Molecular analysis of ammonia-oxidizing bacterial populations in aerated-anoxic Orbal processes

2002 ◽  
Vol 46 (1-2) ◽  
pp. 273-280 ◽  
Author(s):  
H.-D. Park ◽  
J.M. Regan ◽  
D.R. Noguera
Keyword(s):  
Fragment Length Polymorphism ◽  
Ammonia Oxidation ◽  
Wastewater Treatment Plants ◽  
Phylogenetic Analyses ◽  
Amoa Gene ◽  
Ammonia Oxidizing Bacteria ◽  
Ammonia Monooxygenase ◽  
Anoxic Zone ◽  
Bacterial Populations ◽  
Nitrosomonas Oligotropha

Aerated-anoxic processes operate under the principle that small additions of oxygen to an anoxic reactor induce simultaneous nitrification and denitrification. In these systems, ammonia oxidation in the anoxic zone can easily account for 30–50% of the total nitrification in the reactor, even though the dissolved oxygen concentration is usually below detection limit. To investigate whether the nitrification efficiency in aerated-anoxic processes was due to the presence of specialized ammonia-oxidizing bacteria (AOB), an analysis of the AOB population in an aerated-anoxic Orbal process and a conventional nitrogen removal process was carried out using phylogenetic analyses based on the ammonia monooxygenase A (amoA) gene. Terminal restriction fragment length polymorphism (TRFLP) analyses revealed that Nitrosospira-like organisms were one of the major contributors to ammonia oxidation in a full-scale aerated-anoxic Orbal reactor. However, the relative populations of Nitrosospira-like and Nitrosomonas-like AOB were not constant and appeared to have seasonal variability. Cloning and sequence comparison of amoA gene fragments demonstrated that most of the AOB in the aerated-anoxic Orbal process belonged to the Nitrosospira sp. and Nitrosomonas oligotropha lineages. The abundance of Nitrosospira-like organisms in aerated-anoxic reactors is significant, since this group of AOB has not been usually associated with nitrification in wastewater treatment plants.

scholarly journals Effects of Zeolite and Biochar Addition on Ammonia-Oxidizing Bacteria and Ammonia-Oxidizing Archaea Communities during Agricultural Waste Composting

2020 ◽  
Vol 12 (16) ◽  
pp. 6336 ◽  
Author(s):  
Xin Wu ◽  
Liheng Ren ◽  
Jiachao Zhang ◽  
Hui Peng
Keyword(s):  
Enzyme Activity ◽  
Ammonia Oxidation ◽  
Agricultural Waste ◽  
Amoa Gene ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ammonia Oxidizing Bacteria ◽  
Ammonia Monooxygenase ◽  
Gene Abundance ◽  
Ammonia Oxidizing Archaea ◽  
The Relationship

The effects of zeolite and biochar addition on ammonia-oxidizing bacteria (AOB) and archaea (AOA) communities during agricultural waste composting were determined in this study. Four treatments were conducted as follows: Treatment A as the control with no additive, Treatment B with 5% of zeolite, Treatment C with 5% of biochar, and Treatment D with 5% of zeolite and 5% biochar, respectively. The AOB and AOA amoA gene abundance as well as the ammonia monooxygenase (AMO) activity were estimated by quantitative PCR and enzyme-linked immunosorbent assay, respectively. The relationship between gene abundance and AMO enzyme activity was determined by regression analysis. Results indicated that the AOB was more abundant than that of AOA throughout the composting process. Addition of biochar and its integrated application with zeolite promoted the AOB community abundance and AMO enzyme activity. Significant positive relationships were obtained between AMO enzyme activity and AOB community abundance (r2 = 0.792; P < 0.01) and AOA community abundance (r2 = 0.772; P < 0.01), indicating that both bacteria and archaea played significant roles in microbial ammonia oxidation during composting. Using biochar and zeolite might promote the nitrification activity by altering the sample properties during agricultural waste composting.

scholarly journals Ecophysiological Characterization of Ammonia-Oxidizing Archaea and Bacteria from Freshwater

2012 ◽  
Vol 78 (16) ◽  
pp. 5773-5780 ◽  
Author(s):  
Elizabeth French ◽  
Jessica A. Kozlowski ◽  
Maitreyee Mukherjee ◽  
George Bullerjahn ◽  
Annette Bollmann
Keyword(s):  
Growth Rates ◽  
Ammonia Oxidation ◽  
Light Exposure ◽  
Enrichment Culture ◽  
Ammonia Oxidizing Bacteria ◽  
Content Type ◽  
Ammonia Oxidizing Archaea ◽  
Group I ◽  
Nitrosomonas Oligotropha

