Indicator organisms like coliforms, E. coli and enterococci are normally associated with the intestinal tract of warm-blooded animals, and can thus be used for indicating fecal contamination of water. This study investigated the removal efficiency of indicator organisms in a free water surface constructed wetland used for additional treatment of municipal wastewater. The species composition of enterococci and the prevalence of the tetracycline resistance gene tetM were also investigated. Samplings were done at five different sites: at the inlet, within and at the outlet of the wetland, as well as upstream and downstream of the wetland outlet into the adjoining river. A total of 86 enterococci isolates were characterized as E. faecalis, E. faecium or other Enterococcus spp., and the two former were also screened for the tetM gene. The investigated wetland showed a high removal rate of indicator organisms, with a removal of approximately 98%. E. faecium was identified as the predominant species in the wetland at a mean of 55.8%. In comparison, only 9.3% were E. faecalis, while other Enterococcus spp. were found to comprise 34.9%. Of the 56 isolates identified as E. feacalis or E. faecium 10.7% were positive for the tetM gene. The high removal of bacteria shown by this investigation provides further evidence of the benefits of constructed wetlands on wastewater treatment. It also offers a first indication of the Enterococcus spp. composition in Ekeby wetland, Eskilstuna, Sweden, and shows that the prevalence of the tetM gene is relatively low at this point in time.

Doctor of Philosophy / Department of Food Science / Daniel Y.C. Fung / In the food industry, coliform testing is traditionally done by the time consuming and labor intensive plate count method or tube enumeration methods. The TEMPO® system (bioMérieux, Inc.) was developed to improve laboratory efficiency and to replace traditional methods. It uses a miniaturization of the Most Probable Number (MPN) method with 16 tubes with 3 dilutions in one single disposable card. It utilizes two stations: the TEMPO® Preparation station and the TEMPO® Reading station. In this study, the Oxyase® (Oxyase®, Inc.) enzyme was added to TEMPO® CC (Coliforms Count), TEMPO® AC (aerobic colony count) and TEMPO® EC (E. coli Count) methods. Water samples of 1 ml with 0.1 ml of Oxyase® enzyme were compared to samples without the Oxyase® enzyme using the TEMPO® system. Samples were spiked with different levels of coliforms (10, 102, 103 and 104 CFU/ml), stomached (20 sec), and pipetted into the three different TEMPO® media reagents (4 ml) in duplicate and then automatically transferred into the corresponding TEMPO® cards by the TEMPO® preparation station. Counts were obtained using the TEMPO® reading station after 8, 12, 16, 22 and 24 hours at an incubation temperature of 35°C. Results from 20 replicates were compared statistically. Using TEMPO® tests, high counts in food samples (>6 log 10 CFU/ml) can be read in 6±2 hours of incubation using the time-to-detection calibration curve. The TEMPO® system reduces reading time (reading protocol should be changed). There is no need to wait for 22 hours of incubation only 12 hours is required. Oxyrase® enzyme is not needed for the TEMPO® system.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2006. / Härtill 5 uppsatser.

In Virginia, over one million households rely on private water supplies (e.g. well, spring, cistern). Previous literature acknowledges bacterial contamination in private water supplies as a significant public health concern in the United States. The present study tested private wells and springs in 20 Virginia counties for total coliforms (TC) and E. coli (EC) along with a suite of chemical contaminants. Sample collection was organized by the Virginia Household Water Quality Program (VAHWQP), a Virginia Cooperative Extension effort managed by faculty in the Biological Systems Engineering Department. Microbial and chemical source tracking were used to identify possible sources of contamination. A logistic regression was employed to investigate potential correlations between TC contamination and chemical parameters (e.g. NO3-, turbidity) as well as homeowner provided survey data describing system characteristics and perceived water quality. TC and EC contamination were quantified via the Colilert (www.idexx.com) defined substrate method for most probable number (MPN) of EC and TC per 100 mL of water. Of the 538 samples collected, 41% (n=221) were positive for TC and 10% (n=53) for EC. Chemical parameters were not statistically predictive of microbial contamination. Well depth, water treatment, and farm location proximate to the water supply were factors in a regression model that predicted presence/absence of TC with 74% accuracy. Microbial and chemical source tracking techniques (Polymerase Chain Reaction (PCR) and fluorometry, respectively) identified 4 of 26 samples as likely contaminated with human wastewater. Application of these source-tracking analyses on a larger scale will prove useful in defining remediation strategies. / Master of Science

