Biomedical subjects

T Mino

Publications and source records attributed to T Mino.

At least 19 recordsLinked to original sources

PubMed2006

Isolation, characterization of bacteriophages specific to Microlunatus phosphovorus and their application for rapid host detection.

AIMS: To isolate and characterize lytic-bacteriophages specific to Microlunatus phosphovorus, and prepare fluorescently labelled phages (FLPs) for the rapid detection of the host bacterium in activated sludge. METHODS AND RESULTS: Isolation of bacteriophages lytic to M. phosphovorus was attempted by applying supernatants of activated sludge processes on the lawn of M. phosphovorus JCM9379 for plaque formation. Thirteen bacteriophage isolates were obtained. The restriction fragment length polymorphism analysis distinguished them into two different bacteriophages designated as phiMP1 and phiMP2. They were found to possess double-stranded DNA and host specificity. Morphological observations were done by electron microscopy. The bacteriophage particles stained by SYBR Green I was shown to be applicable to detect their host bacterial cells mixed with activated sludge. CONCLUSIONS: Two M. phosphovorus-specific bacteriophages were isolated and classified as Siphoviridae. FLPs of them were prepared, and successfully applied to detect the host bacterium added into the activated sludge. SIGNIFICANCE AND IMPACT OF THE STUDY: At least some of bacteria in activated sludge are susceptible to their related bacteriophages. Bacteriophages lytic to activated sludge bacteria could be affecting the bacterial population in activated sludge. The FLPs could be used for the easy-rapid detection of their host bacterium in activated sludge.

Bacteriolysis↗

PubMed2006

Sludge-sludge interaction in the enhanced biological phosphorus removal process.

Metabolisms related to enhanced biological phosphorus removal (EBPR) were found to be affected when two activated sludges with different EBPR activities were mixed together. In the present study, two laboratory scale EBPR processes were operated in parallel, one of them with higher and another with lower EBPR activities. The activated sludges from the two reactors were mixed together at different mixing ratios. The supernatant was made the same for all mixing ratios, anaerobic-aerobic batch experiments were performed, and acetate uptake rate and phosphate release rate under anaerobic conditions and phosphate uptake rate under aerobic condition were determined. The metabolic rates measured were expected to be linear to the mixing ratios, as the supernatant was the same for all mixing ratios, whereas the metabolic rates were either promoted or inhibited by mixing of sludges. As an indicator for the sludge mixing effect on the metabolic rates, mixing effect intensity (MEI) was introduced. Chemical substances that are produced by microorganisms in activated sludge are proposed to be one of the possible causes of the sludge mixing effect.

Anaerobiosis↗

PubMed2006

Examining substrate uptake patterns of Rhodocyclus-related PAO in full-scale EBPR plants by using the MAR-FISH technique.

While recognised as the important population responsible for enhanced biological phosphorus removal (EBPR), detailed knowledge on the physiology of Rhodocyclus-related polyphosphate accumulating organisms (PAO) has yet to be grasped. The objective of this study was to examine the in situ substrate uptake patterns of Rhodocyclus-related PAO present in full-scale EBPR plants by the combined technique of microautoradiography-fluorescent in situ hybridization (MAR-FISH). The presence of these PAO in the four investigated plants was confirmed by FISH and they constituted 17%, 9%, 8%, and 7% of the sludge community. By using MAR-FISH technique, Rhodocyclus-related PAO in all the plants demonstrated similar anaerobic substrate uptake patterns. They were capable of assimilating acetate, aspartate and glutamate under anaerobic condition but they showed negative uptake with palmitate. A significant fraction of the MAR-positive cells assimilated acetate, aspartate or glutamate was found to be Rhodocyclus-related PAO. Dual staining with DAPI and FISH showed that these PAO also accumulated polyphosphate aerobically with aspartate and glutamate as carbon source. The ability of assimilating amino acids besides acetate strongly indicates the versatile physiology of Rhodocyclus-related PAO, which could benefit them to achieve predominance in EBPR activated sludge.

Environmental Restoration and Remediation↗

PubMed2006

Microbial community of biological phosphorus removal process fed with municipal wastewater under different electron acceptor conditions.

