallwirtschaft--Gesundheit: Eine Longitudinalstudie.

Altogether 256 (148 men and 108 women) employees from manual sorting facilities and 120 (98 men and 22 women) employees from compost facilities were examined The main focus of the investigations was in the assessment of the lung-function and the immune-system. A part of employees (87 in sorting and 34 in compost facilities) was observed over a period of 3 years. The lung-function was in the area of the expected values, decreased however from year to year. The concentration of total-IgE increased over the period of 3 years. The observation is from great interest that the difference of the concentration of the total-IgE fluctuated within the 3 years at the employees of manual sorting facilities from +200% to -100% while it amounted to 0% at employees of the compost facilities.

[Comparative studies of airborne, cultivatable microorganisms at selected sites of waste managements, domestic animal husbandry and in the surrounding residential area]. / Vergleichende Untersuchungen luftgetragener, kultivierbarer Mikroorganismen an ausgewählten Standorten in der Abfallwirtschaft, der Nutztierhaltung und im Anwohnerbereich.

During the last years, sampling of airborne microorganisms (including mesophilic bacteria, moulds, thermophilic bacteria and A. fumigatus) in waste disposal, recycling industry and control sampling locations in the urban and rural districts of Graz was conducted using Andersen 6-stage viable cascade impactors. In the present study additional sampling in livestock buildings has been conducted. Except mesophilic bacteria the emission in the area of waste disposal and recycling industry was significantly higher than in livestock buildings. In the surroundings of the livestock buildings the number of microorganisms was not increased, while the normal background level in the surroundings of the waste proceeding plants was reached from a distance of 150 to 300 meters.

Diarrhea in returning Austrian tourists: epidemiology, etiology, and cost-analyses.

BACKGROUND: Between 1995 and 1997, stool samples of 322 Austrian tourists returning from abroad with diarrhea were examined for bacteria, parasites and viruses. METHODS: Epidemiologic data were collected from information furnished by physicians and hospitals and from questionnaires. Moreover, testing expenses and additional cost for treated cases were evaluated. RESULTS: In 97 of 322 patients examined (30%), one or more pathogens were detected in the stool. Bacteria were found in 38 patients (39%), parasites in 33 patients (34%) and viruses in 26 patients (27%). In 6 patients, mixed infections with parasites and viruses were detected and in 5 patients with bacteria and viruses. Among bacteria, Campylobacter jejuni was most frequent; among parasitic infections, Giardia lamblia. Significant correlations were established between the country of destination, age, travel style and length of stay. Forty-four percent of all patients visited Asia (including Turkey), 27% Africa, 18% Latin America, and only 10% southern Europe. The group between 20 and 29 years of age was most frequently affected (p<.001), the group between 0 and 19 years of age least. Fifty-seven percent stayed in a hotel without frequent changes of location; 43% undertook a trekking trip; and of those, 75% belonged to the group aged between 20 and 39. In terms of the correlation between travel style and pathogen, it was found that 74% of patients with bacterial infections stayed in a hotel (avg. 57.9%; p<.05) whereas 64% of all patients with parasitic infections undertook a trekking trip (avg. 42%; p<.001). Thirty-six percent of all patients with parasitic infections spent their vacation in India (avg. 13%; p<.001). The length of stay of patients with bacterial infections was shorter than average (72% spent between 1 and 2 weeks abroad, avg. 49.8%). Patients with parasitic infections spent significantly more time abroad than average (42% more than 2 months; avg. 17.7%; p<.001). Average cost of specific antimicrobial therapy was U.S.$31 whereas the average cost of identifying a patient needing such treatment was almost U.S.$580. CONCLUSION: Optimal detection rate and cost reduction for the diagnosis require precise history, adequate collection of samples using adequate transport media, and rapid transfer to the laboratory.

Air-borne microorganisms in the metropolitan area of Graz, Austria.

Urban and rural regions are affected by different microorganism loads depending on their structure and utilization. At 7 sampling sites in the metropolitan area of Graz, counts of airborne bacteria as well as yeasts and molds were conducted over a one-year period at two-week intervals. Bacteria and yeasts/molds counts in a village area to the South of Graz dominated by agriculture exceeded the corresponding counts in a suburban residential area fourfold (327 CFU/m3 air-bacteria) and twofold (185 CFU/m3 air-yeasts/molds) respectively. In the vicinity of a composting facility located in the same residential area, microorganism counts exceeded those of the neighboring "unaffected" area by 29% in the case of bacteria and by 54% in the case of yeasts/molds. At an industrial and business site with heavy traffic, the counts are twice that of the area affected by the composting facility (146 CFU/m3 for bacteria and 168 CFU/m3 for yeasts/molds). The proportion of Aspergillus fumigatus is highest in the village area with 23%, compared to 10% in the open land. 49% of the bacteria and 54% of the yeasts and molds can be shown on stages 4-6 of the Andersen-Volumetric-Sampler registering the respirable particle sizes.

