scholarly journals In Situ Activity and Spatial Organization of Anaerobic Ammonium-Oxidizing (Anammox) Bacteria in Biofilms

2007 ◽  
Vol 73 (15) ◽  
pp. 4931-4939 ◽  
Author(s):  
Tomonori Kindaichi ◽  
Ikuo Tsushima ◽  
Yuji Ogasawara ◽  
Masaki Shimokawa ◽  
Noriatsu Ozaki ◽  
...  
Keyword(s):  
Organic Compounds ◽  
Spatial Organization ◽  
Flow Direction ◽  
Anammox Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Reactor Performance ◽  
Inlet Port ◽  
Anammox Activity ◽  
Total Bacteria

ABSTRACT We investigated autotrophic anaerobic ammonium-oxidizing (anammox) biofilms for their spatial organization, community composition, and in situ activities by using molecular biological techniques combined with microelectrodes. Results of phylogenetic analysis and fluorescence in situ hybridization (FISH) revealed that “Brocadia”-like anammox bacteria that hybridized with the Amx820 probe dominated, with 60 to 92% of total bacteria in the upper part (<1,000 μm) of the biofilm, where high anammox activity was mainly detected with microelectrodes. The relative abundance of anammox bacteria decreased along the flow direction of the reactor. FISH results also indicated that Nitrosomonas-, Nitrosospira-, and Nitrosococcus-like aerobic ammonia-oxidizing bacteria (AOB) and Nitrospira-like nitrite-oxidizing bacteria (NOB) coexisted with anammox bacteria and accounted for 13 to 21% of total bacteria in the biofilms. Microelectrode measurements at three points along the anammox reactor revealed that the NH4 + and NO2 − consumption rates decreased from 0.68 and 0.64 μmol cm−2 h−1 at P2 (the second port, 170 mm from the inlet port) to 0.30 and 0.35 μmol cm−2 h−1 at P3 (the third port, 205 mm from the inlet port), respectively. No anammox activity was detected at P4 (the fourth port, 240 mm from the inlet port), even though sufficient amounts of NH4 + and NO2 − and a high abundance of anammox bacteria were still present. This result could be explained by the inhibitory effect of organic compounds derived from biomass decay and/or produced by anammox and coexisting bacteria in the upper parts of the biofilm and in the upstream part of the reactor. The anammox activities in the biofilm determined by microelectrodes reflected the overall reactor performance. The several groups of aerobic AOB lineages, Nitrospira-like NOB, and Betaproteobacteria coexisting in the anammox biofilm might consume a trace amount of O2 or organic compounds, which consequently established suitable microenvironments for anammox bacteria.

scholarly journals Potential Interactions of Particle-Associated Anammox Bacteria with Bacterial and Archaeal Partners in the Namibian Upwelling System

2007 ◽  
Vol 73 (14) ◽  
pp. 4648-4657 ◽  
Author(s):  
Dagmar Woebken ◽  
Bernhard M. Fuchs ◽  
Marcel M. M. Kuypers ◽  
Rudolf Amann
Keyword(s):  
16S Rrna ◽  
Free Water ◽  
Water Phase ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Anammox Bacteria ◽  
Upwelling System ◽  
Group I ◽  
Anammox Activity

ABSTRACT Recent studies have shown that the anaerobic oxidation of ammonium by anammox bacteria plays an important role in catalyzing the loss of nitrogen from marine oxygen minimum zones (OMZ). However, in situ oxygen concentrations of up to 25 μM and ammonium concentrations close to or below the detection limit in the layer of anammox activity are hard to reconcile with the current knowledge of the physiology of anammox bacteria. We therefore investigated samples from the Namibian OMZ by comparative 16S rRNA gene analysis and fluorescence in situ hybridization. Our results showed that “Candidatus Scalindua” spp., the typical marine anammox bacteria, colonized microscopic particles that were likely the remains of either macroscopic marine snow particles or resuspended particles. These particles were slightly but significantly (P < 0.01) enriched in Gammaproteobacteria (11.8% ± 5.0%) compared to the free-water phase (8.1% ± 1.8%). No preference for the attachment to particles could be observed for members of the Alphaproteobacteria and Bacteroidetes, which were abundant (12 to 17%) in both habitats. The alphaproteobacterial SAR11 clade, the Euryarchaeota, and group I Crenarchaeota, were all significantly depleted in particles compared to their presence in the free-water phase (16.5% ± 3.5% versus 2.6% ± 1.7%, 2.7% ± 1.9% versus <1%, and 14.9% ± 4.6% versus 2.2% ± 1.8%, respectively, all P < 0.001). Sequence analysis of the crenarchaeotal 16S rRNA genes showed a 99% sequence identity to the nitrifying “Nitrosopumilus maritimus.” Even though we could not observe conspicuous consortium-like structures of anammox bacteria with particle-enriched bacterioplankton groups, we hypothesize that members of Gammaproteobacteria, Alphaproteobacteria, and Bacteroidetes play a critical role in extending the anammox reaction to nutrient-depleted suboxic water layers in the Namibian upwelling system by creating anoxic, nutrient-enriched microniches.

