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.

Aeration of anaerobically digested sewage sludge for COD and nitrogen removal: optimization at large-scale.

The paper will report about the experiences at an Austrian large wastewater treatment plant of 720,000 population equivalents, where anaerobically digested sewage sludge is further stabilised under aerobic conditions. Enhanced stabilisation of the anaerobically digested sludge was required at the plant in order to get a permit for landfill disposal of the dewatered stabilized sludge. By implementing a post-aeration treatment (SRT approximately 6d; 36 degrees C) after anaerobic digestion the organic content of the anaerobically well digested sludge can be decreased by 16%. Investigations on site showed that during digested sludge post-aeration anoxic phases for denitrification are needed to provide stable process conditions. In this way the pH value can be kept in a more favourable range for micro-organisms and concrete structures. Additionally, inhibition of the biological process due to nitrite accumulation can be avoided. By optimising the aeration/pause ratio approximately 45% of total nitrogen in digested sludge can be removed. This significantly improves nitrogen removal efficiency at the wastewater treatment plant. NH(4)-removal occurs mainly through nitritation and denitritation with an efficiency of 98%. The costs/benefit analysis shows that post-aeration of digested sludge results in an increase of total annual costs for wastewater treatment of only 0.84%, corresponding to 0.19 Euro/pe/a. Result of molecular biological analyses (DGGE) indicate that all four ammonium-oxidizing bacteria species present in activated sludge can survive anaerobic digestion, but only two of them can adapt in the digested sludge post-aeration tanks. Additionally, in the post-aerated digested sludge a further ammonium-oxidizing bacteria species was identified.

Diversity of ammonia oxidising bacteria in a vertical flow constructed wetland.

Vertical flow constructed wetlands (VFCWs) with intermittent loading are very suitable for nitrification. Ammonia oxidising bacteria (AOB) are the limiting step of nitration. Therefore the AOB community of a full-scale VFCW, receiving municipal wastewater, was investigated within this study. The diversity of the functional gene encoding the alpha-subunit of the ammonia monooxygenase (amoA), present only in AOB, was assessed by denaturing gradient gel electrophoresis (DGGE). Only very few amoA sequence types dominated the wetland filter substrate; nevertheless a stable nitrification performance could be observed. During the cold season the nitrification was slightly reduced, but it has been shown that the same AOB could be identified. No spatial AOB pattern could be observed within the filter body of the VFCW. The most prominent bands were excised from DGGE gels and sequenced. Sequence analyses revealed two dominant AOB lineages: Nitrosomonas europaea/"Nitrosococcus mobilis" and Nitrosospira. Species of the Nitrosomonas lineage are commonly found in conventional wastewater treatment plants (WWTPs). In contrast, members of the Nitrosospira lineage are rarely present in WWTPs. Our observations indicate that the AOB community in this VFCW is similar to that found in horizontal flow constructed wetlands, but differs from common WWTPs regarding the presence of Nitrosospira.

Molecular biological methods (DGGE) as a tool to investigate nitrification inhibition in wastewater treatment.

Incomplete nitrification at an activated sludge plant for biological pre-treatment of rendering plant effluents led to a detailed investigation on the origin and solution of this problem. Preliminary studies revealed that an inhibition of ammonia oxidising microorganisms (AOM) by process waters of the rendering plant was responsible for the situation. We were able to show a correlation between the existence of specific AOM and nitrification capacity expressed as oxygen uptake rate for maximal nitrification (OURNmax). Only Nitrosospira sp. was found in the activated sludge of the rendering plant and another industrial wastewater treatment plant with problems in nitrification, while reference plants without nitrification problems showed Nitrosomonas spp. as the predominant ammonia oxidising bacteria. By accompanying engineering investigations and experiments (cross-feeding experiments, operation of a two-stage laboratory plant) with molecular biological methods (DGGE--Denaturing Gradient Gel Electrophoresis) we were able to elaborate an applicable solution for the rendering plant. Laboratory experiments with a two-stage process layout finally provided complete nitrification overcoming the inhibiting nature of process waters from the rendering plant. DGGE analysis of the second stage activated sludge from the laboratory plant showed a shift in population structure from Nitrosospira sp. towards Nitrosomonas spp. simultaneous to the increase of nitrification capacity. Nitrification capacities comparable to full-scale municipal wastewater treatment plants could be maintained for more than two months. As the design of wastewater treatment plants for nitrification is linked to the growth characteristics of Nitrosomonas spp., established criteria can be applied for the redesign of the full-scale plant.