The microbial community in a biological phosphorus removal process under different electron acceptor conditions was estimated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) assay and principal-component analysis (PCA). For this purpose, a lab-scale sequencing batch reactor (SBR) fed with municipal wastewater was operated under anaerobic-aerobic, anaerobic-anoxic-aerobic and anaerobic-anoxic conditions. The results of PCR-DGGE targeting the 16S rRNA gene indicated a significant shift in the microbial community with electron acceptor conditions. From the 16S rRNA-based PCA, the microbial shift implies that little oxygen supply caused the deterioration of aerobic bacteria, including aerobic polyphosphate-accumulating organisms (PAOs). Moreover, it also reflects the existence of nitrate-utilizing denitrifiers. On the other hand, although the band patterns of DGGE targeting a functional gene of denitrification (nirS) also showed the microbial shift, the result of PCA differed from that of 16S rRNA-based analysis. There is no conclusive proof that the bacteria represented as the dominant bands detected in the present study are denitrifying-PAOs so far, it should be worthwhile to identify the detected bacteria and to examine their traits as new denitrifying-PAO candidates.

Bacteria↗

PubMed2006

Influence of flocculation and settling properties of activated sludge in relation to secondary settler performance.

Floc characteristics were studied at a full scale activated sludge treatment plant with a unique process solution incorporating pre-denitrification with post-nitrification in nitrifying trickling filters. Since greater nitrogen removal is achieved when more secondary settled wastewater is recirculated to the trickling filters, the secondary settlers are always operated close to their maximal capacity. The flocculation and settling properties are therefore crucial and have an effect on the overall plant performance. Since the plant is operated at a short sludge age, these properties change quickly, resulting in variable maximal secondary settler capacity. The dynamics in floc structure and microbial community composition were studied and correlated to the secondary settler performance. Fluorescence in situ hybridisation was used to investigate the microbial community structure and their spatial distribution. The floc structure could to some extent be related to the flocculation and settling properties of the sludge. Even small differences had an influence suggesting that colloidal properties also play a significant role in determining the floc properties. No correlation between microbial community composition and settling properties could be established with the group-specific probes investigated.

Environmental Restoration and Remediation↗

PubMed2006

Rain events and their effect on effluent quality studied at a full scale activated sludge treatment plant.

The effect of rain events on effluent quality dynamics was studied at a full scale activated sludge wastewater treatment plant which has a process solution incorporating pre-denitrification in activated sludge with post-nitrification in trickling filters. The incoming wastewater flow varies significantly due to a combined sewer system. Changed flow conditions have an impact on the whole treatment process since the recirculation to the trickling filters is set by the hydraulic limitations of the secondary settlers. Apart from causing different hydraulic conditions in the plant, increased flow due to rain or snow-melting, changes the properties of the incoming wastewater which affects process performance and effluent quality, especially the particle removal efficiency. A comprehensive set of on-line and laboratory data were collected and analysed to assess the impact of rain events on the plant performance.

Nitrogen↗

PubMed2006

Filter clogging in coarse pore filtration activated sludge process under high MLSS concentration.

Coarse pore filtration activated sludge process is a type of hybrid process in which the secondary settling tank of the conventional activated sludge process is replaced by non- woven and coarse pore filter modules. The filter has pores, which are irregular in shape, and much bigger than micro-filtration membrane pores in size. The objective of the study is to find out the effect of the microbial community structure on filter clogging in the coarse pore filtration activated sludge process under high MLSS concentration in aerobic and anoxic/aerobic (A/A) conditions. Filter clogging started from day 65 and 70 in the A/A and aerobic process, respectively, but it was more severe in the A/A process compared to that in the aerobic process. EPS contents of sludge did not change significantly during the operation in both processes, and did not have a crucial effect on the observed filter clogging. There was no strong evidence for direct effect of the type and number of metazoa on filter clogging. The main difference between aerobic sludge and A/A sludge during the filter clogging period was the relative abundance of filamentous bacteria. According to the obtained results, it can be concluded that a higher presence of filamentous bacteria could reduce the severity of filter clogging in a coarse pore filtration activated sludge process.

Bacteria, Aerobic↗

PubMed2004

The large PAO cells in full-scale EBPR biomass samples are not yeast spores but possibly novel members of the beta-Proteobacteria.

