The impact of cyanobacteria on growth and death of opportunistic pathogenic bacteria.

Climate change may cause increased microbial growth in water sources and more knowledge is required on how this may affect the hygienic water quality, i.e., whether increased occurrence of cyanobacteria and algae may stimulate the growth rate of opportunistic pathogenic bacteria. Laboratory experiments were performed to investigate if the presence of the cyanobacteria Anabanea lemmermannii and Microcystis aeruginosa affected the survival and growth rate of the opportunistic pathogenic bacteria Aeromonas hydrophila and Pseudomonas aeruginosa, and the faecal indicators Escherichia coli and coliforms. Cyanobacteria were cultured in bottles containing the nutrient-poor medium 02. Sewage, A. hydrophila or P. aeruginosa was added to cyanobacterial cultures and the bacterial growth and survival was followed. E. coli and coliforms from sewage died within few days and the decay rate was not affected by the presence of cyanobacteria. The presence of Anabaena stimulated the growth rate of P. aeruginosa, but had no effect on the growth rate of A. hydrophila. Microcystis had no effect on the growth rate of P. aeruginosa and an inhibiting effect on the growth rate of A. hydrophila.

Application of rapid enzyme assay techniques for monitoring of microbial water quality.

Rapid enzyme assay techniques based on direct measurement of beta-d-galactosidase (GALase) or beta-d-glucuronidase (GLUase) activity without selective cultivation are used for rapid estimation of the level of coliform bacteria and Escherichia coli in water samples. Reported detection limits using fluorogenic substrates correspond to culturable target bacteria concentrations that can be appropriate within present guidelines for recreational waters. The rapidity, that is detection within one hour, compromises the specificity of the assay; enzyme activity contributions from other than target bacteria need to be considered, particularly at low levels of target bacteria. Enzyme activities are more persistent than the culturability of target bacteria to environmental and disinfection stress, thus water samples may express enzyme activities of both culturable and viable non-culturable cells.

Quantitative microbial risk assessment combined with hydrodynamic modelling to estimate the public health risk associated with bathing after rainfall events.

This study investigated the public health risk from exposure to infectious microorganisms at Sandvika recreational beaches, Norway and dose-response relationships by combining hydrodynamic modelling with Quantitative Microbial Risk Assessment (QMRA). Meteorological and hydrological data were collected to produce a calibrated hydrodynamic model using Escherichia coli as an indicator of faecal contamination. Based on average concentrations of reference pathogens (norovirus, Campylobacter, Salmonella, Giardia and Cryptosporidium) relative to E. coli in Norwegian sewage from previous studies, the hydrodynamic model was used for simulating the concentrations of pathogens at the local beaches during and after a heavy rainfall event, using three different decay rates. The simulated concentrations were used as input for QMRA and the public health risk was estimated as probability of infection from a single exposure of bathers during the three consecutive days after the rainfall event. The level of risk on the first day after the rainfall event was acceptable for the bacterial and parasitic reference pathogens, but high for the viral reference pathogen at all beaches, and severe at Kalvøya-small and Kalvøya-big beaches, supporting the advice of avoiding swimming in the day(s) after heavy rainfall. The study demonstrates the potential of combining discharge-based hydrodynamic modelling with QMRA in the context of bathing water as a tool to evaluate public health risk and support beach management decisions.

Diverse Land Use and the Impact on (Irrigation) Water Quality and Need for Measures - A Case Study of a Norwegian River.

Surface water is used for irrigation of food plants all over the World. Such water can be of variable hygienic quality, and can be contaminated from many different sources. The association of contaminated irrigation water with contamination of fresh produce is well established, and many outbreaks of foodborne disease associated with fresh produce consumption have been reported. The objective of the present study was to summarize the data on fecal indicators and selected bacterial pathogens to assess the level of fecal contamination of a Norwegian river used for irrigation in an area which has a high production level of various types of food commodities. Sources for fecal pollution of the river were identified. Measures implemented to reduce discharges from the wastewater sector and agriculture, and potential measures identified for future implementation are presented and discussed in relation to potential benefits and costs. It is important that the users of the water, independent of intended use, are aware of the hygienic quality and the potential interventions that may be applied. Our results suggest that contamination of surface water is a complex web of many factors and that several measures and interventions on different levels are needed to achieve a sound river and safe irrigation.

On-line monitoring of Escherichia coli in raw water at Oset drinking water treatment plant, Oslo (Norway).

The fully automated Colifast ALARM™ has been used for two years for daily monitoring of the presence/absence of Escherichia coli in 100 mL raw water at Oset drinking water treatment plant in Oslo, Norway. The raw water is extracted from 35 m depth from the Lake Maridalsvannet. E. coli was detected in 18% of the daily samples. In general, most samples positive for E. coli were observed during the autumn turnover periods, but even in some samples taken during warm and dry days in July, with stable temperature stratification in the lake, E. coli was detected. The daily samples gave useful additional information compared with the weekly routine samples about the hygienic raw water quality and the hygienic barrier efficiency of the lake under different weather conditions and seasons. The winter 2013/2014 was much warmer than the winter 2012/2013. The monitoring supported the hypothesis that warmer winters with shorter periods with ice cover on lakes, which may be a consequence of climate changes, may reduce the hygienic barrier efficiency in deep lakes used as drinking water sources.

