Development of a super high-rate Anammox reactor and in situ analysis of biofilm structure and function

2007 ◽  
Vol 55 (8-9) ◽  
pp. 9-17 ◽  
Author(s):  
Ikuo Tsushima ◽  
Yuji Ogasawara ◽  
Masaki Shimokawa ◽  
Tomonori Kindaichi ◽  
Satoshi Okabe

The anaerobic ammonium oxidation (Anammox) process is a new efficient and cost effective method of ammonium removal from wastewater. Under strictly anoxic condition, ammonium is directly oxidised with nitrite as electron acceptor to dinitrogen gas. However, it is extremely difficult to cultivate Anammox bacteria due to their low growth rate. This suggests that a rapid and efficient start-up of Anammox process is the key to practical applications. To screen appropriate seeding sludge with high Anammox potential, a real-time quantitative PCR assay with newly designed primers has been developed. Thereafter, the seeding sludge with high abundance of Anammox bacteria (1.7 × 108 copies/mg-dry weight) was selected and inoculated into an upflow anaerobic biofilters (UABs). The UABs were operated for more than 1 year and the highest nitrogen removal rate of 24.0 kg-N m−3 day−1 was attained. In addition, the ecophysiology of Anammox bacteria (spatial distribution and in situ activity) in biofilms was analysed by combining a full-cycle 16S rRNA approach and microelectrodes. The microelectrode measurement clearly revealed that a successive vertical zonation of the partial nitrification (NH4+ to NO2−), Anammox reaction and denitrification was developed in the biofilm in the UAB. This result agreed with the spatial distribution of corresponding bacterial populations in the biofilm. We linked the micro-scale information (i.e. single cell and/or biofilm levels) with the macro-scale information (i.e. the reactor level) to understand the details of Anammox reaction occurring in the UABs.

2012 ◽  
Vol 66 (5) ◽  
pp. 958-964 ◽  
Author(s):  
Takanori Awata ◽  
Katsuichiro Tanabe ◽  
Tomonori Kindaichi ◽  
Noriatsu Ozaki ◽  
Akiyoshi Ohashi

Anaerobic ammonium oxidation (anammox) is a type of biological oxidation mediated by a group of Planctomycete-like bacteria. Members of the genus Candidatus Scalindua are mainly found in marine environments, but not exclusively. This group is cultured using different inoculums and conditions; however, its optimal growth conditions are not clear. Additionally, little information is known about the factors that influence the activity and the selection of a population of marine anammox bacteria. This study was conducted to investigate the influence of temperature and salinity on the marine anammox community. To accomplish this, an up-flow fixed-bed column reactor was operated, and quantitative fluorescence in situ hybridization (FISH) with probes specific to dominant marine anammox bacteria was conducted. Anammox activity was observed at 20 and 30 °C, but not at 10 °C. A nitrogen removal rate of 0.32 kg TN m–3 day–1 was obtained at 20 °C. These results suggest that temperature affects the activity (nitrogen removal rate) of anammox bacteria, while salinity does not affect the activity in the marine anammox biofilm.


2008 ◽  
Vol 58 (5) ◽  
pp. 1113-1120 ◽  
Author(s):  
S. E. Vlaeminck ◽  
L. F. F. Cloetens ◽  
M. Carballa ◽  
N. Boon ◽  
W. Verstraete

A novel and efficient way of removing nitrogen from wastewater poor in biodegradable organic carbon, is the combination of partial nitritation and anoxic ammonium oxidation (anammox), as in the one-stage oxygen-limited autotrophic nitrification/denitrification (OLAND) process. Since anoxic ammonium-oxidizing bacteria grow very slowly, maximum biomass retention in the reactor is required. In this study, a lab-scale sequencing batch reactor (SBR) was used to develop granular, rapidly settling biomass. With SBR cycles of one hour and a minimum biomass settling velocity of 0.7 m/h, OLAND granules were formed in 1.5 months and the nitrogen removal rate increased from 50 to 450 mg N L−1 d−1 in 2 months. The granules had a mean diameter of 1.8 mm and their aerobic and anoxic ammonium-oxidizing activities were well equilibrated to perform the OLAND reaction. Fluorescent in-situ hybridization (FISH) demonstrated the presence of both β-proteobacterial aerobic ammonium oxidizers and planctomycetes (among which anoxic ammonium oxidizers) in the granules. The presented results show the applicability of rapidly settling granular biomass for one-stage partial nitritation and anammox.