Transition of bacterial spatial organization in a biofilm monitored by FISH and subsequent image analysis

2004 ◽  
Vol 49 (11-12) ◽  
pp. 365-370 ◽  
Author(s):  
Y. Aoi ◽  
S. Tsuneda ◽  
A. Hirata
Keyword(s):  
Image Analysis ◽  
Outer Layer ◽  
Spatial Organization ◽  
Heterotrophic Bacteria ◽  
Great Influence ◽  
Ammonia Oxidizing Bacteria ◽  
Nitrite Oxidizing Bacteria ◽  
Growth Activity ◽  
Do Concentration

The dynamic transition of bacterial community structure in a biofilm was monitored by the fluorescence in situ hybridization (FISH) technique and subsequent image analysis. Heterotrophic bacteria that had occupied the outer layer were gradually decreased whereas ammonia-oxidizing bacteria (AOB) gradually increased their growth activity and extended their existence area to the outer layer of the biofilm through the gradual reduction of the C/N ratio. The spatial organization of AOB in the biofilm dynamically changed responding to the environmental conditions such as pH fluctuation and lack of dissolved oxygen (DO) and had great influence on the nitrification activity. The accumulation of nitrite was observed at lower DO concentration, which might be due to the property that nitrite-oxidizing bacteria (NOB) possess of higher Km values for oxygen than AOB.

scholarly journals Short- and long-term orange dye effects on ammonium oxidizing and anammox bacteria activities

2017 ◽  
Vol 76 (1) ◽  
pp. 79-86 ◽  
Author(s):  
A. Val del Río ◽  
A. Stachurski ◽  
R. Méndez ◽  
J. L. Campos ◽  
J. Surmacz-Górska ◽  
...  
Keyword(s):  
Azo Dye ◽  
Specific Activity ◽  
Dye Adsorption ◽  
Anammox Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Continuous Stirred Tank Reactor ◽  
Partial Nitritation ◽  
Anammox Activity ◽  
Anammox Process

The effects of orange azo dye over ammonia oxidizing bacteria (AOB) and anammox bacteria activities were tested. Performed batch tests indicated that concentrations lower than 650 mgorange/L stimulated AOB activity, while anammox bacteria activity was inhibited at concentrations higher than 25 mgorange/L. Long-term performance of a continuous stirred tank reactor (CSTR) for the partial nitritation and a sequencing batch reactor (SBR) for the anammox process was tested in the presence of 50 mgorange/L. In the case of the partial nitritation process, both the biomass concentration and the specific AOB activity increased after 50 days of orange azo dye addition. Regarding the anammox process, specific activity decreased down to 58% after 12 days of operation with continuous feeding of 50 mgorange/L. However, the anammox activity was completely recovered only 54 days after stopping the dye addition in the feeding. Once the biomass was saturated the azo dye adsorption onto the biomass was insignificant in the CSTR for the partial nitritation process fed with 50 mgorange/L. However, in the SBR the absorption was determined as 6.4 mgorange/g volatile suspended solids. No biological decolorization was observed in both processes.

scholarly journals Effect of temperature shifts and anammox biomass immobilization on sequencing batch reactor performance and bacterial genes abundance

2020 ◽  
Author(s):  
A. Banach-Wiśniewska ◽  
M. Ćwiertniewicz-Wojciechowska ◽  
A. Ziembińska-Buczyńska
Keyword(s):  
Wastewater Treatment ◽  
Removal Efficiency ◽  
Nitrogen Removal ◽  
Batch Reactor ◽  
Effect Of Temperature ◽  
Anammox Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Ammonium Oxidation ◽  
Reactor Performance ◽  
Gene Abundance

