[Detection of viruses in raw water as a basic tool for risk assessment].

Human pathogenic viruses may end up in surface waters by fecal contamination. However, the German drinking water ordinance requests that pathogens in drinking water should not be present in concentrations constituting a potential danger to human health. Since many viruses do have a very low dose of infection, they have to be sufficiently eliminated in the process of drinking water purification.

[Viruses in drinking water].

Viruses in drinking water can cause infectious diseases. In the past, hepatitis A and E were the most frequently observed drinking- water-borne viral infections, but in recent years several small- and large-scale norovirus epidemics have been described, even in Europe. All virus species spread via drinking water are of fecal origin.

A randomized controlled trial assessing infectious disease risks from bathing in fresh recreational waters in relation to the concentration of Escherichia coli, intestinal enterococci, Clostridium perfringens, and somatic coliphages.

We performed epidemiologic studies at public freshwater bathing sites in Germany to provide a better scientific basis for the definition of recreational water quality standards. A total of 2,196 participants were recruited from the local population and randomized into bathers and nonbathers. Bathers were exposed for 10 min and had to immerse their head at least three times.

Development and test for long-term stability of a synthetic standard for a quantitative Cryptosporidium parvum LightCycler PCR assay.

A recently described quantitative rapid cycle real time PCR (LightCycler) assay detects Cryptosporidium parvum after in vitro excystation, which is a surrogate marker for the viability of the organisms. In the original assay the quantification standard is a dilution series of C. parvum oocysts with a microscopically determined excystation rate.

Effect of comprehensive validation of the template isolation procedure on the reliability of bacteraemia detection by a 16S rRNA gene PCR.

The influence of the DNA extraction method on the sensitivity and specificity of bacteraemia detection by a 16S rRNA gene PCR assay was investigated. The detection limit of the assay was 5 fg with purified DNA from Escherichia coli or Staphylococcus aureus, corresponding to one bacterial cell. However, with spiked blood samples, the detection limits were 10(4) and 10(6) CFU/mL, respectively.

Improved detection of Salmonella spp. in foods by fluorescent in situ hybridization with 23S rRNA probes: a comparison with conventional culture methods.

This report describes a new technique for the detection and identification of Salmonella species in food with the use of fluorescent in situ hybridization (FISH) with 23S rRNA-targeted oligonucleotide probes. Two species-specific 23S rRNA-targeted oligonucleotide probes (Sal-1 and Sal-3) were selected, and one (Sal-544) was newly designed. The relative specificities of these probes were compared with those of bacterial 23S rRNA sequences from the GenBank database and tested by in situ hybridization with bacterial cell smears of pure cultures.

DNase pretreatment of master mix reagents improves the validity of universal 16S rRNA gene PCR results.

DNase I pretreatment of 16S rRNA gene PCR reagents was tested. The DNase I requirement for the elimination of false-positive results varied between 0.1 and 70 IU per master mix depending on the applied Taq polymerase.

A case report of false negative Legionella test results in a chlorinated public hot water distribution system due to the lack of sodium thiosulfate in sampling bottles.

We examined samples from the showers and the central water distribution system of a public building with an indoor swimming pool. The pool was used for school and recreational activities and as a sports therapy facility for patients with coronary heart disease. The building's hot water system was contaminated with Legionella pneumophila.

Effects of reduced mucus oxygen concentration in airway Pseudomonas infections of cystic fibrosis patients.

Current theories of CF pathogenesis predict different predisposing "local environmental" conditions and sites of bacterial infection within CF airways. Here we show that, in CF patients with established lung disease, Pseudomonas aeruginosa was located within hypoxic mucopurulent masses in airway lumens. In vitro studies revealed that CF-specific increases in epithelial O(2) consumption, linked to increased airway surface liquid (ASL) volume absorption and mucus stasis, generated steep hypoxic gradients within thickened mucus on CF epithelial surfaces prior to infection.

The effect of human serum DNAases on the ability to detect antibiotic-killed Escherichia coli in blood by PCR.

PCR has proved superior to conventional blood culture for diagnosing bacteraemia in the presence of antibiotics. Nevertheless, even PCR might yield false-negative results if the template DNA were to be cleaved by serum DNAases after antibiotics had induced bacterial death. To evaluate the cleavage of bacterial template DNA by human serum DNAase I, serum samples inoculated with purified Escherichia coli DNA were incubated with increasing amounts of recombinant human DNAase (rhDNAase) and then examined by a PCR specific for E.

Effects of amoxicillin, gentamicin, and moxifloxacin on the hemolytic activity of Staphylococcus aureus in vitro and in vivo.

In Staphylococcus aureus infection hemolysis caused by the extracellular protein alpha-toxin encoded by hla is thought to contribute significantly to its multifactorial virulence. In vitro, subinhibitory concentrations of beta-lactam antibiotics and fluoroquinolones increase the levels of hla and alpha-toxin expression, whereas aminoglycosides decrease the levels of hla and alpha-toxin expression. In the present study we investigated the effects of subinhibitory concentrations of amoxicillin, gentamicin, and moxifloxacin on hla and alpha-toxin expression and total hemolysis of S.