2021 ◽  
Vol 26 (1) ◽  
pp. 25
Author(s):  
Zulkarnaini Zulkarnaini ◽  
Puti Sri Komala ◽  
Arief Almi

The anaerobic ammonium oxidation (anammox) biofilm process commonly uses various inorganic carriers to enhance nitrogen removal under anaerobic conditions. This study aims to analyze the performance of nitrogen removal in anammox process using sugarcane bagasse as an organic carrier. The experiment was carried out by using an up‐flow anaerobic sludge blanket (UASB) reactor for treating artificial wastewater at room temperature. The reactor was fed with ammonium and nitrite with the concentrations of 70‐150 mg–N/L and variations in the hydraulic retention time of 24 and 12 h. The granular anammox belongs to the genus Candidatus Brocadia sinica that was added as an inoculum of the reactor operation. The experimental stoichiometric of anammox for ΔNO2‐–N: ΔNH4+–N and ΔNO3‐: ΔNH4+ were 1.24 and 0.18, respectively, which is similar to anammox stoichiometry. The maximum Nitrogen Removal Rate (NRR) has achieved 0.29 kg–N/m3.d at Nitrogen Loading Rate (NLR) 0.6 kg–N/m3.d. The highest ammonium conversion efficiency (ACE) and nitrogen removal efficiency (NRE) were 88% and 85%, respectively. Based on this results, it indicated that sugarcane bagasse as organic carriers could increase the amount of total nitrogen removal by provided of denitrification process but inhibited the anammox process at a certain COD concentration.


2013 ◽  
Vol 67 (5) ◽  
pp. 968-975 ◽  
Author(s):  
C. G. Casagrande ◽  
A. Kunz ◽  
M. C. De Prá ◽  
C. R. Bressan ◽  
H. M. Soares

The anaerobic ammonium oxidation (ANAMMOX) is a chemolithoautotrophic process, which converts NH4+ to N2 using nitrite (NO2−) as the electron acceptor. This process has very high nitrogen removal rates (NRRs) and is an alternative to classical nitrification/denitrification wastewater treatment. In the present work, a strategy for nitrogen removal using ANAMMOX process was tested evaluating their performance when submitted to high loading rates and very short hydraulic retention times (HRTs). An up-flow ANAMMOX column reactor was inoculated with 30% biomass (v v−1) fed from 100 to 200 mg L−1 of total N (NO2−-N + NH4+-N) at 35 °C. After start-up and process stability the maximum NRR in the up-flow anaerobic sludge blanket (UASB) reactor was 18.3 g-N L−1 d−1 operated at 0.2 h of HRT. FISH (fluorescence in situ hybridization) analysis and process stoichiometry confirmed that ANAMMOX was the prevalent process for nitrogen removal during the experiments. The results point out that high NRRs can be obtained at very short HRTs using up-flow ANAMMOX column reactor configuration.


2011 ◽  
Vol 77 (19) ◽  
pp. 6802-6807 ◽  
Author(s):  
Francisca A. Luesken ◽  
Jaime Sánchez ◽  
Theo A. van Alen ◽  
Janeth Sanabria ◽  
Huub J. M. Op den Camp ◽  
...  

ABSTRACTNitrite-dependent anaerobic oxidation of methane (n-damo) and ammonium (anammox) are two recently discovered processes in the nitrogen cycle that are catalyzed by n-damo bacteria, including “CandidatusMethylomirabilis oxyfera,” and anammox bacteria, respectively. The feasibility of coculturing anammox and n-damo bacteria is important for implementation in wastewater treatment systems that contain substantial amounts of both methane and ammonium. Here we tested this possible coexistence experimentally. To obtain such a coculture, ammonium was fed to a stable enrichment culture of n-damo bacteria that still contained some residual anammox bacteria. The ammonium supplied to the reactor was consumed rapidly and could be gradually increased from 1 to 20 mM/day. The enriched coculture was monitored by fluorescencein situhybridization and 16S rRNA andpmoAgene clone libraries and activity measurements. After 161 days, a coculture with about equal amounts of n-damo and anammox bacteria was established that converted nitrite at a rate of 0.1 kg-N/m3/day (17.2 mmol day−1). This indicated that the application of such a coculture for nitrogen removal may be feasible in the near future.


2015 ◽  
Vol 57 (30) ◽  
pp. 13958-13978 ◽  
Author(s):  
Mumtazah Ibrahim ◽  
Norjan Yusof ◽  
Mohd Zulkhairi Mohd Yusoff ◽  
Mohd Ali Hassan

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Chunzhen Zou ◽  
Beibei Guo ◽  
Xuming Zhuang ◽  
Liying Ren ◽  
Shou-Qing Ni ◽  
...  