Large, homogenous clusters of coccobacilli were found to be abundant in the biomasses from a conventional plant at Rosebud, Victoria, Australia. The identity and the in situ physiology of these dominant microorganisms were investigated in this study. These large clustered cells were revealed to be neither Gram positive nor Gram negative bacteria and contain polyP granules. Cells with similar features were also observed in some enhanced biological phosphate removal (EBPR) systems and reported as yeast spores and Rhodocyclus-related polyphosphate accumulating organisms (PAOs). In this study, fluorescent in situ hybridization (FISH) probing showed these cells were prokaryotic and members of the beta-Proteobacteria. However, these large clustered cells did not respond to the PAO mix FISH probes. The in situ physiology of these large cells was studied with FISH in combination with microautoradiography (MAR) in order to understand their substrate assimilation abilities under different conditions as well as their phosphate uptake ability. These cells were able to take up acetate, glutamate and aspartate, but not glucose under both aerobic and anaerobic conditions. Nile Blue A staining in combination with MAR showed that cells incubated under anaerobic conditions contained polyhydroxyalkanoates (PHA) granules. In addition, MAR showed aerobic 33Pi assimilation with all these substrates, consistent with them supporting an EBPR capacity in these large cells. As well as raising doubts about a role for yeasts in EBPR, this study suggests that much still needs to be learned about the identity and level of biodiversity of the PAO in EBPR systems, and emphasizes the benefits of using techniques like FISH/MAR and PHA staining/MAR to resolve the in situ physiology of the populations of interest there.

Australia↗

PubMed2003

Resting T cells negatively regulate osteoclast generation from peripheral blood monocytes.

There is accumulating evidence that T cells may be involved in osteoclastogenesis in a variety of murine systems. However, the precise role of human T cells in the regulation of osteoclast generation is still unclear. To address this issue, we investigated the effect of resting peripheral T cells on receptor activator of NF-kappaB ligand (RANKL)-induced osteoclast generation from human peripheral monocytes. Although osteoclasts were not generated in the culture of human peripheral blood mononuclear cells (PBMC) in the presence of RANKL and macrophage colony-stimulating factor (M-CSF), the addition of cyclosporine A (CsA), a potent inhibitor of T-cell function, resulted in the formation of an increasing number of lacunae resorption on dentine, suggesting T cells may inhibit osteoclast formation. In a coculture of T cells and monocytes, which were isolated from PBMC, T cells inhibited the osteoclast generation from monocytes, as determined by tartrate-resistant acid phosphatase (TRAP) staining and a pit assay using dentine. This inhibition of osteoclast generation by T cells was also observed in a culture of the parathyroid hormone-stimulated SaOS4/3 osteoblast cell line and monocytes. The culture in Transwell plates revealed that the cell-to-cell interaction was not required for the inhibition, suggesting that T-cell cytokines may be responsible for the inhibition. Among inhibitory T-cell cytokines on osteoclastogenesis, granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (IFN-gamma) were actively produced by CD4 T cells but not CD8 T cells in the coculture of T cells with monocytes, and the neutralizing antibodies to these cytokines partially rescued the T-cell-induced inhibition of osteoclast formation. Although CsA did not affect RANKL-induced osteoclast generation in the culture of monocytes alone, it completely rescued the T-cell-induced inhibition of osteoclast formation and strongly inhibited the production of GM-CSF and IFN-gamma. Thus, we demonstrate that resting T cells negatively regulate the osteoclast generation via production of GM-CSF and IFN-gamma by CD4 T cells and that CsA stimulates the osteoclast generation through the inhibition of the production of these cytokines. These findings provide new insight into therapeutic strategies for immunosuppression-induced bone loss in transplant and other diseases.

Bone Remodeling↗

PubMed2003

A quantitative method for measuring the mass concentration of the filamentous bacterium Type 021N in activated sludge using fluorescence in situ hybridization.

AIMS: This study aimed to develop a quantitative method for measuring mass concentrations of Type 021N, a bacterium causing bulking in activated sludge. METHODS AND RESULTS: Fluorescence in situ hybridization was used to determine the relationship between the concentration ratio of the mass of the bacterium Type 021N to mass of activated sludge, and the proportion of fluorescence area imparted by probe G123T specific for Type 021N to that obtained with probe EUB338 for bacteria. A linear relationship existed between the cube root of the mass concentration ratio and square root of this area proportion. CONCLUSIONS: A standard curve was obtained for quantifying Type 021N in activated sludge. SIGNIFICANCE AND IMPACT OF THE STUDY: This method may allow the determination of growth rate constant of filamentous bacteria in activated sludge, information that will help in understanding their ecology.

Bacteria↗

PubMed2003

Quantitative estimation of the role of denitrifying phosphate accumulating organisms in nutrient removal.

