Biomedical subjects

P H Nielsen

Publications and source records attributed to P H Nielsen.

At least 19 recordsLinked to original sources

PubMed2006

Identity, abundance and physiology of Aquaspirillum-related filamentous bacteria in activated sludge.

Microcolony-forming bacteria closely related to the genus Aquaspirillum in the Betaproteobacteria were recently observed to be abundant in many nutrient removal wastewater treatment plants. The developed oligonucleotide probe, Aqs997, however, occasionally also targeted some filamentous bacteria in activated sludge samples when fluorescence in situ hybridization was performed. In this study, the identity, abundance, and ecophysiology of these Aqs997-positive filamentous bacteria were studied in detail. Most of the Aqs997-positive filamentous bacteria could morphologically be identified as either Eikelboom Type 1701, Type 0041/0675 or possibly Type 1851, all characterized by epiphytic growth. They were found in almost all 21 wastewater treatment plants investigated. Two morphotypes were found. Type A filaments, which seemed to be the same genotype as the microcolony-forming bacteria targeted by probe Aqs997.Type B filaments also hybridized with probe GNS941, specific for the Chloroflexi phylum, so the true identity remains unclear. Aqs997-positive filaments usually stained Gram-negative, but Gram-positive filaments were also found, stressing the difficulties in identifying bacteria from morphology and simple staining results. Studies on the ecophysiology by microautoradiography showed that Aqs997-positive filamentous bacteria did not consume acetate and glucose, while some took up butyrate, mannose, and certain amino acids. Most likely, some Aqs997-positive filamentous bacteria were able to perform full denitrification such as the Aqs997-positive microcolony-forming bacteria, and some were able to store polyhydroxyalkanoates under anaerobic conditions, potentially being glycogen accumulating organisms.

Base Sequence↗

PubMed2006

Floc-forming properties of polyphosphate accumulating organisms in activated sludge.

The physico-chemical characteristics of polyphosphate-accumulating organisms (PAO) involved in enhanced biological phosphorus removal (EBPR) was investigated in order to find a novel method for phosphorus recovery. If the physico-chemical characteristics of PAO are different from those of other main floc components, it may be possible to enrich PAO in bulk water or in the floc material for improved recovery of phosphorus. A combination of shear tests, chemical manipulation, and quantification of PAO by fluorescence in situ hybridization was applied. The microcolony strength of both Rhodocyclus-related PAO and Actinobacteria-related PAO was generally high as no treatment could break up more than 20% of all PAO in microcolonies. In contrast, it was possible to remove 20-40% of the organic matter and other bacterial cells by applying a high pH value or adding EDTA. With that a selective enrichment of PAO in the remaining floc material was possible. The feasibility of applying this selective PAO enrichment in flocs remains to be evaluated in full-scale plants for P-recovery.

Actinobacteria↗

PubMed2006

Applicability of experience from laboratory reactors with biological phosphorus removal in full-scale plants.

Enhanced biological phosphorus removal (EBPR) has been used at many wastewater treatment plants all over the world for many years. In this study a full-scale sludge with good EBPR was tested with P-release batch tests and combined FISH/MAR (fluorescence in situ hybridisation and microautoradiography). Proposed models of PAOs and GAOs (polyphosphate- and glycogen-accumulating organisms) and microbial methods suggested from studies of laboratory reactors were found to be applicable also on sludge from full-scale plants. Dependency of pH and the uptake of both acetate and propionate were studied and used for calculations for verifying the models and results from microbial methods. All rates found from the batch tests with acetate were higher than in the batch tests with propionate, which was explained by the finding that only those parts of the bacterial community that were able to take up acetate anaerobically were able to take up propionate anaerobically.

Acetates↗

PubMed2005

Microbial diversity in biofilms from corroding heating systems.

Culture-independent investigations of the bacterial diversity and activity in district heating systems with and without corrosion did not make it possible to relate one group of microorganisms with the observed corrosion. Fluorescence in situ hybridization by oligonucleotide probes revealed the dominance of beta-proteobacteria, sulphate reducing prokaryotes and alpha-proteobacteria. Analysis of a clone library from one Danish heating (DH) system showed that the most sequences formed two clusters within the alpha-proteobacteria affiliated to the families Rhizobiaceae and Acetobacteraceae and two clusters within the beta-proteobacteria belonging to the family Comamonadaceae. Functional groups were determined by microautoradiography showing aerobic and anaerobic bacteria (sulphate reducing and methanogenic bacteria). The corrosion study showed that pitting corrosion rates were five to ten times higher than the general corrosion rates, suggesting the presence of biocorrosion. The results indicate that several bacterial groups could be involved in corrosion of DH system piping including sulphate reducing prokaryotes, Acidovorax (within the beta-proteobacteria), methanogenic bacteria and others.