ABSTRACTAerobic biological ammonia oxidation is carried out by two groups of microorganisms, ammonia-oxidizing bacteria (AOB) and the recently discovered ammonia-oxidizing archaea (AOA). Here we present a study using cultivation-based methods to investigate the differences in growth of three AOA cultures and one AOB culture enriched from freshwater environments. The strain in the enriched AOA culture belong to thaumarchaeal group I.1a, with the strain in one enrichment culture having the highest identity with “CandidatusNitrosoarchaeum koreensis” and the strains in the other two representing a new genus of AOA. The AOB strain in the enrichment culture was also obtained from freshwater and had the highest identity to AOB from theNitrosomonas oligotrophagroup (Nitrosomonascluster 6a). We investigated the influence of ammonium, oxygen, pH, and light on the growth of AOA and AOB. The growth rates of the AOB increased with increasing ammonium concentrations, while the growth rates of the AOA decreased slightly. Increasing oxygen concentrations led to an increase in the growth rate of the AOB, while the growth rates of AOA were almost oxygen insensitive. Light exposure (white and blue wavelengths) inhibited the growth of AOA completely, and the AOA did not recover when transferred to the dark. AOB were also inhibited by blue light; however, growth recovered immediately after transfer to the dark. Our results show that the tested AOB have a competitive advantage over the tested AOA under most conditions investigated. Further experiments will elucidate the niches of AOA and AOB in more detail.

Microbial community structure and occurrence of diverse autotrophic ammonium oxidizing microorganisms in the anammox process

2010 ◽  
Vol 61 (11) ◽  
pp. 2723-2732 ◽  
Author(s):  
H. Bae ◽  
Y.-C. Chung ◽  
J.-Y. Jung
Keyword(s):  
Community Structure ◽  
Microbial Community ◽  
Microbial Community Structure ◽  
Fragment Length Polymorphism ◽  
Amoa Gene ◽  
Continuous Operation ◽  
Anaerobic Sludge ◽  
Anammox Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Anammox Process

The enrichment of anaerobic ammonium oxidizing (anammox) bacteria using an upflow anaerobic sludge bioreactor was successfully conducted for 400 days of continuous operation. The bacterial community structure of anammox bioreactor included Proteobacteria (42%), Chloroflexi (22%), Planctomycetes (20%), Chlorobi (7%), Bacteroidetes (5%), Acidobacteria (2%), and Actinobacteria (2%). All clones of Planctomycetes were affiliated with the anammox bacteria, Planctomycete KSU-1 (AB057453). The presence and diversity of ammonia oxidizing bacteria (AOB) and archaea (AOA) were identified by terminal restriction fragment length polymorphism (T-RFLP) based on the amoA gene sequences. The AOB in anammox bioreactor were affiliated with the Nitrosomonas europaea cluster. The T-RFLP result of AOA showed the diverse microbial community structure of AOA with three terminal restriction fragments (T-RFs).

Quantification of amoA gene abundance and their amoA mRNA levels in activated sludge by real-time PCR

2004 ◽  
Vol 50 (8) ◽  
pp. 1-8 ◽  
Author(s):  
N. Araki ◽  
T. Yamaguchi ◽  
S. Yamazaki ◽  
H. Harada
Keyword(s):  
16S Rrna ◽  
Dissolved Oxygen ◽  
Real Time ◽  
Real Time Pcr ◽  
Ammonia Oxidation ◽  
Amoa Gene ◽  
Ammonia Concentration ◽  
Ammonia Oxidizing Bacteria ◽  
Mrna Levels ◽  
Transcription Level

The transcription level of amoA mRNA encoding a subunit of ammonia monooxygenase (AMO) in ammonia-oxidizing bacteria (AOB) was quantified by reverse transcription-polymerase chain reaction (RT-PCR) methods in combination with real-time PCR technology. The effects of ammonia concentration and dissolved oxygen (DO) on the transcription levels of amoA mRNA and 16S rRNA in AOB were evaluated in batch experiments with nitrifying sludge taken from a lab-scale reactor treating artificial wastewater. A batch incubation without ammonia resulted in a rapid decrease, within four hours, in the transcription level of amoA mRNA to as low as 1/10 of that at the beginning of the experiment, while the 16S rRNA level in AOB was almost constant. After subsequent incubation with 3 mM ammonia for eight hours, a small increase in the transcription level of amoA mRNA occurred, but ammonia oxidation proceeded in the interim. Copy numbers of amoA mRNA showed an almost fixed value for over eight hours in the absence of dissolved oxygen.

