Isolation and characterization of a Gram-positive polyphosphate-accumulating bacterium.

Onda, Shin; Takii, Susumu

doi:10.2323/jgam.48.125

Cited by 18 publications

(14 citation statements)

References 25 publications

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“…Some Actinobacteria have been suggested to be potential PAO, based on laboratory-scale reactor studies and on physiological investigations of pure cultures (11,19,29), but this is the first time they have been identified and found in significant amounts and exhibiting PAO behavior in full-scale EBPR systems. The polyphosphate-accumulating capability was evaluated with MAR-FISH and FISH combined with Neisser staining, and the APAO found in this study demonstrated the ability to take up P i and accumulate poly(P) using oxygen or nitrate as an electron acceptor after anaerobic uptake of organic substrates.…”

Section: Discussionmentioning

confidence: 99%

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Identity and Ecophysiology of Uncultured Actinobacterial Polyphosphate-Accumulating Organisms in Full-Scale Enhanced Biological Phosphorus Removal Plants

Kong

Nielsen

2005

Appl Environ Microbiol

249

238

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Microautoradiography combined with fluorescence in situ hybridization (MAR-FISH) was used to screen for potential polyphosphate-accumulating organisms (PAO) in a full-scale enhanced biological phosphorus removal (EBPR) plant. The results showed that, in addition to uncultured Rhodocyclus-related PAO, two morphotypes hybridizing with gene probes for the gram-positive Actinobacteria were also actively involved in uptake of orthophosphate (P i ). Clone library analysis and further investigations by MAR-FISH using two new oligonucleotide probes revealed that both morphotypes, cocci in clusters of tetrads and short rods in clumps, were relatively closely related to the genus Tetrasphaera within the family Intrasporangiaceae of the Actinobacteria (93 to 98% similarity in their 16S rRNA genes). FISH analysis of the community biomass in the treatment plant investigated showed that the short rods (targeted by probe Actino-658) were the most abundant (12% of all Bacteria hybridizing with general bacterial probes), while the cocci in tetrads (targeted by probe Actino-221) made up 7%. Both morphotypes took up P i aerobically only if, in a previous anaerobic phase, they had taken up organic matter from wastewater or a mixture of amino acids. They could not take up short-chain fatty acids (e.g., acetate), glucose, or ethanol under anaerobic or aerobic conditions. The storage compound produced during the anaerobic period was not polyhydroxyalkanoates, as for Rhodocyclus-related PAO, and its identity is still unknown. Growth and uptake of P i took place in the presence of oxygen and nitrate but not nitrite, indicating a lack of denitrifying ability. A survey of the occurrence of these actinobacterial PAO in 10 full-scale EBPR plants revealed that both morphotypes were widely present, and in several plants more abundant than the Rhodocyclus-related PAO, thus playing a very important role in the EBPR process.In the wastewater treatment industry, enhanced biological phosphorus removal (EBPR) has been widely used to remove orthophosphate (P i ) from wastewater to protect the receiving water bodies against eutrophication. In EBPR processes, the principle is to enrich microorganisms that can accumulate excessive amounts of intracellular polyphosphate [poly(P)] by using sequential anaerobic-aerobic and/or anaerobic-denitrifying conditions (37). The understanding of the biochemical pathways of these poly(P)-accumulating organisms (PAO) is based mainly on analysis of chemical transformations in enriched laboratory-scale EBPR systems, because no pure cultures are available (25, 34, 37). The generally accepted hypothesis (25, 34) proposes that PAO take up organic matter (usually assumed to be acetate) during the anaerobic period by using poly(P) as an energy source and sequester the acetate taken up into polyhydroxyalkanoates (PHA). The reducing power to form PHA is provided by hydrolysis of an intracellular glycogen pool through the glycolytic pathway. The PHA are used as energy and carbon sources in the following aerobic or denitrifying...

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Section: Discussionmentioning

confidence: 99%

“…These bacteria include Microlunatus phosphovorus (32) and Tetrasphaera elongata (strain ASP12) (29). However, their aerobic P i uptake depends on their anaerobic uptake of glucose (for M. phosphovorus) or an amino acid mixture (Casamino Acids) (for T. elongata) instead of short-chain fatty acids, as for the RPAO.…”

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confidence: 99%

Identity and Ecophysiology of Uncultured Actinobacterial Polyphosphate-Accumulating Organisms in Full-Scale Enhanced Biological Phosphorus Removal Plants

Kong

Nielsen

2005

Appl Environ Microbiol

249

238

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“…That for the quantification of APAOs was designed to be specific for the sequences of some Tetrasphaera spp. 6,17,22) and the Tetrasphaera-related clones 9) (Fig. 1B).…”

