Regina Sommer

UV inactivation, liquid-holding recovery, and photoreactivation of Escherichia coli O157 and other pathogenic Escherichia coli strains in water.

Drinking water, water used in food production and for irrigation, water for fish farming, waste water, surface water, and recreational water have been recently recognized as a vector for the transmission of pathogenic Escherichia coli, especially serotype O157:H7. We investigated the UV (253.7 nm) inactivation behavior and the capability of dark repair (liquid-holding recovery) and photoreactivation of seven pathogenic (including three enterohemorrhagic E. coli) strains and one nonpathogenic strain of E. coli (ATCC 11229) with respect to the use of UV light for water disinfection purposes. Because most bacteria and yeast are known to be able to repair UV damage in their nucleic acids, repair mechanisms have to be considered to ensure safe water disinfection. We found a wide divergence in the UV susceptibility within the strains tested. A 6-log reduction of bacteria that fulfills the requirement for safe water disinfection was reached for the very most susceptible strain O157:H7 (CCUG 29199) at a UV fluence of 12 J/m2, whereas for the most resistant strain, O25:K98:NM, a UV fluence of about 125 J/m2 was needed. Except for one strain (O50:H7) liquid-holding recovery did not play an important role in recovery after UV irradiation. By contrast, all strains, particularly strains O25:K98:NM, O78:K80:H12, and O157:H7 (CCUG 29193), demonstrated photorepair ability. For a 6-log reduction of these strains, a UV fluence (253.7 nm) up to 300 J/m2 is required. The results reveal that the minimum fluence of 400 J/m2 demanded in the Austrian standard for water disinfection is sufficient to inactivate pathogenic E. coli. A fluence of 160 J/m2 (recommendation in Norway) or 250 J/m2 (recommendation in Switzerland) cannot be regarded as safe in that respect.

Calicivirus Inactivation by Nonionizing (253.7-Nanometer-Wavelength [UV]) and Ionizing (Gamma) Radiation

ABSTRACT Noroviruses (previously Norwalk-like viruses) are the most common viral agents associated with food- and waterborne outbreaks of gastroenteritis. In the absence of culture methods for noroviruses, animal caliciviruses were used as model viruses to study inactivation by nonionizing (253.7-nm-wavelength [UV]) and ionizing (gamma) radiation. Here, we studied the respiratory feline calicivirus (FeCV) and the presumed enteric canine calicivirus (CaCV) and compared them with the well-studied bacteriophage MS2. When UV irradiation was used, a 3-log10 reduction was observed at a fluence of 120 J/m2 in the FeCV suspension and at a fluence of 200 J/m2 for CaCV; for the more resistant phage MS2 there was a 3-log10 reduction at a fluence of 650 J/m2. Few or no differences were observed between levels of UV inactivation in high- and low-protein-content virus stocks. In contrast, ionizing radiation could readily inactivate MS2 in water, and there was a 3-log10 reduction at a dose of 100 Gy, although this did not occur when the phage was diluted in high-protein-content stocks of CaCV or FeCV. The low-protein-content stocks showed 3-log10 reductions at a dose of 500 Gy for FeCV and at a dose of 300 for CaCV. The inactivation rates for both caliciviruses with ionizing and nonionizing radiation were comparable but different from the inactivation rates for MS2. Although most FeCV and CaCV characteristics, such as overall particle and genome size and structure, are similar, the capsid sequences differ significantly, making it difficult to predict human norovirus inactivation. Adequate management of UV and gamma radiation processes for virus inactivation should limit public health risks.

Occurrence and levels of indicator bacteriophages in bathing waters throughout Europe

