Microbial Ecology of Nitrifying Bacteria in Wastewater Treatment Process Examined by Fluorescence In Situ Hybridization.

AOI, YOSHITERU; Miyoshi, Tomoko; Okamoto, T.; Tsuneda, Satoshi; Hirata, A.; Kitayama, Atsushi; Nagamune, Teruyuki

doi:10.1263/jbb.90.234

Cited by 13 publications

(8 citation statements)

References 19 publications

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“…The species and characteristics of AOB were found to be strongly affected by the reactor operational conditions such as substrate composition and concentration in a WWTP. The results indicated that effective control of the microbial ecology by controlling the conditions of the reactor operation could be applied to engineered systems with direct monitoring using FISH (Aoi et al 2000). During experiments carried out with freshwater aquaria, FISH analysis revealed that Nitrosomonas marina-like AOB may outcompete Nitrosomonas tenius-like AOB in the low-ammonia concentration environment of an aquarium, whereas Nitrosomonas europea-like AOB were found only in reactors with a history of high ammonia concentrations (Burrell et al 2001).…”

Section: Fluorescence In Situ Hybridization Techniquementioning

confidence: 99%

Partial nitrification—operational parameters and microorganisms involved

Sinha

Annachhatre

2006

Rev Environ Sci Biotechnol

163

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Nitrite is a common intermediate in at least three different oxidative or reductive biochemical pathways that occur in nature (nitrification, denitrification and dissimilatory or assimilatory nitrate reduction). Nitrite accumulation or partial nitrification has been reported in literature for decades. In engineered systems, partial nitrification is of interest as it offers cost savings in aeration as well as in the form of lesser need for addition of organic carbon as compared to the conventional denitrification. A broad range of operating parameters and factors has been reviewed in this paper which are essential for achieving partial nitrification. Of these, pH, dissolved oxygen (DO), temperature, free ammonia (FA) and nitrous acid concentrations, inhibitory compounds are important factors in achieving partial nitrification.Two groups of bacteria, namely ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) are involved in nitrification. Chemolitho-autotrophic AOB are responsible for the rate-limiting step of nitrification in a wide variety of environments, making them important in the global cycling of nitrogen. Characterization

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Section: Fluorescence In Situ Hybridization Techniquementioning

confidence: 99%

Partial nitrification—operational parameters and microorganisms involved

Sinha

Annachhatre

2006

Rev Environ Sci Biotechnol

163

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“…Nitrosomonas traditionally have been considered the most important bacteria for NH 3 oxidation [23]. Furthermore, Nitrosospira and Nitrosomonas have both been recently found in activated sludge [1], freshwater lakes [30] and estuaries [5]. There is culturebased evidence that Nitrosospira strains may be more common in soils [2].…”

Section: Introductionmentioning

confidence: 99%

Diversity and abundance of ammonia-oxidizing bacteria in eutrophic and oligotrophic basins of a shallow Chinese lake (Lake Donghu)

Chen

Qiu

Zhou

2009

Research in Microbiology

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“…The probe concentration was 0.5 ng/l. Hybridization was followed by a stringent washing step at 48°C for 20 min in a washing buffer containing Tris-HCl (20 mM, pH 7.4), NaCl (35 mM) (5,16), and sodium dodecyl sulfate. The washing buffer was removed by rinsing the slides with ddH 2 O.…”

Section: Methodsmentioning

confidence: 99%

“…In particular, in situ hybridization with fluorescencelabeled oligonucleotide probes has been widely used for in situ analysis of microbial communities, such as a biofilm in a wastewater treatment process (5,18,22,29). This method relies on the presence of many target sequences within an individual cell.…”

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confidence: 99%

Direct Detection by In Situ PCR of the amoA Gene in Biofilm Resulting from a Nitrogen Removal Process

Hoshino

Noda

Tsuneda

et al. 2001

Appl Environ Microbiol

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Ammonia oxidation is a rate-limiting step in the biological removal of nitrogen from wastewater. Analysis of microbial communities possessing the amoA gene, which is a small subunit of the gene encoding ammonia monooxygenase, is important for controlling nitrogen removal. In this study, the amoA gene present in Nitrosomonas europaea cells in a pure culture and biofilms in a nitrifying reactor was amplified by in situ PCR. In this procedure, fixed cells were permeabilized with lysozyme and subjected to seminested PCR with a digoxigenin-labeled primer. Then, the amplicon was detected with an alkaline phosphatase-labeled antidigoxigenin antibody and HNPP (2-hydroxy-3-naphthoic acid-2-phenylanilide phosphate), which was combined with Fast Red TR, and with an Alexa Fluor 488-labeled antidigoxigenin antibody. The amoA gene in the biofilms was detected with an unavoidable nonspecific signal when the former method was used for detection. On the other hand, the amoA gene in the biofilms was detected without a nonspecific signal, and the cells possessing the amoA gene were clearly observed near the surface of the biofilm when Alexa Fluor 488-labeled antidigoxigenin antibody was used for detection. Although functional gene expression was not detected in this study, detection of cells in a biofilm based on their function was demonstrated.The biological removal of nitrogen compounds is an integral part of most modern wastewater treatment facilities to preserve environmental water resources. In this process, ammonia oxidation is a rate-limiting step, where autotrophic ammonia conversion into hydroxylamine is catalyzed by ammonia monooxygenase. Thus, analysis of microbial communities possessing the amoA gene, which encodes ammonia monooxygenase, is important for controlling nitrogen removal.On the other hand, fluorescent in situ hybridization (FISH) (4, 9) and denaturing gradient gel electrophoresis (10, 17) based on 16S ribosomal DNA and rRNA for molecular analysis have been used in various fields to determine the genetic diversity of a microbial community and to identify individual members. In particular, in situ hybridization with fluorescencelabeled oligonucleotide probes has been widely used for in situ analysis of microbial communities, such as a biofilm in a wastewater treatment process (5,18,22,29). This method relies on the presence of many target sequences within an individual cell. Therefore, bacterial cells containing insufficient rRNA cannot be detected by this approach. Moreover, this taxonomic identification approach cannot be used to detect the presence of single-copy functional genes or their expression at the single-cell level. Hence, in situ hybridization cannot estimate a specific metabolic activity such as ammonia oxidation.Recently, in situ PCR was developed to amplify and detect functional genes and their expression inside a single cell, thus making it possible to detect a single copy of a functional gene. This method was first developed to amplify and detect a DNA virus inside a cell (11), and Nuovo et ...

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Microbial Ecology of Nitrifying Bacteria in Wastewater Treatment Process Examined by Fluorescence In Situ Hybridization.

Cited by 13 publications

References 19 publications

Partial nitrification—operational parameters and microorganisms involved

Partial nitrification—operational parameters and microorganisms involved

Diversity and abundance of ammonia-oxidizing bacteria in eutrophic and oligotrophic basins of a shallow Chinese lake (Lake Donghu)

Direct Detection by In Situ PCR of the amoA Gene in Biofilm Resulting from a Nitrogen Removal Process

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