Section: Primer Specificitymentioning

confidence: 99%

“…Because PAOs contain a large amount of phosphorus as polyphosphate, they are considered to play an important role in EBPR processes 18) . There have been many attempts to isolate major PAOs in order to understand the population of PAOs and optimize EBPR processes 6,17,20,22) . However, there have been no reports on the isolation of the major PAOs in EBPR sludge, and little is known about the microbiology of major PAOs using culture-dependent methods.…”

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confidence: 99%

Quantification of Rhodocyclus-Related and Actinobacterial Polyphosphate-Accumulating Organisms in an Enhanced Biological Phosphorus Removal Process Using Quenching Probe PCR

et al. 2007

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Quenching probe (QProbe) PCR-based methods for quantifying the abundance of Rhodocyclus-related and actinobacterial polyphosphate-accumulating organisms (PAOs) in an enhanced biological phosphorus removal (EBPR) process are described. PCR primer-QProbe sets specific for the 16S rRNA genes of Rhodocyclus-related PAOs (RPAOs) and actinobacterial PAOs (APAOs) were newly designed, and their specificity tested using database searches, a selection of pure cultures, and sludge DNA. The sets were found to be specific for most of the target sequences. QProbe PCR with the newly designed sets was used to enumerate the 16S rRNA genes of RPAOs and APAOs in a laboratory-scale EBPR sludge. The copy number of 16S rRNA genes of RPAOs was larger than that of APAOs before the phosphorus removal performance of the reactor deteriorated, and the amount of both PAOs decreased in the deterioration period. The approaches developed in this study may become a powerful tool for a high-throughput analysis of PAO abundance in EBPR processes.

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“…Currently, the genus Tetrasphaera consists of six species cultured from activated sludge; T. australiensis (strains Ben 109 and Ben 110), T. japonica (strain T1-X7) (Maszenan et al, 2000), T. elongata (strain Lp2) (Hanada et al, 2002), T. elongata (strain ASP12) (Onda and Takii, 2002), T. jenkinsii, T. veronensis and T. vanveenii (McKenzie et al, 2006), but no genome sequence data exist for any of these isolates. As these organisms seem to have an important role in phosphorus removal in EBPR plants (Kong et al, 2005;Nguyen et al, 2011), a metabolic model for Tetrasphaera is needed to clarify what their function in these systems is, and how their ecophysiology might differ from that of Accumulibacter.…”

Section: Introductionmentioning

confidence: 99%

A metabolic model for members of the genus Tetrasphaera involved in enhanced biological phosphorus removal

Kristiansen

Nguyen²,

Saunders³

et al. 2012

The ISME Journal

202

154

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Members of the genus Tetrasphaera are considered to be putative polyphosphate accumulating organisms (PAOs) in enhanced biological phosphorus removal (EBPR) from wastewater. Although abundant in Danish full-scale wastewater EBPR plants, how similar their ecophysiology is to 'Candidatus Accumulibacter phosphatis' is unclear, although they may occupy different ecological niches in EBPR communities. The genomes of four Tetrasphaera isolates (T. australiensis, T. japonica, T. elongata and T. jenkinsii) were sequenced and annotated, and the data used to construct metabolic models. These models incorporate central aspects of carbon and phosphorus metabolism critical to understanding their behavior under the alternating anaerobic/aerobic conditions encountered in EBPR systems. Key features of these metabolic pathways were investigated in pure cultures, although poor growth limited their analyses to T. japonica and T. elongata. Based on the models, we propose that under anaerobic conditions the Tetrasphaerarelated PAOs take up glucose and ferment this to succinate and other components. They also synthesize glycogen as a storage polymer, using energy generated from the degradation of stored polyphosphate and substrate fermentation. During the aerobic phase, the stored glycogen is catabolized to provide energy for growth and to replenish the intracellular polyphosphate reserves needed for subsequent anaerobic metabolism. They are also able to denitrify. This physiology is markedly different to that displayed by 'Candidatus Accumulibacter phosphatis', and reveals Tetrasphaera populations to be unusual and physiologically versatile PAOs carrying out denitrification, fermentation and polyphosphate accumulation.

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Isolation and characterization of a Gram-positive polyphosphate-accumulating bacterium.

Cited by 18 publications

References 25 publications

Identity and Ecophysiology of Uncultured Actinobacterial Polyphosphate-Accumulating Organisms in Full-Scale Enhanced Biological Phosphorus Removal Plants

Identity and Ecophysiology of Uncultured Actinobacterial Polyphosphate-Accumulating Organisms in Full-Scale Enhanced Biological Phosphorus Removal Plants

Quantification of Rhodocyclus-Related and Actinobacterial Polyphosphate-Accumulating Organisms in an Enhanced Biological Phosphorus Removal Process Using Quenching Probe PCR

A metabolic model for members of the genus Tetrasphaera involved in enhanced biological phosphorus removal

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