Bacteria↗

PubMed2004

Micromanipulation and further identification of FISH-labelled microcolonies of a dominant denitrifying bacterium in activated sludge.

The activated sludge process relies on the formation of strong microbial flocs. The knowledge about dominant floc-forming bacteria is at present very limited, especially from a phylogenetic perspective. In this study, numerous microcolonies in the activated sludge flocs were found to be targeted by a Betaproteobacteria-group-specific oligonucleotide probe using fluorescence in situ hybridization (FISH). Some of these were micromanipulated and further identified by reverse transcriptase polymerase chain reaction (RT-PCR) and sequencing to belong to the Aquaspirillum genus in the Neisseriaceae family. A specific oligonucleotide probe, Aqs997, was designed to target the identified bacteria. A survey in nine different wastewater treatment plants with nutrient removal (WWTP) showed a high abundance of bacteria hybridizing to the oligonucleotide probe developed. Microautoradiography (MAR) combined with FISH on activated sludge incubated with radiolabelled substrate showed uptake of substrate with oxygen, nitrate and nitrite as electron acceptor demonstrating a denitrifying potential of the bacteria investigated. The Aquaspirillum-related bacteria seemed to be abundant denitrifiers in WWTPs with nitrogen removal and they were particularly numerous in plants mainly receiving domestic wastewater, where they constituted up to 30% of all bacteria.

Bacteria↗

PubMed2004

MIC mitigation in a 100 MW district heating peak load unit.

During inspection of AISI316 stainless steel plate heat exchangers in a district heating peak load unit, localised corrosion attacks along with indications of microbiological activity were found on the boiler side beneath patches of sturdy black deposits. Bacteria and sulphide were detected within black deposits. Thorough investigation of the boiler system revealed several incidents of localised corrosion on low alloy steel along with deposits of organic matter and bacteria primarily in places with stagnant water or places operating at a low flow rate. A relatively large amount of bacteria was detected within the system, primarily in deposits and around corrosion sites. The observations suggested the combination of deposits and bacterial activity, being the major reason for the observed corrosion. Prior to the investigation, the boiler system had operated with cat-/anion-exchanged, de-aerated water for 3 years, during which the water fulfilled strict chemical limits set to minimise corrosion. Based on these findings, the system has been modified in order to minimise the risk of microbiologically influenced corrosion and a monitoring program for fouling and corrosion has been established.

Bacteria↗

PubMed2004

Bacterial composition of activated sludge--importance for floc and sludge properties.

Activated sludge flocs consist of numerous constituents which, together with other factors, are responsible for floc structure and floc properties. These properties largely determine the sludge properties such as flocculation, settling and dewaterability. In this paper we briefly review the present knowledge about the role of bacteria in relation to floc and sludge properties, and we present a new approach to investigate the identity and function of the bacteria in the activated sludge flocs. The approach includes identification of the important bacteria and a characterization of their physiological and functional properties. It is carried out by use of culture-independent molecular biological methods linked with other methods to study the physiology and function, maintaining a single cell resolution. Using this approach it was found that floc-forming properties differed among the various bacterial groups, e.g. that different microcolony-forming bacteria had very different sensitivities to shear and that some of them deflocculated under anaerobic conditions. In our opinion, the approach to combine identity with functional analysis of the dominant bacteria in activated sludge by in situ methods is a very promising way to investigate correlations between presence of specific bacteria, and floc and sludge properties that are of interest.

Bacteria↗

PubMed2003

Monitoring and characterisation of bacteria in corroding district heating systems using fluorescence in situ hybridisation and microautoradiography.

Presence of biofilm and biocorrosion has been observed in Danish district heating (DH) systems despite very good water quality that was expected to prevent significant microbial growth. The microbiological water quality was investigated in order to identify the dominating bacterial groups on surfaces with corrosion problems. Water samples from 29 DH systems were investigated for the total number of bacteria and presence of sulphate reducing bacteria (SRBs). SRBs were found to be present in more than 80% of the DH systems. The microbial population in samples from 2 DH system (biofilm from a test coupon and an in situ sample from a heat exchanger) was investigated with fluorescence in situ hybridisation, and the results showed significant differences in population composition. Betaproteobacteria was the dominant population in both samples. SRBs were present in both samples but were most numerous in the biofilm from the test coupon. Examination of functional groups based on uptake of radiolabelled acetate (microautoradiography) showed presence of both aerobic and anaerobic bacteria despite the fact that oxygen is not anticipated in DH systems.