Abstract Implementation of anaerobic ammonium oxidation (anammox) below its optimal temperature, known as “cold anammox”, may lead to its common use in wastewater treatment plants, reducing the operational costs of wastewater treatment. Thus, we investigated the effects of immobilization in polyvinyl alcohol–sodium alginate gel beads on anammox performance at temperatures of 30 °C, 23 °C, and 15 °C in laboratory-scale sequencing batch reactors. We determined the relative gene abundance of the nitrogen removal bacterial groups, which are considered as the key functional microbes of nitrogen cycle in activated sludge: denitrifies, ammonia-oxidizing bacteria, nitrite-oxidizing bacteria, and anammox bacteria. Nitrogen removal efficiency was higher for immobilized anammox sludge in comparison with non-immobilized anammox biomass at each investigated temperature. At 30 °C, nitrogen removal efficiency was 83.7 ± 6.46% for immobilized reactor, and 79.4 ± 7.83% for the control reactor, while at 15 °C was remained at the level of 50 ± 2.5% for immobilized reactor, and fluctuated from 13.2 to 45.3% for the control one. During temperature shifts, the process was also more stable in the case of the reactor with immobilized biomass. A statistically significant correlation was found between nitrogen removal efficiency and hydrazine oxidoreductase gene abundance.

scholarly journals Biomass aggregation influences NaN3 short-term effects on anammox bacteria activity

2016 ◽  
Vol 75 (5) ◽  
pp. 1007-1013 ◽  
Author(s):  
A. Pedrouso ◽  
A. Val del Río ◽  
J. L. Campos ◽  
R. Méndez ◽  
A. Mosquera-Corral
Keyword(s):  
Inhibitory Effect ◽  
Process Efficiency ◽  
Anammox Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Long Term Stability ◽  
Anammox Activity ◽  
Nitrite Oxidizing Bacteria ◽  
Biomass Activity ◽  
Nitrogen Concentrations ◽  
Granular Biomass

The main bottleneck to maintain the long-term stability of the partial nitritation-anammox processes, especially those operated at low temperatures and nitrogen concentrations, is the undesirable development of nitrite oxidizing bacteria (NOB). When this occurs, the punctual addition of compounds with the capacity to specifically inhibit NOB without affecting the process efficiency might be of interest. Sodium azide (NaN3) is an already known NOB inhibitor which at low concentrations does not significantly affect the ammonia oxidizing bacteria (AOB) activity. However, studies about its influence on anammox bacteria are unavailable. For this reason, the objective of the present study was to evaluate the effect of NaN3 on the anammox activity. Three different types of anammox biomass were used: granular biomass comprising AOB and anammox bacteria (G1), anammox enriched granules (G2) and previous anammox granules disaggregated (F1). No inhibitory effect of NaN3 was measured on G1 sludge. However, the anammox activity decreased in the case of G2 and F1. Granular biomass activity was less affected (IC50 90 mg/L, G2) than flocculent one (IC50 5 mg/L, F1). Summing up, not only does the granular structure protect the anammox bacteria from the NaN3 inhibitory effect, but also the AOB act as a barrier decreasing the inhibition.

scholarly journals Sustained nitrogen loss in a symbiotic association of Comammox Nitrospira and Anammox bacteria

2020 ◽  
Author(s):  
Ekaterina Y. Gottshall ◽  
Sam J. Bryson ◽  
Kathryn I. Cogert ◽  
Matthieu Landreau ◽  
Christopher J. Sedlacek ◽  
...  
Keyword(s):  
Spatial Organization ◽  
Nitrogen Loss ◽  
Anammox Bacteria ◽  
Symbiotic Association ◽  
Ammonia Oxidizing Bacteria ◽  
Ammonia Oxidizing Archaea ◽  
Substrate Affinities ◽  
Global Nitrogen Cycle ◽  
Aerobic And Anaerobic ◽  
Reproducible Manner

