• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 2000.10a
• /
• pp.109-117
• /
• 2000

Bacterial community structure composing enhanced biological phosphorus removal (EBPR) activated sludge was analyzed phylogenetically by cloning 165 rDNA after direct DNA extraction. Then, this result was compared with 165 rDNA sequences of randomly isolated bacterial species. The results clearly showed that there are no coincidence between the sequences retrieved directly from activated sludge and those of isolated strains, suggesting that many important bacteria are hidden in activated sludge because of the difficulty in isolation and culture of them.

• Journal of Korean Society on Water Environment
• /
• v.27 no.6
• /
• pp.754-759
• /
• 2011

A sequencing batch reactor (SBR) was operated under different pH conditions to better understand the influence of pH to granulation in enhanced biological phosphorus removal systems. Granules from the SBR were also investigated using scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX). Considerable decreases in the amount of phosphorus released per substrate provided under anaerobic conditions and the content of biomass polyphosphate under aerobic conditions were observed when pH was changed from 7.5 to 7.0, followed by 6.5. Aerobic granulation was also observed at pH 7.0. A number of bacteria with the typical morphological traits of tetrad-forming organisms (TFOs) were observed at pH 7.0, including large members of cluster. Filamentous bacteria were also there in large numbers. The occurrence and growth of granules were further enhanced at pH 6.5. A SEM analysis showed that the aerobic granules had a compact microbial structure with shaperical shape and morphologically consisted of aggregates of small coccoid bacteria and filamentous bacteria encapsulated by extracellular polymeric substance. The main material ions identified by EDX moreover revealed that the structural materials for polyphosphate in the granules include phosphorus, potassium and calcium. Therefore, these results strongly suggested that PAOs are a dominant population in the microbial community of the aerobic granules.

• 한국생물공학회:학술대회논문집
• /
• 2000.11a
• /
• pp.77-80
• /
• 2000

Various analytical methods such as electron microscopy, quinone analysis, and 16S rDNA sequencing studies were used to investigate the microbial communities and to identify the microorganisms responsible for enhanced biological phosphorus removal (EBPR) in an anaerobic/aerobic sequencing batch reactor (SBR) fed with acetate. Electron photomicrographs showed that oval-shaped microorganisms of about $0.7\;{\sim}\;1\;{\mu}m$ in diameter dominated the microbial sludge. These microorganisms contained polyphosphate granules and glycogen inclusions, which suggests that they are a kind of phosphorus accumulating organism. Quinone and 16S rRNA sequence analyses showed that the members of Proteobacteria beta subclass were the most abundant species, which were affiliated with the Rhodocyclus-likes group. Phylogenetic analysis revealed that the two dominating clones of the beta subclass were most distantly related to Propionivibrio dicarboxylicus DSM 5885 and Rhodocyclus tenuis DSM 109 with about 95% and 96% sequence similarity, respectively. Therefore, it was concluded that the oval-shaped organisms related to the Rhodocyclus-likes group are likely to be responsible for biological phosphorus removal in SBR operation supplied with acetate.

• Journal of Korean Society of Water and Wastewater
• /
• v.34 no.6
• /
• pp.437-443
• /
• 2020

Biological phosphorus removal is accomplished by exposing PAO(phosphorus accumulating organisms) to anaerobic-aerobic conversion conditions. In the anaerobic condition, PAO synthesize PHB(polyhydroxybutyrate) and simultaneously hydrolysis of poly-p resulting phosphorus(Pi) release. In aerobic condition, PAO uptake phosphorus(Pi) more than they have released. In this study, cyanobacteria Synechococcus sp., which is known to be able to synthesize PHB like PAO, was exposed to anaerobic-aerobic conversion. If Synechococcus sp. can remove excess phosphorus by the same mechanism as PAO, synergistic effects can occur through photosynthesis. Moreover, Synechococcus sp. is known to be capable of synthesizing PHB using inorganic carbon as well as organic carbon, so even if the available capacity of organic carbon decreases, it was expected to show stable phosphorus removal efficiency. In 6 hours of anaerobic condition, phosphorus release occurred in both inorganic and organic carbon conditions but SPRR(specific phosphorus release rate) of both conditions was 10 mg-P/g-MLSS/day, which was significantly lower than that of PAO. When converting to aerobic conditions, SPUR(specific phosphorus uptake rate) was about 9 mg-P/g-MLSS/day in both conditions, showing a higher uptake rate than the control condition showing SPUR of 6.4 mg-P/g-MLSS/day. But there was no difference in terms of the total amount of removal. According to this study, at least, it seems to be inappropriate to apply Synechococcus sp. to luxury uptake process for phosphorus removal.