Nitrous oxide production by lithotrophic ammonia-oxidizing bacteria and implications for engineered nitrogen-removal systems

2011 ◽  
Vol 39 (6) ◽  
pp. 1832-1837 ◽  
Author(s):  
Kartik Chandran ◽  
Lisa Y. Stein ◽  
Martin G. Klotz ◽  
Mark C.M. van Loosdrecht
Keyword(s):  
Nitrous Oxide ◽  
Nitrogen Removal ◽  
Ammonia Oxidation ◽  
Wastewater Treatment Plants ◽  
N2o Emissions ◽  
Ammonia Oxidizing Bacteria ◽  
N2o Production ◽  
Nitrogen Emissions ◽  
Greenhouse Gas Inventories ◽  
Nitrogen Concentrations

Chemolithoautotrophic AOB (ammonia-oxidizing bacteria) form a crucial component in microbial nitrogen cycling in both natural and engineered systems. Under specific conditions, including transitions from anoxic to oxic conditions and/or excessive ammonia loading, and the presence of high nitrite (NO2−) concentrations, these bacteria are also documented to produce nitric oxide (NO) and nitrous oxide (N2O) gases. Essentially, ammonia oxidation in the presence of non-limiting substrate concentrations (ammonia and O2) is associated with N2O production. An exceptional scenario that leads to such conditions is the periodical switch between anoxic and oxic conditions, which is rather common in engineered nitrogen-removal systems. In particular, the recovery from, rather than imposition of, anoxic conditions has been demonstrated to result in N2O production. However, applied engineering perspectives, so far, have largely ignored the contribution of nitrification to N2O emissions in greenhouse gas inventories from wastewater-treatment plants. Recent field-scale measurements have revealed that nitrification-related N2O emissions are generally far higher than emissions assigned to heterotrophic denitrification. In the present paper, the metabolic pathways, which could potentially contribute to NO and N2O production by AOB have been conceptually reconstructed under conditions especially relevant to engineered nitrogen-removal systems. Taken together, the reconstructed pathways, field- and laboratory-scale results suggest that engineering designs that achieve low effluent aqueous nitrogen concentrations also minimize gaseous nitrogen emissions.

scholarly journals Flavobacteria Blooms in Four Eutrophic Lakes: Linking Population Dynamics of Freshwater Bacterioplankton to Resource Availability

2007 ◽  
Vol 73 (11) ◽  
pp. 3511-3518 ◽  
Author(s):  
Alexander Eiler ◽  
Stefan Bertilsson
Keyword(s):  
Resource Availability ◽  
Bacterial Production ◽  
Fragment Length Polymorphism ◽  
Heterotrophic Bacteria ◽  
Phylogenetic Analyses ◽  
Polymorphism Analysis ◽  
Eutrophic Lakes ◽  
Specific Primers ◽  
Bacterial Populations ◽  
Bacterial Succession

ABSTRACT Heterotrophic bacteria are major contributors to biogeochemical cycles and influence water quality. Still, the lack of representative isolates and the few quantitative surveys leave the ecological role and significance of single bacterial populations to be revealed. Here we analyzed the diversity and dynamics of freshwater Flavobacteria populations in four eutrophic temperate lakes. From each lake, clone libraries were constructed using primers specific for either the class Flavobacteria or Bacteria. Sequencing of 194 Flavobacteria clones from 8 libraries revealed a diverse freshwater Flavobacteria community and distinct differences among lakes. Abundance and seasonal dynamics of Flavobacteria were assessed by quantitative PCR with class-specific primers. In parallel, the dynamics of individual populations within the Flavobacteria community were assessed with terminal restriction fragment length polymorphism analysis using identical primers. The contribution of Flavobacteria to the total bacterioplankton community ranged from 0.4 to almost 100% (average, 24%). Blooms where Flavobacteria represented more than 30% of the bacterioplankton were observed at different times in the four lakes. In general, high proportions of Flavobacteria appeared during episodes of high bacterial production. Phylogenetic analyses combined with Flavobacteria community fingerprints suggested dominance of two Flavobacteria lineages. Both drastic alterations in total Flavobacteria and in community composition of this class significantly correlated with bacterial production, emphasizing that resource availability is an important driver of heterotrophic bacterial succession in eutrophic lakes.

scholarly journals Agreement between Theory and Measurement in Quantification of Ammonia-Oxidizing Bacteria

