[논문]Nitrifying Bacterial Community Structure of a Full-Scale Integrated Fixed-Film Activated Sludge Process as Investigated by Pyrosequencing

Kim, Taek-Seung; Kim, Han-Shin; Kwon, Soon-Dong; Park, Hee-Deung

doi:10.4014/jmb.1009.09042

Nitrifying Bacterial Community Structure of a Full-Scale Integrated Fixed-Film Activated Sludge Process as Investigated by Pyrosequencing 원문보기

Journal of microbiology and biotechnology, v.21 no.3, 2011년, pp.293 - 298

Kim, Taek-Seung (School of Civil, Environmental and Architectural Engineering, Korea University) , Kim, Han-Shin (School of Civil, Environmental and Architectural Engineering, Korea University) , Kwon, Soon-Dong (School of Civil, Environmental and Architectural Engineering, Korea University) , Park, Hee-Deung (School of Civil, Environmental and Architectural Engineering, Korea University)

Abstract ▼ AI-Helper

Nitrifying bacterial community structures of suspended and attached biomasses in a full-scale integrated fixed-film activated sludge process were investigated by analyzing 16S rRNA gene sequences obtained from pyrosequencing. The suspended biomass had a higher number of ammonia-oxidizing bacterial sequences (0.8% of total sequences) than the attached biomass (0.07%), although most of the sequences were within the Nitrosomonas oligotropha lineage in both biomasses. Nitrospira-like nitrite-oxidizing bacterial sequences were retrieved in the suspended biomass (0.06%), not in the attached biomass, whereas the existence of Nitrobacter-like sequences was not evident. The suspended biomass had higher nitrification activity (1.13 mg N/TSS/h) than the attached biomass (0.07 mg N/TSS/h). Overall, the results made it possible to conclude the importance of the suspended biomass, rather than the attached biomass, in nitrification in the wastewater treatment process studied.

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문제 정의

An investigation of the diversity and compositions of nitrifying organisms is also required, which would be the basis for a better understanding of nitrification in the IFAS processes. Thus, the main objective of this study was to analyze nitrifying bacterial communities in a full-scale IFAS process and to characterize them by comparing with the nitrifying bacterial communities that are found in conventional activated sludge systems.

제안 방법

These modifications should be studied in the future. Furthermore, this study used sequences obtained from pyrosequencing instead of clone sequences for the phylogenetic study of nitrifying bacteria, and this involved skipping the cloning step and possibly omitted a bias conferred by cloning.
In order to investigate AOB communities in the Guri WWTP, AOB-like sequences were retrieved from total bacterial 16S rRNA gene sequences obtained in our previous study using a high-throughput pyrosequencing in the Guri WWTP [6]. Suspended and attached biomass samples were collected in June 2009 by grab sampling at three different points in the aerobic tank of the Guri WWTP and pooled together for the sequencing experiments, respectively.
This study reports the diversity and phylogenetic affiliation of AOB-like and NOB-like sequences as well as nitrification rates of the suspended and attached biomasses in a full-scale IFAS process. The information made it possible to deduce the relative importance of the two biomasses in the studied WWTP.

대상 데이터

The Guri wastewater treatment plant (WWTP) located in Guri, Gyeonggi-Do, Korea, was used in this study. The Guri WWTP has a capacity to treat 160,000 m3 of domestic wastewater per day and operates using an IFAS process.

성능/효과

2) demonstrated that all of the Nitrospira-like sequences were grouped in Cluster 1, which included Candidatus Nitrospira defluvii [18] and sequences retrieved from the activated sludge, biofilm, and anammox reactor. A total of 26 Nitrobacter-like sequences (3 in the suspended sample and 23 in the attached sample) were identified within genus Nitrobacter (50% confidence level) by the RDP Classifier, but all of the sequences were not closely affiliated with known Nitrobacter sequences phylogenetically, including N. hamburgensis, N. wibogradsky, N. alkalicus, and N. vulgaris (Supplemental Fig. 2). Although those sequences might be related to unknown Nitrobacter species, it appears that those sequences were originated from different species irrelevant to NOB, based on clustering distinct from the Nitrobacter cluster (bootstrap value distinguishing the cluster = 100%) and low sequence identities with pureculture Nitrobacter species (95-97%).
2). Although those sequences might be related to unknown Nitrobacter species, it appears that those sequences were originated from different species irrelevant to NOB, based on clustering distinct from the Nitrobacter cluster (bootstrap value distinguishing the cluster = 100%) and low sequence identities with pureculture Nitrobacter species (95-97%). Therefore, NOB within the genus Nitrobacter appear to be unimportant in the Guri WWTP.
After constructing a phylogenetic tree, sequences not affiliated with AOB were removed for further analysis. As a result, a total of 189 AOB-like sequences were retrieved out of 23,536 sequences (0.80%) in the suspended sample, whereas only 31 sequences out of 44,003 sequences (0.07%) showed affinity with AOB in the attached sample. The AOB sequences retrieved from the suspended sample were more diverse than those retrieved from the attached sample: for the suspended and attached samples, 26 and 3 unique operational taxonomic units (OTUs), respectively, were observed (3% cutoff) and 31 and 3 OTUs, respectively, were estimated by Chao 1 richness (3% cutoff), as determined by Mothur[16].
Guri WWTP sequences retrieved from the suspended biomass are indicated as Guri S, and those retrieved from the attached biomass are indicated as Guri A. Sequences showing >98% identities are indicated as a representative sequence.
1. Most of the sequences retrieved from the suspended sample (186 sequences) showed affinity with the Nitrosomonas oligotropha lineage, and only three sequences were within the N. marina lineage. N.

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