Bacteria, Aerobic↗

PubMed2002

Effect of solids retention time and wastewater characteristics on biological phosphorus removal.

The paper deals with the effect of wastewater, plant design and operation in relation to biological nitrogen and phosphorus removal and the possibilities to model the processes. Two Bio-P pilot plants were operated for 2.5 years in parallel receiving identical wastewater. The plants had SRT of 4 and 21 days, the latter had nitrification and denitrification. The plant with 4 days SRT had much more variable biomass characteristics, than the one with the high SRT. The internal storage compounds, PHA, were affected significantly by the concentration of fatty acids or other easily degradable organics in the wastewater, and less by the plant lay-out. The phosphorus removal is mainly dependent on availability in the wastewater of fatty acids but also by the suspended solids in the effluent, which is higher in the plant with nitrification-denitrification, probably due to a higher SVI or denitrification in the settler. The addition of glucose to the influent seems to have an effect on the performance of the plants similar to that of acetic acid. In spite of great load variations over time to the pilot plants and the different operational modes, the study of population dynamics showed less significant variations with time which has importance in relation to modelling. The overall conclusion of the comparison between the two plants is that the biological phosphorus removal efficiency under practical operating conditions is affected by the SRT in the plant and the wastewater composition. Thus great care should be taken when extrapolating results from one type of plant to another. Indirectly the experiments confirm that results from lab-experiments with artificial wastewater are difficult to extrapolate through modelling to real life wastewater and conditions. The 2.5 years time series can be valuable in verification of models for Nitrogen and Enhanced Biological Phosphorus Removal.

Bacteria↗

PubMed2002

Use of phosphorus release batch tests for modelling an EBPR pilot plant.

The aim of this study was to evaluate how routinely performed phosphorus release tests could be used when modelling enhanced biological phosphorus removal (EBPR) using activated sludge models such as ASM2d. A pilot plant with an extensive analysis programme was used as basis for the simulations. Without any calibration the prediction of phosphorus removal was poor and the initial release rates from the simulations were not similar to those found from the laboratory tests. A period with low organic loading was chosen as a calibration period. In this period averages of daily influent measurements were used as influent parameters. First, calibration was performed in order to fit effluent COD and MLVSS in the sludge. Next, the phosphorus content in the sludge was decreased to the measured level by decreasing the fermentation rate. Finally, the initial phosphorus release rate was calculated from a simulated batch test and the PHA uptake rate was increased to fit this release rate with the average initial rates from laboratory batch tests performed during the period. The calibrated model was verified with data from the subsequent period where acetate was dosed.

Calibration↗

PubMed2002

Population dynamics in wastewater treatment plants with enhanced biological phosphorus removal operated with and without nitrogen removal.

The population dynamics of activated sludge in a pilot plant with two activated sludge systems, both designed for enhanced biological phosphorus removal (EBPR), but one of them with (BNP) and the other without (BP) nitrogen removal, was monitored during a period of 2.5 years. The influent water to the pilot plant was periodically manipulated by external addition of phosphorus (P), acetate and glucose, respectively. The population dynamics and the in situ physiology were monitored by quantitative fluorescence in situ hybridization (FISH) and microautoradiography. Significant P removal was observed in both systems throughout the whole period, with significant increases of the P removal when substrates were dosed. The activated sludge in both systems contained large amounts of dense clusters of gram-negative, methylene-blue staining coccoid rods during the whole period. A large part of the clusters belonged to the beta Proteobacteria, whereas the rest of the clusters belonged either to the Actinobacteria or to the alpha Proteobacteria. The relative abundance of Rhodocyclus-related bacteria in the activated sludge varied significantly in both systems during the whole period (from 6 to 18% in BNP, and from 4 to 28% in BP). However, no statistically significant correlation of the Rhodocyclus-related nor any of the other investigated bacterial groups to the P content of the activated sludge (correlation for all groups investigated was always < 0.5) was observed. A significant 33Pi uptake was observed by the beta Proteobacteria (part of them Rhodocyclus-related, the identity of the rest unknown) and the Actinobacteria. However, not all of the Rhodocyclus-related bacteria showed 33Pi uptake. The P removal in the investigated plants is thus believed to be mediated by a mixed population consisting of a part of the Rhodocyclus-related bacteria, the Actinobacteria and other, yet unidentified bacteria.

Autoradiography↗

PubMed2002

Quantification of functional groups in activated sludge by microautoradiography.