It has been reported that a proportion of polyphosphate-accumulating organisms (PAOs) can denitrify or utilize nitrate as an electron acceptor. The usage of denitrifying-PAO (DN-PAO) can relieve the competition for COD between denitrification and phosphorus removal because they can treat nitrate and phosphate by using the same carbon source. To effectively use DN-PAO for biological nutrient removal (BNR), a new system was proposed in which an anaerobic phase is placed at the influent end, followed by he anoxic and external nitrification phase. In this study, the lab-scale proposed system (A2N system) was operated with a municipal wastewater 1) to confirm stable settlement of DN-PAO in the proposed system, 2) to quantitatively estimate the fraction of different groups of organisms like denitrifiers without polyphosphate accumulating capability, aerobic-PAO and DN-PAO and 3) to estimate the advantages of DN-PAO's presence in the system. Moreover, batch experiments in which anoxic and aerobic phosphate uptake rates (PUR) were measured were also carried out. The activity of DN-PAO was observed throughout the experimental period by the batch experiment. From the results of the calculation of COD utilized by each group of organisms, it was concluded that the proposed system could accumulate much more PAO (as DN-PAO) than conventional BNR systems. Moreover, they were responsible for both EBPR and denitrification.

Bacteria, Anaerobic↗

PubMed2003

Performance and microbial dynamics in the coarse pore filtration activated sludge process at different SRTs (solids retention times).

In this research, three SRTs (about 10, 30 and 75 days (without wasting the sludge except for sampling)) were applied to three reactors equipped with non-woven and coarse pore filter modules. The flux was adjusted to about 1 m/d during operation. The main objective of the study was to compare the performance and microbial population dynamics under different SRTs in this process. The results of reactors with SRTs of about 10 and 30 days have shown very good effluent quality without any clogging problem for more than 4 months of operation. For the reactor with long SRT (75 days), the filter clogging was observed after about 80 days of operation and caused an increase in the operation pressure and deterioration effluent quality on some days. Excessive abundance of filamentous bacteria was observed in the reactor with SRT of about 10 days, which had the best effluent quality. According to the FISH results, type 021N was predominant in the reactor with long SRT, which had the clogging problem. On the other hand, other reactors (with SRTs of about 10 and 30 days) did not contain much type 021N, but some other filamentous bacteria dominated. Maximum EPS concentration (as mg/L) was observed in the reactor with long SRT. Also the abundance of two types of metazoa (Pristina sp. and tardigrades) was observed in the reactor with long SRT, which had the clogging problem and poor effluent quality.

Animals↗

PubMed2002

Measurement of microorganisms with PHA production capability in activated sludge and its implication in activated sludge model no. 3.

In Activated Sludge Model No.3 (ASM3), it is hypothesized that all heterotrophic microorganisms (X(H)) can store substrate. However, in reality, both microorganisms with and without substrate storage capability (X(H/STO) and X(H/S), respectively) could exist. If the ratio of X(H/STO) in activated sludge is influenced by operational and environmental conditions, kSTO (storage rate constant of heterotrophic microorganisms which is defined in ASM3) may not be a universal parameter and can change from case to case. In this study, polyhydroxyalkanoate (PHA) is assumed as the principal storage product, and the ratio of microorganisms with PHA production capability (X(H/PHA)) in various activated sludges was estimated by the dual staining of Nile Blue A (NB) and DAPI. It was shown that the ratio of X(H/PHA) in sludge varied among different municipal and laboratory activated sludges.

Environmental Monitoring↗

PubMed2002

PHA (polyhydroxyalkanoate) production potential of activated sludge treating wastewater.

The main purposes of wastewater treatment systems are to remove organic pollutants, but it would be very attractive if there were a way to recover the organic pollutants as valuable organic materials. One of the possible ways to recover organic pollutants in wastewater is to convert them into polyhydroxyalkanoates (PHAs), which are biodegradable plastics. In this study, 18 activated sludge samples in 4 wastewater treatment plants (WWTPs) in Tokyo, Japan, were evaluated for their potential to produce PHAs by aerobic batch experiments with excess supply of acetate as the sole carbon source. The activated sludge samples tested had the capability to accumulate PHA up to 18.8% of dry cell weight on average, with the range of 6.0% to 29.5%. The results showed that the maximum PHA content was dependent on the influent more than on the operational conditions of the activated sludge, and that conventional activated sludge produced PHA as much as anaerobic-aerobic activated sludge did. The PHA content achieved in this study is still low, and further improvement is needed to put into practice the recovery process of organic pollutants as biodegradable plastics by activated sludge.

Alkanes↗

PubMed2002

Analysis of microbial community that performs enhanced biological phosphorus removal in activated sludge fed with acetate.