Abstract The effects of FeS on nitrogen removal performance and microbial community of anammox process were studied. During the start-up period, the removal efficiencies of nitrite and total nitrogen were significantly improved by FeS. The addition of FeS increased the content of iron ions in the reactor and promoted the synthesis of heme c, which was involved in the formation of various enzymes. Compared with the control, the abundance of anammox bacteria in the FeS reactor was increased by 29%, and the expression level of the nirS gene (encoding cd1 type nitrite reductase containing heme) was nearly doubled. The content of nitrite reductase (ammonia-forming) in the community was increased by 26.4%. The difference in functional bacteria and enzyme contents in the microbial community resulted in a difference in nitrogen removal rate (NRR) between the two reactors. High-throughput results indicated that FeS increased the richness and diversity of microbial community and enhanced the metabolic function of the microbial community. The addition of FeS did not change the dominant position of Ca. Kuenenia in both reactors. But the relative abundance of heterotrophic denitrifying bacteria was reduced with FeS, which may be related to the inhibition effect of S2− produced by FeS.


Microbiology ◽  
2011 ◽  
Vol 157 (6) ◽  
pp. 1706-1713 ◽  
Author(s):  
Mamoru Oshiki ◽  
Masaki Shimokawa ◽  
Naoki Fujii ◽  
Hisashi Satoh ◽  
Satoshi Okabe

The present study investigated the phylogenetic affiliation and physiological characteristics of bacteria responsible for anaerobic ammonium oxidization (anammox); these bacteria were enriched in an anammox reactor with a nitrogen removal rate of 26.0 kg N m−3 day−1. The anammox bacteria were identified as representing ‘Candidatus Brocadia sinica’ on the basis of phylogenetic analysis of rRNA operon sequences. Physiological characteristics examined were growth rate, kinetics of ammonium oxidation and nitrite reduction, temperature, pH and inhibition of anammox. The maximum specific growth rate (μmax) was 0.0041 h−1, corresponding to a doubling time of 7 days. The half-saturation constants (K s) for ammonium and nitrite of ‘Ca. B. sinica’ were 28±4 and 86±4 µM, respectively, higher than those of ‘Candidatus Brocadia anammoxidans’ and ‘Candidatus Kuenenia stuttgartiensis’. The temperature and pH ranges of anammox activity were 25–45 °C and pH 6.5–8.8, respectively. Anammox activity was inhibited in the presence of nitrite (50 % inhibition at 16 mM), ethanol (91 % at 1 mM) and methanol (86 % at 1 mM). Anammox activities were 80 and 70 % of baseline in the presence of 20 mM phosphorus and 3 % salinity, respectively. The yield of biomass and dissolved organic carbon production in the culture supernatant were 0.062 and 0.005 mol C (mol NH 4 + )−1, respectively. This study compared physiological differences between three anammox bacterial enrichment cultures to provide a better understanding of anammox niche specificity in natural and man-made ecosystems.


2005 ◽  
Vol 71 (2) ◽  
pp. 1066-1071 ◽  
Author(s):  
Didem Güven ◽  
Ana Dapena ◽  
Boran Kartal ◽  
Markus C. Schmid ◽  
Bart Maas ◽  
...  

ABSTRACT Anaerobic ammonium oxidation (anammox) is a recently discovered microbial pathway and a cost-effective way to remove ammonium from wastewater. Anammox bacteria have been described as obligate chemolithoautotrophs. However, many chemolithoautotrophs (i.e., nitrifiers) can use organic compounds as a supplementary carbon source. In this study, the effect of organic compounds on anammox bacteria was investigated. It was shown that alcohols inhibited anammox bacteria, while organic acids were converted by them. Methanol was the most potent inhibitor, leading to complete and irreversible loss of activity at concentrations as low as 0.5 mM. Of the organic acids acetate and propionate, propionate was consumed at a higher rate (0.8 nmol min−1 mg of protein−1) by Percoll-purified anammox cells. Glucose, formate, and alanine had no effect on the anammox process. It was shown that propionate was oxidized mainly to CO2, with nitrate and/or nitrite as the electron acceptor. The anammox bacteria carried out propionate oxidation simultaneously with anaerobic ammonium oxidation. In an anammox enrichment culture fed with propionate for 150 days, the relative amounts of anammox cells and denitrifiers did not change significantly over time, indicating that anammox bacteria could compete successfully with heterotrophic denitrifiers for propionate. In conclusion, this study shows that anammox bacteria have a more versatile metabolism than previously assumed.


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