Different functional groups of bacteria in activated sludge from a Danish municipal wastewater treatment plant were investigated and quantified on the basis of their ability to take up acetate under different electron acceptor conditions. The number of bacteria in the different functional groups was quantified by microautoradiography or by fluorescence in situ hybridization (FISH). Uptake of radiolabeled acetate was tested under conditions where oxygen, nitrate, ferric iron, and sulfate served as electron acceptors and under methanogenic conditions. Ammonia-oxidizing bacteria and nitrite-oxidizing bacteria were enumerated by applying oligonucleotide probes (FISH). 80% of the total DAPI count hybridised with a mixture of bacterial probes. Most baceria were able to take up acetate with oxygen or nitrate as electron acceptors (74 and 71% of all DAPI-stained bacteria, respectively). The very similar numbers indicate that the alternating aerobic and anoxic conditions in the wastewater treatment plant investigated favoured the presence of facultative aerobic and denitrifying heterotrophic bacteria. The number of bacteria able to take up acetate under anaerobic conditions was around 8% of the total DAPI count. About half of these were able to take up acetate and store it and were thus probably phosphate-accumulating organisms. The remaining 4% of anaerobic acetate-consuming bacteria were mainly iron reducers. By incubating the sludge samples with specific inhibitors against sulfate reduction and methanogenic activity, it was found that sulfate reducers and methanogenic bacteria constituted approximately 1% and less than 0.5% of the total DAPI count, respectively.

Acetates↗

PubMed2002

Microthrix parvicella, a specialized lipid consumer in anaerobic-aerobic activated sludge plants.

The in situ physiology of the filamentous bacterium Microthrix parvicella was investigated in anaerobic-aerobic dynamic phases in activated sludge with focus on the uptake of long chain fatty acids (LCFA) and growth. When 14C-labeled oleic acid was added to activated sludge with an excessive growth of M. parvicella, only little 14C-CO2 was produced under anaerobic conditions while a lot was produced under aerobic conditions. Microautoradiographic studies revealed that M. parvicella took up oleic acid under both anaerobic and aerobic conditions, while only a few floc formers were able to take it up under anaerobic conditions. Extraction and separation of the radioactive biomass into different lipid fractions showed that the oleic acid was stored mainly as neutral lipids under anaerobic conditions, whereas conversion to membrane phospholipids occurred almost exclusively under aerobic conditions, indicating growth. The surface properties of M. parvicella and other bacteria were characterized by hydrophobic fluorescent microspheres, which showed that M. parvicella was relatively hydrophobic. Furthermore, a surface-associated extracellular lipase activity was observed, indicating the ability of M. parvicella to degrade lipids near the filament surface. The results support the hypothesis that uptake and storage of LCFA as lipids under anaerobic conditions provide an effective competition strategy against bacteria that can only take up LCFA under aerobic conditions. Thus, M. parvicella seems to be a specialized lipid consumer with a physiological potential analogous to PAOs and GAOs being able to take up LCFA (but not short chain fatty acids or glucose) under anaerobic conditions and subsequently use the storage material for growth when nitrate or oxygen are available as electron acceptors.

Actinobacteria↗

PubMed2001

Comparison of methods for determination of microbial biomass in wastewater.

Microbial biomass in wastewater was determined by methods used in environmental microbiology and by a method used in wastewater engineering based on a conceptual model simulating fundamental microbial processes in wastewater from measured oxygen uptake rates. The methods originating from environmental microbiology are based on staining and counting of cells for the determination of total cell biomass (acridine orange and DAPI), physiological state of cells (LIVE/DEAD BacLight) and activity of cells (reduction of the redox dye CTC and microautoradiography). Depending on the staining method applied, cell biomasses yielded 15-86% of the biomass defined by the model, and good correlations between cell biomass and model biomass were found. Cell biomass, oxygen uptake and acetate uptake were measured in wastewater, where acetate was added. Substrate uptake rates were found not to be proportional to the increases in cell biomass, suggesting that only a small fraction of the cell biomass was responsible for the main part of the substrate uptake. Despite the differences found between cell biomass and model biomass, it was recommended to use the conceptual model as an engineering tool for simulation of microbial processes and wastewater quality changes. However, there should be a clear distinction between the terms 'model biomass', 'cell biomass' and different activity measurements of cells.

Bacteria↗

PubMed2001

In situ characterization of Nitrospira-like nitrite-oxidizing bacteria active in wastewater treatment plants.

