Water column anammox and denitrification in a temperate permanently stratified lake (Lake Rassnitzer, Germany).

We studied microbial N(2) production via anammox and denitrification in the anoxic water column of a restored mining pit lake in Germany over an annual cycle. We obtained high-resolution hydrochemical profiles using a continuous pumping sampler. Lake Rassnitzer is permanently stratified at ca. 29m depth, entraining anoxic water below a saline density gradient. Mixed-layer nitrate concentrations averaged ca. 200 micromol L(-1), but decreased to zero in the anoxic bottom waters. In contrast, ammonium was <5 micromol L(-1) in the mixed layer but increased in the anoxic waters to ca. 600 micromol L(-1) near the sediments. In January and October, (15)N tracer measurements detected anammox activity (maximum 504 nmol N(2)L(-1)d(-1) in (15)NH(4)(+)-amended incubations), but no denitrification. In contrast, in May, N(2) production was dominated by denitrification (maximum 74 nmol N(2)L(-1)d(-1)). Anammox activity in May was significantly lower than in October, as characterized by anammox rates (maximum 6 vs. 16 nmol N(2)L(-1)d(-1) in incubations with (15)NO(3)(-)), as well as relative and absolute anammox bacterial cell abundances (0.56% vs. 0.98% of all bacteria, and 2.7x10(4) vs. 5.2x10(4)anammox cells mL(-1), respectively) (quantified by catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH) with anammox bacteria-specific probes). Anammox bacterial diversity was investigated with anammox bacteria-specific 16S rRNA gene clone libraries. The majority of anammox bacterial sequences were related to the widespread Candidatus Scalindua sorokinii/brodae cluster. However, we also found sequences related to Candidatus S. wagneri and Candidatus Brocadia fulgida, which suggests a high anammox bacterial diversity in this lake comparable with estuarine sediments.

Particulates, not plants, dominate nitrogen processing in a septage-treating aerated pond system.

In pond and wetland systems for wastewater treatment, plants are often thought to enhance the removal of ammonium and nitrogen through the activities of root-associated bacteria. In this study, we examined the role of plant roots in an aerated pond system with floating plants designed to treat high-strength septage wastewater. We performed both laboratory and full-scale experiments to test the effect of different plant root to septage ratios on nitrification and denitrification, and measured the abundances of nitrifying bacteria associated with roots and septage particulates. Root-associated nitrifying bacteria did not play a significant role in ammonium and total nitrogen removal. Investigations of nitrifier populations showed that only 10% were associated with water hyacinth [Eichhornia crassipes (Mart.) Solms] roots (at standard facility plant densities equivalent to 2.2 wet g roots L(-1) septage); instead, nitrifiers were found almost entirely (90%) associated with suspended septage particulates. The role of root-associated nitrifiers in nitrification was examined in laboratory batch experiments where high plant root concentrations (7.4 wet g L(-1), representing a 38% net increase in total nitrifier populations over plant-free controls) yielded a corresponding increase (55%) in the non-substrate-limited nitrification rate (V(max)). However, within the full-scale septage-treating pond system, nitrification and denitrification rates remained unchanged when plant root concentrations were increased to 7.1 g roots L(-1) (achieved by increasing the surface area available for plants while maintaining the same tank volume). Under normal facility operating conditions, nitrification was limited by ammonium concentration, not nitrifier availability. Maximizing plant root concentrations was found to be an inefficient mechanism for increasing nitrification in organic particulate-rich wastewaters such as septage.

Control of denitrification in a septage-treating artificial wetland: the dual role of particulate organic carbon.

We examined the factors controlling organic carbon (C) cycling and its control of nitrogen (N) removal via denitrification in an aerated artificial wetland treating highly concentrated wastewater to nutrient-removal standards. Processing of organic material by the septage-treating wetland affected the biological reactivity (half-life, or t1/2) of organic C pools through microbial degradation and gravity fractionation of the influent septage. Primary sedimentation fractionated the initial septage material (t1/2 = 8.4d) into recalcitrant waste solids (t1/2 = 16.7d) and highly labile supernatant (t1/2 = 5.0d), allowing this reactive fraction to be further degraded during treatment in aerobic wetland tanks until a less labile material (t1/2 = 7.3d) remained. Organic C contributions from in situ fixation by nitrifying bacteria or algae in these tanks were small, about 1% of the C degradation rate. In the aerated tanks, denitrification was correlated with particulate organic C loading rates, although the average C required (0.35 mg C L(-1)h(-1)) to support denitrification was only 12% of the total C respiration rate (2.9 mg C L(-1)h(-1)). Additions of plant litter (2.5g C L(-1)) to the aerated tanks under normal operating conditions doubled denitrification rates to 0.58 mg N L(-1)h(-1), and reduced effluent nitrate levels by half, from 12.7 to 6.4 mg N L(-1). However, C degradation within the plant litter (0.15mg C L(-1)h(-1)) was sufficient to have accounted for only 35% of the additional denitrification. Evidence from laboratory and full-scale plant litter additions as well as process monitoring indicates that the stimulation of denitrification is due to the respiration-driven formation of anaerobic microsites within particulate organic C. In this aerated highly C-loaded septage-treating wetland, anaerobic microsite, rather than C substrate availability limits denitrification.