Rapid detection of Legionella species in bronchoalveolar lavage fluids with the EnviroAmp Legionella PCR amplification and detection kit.

A molecular assay based on a rapid DNA extraction protocol and the EnviroAmp Legionella Kits was used to detect Legionella species in bronchoalveolar fluid specimens. All Legionella strains isolated from tap water in hospitals could be detected distinctly. Both sensitivity and specificity were tested. In a prospective study, bronchoalveolar lavage fluids obtained from patients with atypical pneumonia were investigated. Three positive samples were detected with the molecular techniques and were subsequently confirmed by culture. Application of the system described may lead to safe and early diagnosis of Legionnaires' disease in patients with atypical pneumonia.

Comparative study of procedures for isolation and cultivation of Legionella pneumophila from tap water in hospitals.

For the isolation and cultivation of Legionella pneumophila from tap water in hospitals, we compared different media and selection techniques. A second part of the study compared the L. pneumophila yields from different water samples at identical sites. A total of 210 water samples (500 ml each) were collected from two selected sites in each of 21 hospitals. Warm water samples were collected after flow times of 0, 5, 10, and 15 min; in addition, one cold water sample was collected. Filtration was used to concentrate all samples. Following filtration, 0.1 and 1 ml each of untreated samples, heat-treated samples (3 min, 59 degrees C), and acid-treated samples (pH 2.2, 15 min) were spread onto the selective media MWY (SR 118; Oxoid) and BMPA alpha (SR 111; Oxoid), and samples from 12 hospitals were also spread onto GVPC medium (SR 152; Oxoid). A total of 72 (34%) of the 210 samples from 12 hospitals were positive. With respect to the positive Legionella cultures, there was no significant difference between the selective media MWY, BMPA alpha, and GVPC. With the BMPA alpha supplement, more samples were positive following heat treatment (P < 0.05) or acid treatment (P < 0.05) than without any further treatment. For the maximum yield of Legionella colonies with minimum additional microbial flora, acid treatment was the most effective, and by all methods, the GVPC supplement was the most selective. For routine water tests in hospitals for differentiating between systemic and local contamination, acid treatment of the concentrated samples, the use of different selective media, and the correct selection of sampling sites are recommended.

[Detection of legionellae in thermal water]. / Nachweis von Legionellen im Thermalwasser.

Water samples taken from hot water springs, from the water distribution system and from various places of consumption of thermal water were collected in an Austrian spa. 56.5% of the samples contained Legionella; serogroups 1, 3 and 5 of Legionella pneumophila were most frequently identified. The large number of positive Legionella samples indicates a potential risk of infection to patients, especially those undergoing inhalation treatment with thermal water, or those using a whirlpool or taking a shower. In those inhalators actually in use, no Legionella could be detected. Thus they are no source of infection when they are maintained and serviced properly. Thermal disinfection and service checks at regular intervals are suggested for the contaminated systems.

Hygienic investigation of coastal waters of the upper Adriatic Sea.

In the course of the bathing season of 1989, investigations of bathing waters were carried out in two-week intervals. From the point of view of public health, the chemical-physical and microbiological results do not suggest objections against bathing at the beach areas investigated (Grado, Lignano). However, these favorable results do not imply intact ecological conditions. National and international standards are designed for humans and allow only very limited conclusions about the living conditions of the marine ecosystem. This also means that ecological investigations are not sufficient to permit conclusions about hygienic conditions. In spite of this seeming contradiction, hygienic and ecological concerns are clearly identical. Hygienic measures must not be limited to local "cosmetic" corrections but must target foremost unfavorable basic conditions.

Cryptosporidiosis in children in Idukki District in southern India.

Concentrated stool samples from 560 children, aged from two weeks to 10 years, from Idukki District, southern India, were examined to detect Cryptosporidium oocysts, using a phenol-auramin stain and fluorescent microscopy. Smears of stool samples were also stained by a modified Ziehl-Neelsen method and were examined by light-microscopy. One sample was collected from each of 266 children with diarrhoea and 294 age-matched children without diarrhoea. Cryptosporidium oocysts were detected in 4.5% of all stools, in 6% of diarrhoeal stools and in 3% of stools from healthy controls. Results of the study show that Cryptosporidium infection is common in children with diarrhoea in southern India, and there is also a high frequency of asymptomatic cyst-passers in this area.