Fecal indicator and pathogen reduction in vegetation microcosms

Municipal wastewater has been applied as a fertilizer to coniferous forest in northern Sweden with the additional aims to purify the wastewater and reduce the treatment costs. This practice has the potential to introduce pathogenic microorganisms from the applied wastewater or sludge to the environment, with a subsequent risk of transmission to exposed humans or animals. Seasonality and climatological conditions, in particular low temperature, humidity and low sun intensity will furthermore affect the persistence of pathogens in the forest in northern Scandinavia and may therefore enhance the survival and thereby the exposure risk. The survival of two pathogens (Salmonella senftenberg and Campylobacter coli) and four fecal indicator organisms; bacteria strains (E. coli and E. fecalis) and bacteriophages (S. typhimurium phage 28 B and phi x174), used here as viral models, was evaluated in microcosms (60 Litre terrariums) with two vegetation regimes (characterized as moss and lichen) at selected temperatures (+3 degrees C to +27 degrees C) and artificial light exposure (light and dark). Salmonella persisted in high numbers after 35 days in the vegetation, indicating a potential risk for further transmission. Neither the vegetation regime nor temperature or light exposure as independent factors governed the reduction of the indicator bacteria and the bacteriophages. The persistence of the indicator organisms was generally best supported in the moss vegetation at low temperature in the dark, where the time for a 90% reduction (T-90) was in the range from 8 days (E. coli) to 31 days (Salmonella phage). The die-off characteristics found in this study provide baseline values for further validations in the field and verify that the purification and reuse of municipal wastewater in coniferous forest could potentially be responsible for transmission of fecal pathogens to humans and animals.