Somatic coliphages, F-specific RNA bacteriophages, bacteriophages infecting Bacteroides fragilis, Escherichia coli and enterococci were counted in bathing waters in the late spring and summer. We tested fresh and marine bathing waters from North, South, East and West Europe expected to contain between 100 and 500 E. coli per 100 ml, although wider ranges were sometimes found. Bacteriophages were counted after concentration, since a preliminary study proved that this step was necessary to obtain positive counts. During monitoring, a first-line quality control with reference materials for bacteria and bacteriophages was performed by all the laboratories participating in the study. The same microbes were also counted in raw sewage samples from various areas in Europe, where the bacterial indicators and the three groups of bacteriophages were detected in roughly the same numbers. All groups of bacteriophages were detected in both fresh and marine bathing waters throughout Europe. Reliable and complete results from 147 samples showed that for log-transformed values, E. coli and bacteriophages were slightly correlated. However, the slope of the regression line changed according to E. coli concentration and the correlation diminished when this concentration was close to zero per 100 ml. The ratios between E. coli and phages in bathing waters differed significantly from those in sewage. The relative amounts of bacteriophages, mainly somatic coliphages and phages infecting Bact. fragilis RYC2056, increased in bathing waters with low E. coli concentration, especially in seawater samples containing < 100 E. coli per 100 ml. The relationship of bacteriophages with respect to enterococci paralleled that of bacteriophages with respect to E. coli. Somatic coliphages and bacteriophages infecting Bact. fragilis are useful to predict the presence of some pathogens with the same origin as present bacterial indicators but with higher survival rates.

Inactivation of bacteriophages in water by means of non-ionizing (uv-253.7 nm) and ionizing (gamma) radiation: a comparative approach

Thc inactivation behaviour of the bacteriophages PHI X 174 (ssDNA virus). MS2 (ssRNA virus) and B40-8 (dsDNA) toward non-ionizing (UV-253.7 nm) as well as to ionizing radiation (gamma radiation) was studied in order to evaluate their potential as viral indicators for water disinfection by irradiation. Previous findings of the high UV-253.7 nm resistance of MS2 were confirmed whereas an unexpected high sensitivity to gamma radiation compared to the two other phages was found. On the other hand, PHI X 174 revealed an enhanced UV sensitivity but a high resistance to ionizing radiation. B40-8 had an intermediate position between the other two bacteriophages relative to both types of radiation. As expected, the data of E. coli reconfirmed the unreliability of fecal indicator bacteria for the purpose of predicting responses of viruses to water treatment. In UV disinfection the influence of water matrix may be adequately controlled by considering the UV (253.7 nm) absorption of the water whereas so far no such parameter has existed for the influence of the water quality on ionizing irradiation with respect to the scavenger concentration.

Quantitative microbial faecal source tracking with sampling guided by hydrological catchment dynamics

The impairment of water quality by faecal pollution is a global public health concern. Microbial source tracking methods help to identify faecal sources but the few recent quantitative microbial source tracking applications disregarded catchment hydrology and pollution dynamics. This quantitative microbial source tracking study, conducted in a large karstic spring catchment potentially influenced by humans and ruminant animals, was based on a tiered sampling approach: a 31-month water quality monitoring (Monitoring) covering seasonal hydrological dynamics and an investigation of flood events (Events) as periods of the strongest pollution. The detection of a ruminant-specific and a human-specific faecal Bacteroidetes marker by quantitative real-time PCR was complemented by standard microbiological and on-line hydrological parameters. Both quantitative microbial source tracking markers were detected in spring water during Monitoring and Events, with preponderance of the ruminant-specific marker. Applying multiparametric analysis of all data allowed linking the ruminant-specific marker to general faecal pollution indicators, especially during Events. Up to 80% of the variation of faecal indicator levels during Events could be explained by ruminant-specific marker levels proving the dominance of ruminant faecal sources in the catchment. Furthermore, soil was ruled out as a source of quantitative microbial source tracking markers. This study demonstrates the applicability of quantitative microbial source tracking methods and highlights the prerequisite of considering hydrological catchment dynamics in source tracking study design.

Microbiological water quality along the Danube River: Integrating data from two whole-river surveys and a transnational monitoring network

The River Danube is, with 2780 km, the second longest river in Europe. Its catchment area covers 801 500 km(2), with approximately 81 million inhabitants in 19 countries. River water for anthropogenic use, transportation and recreation is of major importance in all of these countries. Microbiological contamination from faecal pollution by anthropogenic sources is considered to be a crucial problem throughout the Danube River basin. Thus, detailed knowledge on the extent and the origin of microbial pollution is essential for watershed management. The determination of faecal indicator concentrations along the Danube and its major tributaries during two whole-river surveys and 16 permanent stations allowed for the first time to draw a clear picture of the faecal pollution patterns along the whole longitudinal profile of this important international river. By including a variety of environmental variables in statistical analysis, an integrative picture of faecal pollution in the Danube River basin could be evolved. Four hot spots and six stretches of differing faecal pollution were identified, mainly linked with input from large municipalities. Significant decline of microbiological pollution was observed in the upper and lower Danube stretches over the investigation period. In contrast, a significant increase in the middle part was evident. The planned implementation of new wastewater treatment plants and advanced wastewater treatment measures according to the European Union urban wastewater directive will have a great potential to reduce microbial faecal pollution in the Danube and thus improving water quality.