Enhanced biological phosphorus removal (EBPR) activated sludge was operated in a laboratory-scale sequencing batch reactor (SBR) fed with acetate as the sole carbon source. The microbial community of the sludge was analyzed using the polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) method for about 2 months of start-up period. As a result, the number of major bands decreased during the enrichment, indicating that the microbial community structure was getting simpler. Since the phosphate removal activity was maintained at a high level, the bacteria which still remained at the end can be considered as the important bacteria playing key roles in the present EBPR sludge, maybe polyphosphate accumulating organisms (PAOs). The dominant band in the last sample on the DGGE gel was excised and the DNA recovered from it was sequenced. The sequence was closely related to one of the putative PAOs group which Crocetti et al. (2000) and Hesselmann et al. (1999) have proposed. This PAOs group is closely related to the Rhodocyclus group (b-Proteobacteria). The fluorescence in situ hybridization (FISH) method with the probe specific for this PAOs group and the DAPI staining at a phosphate-probing concentration indirectly showed that these Rhodocyclus related bacteria really accumulated polyphosphate.

Acetates↗

PubMed2002

Density separation and molecular methods to characterize enhanced biological phosphorus removal system populations.

A novel approach to the identification of microorganisms that accumulate high density microbial storage products based on density separation, denaturing gradient gel electrophoresis (DGGE), and DNA sequencing was developed and applied to bench and pilot scale enhanced biological phosphorus removal (EBPR) systems. Polyphosphate (PP), glycogen, and polyhydroxyalkanoates (PHAs), are all of higher density than a typical bacterial cell. PP-accumulating organisms (PAOs), the organisms responsible for EBPR, accumulate all three of these storage products. Density separation in a homogenous solution of Percoll produced a high-density biomass fraction with a relatively high concentration of PAOs, as determined by Neisser staining. DNA was extracted from these fractions, amplified, and separated by DGGE. DGGE profiles demonstrated some bacterial strains were present at a greater concentration in the high density fractions than in low density fractions. These strains were considered PAO candidates. 5 of 12 PAO candidates from high density fractions were gamma Proteobacteria and only 1 was a beta Proteobacterium. 2 PAO candidates were most similar to recently identified gamma Proteobacteria sequences obtained by DGGE analysis of a deteriorated benchtop EBPR system.

Biomass↗

PubMed2002

Bacteriophage-host interaction in the enhanced biological phosphate removing activated sludge system.

Bacteriophages were isolated from a laboratory scale enhanced biological phosphate removing (EBPR) activated sludge process, and their host range was examined. Bacterial isolates to host the bacteriophages were isolated from the EBPR activated sludge process. Bacteriophages were eluted from the EBPR activated sludge, enriched by incubation with the bacterial isolates, and then tested for plaque formation on each of the bacterial isolates. Out of 12 bacterial isolates isolated, 4 supported plaque formation. Four bacteriophages were obtained from the plaques. The host range test was conducted with the combination of the bacteriophage isolates and the bacterial isolates. Three of the bacteriophages were found to form plaques on more than one host, and one of them formed plaques on both gram +ve and gram -ve bacterial isolates. Two of the four bacteriophages failed to form plaques on their original bacterial host, indicating the existence of mutation on either both or one of the host and the bacteriophage. This study strongly suggests that bacteriophages are an active part of the activated sludge microbial ecosystem, having very complex interaction with their host bacteria.

Bacteriophages↗

PubMed2002

Effect of important operational parameters on performance of coarse pore filtration activated sludge process.

A coarse pore filter can be applied inside the aeration tank instead of sedimentation tank for liquid separation from the sludge. It has pores, which are irregular in shape, and much bigger than micro-filtration membrane pores in size. The objective of the study was to investigate the effect of important operational parameters such as flux, aeration intensity, and solid retention time (SRT) on the performance of the coarse pore filtration activated sludge process. The effect of these parameters was studied in laboratory scale experiments. It was found that the flux had a significant role in the effluent quality of this system. The effluent SS and turbidity were not changed significantly at different aeration intensities. Three SRTs, 10, 30 and longer days (without excess sludge) were used for three reactors to check the effect of this parameter on the system performance. The results of the reactors with SRTs about 10 and 30 days have shown very good effluent quality without any filter clogging for more than 4 months operation. For the reactor with long SRT, the filter clogging was observed after about 80 days of operation, which caused the increase of the operation pressure and deterioration in the effluent quality for a few days.

Filtration↗