Gastropods as a source for fecal indicator bacteria in drinking water.

Microbial water quality is routinely examined using the fecal indicator bacteria Escherichia coli, coliform bacteria and enterococci. Several practical cases in German drinking water distribution systems indicated invertebrates such as insects or gastropods as a source for the microbiological deterioration. Therefore, we examined three genera of Gastropoda (Arion, Helix and Cepaea) for the presence of fecal indictor bacteria in excreta using standard methods. Enterococci and coliform bacteria were detected in high concentrations (mean values of 1.5 × 106 and 6.3 × 106 per gram feces, respectively). E. coli was also detected, still specification revealed that what was assigned by standard ISO-methods to be E. coli was indeed a novel species of Buttiauxella, exhibiting ß-D-glucuronidase activity, thus, explaining the false-positive results. Microbiome analyses confirmed the cultural results. Enterobacteriaceae were dominant in the samples, yet only very few sequences could be assigned to Escherichia. Our study suggests, that enterococci and coliform bacteria are an integral component in the gastropod microbiome, whereas E. coli might be derived from other sources with gastropods being a vector. The results further indicate, that the current concept of fecal indicator bacteria needs to be extended, as not only humans and homeothermic animals could be a source for fecal indictor bacteria, but also gastropods need to be taken into consideration.

Assessing and modeling biocatalysis in field denitrification beds reveals key influencing factors for future constructions.

Environmental contamination with fertilizers is threatening biodiversity in many ecosystems due to nitrate-based eutrophication. One opportunity for a cost-efficient nitrate elimination are denitrification beds in which a microbial community thrives under anoxic conditions with polymeric plant material as a carbon and an electron source. Incoming nitrate is used as electron acceptor and reduced to molecular nitrogen. Projects realizing denitrification beds in field scale are sparse and robust data on their efficiency throughout the year mostly not available. This study analyzed the nitrate elimination efficiency and microbiology of a 216 m3 denitrification bed over the time course of more than three years. Phylogenetic as well as transcriptomic analysis revealed that the reactor contained a biofilm community growing on the surface of the wood chips and a planktonic community. Both differed in composition but their variance was affected only to a minor extend by seasonal temperature changes. Cellulose degradation was mainly conducted by the biofilm population while denitrification was mostly conducted by the planktonic community. Methanogens were detectable only to a very minor extend. Using online data from the nitrate concentration of in- and outflowing water as well as a hydrological model to predict the water inflow, it was possible to establish a process model that sufficiently describes the denitrification process. This model clearly indicates that the denitrification efficiency is mostly impacted by temperature and hydraulic retention time. It also suggests that the simple design of the denitrification bed most likely leads to different flow paths through the reactor depending on the volumetric flow rate. This study allows for the first time a robust estimation of the necessary reactor size for nitrate removal in a moderate continental climate setting. It also suggests how future denitrification beds could be improved for better performance.

NO3 - removal efficiency in field denitrification beds: key controlling factors and main implications.

Nitrate pollution is a growing environmental threat that affects both ground- and surface-water. The typically used technique for nitrate elimination in wastewater treatment plants cannot be applied for all water streams as it necessitates a highly developed technical infrastructure. Field denitrification beds comprise one strategy to treat surface water containing high nitrate loads, which typically is due to the increasing agricultural land use. Here, the water passes through a basin containing a cheap carbon material as electron donor that provides the environmental niche for a complex microbial biocenosis. The microorganisms catalyse the hydrolysis of the polymeric organic carbon substrate and a variety of fermentative and respiratory pathways that are in the end supposed to lead to an efficient denitrification process. This review article integrates our current knowledge on environmental and operating parameters of and within denitrification beds including biotic and abiotic factors influencing the nitrate removal efficiency. Steering of these two factors can allow to minimise pollution swapping and the formation of greenhouse gases.

Influence of the Potential Carbon Sources for Field Denitrification Beds on Their Microbial Diversity and the Fate of Carbon and Nitrate.

Nitrogen based eutrophication of ecosystems is a global problem that gains momentum through a growing global population. The water quality of nitrate or ammonium contaminated rivers and streams cannot always be amended in centralized waste water treatment plants. Field denitrification plants were suggested as a solution for a decentralized reduction of nitrate to dinitrogen. Here, stable and cheap organic carbon sources serve as carbon and electron source for a microbial community. Still, our knowledge on the impact of these organic carbon sources on the development and diversity of these cultures is sparse. Moreover, the stability of these denitrification plants at different nitrate loading rates especially in the higher concentration regime were not tested so far. In this study, we compare the fate of carbon and nitrogen as well as the microbial community of wood pellet (WP) (pressed sawdust), wheat straw, and wood chips (WC) based laboratory denitrification reactors. Our study reveals that the diversity and composition of the community is strongly dependent on the carbon source. The diversity decreased in the order WC, wheat straw, and WPs. The three reactor types were characterized by different nitrate reduction kinetics and were affected differently by high nitrate loading rates. While the nitrate reduction kinetics were negatively influenced by higher nitrate doses in the wheat straw reactors, WPs as carbon source sustained the opposite trend and WC lead to an overall slower but concentration independent nitrate reduction rate. Counterintuitively, the concentration of soluble organic carbon was highest in the WP reactors but methane emission was not detectable. This is corroborated by the microbial diversity data in which methanogenic species were highly underrepresented compared to the other two reactor types. In contrary, the methane emissions in the wheat straw and WC reactors were comparable to each other.

Investigation of different nitrogen reduction routes and their key microbial players in wood chip-driven denitrification beds.

Field denitrification beds containing polymeric plant material are increasingly used to eliminate nitrate from agricultural drainage water. They mirror a number of anoxic ecosystems. However, knowledge of the microbial composition, the interaction of microbial species, and the carbon degradation processes within these denitrification systems is sparse. This study revealed several new aspects of the carbon and nitrogen cycle, and these findings can be correlated with the dynamics of the microbial community composition and the activity of key species. Members of the order Pseudomonadales seem to be important players in denitrification at low nitrate concentrations, while a switch to higher nitrate concentrations seems to select for members of the orders Rhodocyclales and Rhizobiales. We observed that high nitrate loading rates lead to an unpredictable transition of the community's activity from denitrification to dissimilatory reduction of nitrate to ammonium (DNRA). This transition is mirrored by an increase in transcripts of the nitrite reductase gene nrfAH and the increase correlates with the activity of members of the order Ignavibacteriales. Denitrification reactors sustained the development of an archaeal community consisting of members of the Bathyarchaeota and methanogens belonging to the Euryarchaeota. Unexpectedly, the activity of the methanogens positively correlated with the nitrate loading rates.