2005 ◽  
Vol 71 (10) ◽  
pp. 6325-6334 ◽  
Author(s):  
Gulnur Coskuner ◽  
Stuart J. Ballinger ◽  
Russell J. Davenport ◽  
Rheanne L. Pickering ◽  
Rosario Solera ◽  
...  
Keyword(s):  
Ammonia Oxidation ◽  
Wastewater Treatment Plants ◽  
Process Variable ◽  
Confocal Scanning Laser Microscopy ◽  
Ammonia Oxidizing Bacteria ◽  
Cell Numbers ◽  
Oxidation Rates ◽  
Cell Counts ◽  
Confocal Scanning Laser ◽  
Log Normal

ABSTRACT Autotrophic ammonia-oxidizing bacteria (AOB) are of vital importance to wastewater treatment plants (WWTP), as well as being an intriguing group of microorganisms in their own right. To date, corroboration of quantitative measurements of AOB by fluorescence in situ hybridization (FISH) has relied on assessment of the ammonia oxidation rate per cell, relative to published values for cultured AOB. Validation of cell counts on the basis of substrate transformation rates is problematic, however, because published cell-specific ammonia oxidation rates vary by over two orders of magnitude. We present a method that uses FISH in conjunction with confocal scanning laser microscopy to quantify AOB in WWTP, where AOB are typically observed as microcolonies. The method is comparatively simple, requiring neither detailed cell counts or image analysis, and yet it can give estimates of either cell numbers or biomass. Microcolony volume and diameter were found to have a log-normal distribution. We were able to show that virtually all (>96%) of the AOB biomass occurred as microcolonies. Counts of microcolony abundance and measurement of their diameter coupled with a calibration of microcolony dimensions against cell numbers or AOB biomass were used to determine AOB cell numbers and biomass in WWTP. Cell-specific ammonia oxidation rates varied between plants by over three orders of magnitude, suggesting that cell-specific ammonia oxidation is an important process variable. Moreover, when measured AOB biomass was compared with process-based estimates of AOB biomass, the two values were in agreement.

scholarly journals Effects of sea animal colonization on the coupling between dynamics and activity of soil ammonia-oxidizing bacteria and archaea in maritime Antarctica

2019 ◽  
Vol 16 (20) ◽  
pp. 4113-4128 ◽  
Author(s):  
Qing Wang ◽  
Renbin Zhu ◽  
Yanling Zheng ◽  
Tao Bao ◽  
Lijun Hou
Keyword(s):  
Ammonia Oxidation ◽  
Amoa Gene ◽  
Ammonia Oxidizing Bacteria ◽  
Nitrogen Transformations ◽  
Tundra Soils ◽  
Maritime Antarctica ◽  
Soil C ◽  
Oxidation Rates ◽  
Ammonia Oxidizing Archaea ◽  
Tundra Ecosystem

Abstract. The colonization by a large number of sea animals, including penguins and seals, plays an important role in the nitrogen cycle of the tundra ecosystem in coastal Antarctica. However, little is known about the effects of sea animal colonization on ammonia-oxidizing archaea (AOA) and bacteria (AOB) communities involved in nitrogen transformations. In this study, we chose active seal colony tundra soils (SSs), penguin colony soils (PSs), adjacent penguin-lacking tundra soils (PLs), tundra marsh soils (MSs), and background tundra soils (BSs) to investigate the effects of sea animal colonization on the abundance, activity, and diversity of AOA and AOB in maritime Antarctica. Results indicated that AOB dominated over AOA in PS, SS, and PL, whereas AOB and AOA abundances were similar in MS and BS. Penguin or seal activities increased the abundance of soil AOB amoA genes but reduced the abundance of AOA amoA genes, leading to very large ratios (1.5×102 to 3.2×104) of AOB to AOA amoA copy numbers. Potential ammonia oxidation rates (PAORs) were significantly higher (P=0.02) in SS and PS than in PL, MS, and BS and were significantly positively correlated (P<0.001) with AOB amoA gene abundance. The predominance of AOB over AOA and their correlation with PAOR suggested that AOB play a more important role in the nitrification in animal colony soils. Sequence analysis for gene clones showed that AOA and AOB in tundra soils were from the Nitrososphaera and Nitrosospira lineages, respectively. Penguin or seal activities led to a predominance of AOA phylotypes related to Nitrososphaera cluster I and AOB phylotypes related to Nitrosospira clusters I and II but very low relative abundances in AOA phylotypes related to cluster II, and AOB phylotypes related to clusters III and IV. The differences in AOB and AOA community structures were closely related to soil biogeochemical processes under the disturbance of penguin or seal activities: soil C : N alteration and sufficient input of NH4+–N and phosphorus from animal excrements. The results significantly enhanced the understanding of ammonia-oxidizing microbial communities in the tundra environment of maritime Antarctica.