Microbiological water quality monitoring of bathing waters does not account for faecal bacteria in sediments. Intertidal deposits are a significant reservoir of faecal bacteria and this indicates there is a risk to human health through direct contact with the sediment, or through the resuspension of bacteria to the water column. This project investigated factors influencing the relative abundance of faecal indicator organisms (FIOs) in intertidal estuarine sediments. The effects of physical, biogeochemical and biological sediment characteristics, environmental variables and native microbial communities were explored through field campaigns on the Ythan and Eden estuaries, Scotland. The contributory role of sediments to adverse water quality was investigated by combining FIO abundance and measurements of sediment stability. The importance of strain and sediment characteristics in the adhesion of E. coli to suspended sediments was also examined using laboratory experiments. E. coli concentrations up to 5.9 log₁₀ CFU 100 g dry wt⁻¹ were observed, confirming that intertidal sediments are an important reservoir of faecal bacteria. The variability of E. coli abundance in estuarine sediments was successfully explained with multiple stepwise linear regression (Adjusted R² up to 87.4) using easily-obtainable measurements of sediment characteristics and environmental variables, with variability most heavily influenced by salinity and particle size gradients. Native microbial community population metrics and community constituent composition correlated with environmental gradients, but did not influence FIO abundance. The amount of E. coli adhering to suspended sediments ranged from 0.02 to 0.74 log₁₀ CFU ml⁻¹, and was dependant on strain characteristics and sediment type rather than zeta potential, with higher cell-particle adhesion at 2 and 3.5 PSU than 0 and 5 PSU. Monitoring of sediment characteristics will lead to more informed bathing water quality advisories to protect public health. Future research should focus on applying the findings here to the modelling of bacterial fate and transport on a catchment scale.

The Water Framework Directive requires reduced environmental impacts from human activities and for the assessment of the non-market benefits of pollution remediation schemes. This policy shift has exacerbated the research problems surrounding the physical, social and economic consequences of the relationship between land use and water quality. This research seeks to quantify the major socio-economic and environmental benefits for people which may arise as riverine pollution is reduced. To achieve these aims this research integrates primary data analyses combining choice experiment techniques with geographical information system based analyses of secondary data concerning the spatial distributions of riverine pollution. Current knowledge on the microbial quality of river water, measured by faecal indicator organism (FIO) concentrations and assessed at catchment scale, is inadequate. This research develops generic regression models to predict base- and high-flow faecal coliform (FC) and enterococci (EN) concentrations, using land cover and population (human and livestock) variables. The resulting models are then used both to predict FIO concentrations in unmonitored watercourses and to evaluate the likely impacts of different land use scenarios, enabling insights into the optimal locations and cost-effective mix of implementation strategies. Valuation experiments frequently conflate respondents’ preferences for different aspects of water quality. This analysis uses stated preference techniques to disaggregate the values of recreation and ecological attributes of water quality, thereby allowing decision makers to better understand the consequences of adopting alternative investment strategies which favour either ecological, recreational or a mix of benefits. The results reveal heterogeneous preferences across society; specifically, latent class analysis identifies three distinct groups, holding significantly different preferences for water quality. From a methodological perspective this research greatly enhances the ongoing synthesis of geographic and economic social sciences and addresses important policy questions which are of interest to a variety of stakeholders, including government departments and the water industry.