ABSTRACTThe discovery of complete aerobic and anaerobic ammonia-oxidizing bacteria (Comammox and Anammox) significantly altered our understanding of the global nitrogen cycle. A high affinity for ammonia (Km(app),NH3 ≈ 63nM) and oxygen place the first described isolate, Comammox Nitrospira inopinata in the same trophic category as organisms such as some ammonia-oxidizing archaea. However, N. inopinata has a relatively low affinity for nitrite (Km,NO2 ≈ 449.2μM) suggesting it would be less competitive for nitrite than other nitrite-consuming aerobes and anaerobes. We examined the ecological relevance of the disparate substrate affinities by coupling it with Anammox (Nitrospira inopinata and Brocadia anammoxidans, respectively). Synthetic communities were established in hydrogel granules in which Comammox grew in the aerobic outer layer to provide Anammox with nitrite in the inner anoxic core to form dinitrogen gas. This spatial organization was confirmed with FISH imaging, supporting a mutualistic or commensal relationship. Successful co-habitation of Comammox N. inopinata and Anammox in synthetic granules broadens our limited understanding of the interplay between these two species and offers potential biotechnological applications to study any type of bacterial pairings in a systematic and reproducible manner.

Structure and function of nitrifying biofilms as determined by in situ hybridization and the use of microelectrodes

2000 ◽  
Vol 42 (12) ◽  
pp. 21-32 ◽  
Author(s):  
S. Okabe ◽  
Y. Watanabe
Keyword(s):  
In Situ Hybridization ◽  
Spatial Organization ◽  
Settling Tank ◽  
Outer Part ◽  
Domestic Wastewater ◽  
Nitrifying Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Bacterial Populations ◽  
Nitrite Oxidizing Bacteria

Time dependent development of the spatial organization of NH4+- and NO2−-oxidizing bacterial populations in a domestic wastewater biofilm and in an autotrophic nitrifying biofilm were investigated by fluorescent in situ hybridization (FISH) with a set of 16S rRNA-targeted oligonucleotide probes. Population dynamics of nitrifying bacteria in the biofilms were correlated with the biofilm performance. In situ hybridization indicated that Nitrosomonas spp. (excluding probe NEU stained NH4+-oxidizing bacteria: i.e., N. marina-lineage, N. europaea-lineage, N. eutropha, and N. halophila) and Nitrospira-like bacteria were the numerically dominant nitrifying species in the domestic wastewater biofilm. However, probe NEU stained NH4+-inoxidizing bacteria became dominant populations in the autotrophic nitrifying biofilm (which were initially cultured with the primary settling tank effluent) after switching to the synthetic media. This population shift might be attributed to the effect of NO2−-–N accumulation and higher growth rates of N. europaea-lineage and N. eutropha, outcompeting other Nitrosomonas spp. in the synthetic medium. This evidence indirectly supports that N. europhaea has been most commonly isolated and studied in most of the previous researches. For the spatial organization of NH4+- and NO2−-oxidizing bacterial populations, bacteria of the genus Nitrobacter could not be detected, instead Nitrospira-like bacteria were found as the main nitrite-oxidizing bacteria in both biofilms. Whereas most of the ammonia-oxidizing bacteria were found throughout the biofilms, the location of nitrite-oxidizing bacteria was restricted to the active nitrite-oxidizing zone, which was detected in the inner part of the biofilms. Microelectrode measurements showed that the active ammonia-oxidizing zone was located in the outer part of a biofilm, whereas the active nitrite-oxidizing zone was located just below the ammonia-oxidizing zone and overlapped the location of NO2−-oxidizing bacteria, as determined with FISH. These observations have considerable significance to our understanding of microbial nitrification occurring in wastewater treatment processes and in the natural environment.

scholarly journals Stark Contrast in Denitrification and Anammox across the Deep Norwegian Trench in the Skagerrak

2013 ◽  
Vol 79 (23) ◽  
pp. 7381-7389 ◽  
Author(s):  
Mark Trimmer ◽  
Pia Engström ◽  
Bo Thamdrup
Keyword(s):  
Organic Matter ◽  
Oxygen Consumption ◽  
Sediment Cores ◽  
Anammox Bacteria ◽  
Ammonium Oxidation ◽  
Oxygen Penetration ◽  
Anammox Activity ◽  
Redfield Ratios ◽  
First Time