• Journal of Korean Society of Environmental Engineers
• /
• v.22 no.1
• /
• pp.141-148
• /
• 2000

In this study, effects of substrates and nitrate on biological phosphorus release in EBPR(enhanced biological phosphorus removal) process were examined using batch test apparatus at anaerobic conditions. The sludge used in this experiments was taken from SBR(sequencing batch reactor) treating swine wastewater at aeration period. Phosphorus release rates obtained with substrates of FSW(fermented swine wastewater), acetate, propionate, domestic wastewater and methanol were 6.19, 5.99, 1.52, 1.2 and $1.03mgP/gVSS{\cdot}hr$, respectively. Those observed with acetate and FSW were 4~5 times greater than those with propionate, methanol and domestic wastewater. Therefore phosphorus release rates were significantly affected by type of substrate added at anaerobic condition. Phosphorus release was greatly affected by concentration of nitrate in anoxic condition. Comparing to acetate, propionate and FSW, phosphorus release was observed after almost completely depletion of nitrate concentration with methanol and domestic wastewater added as substrate. In the cases supplied with acetate, propionate and FSW, phosphorus release rates were less influenced by a nitrate concentration than those with methanol and domestic wastewater.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.7
• /
• pp.1290-1297
• /
• 2008

To investigate the diversities of Accumulibacter phosphatis and its polyhydroxyalkanoate (PHA) synthase gene (phaC) in enhanced biological phosphorus removal (EBPR) sludge, an acetate-fed sequencing batch reactor was operated. Analysis of microbial communities using fluorescence in situ hybridization and 16S rRNA gene clone libraries showed that the population of Accumulibacter phosphatis in the EBPR sludge comprised more than 50% of total bacteria, and was clearly divided into two subgroups with about 97.5% sequence identity of the 16S rRNA genes. PAO phaC primers targeting the phaC genes of Accumulibacter phosphatis were designed and applied to retrieve fragments of putative phaC homologs of Accumulibacter phosphatis from EBPR sludge. PAO phaC primers targeting $G_{1PAO},\;G_{2PAO},\;and\;G_{3PAO}$ groups produced PCR amplicons successfully; the resulting sequences of the phaC gene homologs were diverse, and were distantly related to metagenomic phaC sequences of Accumulibacter phosphatis with 75-98% DNA sequence identities. Degenerate NPAO (non-PAO) phaC primers targeting phaC genes of non-Accumulibacter phosphatis bacteria were also designed and applied to the EBPR sludge. Twenty-four phaC homologs retrieved from NPAO phaC primers were different from the phaC gene homologs derived from Accumulibacter phosphatis, which suggests that the PAO phaC primers were specific for the amplification of phaC gene homologs of Accumulibacter phosphatis, and the putative phaC gene homologs by PAO phaC primers were derived from Accumulibacter phosphatis in the EBPR sludge. Among 24 phaC homologs, a phaC homolog (GINPAO-2), which was dominant in the NPAO phaC clone library, showed the strongest signal in slot hybridization and shared approximately 60% nucleotide identity with the $G_{4PAO}$ group of Accumulibacter phosphatis, which suggests that GINPAO-2 might be derived from Accumulibacter phosphatis. In conclusion, analyses of the 16S rRNA and phaC genes showed that Accumulibacter phosphatis might be phylogenetically and metabolically diverse.

