Fred Sörensson

Nitrous oxide production in a forest soil at low temperatures – processes and environmental controls

Recent investigations have highlighted the relative importance of the winter season for emissions of N(2)O from boreal soils. However, our understanding of the processes and environmental controls regulating these emissions is fragmentary. Therefore, we investigated the potential for, and relative importance of, N(2)O formation at temperatures below 0 degrees C in laboratory experiments involving incubations of a Swedish boreal forest soil. Our results show that frozen soils have a high potential for N(2)O formation and subsequent emission. Net N(2)O production rates at -4 degrees C equaled those observed at +10 to +15 degrees C at moisture contents >60% of the soils water-holding capacity. The source of this N(2)O was found to be denitrification occurring in anoxic microsites in the frozen soil and temperature per se did not control the denitrification rates at temperatures around 0 degrees C. Furthermore, both net nitrogen-mineralisation and nitrification were observed in the frozen soil samples. Based on these findings we propose a conceptual model for the temperature response of N(2)O formation in soils at low temperatures.

Community survey of ammonia-oxidizing bacteria in full-scale activated sludge processes with different solids retention time

Aims:  To study the effects of different solids retention time (SRT) on the nitrification activity and community composition of ammonia‐oxidizing bacteria (AOB) in two full‐scale activated sludge processes during a 5‐month period.

Variation of nitrous oxide formation in the denitrification basin in a wastewater treatment plant with nitrogen removal

The influence of several environmental factors on the nitrous oxide production and denitrification in anoxic activated sludge has been studied in a Swedish wastewater treatment plant. Changes in temperature and in the concentrations of carbon source, nitrate or nitrite showed no significant influence on nitrous oxide production. Variations in pH correlated with changes in the amount of nitrous oxide produced. This correlation was found both with non-manipulated samples at different naturally occurring pH, and also in samples where the pH in activated sludge from the anoxic basin was changed experimentally. In non-manipulated samples, there was no nitrous oxide formation found at pH values above 6.8. The maximum nitrous oxide production was found when the pH was between 5 and 6.

Distribution and activity of ammonia oxidizing bacteria in a large full-scale trickling filter

The biofilm in a full-scale nitrifying trickling filter (NTF) treating municipal wastewater has been investigated with microbiological methods using fluorescence in situ hybridization (FISH) with 16S rRNA oligonucleotide probes in combination with confocal laser scanning microscopy (CLSM) and mathematical modeling using a dynamic multi-species biofilm reactor model. Ammonia oxidizing bacteria (AOB) were found to belong to the genus Nitrosomonas at different depths in the NTF at every sampling occasion, corresponding to different long-term operational conditions for the NTF. Both the measurements and the corresponding simulated predictions showed the same general trend of a decrease with filter depth of the amount of biofilm, the proportion of AOB to all bacteria and the total amount of AOB. The latter decreased by several times from top to bottom of the NTF. Measurements and simulations of potential ammonium oxidizing activity in the biofilm also showed a decreasing activity with depth in the NTF, which generally was operating at close to complete nitrification. However, no difference was observed when the activity was normalized to the amount of biofilm, despite decreasing proportions of AOB to all bacteria with depth in the NTF. This could be explained by diffusion limitations in the biofilm from the upper parts of the NTF according to the biofilm reactor model. The relatively good agreement between the simulations and the measurements shows that the kind of biofilm reactor model used can qualitatively describe an averaged behavior and averaged composition of the biofilm in the reactor.

New Methods for Analysis of Spatial Distribution and Coaggregation of Microbial Populations in Complex Biofilms

ABSTRACT In biofilms, microbial activities form gradients of substrates and electron acceptors, creating a complex landscape of microhabitats, often resulting in structured localization of the microbial populations present. To understand the dynamic interplay between and within these populations, quantitative measurements and statistical analysis of their localization patterns within the biofilms are necessary, and adequate automated tools for such analyses are needed. We have designed and applied new methods for fluorescence in situ hybridization (FISH) and digital image analysis of directionally dependent (anisotropic) multispecies biofilms. A sequential-FISH approach allowed multiple populations to be detected in a biofilm sample. This was combined with an automated tool for vertical-distribution analysis by generating in silico biofilm slices and the recently developed Inflate algorithm for coaggregation analysis of microbial populations in anisotropic biofilms. As a proof of principle, we show distinct stratification patterns of the ammonia oxidizers Nitrosomonas oligotropha subclusters I and II and the nitrite oxidizer Nitrospira sublineage I in three different types of wastewater biofilms, suggesting niche differentiation between the N. oligotropha subclusters, which could explain their coexistence in the same biofilms. Coaggregation analysis showed that N. oligotropha subcluster II aggregated closer to Nitrospira than did N. oligotropha subcluster I in a pilot plant nitrifying trickling filter (NTF) and a moving-bed biofilm reactor (MBBR), but not in a full-scale NTF, indicating important ecophysiological differences between these phylogenetically closely related subclusters. By using high-resolution quantitative methods applicable to any multispecies biofilm in general, the ecological interactions of these complex ecosystems can be understood in more detail.