Rapid identification of Enterobacteriaceae using a novel 23S rRNA-targeted oligonucleotide probe.

The aim of this study was to rapidly identify bacteria of the family of Enterobacteriaceae using fluorescent in situ hybridization (FISH). A comparative sequence analysis was carried out and a 23S rRNA signature sequence for Enterobacteriaceae was identified. A 23S rRNA-targeted oligonucleotide probe (EBAC1790) was constructed and subsequently tested against 40 reference strains.

Adherence of Staphylococcus aureus to endothelial cells: influence of capsular polysaccharide, global regulator agr, and bacterial growth phase.

The adherence of Staphylococcus aureus to human endothelial cells (EC) is probably an important step in the pathogenesis of systemic staphylococcal infections. We examined the influence of type 5 capsular polysaccharide (CP5) production, the global regulator agr, and the bacterial growth phase on S. aureus adherence to EC.

Molecular epidemiology of community-acquired Staphylococcus aureus in families with and without cystic fibrosis patients.

The molecular epidemiology of Staphylococcus aureus nasal commensal strains and community-acquired infecting strains was assessed by comparison of prevalence, persistence, transmission rate, and clonal distribution of S. aureus in families with and without cystic fibrosis (CF) patients. Isolates were typed by pulsed-field gel electrophoresis.

Direct quantitative transcript analysis of the agr regulon of Staphylococcus aureus during human infection in comparison to the expression profile in vitro.

Bacteria possess a repertoire of distinct regulatory systems promoting survival in disparate environments. Under in vitro conditions it was demonstrated for the human pathogen Staphylococcus aureus that the expression of most virulence factors is coordinated by the global regulator agr. To monitor bacterial gene regulation in the host, we developed a method for direct transcript analysis from clinical specimens.

Trefoil factor family domain peptides in the human respiratory tract.

Trefoil factor family domain peptides (TFF) are thought to be involved in mucosal epithelial restitution and wound healing of the gastrointestinal tract and are up-regulated in ulceration and in a variety of solid tumours. It was hypothesized that TFFs are also expressed on mucosal surfaces of the human respiratory tract. Lung tissue, nasal polyps, and sputum samples from seven patients with cystic fibrosis (CF), two with chronic and acute bronchitis, and non-dysplastic material from two cases of bronchial adenocarcinoma were analysed for TFF expression by immunohistochemistry, immunofluorescence, western blot and RT-PCR.

[Comparison of European standards of swimming pool safety].

European countries follow different procedures to monitor the quality of the water of swimming pools open to the public: some use generally accepted technical standards, some officially recommended guidelines, some rules or regulations by local authorities and some have national laws or regulations. These agree insofar as in every country the water of swimming pools must be disinfected. The microbiological limits are to a certain extent identical with drinking water threshold values, e.

PCR and blood culture for detection of Escherichia coli bacteremia in rats.

Critically ill patients often develop symptoms of sepsis and therefore require microbiological tests for bacteremia that use conventional blood culture (BC) techniques. However, since these patients frequently receive early empirical antibiotic therapy before diagnostic procedures are completed, examination by BC can return false-negative results. We therefore hypothesized that PCR could improve the rate of detection of microbial pathogens over that of BC.

[Dust and microorganism count at delivery, sorting and composting of home refuse and home refuse-like industrial waste].

The exposure of employees to airborne dust and microorganisms was assessed in a waste processing plant established to recover reusable materials from unsorted domestic and industrial waste. Exposure criteria considered relevant were the quantity of the individual size-selected particle fractions, the morphological properties of the particles, their heavy metal content, and the degree of their contamination with various microorganisms and mold. In addition, separate microbiological analyses to determine potential pathogen concentrations in the air were made.

Distribution and transmission of Pseudomonas aeruginosa and Burkholderia cepacia in a hospital ward.

Genotyping and antibiotic susceptibility testing were used to analyze Pseudomonas aeruginosa and Burkholderia cepacia strains from sink drain from 14 pediatric patients with cystic fibrosis (CF) and from hospital personnel as part of a 4 week prospective study of strain transmission in a pediatric ward. A total of 87.5% of all washbasin drains were contaminated with P.

Selective detection of viable Cryptosporidium oocysts by PCR.

Although the diagnosis of cryptosporidiosis in diarrheic patients is relatively simple, as large amounts of oocysts are usually shed, environmental samples can contain only few oocysts which have a comparatively high epidemiological relevance. Very sensitive detection methods are therefore requested in environmental hygiene. Additionally these methods should allow a statement about the viability of the detected organisms.

[Occurrence and detection of viruses in drinking water].

Viruses can pass disinfection steps of water treatment plants without being inactivated. Investigations during the last 15 years revealed repeatedly the presence of enteric viruses in finished water meeting standards for coliform bacteria. Methods for the detection of viruses in water which implicate their growth on specific cell cultures are very time consuming and do not cover many viral species.