scholarly journals Effects of Sea Animal Colonization on the Coupling between Dynamics and Activity of Soil Ammonia-oxidizing Bacteria and Archaea in Maritime Antarctica

2019 ◽  
Author(s):  
Qing Wang ◽  
Renbin Zhu ◽  
Yanling Zheng ◽  
Tao Bao ◽  
Lijun Hou
Keyword(s):  
Ammonia Oxidation ◽  
Amoa Gene ◽  
Ammonia Oxidizing Bacteria ◽  
Nitrogen Transformations ◽  
Tundra Soils ◽  
Maritime Antarctica ◽  
Oxidation Rates ◽  
Ammonia Oxidizing Archaea ◽  
Copy Numbers ◽  
Tundra Ecosystem

Abstract. The colonization of a large number of sea animal, including penguins and seals, plays an important role in the nitrogen cycle of the tundra ecosystem in coastal Antarctica. However, little is known about the effects of sea animal colonization on ammonia-oxidizing archaea (AOA) and bacteria (AOB) communities involved in nitrogen transformations. In this study, we chose active seal colony tundra soils (STS), penguin colony soils (PTS), adjacent penguin-lacking tundra soils (PLS), tundra marsh soils (MS), and background tundra soils (BS), to investigate the effects of sea animal colonization on the abundance, activity, and diversity of AOA and AOB in maritime Antarctica. Results indicated that AOB dominated over AOA in PTS, STS, and PLS; whereas AOB and AOA abundances were similar in MS and BS. Penguin or seal activities increases the abundance of soil AOB amoA genes, but reduced the abundance of AOA amoA genes, leading to very large ratios (1.5 × 102 to 3.2 × 104) of AOB to AOA amoA copy numbers. Ammonia oxidation rates were significantly higher (P = 0.02) in STS and PTS than in PLS, MS, and BS, and were significantly positively correlated (P < 0.001) with AOB amoA gene abundance suggesting that AOB are more important in the nitrification in animal colony soils. Sequence analysis for gene clones showed that AOA and AOB in tundra soils were from the Nitrosospira and Nitrososphaera lineages, respectively. Seal or penguin activities led to the predominant existence of AOA phylotypes related to Nitrososphaera cluster I and AOB phylotypes related to Nitrosospira clusters I and II, but very low relative abundances in AOA phylotypes related to cluster II, and AOB phylotypes related to cluster III and IV. The differences in AOB and AOA community structures were closely related to soil biogeochemical processes under the disturbance of penguin or seal activities: soil C:N alteration and sufficient input of NH4+–N and phosphorus from animal excrements. The results provide insights into the mechanisms how microbes drive nitrification in maritime Antarctica.

Molecular assessment of ammonia- and nitrite-oxidizing bacteria in full-scale activated sludge wastewater treatment plants

2003 ◽  
Vol 48 (8) ◽  
pp. 119-126 ◽  
Author(s):  
K.G. Robinson ◽  
H.M. Dionisi ◽  
G. Harms ◽  
A.C. Layton ◽  
I.R. Gregory ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
16S Rdna ◽  
Municipal Wastewater ◽  
Wastewater Treatment Plants ◽  
Amoa Gene ◽  
Full Scale ◽  
Ammonia Oxidizing Bacteria ◽  
Copy Numbers ◽  
Nitrite Oxidizing Bacteria

Nitrification was assessed in two full-scale wastewater treatment plants (WWTPs) over time using molecular methods. Both WWTPs employed a complete-mix suspended growth, aerobic activated sludge process (with biomass recycle) for combined carbon and nitrogen treatment. However, one facility treated primarily municipal wastewater while the other only industrial wastewater. Real time PCR assays were developed to determine copy numbers for total 16S rDNA (a measure of biomass content), the amoA gene (a measure of ammonia-oxidizers), and the Nitrospira 16S rDNA gene (a measure of nitrite-oxidizers) in mixed liquor samples. In both the municipal and industrial WWTP samples, total 16S rDNA values were approximately 2-9 × 1013 copies/L and Nitrospira 16S rDNA values were 2-4 × 1010 copies/L. amoA gene concentrations averaged 1.73 × 109 copies/L (municipal) and 1.06 × 1010 copies/L (industrial), however, assays for two distinct ammonia oxidizing bacteria were required.

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