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Regime shift and microbial dynamics in a sequencing batch reactor for nitrification and anammox treatment of urine.

ABSTRACT: The microbial population and physicochemical process parameters of a sequencing batch reactor for nitrogen removal from urine were monitored over a 1.5-year period. Microbial community fingerprinting (automated ribosomal intergenic spacer analysis), 16S rRNA gene sequencing, and quantitative PCR on nitrogen cycle functional groups were used to characterize the microbial population. The reactor combined nitrification (ammonium oxidation)/anammox with organoheterotrophic denitrification. The nitrogen elimination rate initially increased by 400%, followed by an extended period of performance degradation. This phase was characterized by accumulation of nitrite and nitrous oxide, reduced anammox activity, and a different but stable microbial community. Outwashing of anammox bacteria or their inhibition by oxygen or nitrite was insufficient to explain reactor behavior. Multiple lines of evidence, e.g., regime-shift analysis of chemical and physical parameters and cluster and ordination analysis of the microbial community, indicated that the system had experienced a rapid transition to a new stable state that led to the observed inferior process rates. The events in the reactor can thus be interpreted to be an ecological regime shift. Constrained ordination indicated that the pH set point controlling cycle duration, temperature, airflow rate, and the release of nitric and nitrous oxides controlled the primarily heterotrophic microbial community. We show that by combining chemical and physical measurements, microbial community analysis and ecological theory allowed extraction of useful information about the causes and dynamics of the observed process instability.

SUBMITTER: Burgmann H

PROVIDER: S-EPMC3165423 | biostudies-literature | 2011 Sep

REPOSITORIES: biostudies-literature

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Regime shift and microbial dynamics in a sequencing batch reactor for nitrification and anammox treatment of urine.

Bürgmann Helmut H Jenni Sarina S Vazquez Francisco F Udert Kai M KM

Applied and environmental microbiology 20110701 17

The microbial population and physicochemical process parameters of a sequencing batch reactor for nitrogen removal from urine were monitored over a 1.5-year period. Microbial community fingerprinting (automated ribosomal intergenic spacer analysis), 16S rRNA gene sequencing, and quantitative PCR on nitrogen cycle functional groups were used to characterize the microbial population. The reactor combined nitrification (ammonium oxidation)/anammox with organoheterotrophic denitrification. The nitro ...[more]

PMID: 21724875

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Project description:Elevated nitrogen removal efficiencies from ammonium-rich wastewaters have been demonstrated by several applications, that combine nitritation and anammox processes. Denitrification will occur simultaneously when organic carbon is also present. In this study, the activity of aerobic ammonia oxidizing, anammox and denitrifying bacteria in a full scale sequencing batch reactor, treating digester supernatants, was studied by means of batch-assays. AOB and anammox activities were maximum at pH of 8.0 and 7.8-8.0, respectively. Short term effect of nitrite on anammox activity was studied, showing nitrite up to 42 mg/L did not result in inhibition. Both denitrification via nitrate and nitrite were measured. To reduce nitrite-oxidizing activity, high NH3-N (1.9-10 mg NH3-N/L) and low nitrite (3-8 mg TNN/L) are required conditions during the whole SBR cycle. Molecular analysis showed the nitritation-anammox sludge harbored a high microbial diversity, where each microorganism has a specific role. Using ammonia monooxygenase ?-subunit (amoA) gene as a marker, our analyses suggested different macro- and micro-environments in the reactor strongly affect the AOB community, allowing the development of different AOB species, such as N. europaea/eutropha and N. oligotropha groups, which improve the stability of nitritation process. A specific PCR primer set, used to target the 16S rRNA gene of anammox bacteria, confirmed the presence of the "Ca. Brocadia fulgida" type, able to grow in presence of organic matter and to tolerate high nitrite concentrations. The diversity of denitrifiers was assessed by using dissimilatory nitrite reductase (nirS) gene-based analyses, who showed denitifiers were related to different betaproteobacterial genera, such as Thauera, Pseudomonas, Dechloromonas and Aromatoleum, able to assist in forming microbial aggregates. Concerning possible secondary processes, no n-damo bacteria were found while NOB from the genus Nitrobacter was detected.

Project description:Bacterial community dynamics of the ANAMMOX reactor of an integrated "UASB + SHARON + ANAMMOX" system for treating piggery wastewater were investigated using the Illumina MiSeq method with samples obtained at ~ 2-week intervals during a 314-day period. With aerobic activated sludge as seeds and low content artificial wastewater (NH4+-N 50 mg/L; NO2--N 55 mg/L) as influent for the ANAMMOX reactor, nitrogen removal was initially observed on day 38 with a removal rate 1.3 mg N L-1 day-1, and increased to 90.4 mg N L-1 day-1 on day 55 with almost complete removal of ammonia and nitrite, indicating a successful startup of the reactor. Increasing influent load stepwise to NH4+-N 272.7 mg/L/NO2--N 300 mg/L, nitrogen removal rate increased gradually to 470 mg N L-1 day-1 on day 228, and maintained a stable level (~ 420 mg N L-1 day-1) following introduction of SHARON effluent since day 229. Correlation between microbial community dynamics and nitrogen removal capability was significant (r = 0.489, p < 0.001). Microbial community composition was determined by influent ammonia, influent nitrite, effluent nitrate and some undefined factors. Anammox bacteria, accounting for ~ 98.7% of Planctomycetes, became detectable (0.03% relative abundance) since day 38 and increased to 0.9% on day 58, well consistent with nitrogen removal performance of the reactor. Relative abundance of anammox bacteria gradually increased to 38.4% on day 140 with stepwise increased influent load; decreased to 0.4% on day 169 because of nitrite inhibition; increased to 19.24% on day 233 when the influent load was dropped; kept at ~ 9.0% with SHARON effluent used as influent and dropped to 3.3% finally. Anammox bacteria, only Candidatus Brocadia and Ca. Kuenenia detected, were the most abundant at genus level. Ca. Brocadia related taxa were enriched firstly under low load and detectable during the entire experimental period. Three main groups represented by Ca. Brocadia related OTUs were enriched or eliminated at different loads, but Ca. Kuenenia related taxa were enriched only under high load (NO2--N > 300 mg/L), suggesting their different niches and application for different loads. These findings improve the understanding of relationships among microbial community/functional taxa, running parameters and reactor performance, and will be useful in optimizing running parameters for rapid startup and high, stable efficiency.

Dataset Information

Regime shift and microbial dynamics in a sequencing batch reactor for nitrification and anammox treatment of urine.

Publications

Regime shift and microbial dynamics in a sequencing batch reactor for nitrification and anammox treatment of urine.

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