• Journal of Korean Society of Environmental Engineers
• /
• v.22 no.5
• /
• pp.939-947
• /
• 2000

Enhanced biological phosphorus removal (EBPR) was performed in an anaerobic/aerobic sequencing batch reactor (SBR). Influent was a synthetic wastewater based on acetate as a carbon source. The sludge age and hydraulic retention time were kept at 10 days and 16 hrs, respectively, Phosphate release during the anaerobic period and phosphate uptake in aerobic period were increased gradually with time. and after about 200 days, steady-state operation could be achieved with complete removal of influent phosphate. Number distribution of microbial community in the sludge performing EBPR was investigated during the steady state operation. 17 rRNA targeted oligonucleotide probes were designed and slot hybridization technique was used to determine the number distribution of each microorganism. In the acetate fed SBR, rRNA belonging to the beta subclass of proteobacteria was the most dominant in total rRNA and rRNA matching to CTE probe was the second, rRNAs of Acinetobacter, Aeromonas and Pseudomonas, which are usually thought as phosphorus accumulating organisms in EBPR processes, constituted less than 10% of total rRNA. From this community analysis, it was inferred that microorganisms belong to the beta subclass of proteobacteia (BET) and CTE such as Rhodocyclus group were important in biological phosphorus removal. Therefore, the role of Acinetobacter, Aeromonas and Pseudomonas in the EBPR might have been overestimated.

• Korean Journal of Environmental Agriculture
• /
• v.29 no.2
• /
• pp.189-196
• /
• 2010

The removal of phosphorus (P) in the wastewater is essential for the prevention of eutrophication in the river and stream. This study was initiated to evaluate the effect of the various heavy metals on the growth and P removal capacity of Pseudomonas sp., which was well known as phosphorus accumulating microorganism(PAO's) in the EBPR(Enhanced Biological Phosphorus Removal) process. The five heavy metals used in the study were Cu, As, Zn, Ni, and Cd. The growth rate of Pseudomonas sp. was the greatest at $25^{\circ}C$, but the removal efficiency of P was the highest at $30^{\circ}C$. The $IC_{50}$ (median Inhibition Concentration) values of Pseudomonas sp. for the Cu, As, Zn, Ni, and Cd were 2.35, 11.04, 1.80, 4.92, and 0.24 mg/L, respectively. Therefore, it appears that the sensitivity of the heavy metals to Pseudomonas sp. was in the following order: Cd> Zn> Cu> Ni> AS. Also, the P removal efficiencies by Pseudomonas sp. were correspondingly decreased as the concentrations of heavy metals were increased.

• Journal of Korean Society of Environmental Engineers
• /
• v.29 no.2
• /
• pp.197-204
• /
• 2007

The reciprocal effects towards the enhanced biological phosphorus removal were performed for anaerobic, aerobic and anoxic phases. The batch experiments showed that the p-absorption in the anoxic phase was 50% lower than aerobic phase. The correlation coefficient between p-back-solution and p-absorption was found to be $R^2=0.557$ however, the coefficient b(b = 8.4049) was relatively higher than the other researchers results. The increase and/or acceptance of the $K^+-,\;Mg^{2+}-$ and $NH_4-N$-concentration was proportional to those of the $PO_4-P$-concentration in the batch test. The relationship between $K^+-,\;Mg^{2+}$ and $PO_4-P$ was determined. The average value of this relation-ship agreed with 0.2 mol $K^+Ion$ / mol $PO_4-P$ ion and 0.21 mol $Mg^{2+}Ion$ / moi $PO_4-P$ ion in the anaerobic phase. The absorbed ratio of $K^+$ to $Mg^{2+}$ over $PO_4-P$ was found to be 1 : 5.

• Journal of Korean Society on Water Environment
• /
• v.27 no.5
• /
• pp.646-653
• /
• 2011

The combined system of sequencing batch biofilm reactor (SBBR) and membrane SBR (MSBR) was operated with sewage to evaluate the COD utilization for biological nutrient removal (BNR). The SBBR was operated for nitrification reactor, while denitrifying PAO (dPAO) was cultivated in MSBR with anaerobic-anoxic operation. In the SBBR and MSBR system, the enhanced biological phosphorus removal (EBPR) was successfully achieved with higher N removal. The COD utilization in combined SBBR-MSBR system was significantly reduced compared to ordinary BNR (up to 3.1 g SCOD/g (N+P) and 1.6 g SCOD/g (N+P) with different C/N/P ratio). The results suggest that a dPAO process could effectively reduce carbon energy (=COD) requirement. The combination of oxic-SBBR and anaerobic-anoxic MSBR for dPAO utilization could be an attractive alternative to upgrade the process performance in weak sewage.