ABSTRACTEnvironmental anaerobic ammonium oxidation (anammox) was demonstrated for the first time in 2002, using15N labeling, in homogenized sediment from the Skagerrak, where it accounted for up to 67% of N2production. We returned to some of these original sites in 2010 to make measurements of nitrogen and carbon cycling under conditions more representative of thosein situ, quantifying anammox and denitrification, together with oxygen penetration and consumption, in intact sediment cores. Overall, oxygen consumption and N2production decayed with water depth, as expected, but the drop in N2production was relatively more pronounced. Whereas we confirmed the dominance of N2production by anammox (72% and 77%) at the two deepest sites (∼700 m of water), anammox was conspicuously absent from two shallower sites (∼200 m and 400 m). At the shallower sites, we could measure no anammox activity with either intact or homogeneous sediment, and quantitative PCR (16S rRNA) gave a negligible abundance of anammox bacteria in the anoxic layers. Such an absence of anammox, especially at one locale where it was originally demonstrated, is hard to reconcile. Despite the dominance of anammox at the deepest sites, anammox activity could not make up for the drop in denitrification, and assuming Redfield ratios for the organic matter being mineralized, the estimated retention of fixed N actually increased to 90% to 97% of that mineralized, whereas it was 80% to 86% at the shallower sites.

scholarly journals Nitrogen removal using an anammox membrane bioreactor at low temperature

2015 ◽  
Vol 72 (12) ◽  
pp. 2148-2153 ◽  
Author(s):  
Takanori Awata ◽  
Yumiko Goto ◽  
Tomonori Kindaichi ◽  
Noriatsu Ozaki ◽  
Akiyoshi Ohashi
Keyword(s):  
Low Temperature ◽  
Nitrogen Removal ◽  
Membrane Bioreactor ◽  
Membrane Bioreactors ◽  
Distinct Advantage ◽  
Anammox Bacteria ◽  
Short Term ◽  
Term Operation ◽  
Anammox Activity

Membrane bioreactors (MBRs) have the ability to completely retain biomass and are thus suitable for slowly growing anammox bacteria. In the present study, an anammox MBR was operated to investigate whether the anammox activity would remain stable at low temperature, without anammox biomass washout. The maximum nitrogen removal rates were 6.7 and 1.1 g-N L−1 day−1 at 35 °C and 15 °C, respectively. Fluorescence in situ hybridization and 16S rRNA-based phylogenetic analysis revealed no change in the predominant anammox species with temperature because of the complete retention of anammox biomass in the MBR. These results indicate that the predominant anammox bacteria in the MBR cannot adapt to a low temperature during short-term operation. Conversely, anammox activity recovered rapidly after restoring the temperature from the lower value to the optimal temperature (35 °C). The rapid recovery of anammox activity is a distinct advantage of using an MBR anammox reactor.

scholarly journals Anaerobic Ammonia-Oxidizing Bacteria and Related Activity in Baltimore Inner Harbor Sediment

2005 ◽  
Vol 71 (4) ◽  
pp. 1816-1821 ◽  
Author(s):  
Yossi Tal ◽  
Joy E. M. Watts ◽  
Harold J. Schreier
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Oligonucleotide Primer ◽  
Anammox Bacteria ◽  
Rrna Gene ◽  
Ammonia Oxidizing Bacteria ◽  
Gene Sequences ◽  
16S Rrna Gene Sequences ◽  
Harbor Sediment ◽  
Anammox Activity

ABSTRACT The discovery of bacteria capable of anaerobic ammonia oxidation (anammox) has generated interest in understanding the activity, diversity, and distribution of these bacteria in the environment. In this study anammox activity in sediment samples obtained from the Inner Harbor of Baltimore, Md., was detected by 15N tracer assays. Anammox-specific oligonucleotide primer sets were used to screen a Planctomycetales-specific 16S rRNA gene library generated from sediment DNA preparations, and four new anammox bacterial sequences were identified. Three of these sequences form a cohesive new branch of the anammox group, and the fourth sequence branches separately from this group. Denaturing gradient gel electrophoresis analysis of sediment incubated with anammox-specific media confirmed the presence of the four anammox-related 16S rRNA gene sequences. Evidence for the presence of anammox bacteria in Inner Harbor sediment was also obtained by using an anammox-specific probe in fluorescence in situ hybridization studies. To our knowledge, this is the first report of anammox activity and related bacterial 16S rRNA gene sequences from the Chesapeake Bay basin area, and the results suggest that this pathway plays an important role in the nitrogen cycle of this estuarine environment. Furthermore, the presence of these bacteria and their activity in sediment strengthen the contention that anammox-related Plactomycetales are globally distributed.

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