New method for rapid and sensitive quantification of sulphide-producing bacteria in fish from arctic and temperate waters.

The offensive, fishy, rotten H2S-off-odours in spoiled, aerobically and cold stored fish from arctic and temperate waters are generally caused by sulphide-producing bacteria (SPB), mainly Shewanella putrefaciens. In the present work, a new, rapid, simple and accurate method for estimation of the SPB content in fish from these areas is described. The quantification is based on the formation rate of iron sulphide during growth of SPBs incubated at 30 degrees C in a liquid growth medium containing cysteine, sodium thiosulphate and iron(III)citrate as specific substrates for iron sulphide formation. The iron sulphide turns the medium grey and masks the background fluorescence in the medium when the SPB content in the assay is approximately 10(9) cfu/ml. The fluorescence change could be detected instrumentally and the colour change visually. The method was developed and evaluated in tests with S. putrefaciens CCUG 13452 DT as well as naturally occurring SPBs in cod, salmon, wolf fish and coal fish. A linear correlation between the SPB count and detection time was obtained over the entire range from 1 to 10(9) cfu SPB/g, corresponding to detection times 17 and 1 h, respectively. The correlation is described by the equation: log cfu/g fish= -0.59(+/- 0.17) x DT+ 9.65(+/- 0.09), where DT is the detection time in hours. The model was valid for all the tested fish species and all tested naturally occurring SPBs in these species. The regression coefficients (R2) for cod, coal fish, wolf fish and salmon were 0.99, 0.92, 0.97 and 0.97, respectively. The detection level of the method is 1 SPB per sample tube, corresponding to 16 cfu/g fish. The method could be used to predict the remaining shelf life of the fish for different markets, even when the time-temperature history during storage of the fish is unknown.

Impact of rainfall on the hygienic quality of blue mussels and water in urban areas in the Inner Oslofjord, Norway.

The effects of precipitation on the hygienic quality of water and blue mussels collected from five different localities in the urban areas in the Inner Oslofjord were investigated, with samples analysed for Escherichia coli, Salmonella spp., pathogenic Vibrio spp., Norovirus, Sapovirus, Cryptosporidium spp. and Giardia duodenalis. The sampling sites were located at varying distances from the outlet of combined sewer overflows (CSO)-impacted rivers/streams. In general, 1-3 log10 increases in fecal indicator bacteria and human pathogens were observed after heavy rainfalls. Blue mussels appeared to be a useful indicator of the impact of sewage at these sites, and generally a good correlation was identified between concentrations of E. coli and other human pathogens in the mussels. Provision of general advice to the public of avoiding areas near the outlets of CSO-impacted rivers after heavy rainfall may reduce the risk of gastroenteritis by bathers and others that may swallow water during recreational activities.

Effect of water treatment on the growth potential of Vibrio cholerae and Vibrio parahaemolyticus in seawater.

In laboratory experiments we added Vibrio cholerae and Vibrio parahaemolyticus to bottles with seawater previously treated by filtration, UV, chlorine or ozone. The purpose was to investigate the influence of different treatment techniques on the growth potential of these bacteria in simulated ballast water tanks. Residual oxidants were removed before inoculation, and the bottles were incubated at 21 ± 1 °C. The growth potential of the vibrios was investigated in two different experimental setups, i.e. in presence and absence of added natural microorganisms. In general, V. cholerae and V. parahaemolyticus rapidly lost their culturability after inoculation and storage in untreated seawater, but showed increased survival or growth in the treated water. Highest growth was observed in water previously exposed to high concentrations of ozone. Addition of natural microorganisms reduced the growth of V. cholerae and V. parahaemolyticus.

Real-time PCR and NASBA for rapid and sensitive detection of Vibrio cholerae in ballast water.

Transport of ballast water is one major factor in the transmission of aquatic organisms, including pathogenic bacteria. The IMO-guidelines of the Convention for the Control and Management of Ships' Ballast Water and Sediments, states that ships are to discharge <1 CFU per 100 ml ballast water of toxigenic Vibrio cholerae, emphasizing the need to establish test methods. To our knowledge, there are no methods sensitive and rapid enough available for cholera surveillance of ballast water. In this study real-time PCR and NASBA methods have been evaluated to specifically detect 1 CFU/100ml of V. cholerae in ballast water. Ballast water samples spiked with V. cholerae cells were filtered and enriched in alkaline peptone water before PCR or NASBA detection. The entire method, including sample preparation and analysis was performed within 7 h, and has the potential to be used for analysis of ballast water for inspection and enforcement control.

Different length (DL) qPCR for quantification of cell killing by UV-induced DNA damage.

We describe the different length (DL) qPCR method for quantification of UV induced DNA damage in cell killing. The principle of DL qPCR is that DNA damage inhibits PCR. Applications with different lengths can therefore be used to detect different levels of UV-induced DNA damage. The assay was evaluated on three strains of Escherichia coli exposed to varying levels of ultraviolet (UV) radiation. We show that DL qPCR sensitivity and reproducibility are within the range of practical application to detect the effect of UV cell killing.