Nitrification potential and population dynamics of nitrifying bacterial biofilms in response to controlled shifts of ammonium concentrations in wastewater trickling filters

Nitrogen removal in wastewater treatment is energy consuming and often carried out in biofilm nitrifying trickling filters (NTFs). We investigated nitrification potential and population dynamics of nitrifying bacteria in pilot-plant NTFs fed with full-scale plant wastewater with high (8-9 mg NH(4)(+)l(-1)) or low (<0.5mg NH(4)(+)l(-1)) ammonium concentrations. After ammonium shifts, nitrification potentials stabilized after 10-43 days depending on feed regime. An NTF fed with 3 days of high, and 4 days of low load per week reached a high nitrification potential, whereas a high load for 1 day a week gave a low potential. Nitrosomonas oligotropha dominated the AOB and changes in nitrification potentials were not explained by large population shifts to other AOBs. Although nitrification potentials were generally correlated with the relative amounts of AOB and NOB, this was not always the case. Ammonium feed strategies can be used to optimize wastewater treatment performance.

Three-dimensional stratification of bacterial biofilm populations in a moving bed biofilm reactor for nitritation-anammox.

Moving bed biofilm reactors (MBBRs) are increasingly used for nitrogen removal with nitritation-anaerobic ammonium oxidation (anammox) processes in wastewater treatment. Carriers provide protected surfaces where ammonia oxidizing bacteria (AOB) and anammox bacteria form complex biofilms. However, the knowledge about the organization of microbial communities in MBBR biofilms is sparse. We used new cryosectioning and imaging methods for fluorescence in situ hybridization (FISH) to study the structure of biofilms retrieved from carriers in a nitritation-anammox MBBR. The dimensions of the carrier compartments and the biofilm cryosections after FISH showed good correlation, indicating little disturbance of biofilm samples by the treatment. FISH showed that Nitrosomonas europaea/eutropha-related cells dominated the AOB and Candidatus Brocadia fulgida-related cells dominated the anammox guild. New carriers were initially colonized by AOB, followed by anammox bacteria proliferating in the deeper biofilm layers, probably in anaerobic microhabitats created by AOB activity. Mature biofilms showed a pronounced three-dimensional stratification where AOB dominated closer to the biofilm-water interface, whereas anammox were dominant deeper into the carrier space and towards the walls. Our results suggest that current mathematical models may be oversimplifying these three-dimensional systems and unless the multidimensionality of these systems is considered, models may result in suboptimal design of MBBR carriers.

Assay for determination of α-glucosidase and peptidase activity and location in a nitrifying trickling filter

Abstract. Enzymatic α-glucosidase and peptidase activity in a nitrifying trickling filter (NTF) at the Rya wastewater treatment plant, Göteborg, Sweden, was investigated to evaluate whether these activities can be used as indicators of heterotrophic activity and polymer degradation. Samples of the biofilm were taken from the NTF and incubated in sterile filtered effluent water from the NTF with the addition of soluble starch, peptone, and ammonium chloride. In order to determine the distribution of enzyme activities, the α-glucosidase and peptidase activities were measured in the biofilm samples, in the filtered effluent water from the NTF and in the water phase in which the biofilm was incubated. Activities of both enzymes were found both in the effluent water from the NTF and in the biofilm. The enzyme activities were elevated in the samples when starch and peptone were present. In addition, there was a significant inhibition of ammonium oxidation in samples incubated with starch and peptone. Thus, the presence of starch, peptone and ammonium resulted in increased activity of heterotrophs, which lead to an inhibition of the nitrifiers, probably via competition for available oxygen.