Escherichia coli and enterococci are sensitive and reliable indicators for human, livestock and wildlife faecal pollution in alpine mountainous water resources

Aims:  This study evaluated the applicability of standard faecal indicator bacteria (SFIB) for alpine mountainous water resources monitoring.

Biodosimetry : Model calculations for u.v. water disinfection devices with regard to dose distributions

Abstract The increasing importance of disinfection of drinking water by u.v. radiation makes it necessary to determine the u.v. dose which is applied to the water by u.v.-disinfection plants. In Austria a minimum microbicidal dose of 400 Jm −2 at a wavelength of 253.7 nm is demanded for drinking water, the control of it shall be assured by type testing. A method was developed in using calibrated spores of Bacillus subtilis as biodosimeter which are added to the inflowing water, and after determination of their survival rate in the plant one can deduce the applied dose from it. This procedure is unproblematic as long as all microorganisms receive the same dose on their way through the reactor. But in some cases, and probably this is the normal case, not all microorganisms receive the same dose. It follows that a dose distribution will exist among the test organisms which have passed through the reactor. The dose (reduction-equivalent dose or RED) which is deduced from the survival rate of the microorganisms passing flow-through systems for u.v.-disinfection of wastewater or drinking water in general is different from the arithmetic mean of the dose distribution. The RED depends on the special form of the dose distribution and on the specific shape of the survival curve of the test organisms. The broader the dose distribution and the higher the u.v.-susceptibility of the microorganisms, the lower is the measured RED. But if the survival curve of the microorganisms used would have a shoulder ( D s > D m ), and the dose distribution would overlap with the shoulder the RED may increase. We investigated by calculations and by experiments the influences on the test results obtained by this method.

Performance characteristics of qPCR assays targeting human- and ruminant-associated Bacteroidetes for microbial source tracking across sixteen countries on six continents

Numerous quantitative PCR assays for microbial fecal source tracking (MST) have been developed and evaluated in recent years. Widespread application has been hindered by a lack of knowledge regarding the geographical stability and hence applicability of such methods beyond the regional level. This study assessed the performance of five previously reported quantitative PCR assays targeting human-, cattle-, or ruminant-associated Bacteroidetes populations on 280 human and animal fecal samples from 16 countries across six continents. The tested cattle-associated markers were shown to be ruminant-associated. The quantitative distributions of marker concentrations in target and nontarget samples proved to be essential for the assessment of assay performance and were used to establish a new metric for quantitative source-specificity. In general, this study demonstrates that stable target populations required for marker-based MST occur around the globe. Ruminant-associated marker concentrations were strongly correlated with total intestinal Bacteroidetes populations and with each other, indicating that the detected ruminant-associated populations seem to be part of the intestinal core microbiome of ruminants worldwide. Consequently tested ruminant-targeted assays appear to be suitable quantitative MST tools beyond the regional level while the targeted human-associated populations seem to be less prevalent and stable, suggesting potential for improvements in human-targeted methods.

Hypothesis-Driven Approach for the Identification of Fecal Pollution Sources in Water Resources

Water resource management must strive to link catchment information with water quality monitoring. The present study attempted this for the field of microbial fecal source tracking (MST). A fecal pollution source profile based on catchment data (e.g., prevalence of fecal sources) was used to formulate a hypothesis about the dominant sources of pollution in an Austrian mountainous karst spring catchment. This allowed a statistical definition of methodical requirements necessary for an informed choice of MST methods. The hypothesis was tested in a 17-month investigation of spring water quality. The study followed a nested sampling design in order to cover the hydrological and pollution dynamics of the spring and to assess effects such as differential persistence between parameters. Genetic markers for the potential fecal sources as well as microbiological, hydrological, and chemo−physical parameters were measured. The hypothesis that ruminant animals were the dominant sources of fecal pollution in the catchment was clearly confirmed. It was also shown that the concentration of ruminant markers in feces was equally distributed in different ruminant source groups. The developed approach provides a tool for careful decision-making in MST study design and might be applied on various types of catchments and pollution situations.