Uncultivated Nitrospira-like bacteria in different biofilm and activated-sludge samples were investigated by cultivation-independent molecular approaches. Initially, the phylogenetic affiliation of Nitrospira-like bacteria in a nitrifying biofilm was determined by 16S rRNA gene sequence analysis. Subsequently, a phylogenetic consensus tree of the Nitrospira phylum including all publicly available sequences was constructed. This analysis revealed that the genus Nitrospira consists of at least four distinct sublineages. Based on these data, two 16S rRNA-directed oligonucleotide probes specific for the phylum and genus Nitrospira, respectively, were developed and evaluated for suitability for fluorescence in situ hybridization (FISH). The probes were used to investigate the in situ architecture of cell aggregates of Nitrospira-like nitrite oxidizers in wastewater treatment plants by FISH, confocal laser scanning microscopy, and computer-aided three-dimensional visualization. Cavities and a network of cell-free channels inside the Nitrospira microcolonies were detected that were water permeable, as demonstrated by fluorescein staining. The uptake of different carbon sources by Nitrospira-like bacteria within their natural habitat under different incubation conditions was studied by combined FISH and microautoradiography. Under aerobic conditions, the Nitrospira-like bacteria in bioreactor samples took up inorganic carbon (as HCO(3)(-) or as CO(2)) and pyruvate but not acetate, butyrate, and propionate, suggesting that these bacteria can grow mixotrophically in the presence of pyruvate. In contrast, no uptake by the Nitrospira-like bacteria of any of the carbon sources tested was observed under anoxic or anaerobic conditions.

Autoradiography↗

PubMed2001

Transformation of lipids in activated sludge.

Transformation of lipids in activated sludge treatment plants is of interest for two reasons: lipids contribute 30-40% of the chemical oxygen demand (COD) in wastewater, and they may stimulate the growth of filamentous microorganisms in nutrient removal activated sludge plants. The transformation of lipids was investigated under aerobic and anoxic conditions by measuring the oxygen and nitrate uptake rates (OUR and NUR). The maximal OUR and NUR of long-chain fatty acid was found to be at the same level as acetate indicating that long-chain fatty acid was as easily consumable. However, the adsorption of long-chain fatty acid to surfaces of sludge flocs made it difficult to determine initial uptake rates of long-chain fatty acids. It was not possible to describe the hydrolysis rate of triacylglyceride by OUR and NUR to long-chain fatty acids because the hydrolysis rate was very slow. For a better description of the processes involved in transformation of lipids, a conceptual model was suggested. The processes in the suggested model were the adsorption/desorption of both triacylglyceride, and long-chain fatty acid onto surfaces of sludge flocs, hydrolysis of triacylglyceride by lipases and the uptake of long-chain fatty acid by bacteria under various conditions. The model can be helpful to structure design and evaluation of activated sludge experiment with lipids.

Adsorption↗

PubMed2001

Remember the water--a comment on EPS colligative properties.

The relationship between water and activated sludge components was examined. Reevaluation of published data on freezing point depression, drying rates and dewatering has been performed. The basis of this has been the assumption that the water/sludge relationship is considered to be a colligative effect. Since the results indicate this to be the case, we suggest that the published concepts of "pools of water" are false. Data on swelling properties of EPS as a function of pH suggests that the colligative properties are largely determined by the counterions of charged polymers and surfaces.

Biopolymers↗

PubMed2001

Quantification of the bond energy of bacteria attached to activated sludge floc surfaces.

The great majority of activated sludge bacteria exist incorporated in flocs. The increase in dispersed bacteria when exposed to increasing turbulent shear rates has been successfully modelled by a model assuming that the adhesion and erosion of cells may be considered in analogy to ordinary chemical phase transitions. By this adhesion-erosion model (AE-model), an "enthalpy" of cell adhesion can be estimated, and this value in turn determines the range of shear rates in which erosion of cells predominates. Application of the model has indicated that only a mass fraction less than ca. 6-17% may be released from activated sludge, even when exposed to a severe turbulent environment, i.e. only a small fraction of the flocs is dispersible by means of erosion by turbulence. The shear sensitivity and the dispersible floc fraction were found to depend on the floc composition. A net decrease in the floc EPS content during anaerobic sludge stabilisation causes a dramatic increase in the dispersed fraction, indicating the important role of EPS for the floc strength. It was found also that activated sludge cells do not reflocculate completely after exposure to high shear rates. This may be an indication that the cohesion energy of bacteria growing in colonies is greater than the energy of the more stochastic adhesion of dispersed cells to floc surfaces. It could also be another indicator of the importance of entanglement forces, which do not reform instantly, once broken. When the bond strength of cell attachment to sludge surfaces is altered by changes in the chemical environment, this may cause a change in the dispersible floc fraction as well as a change in the shear range of erosion. When the shear sensitivity constant kss is adopted for sludge characterisation, an increased degree of dispersion under standard test conditions will result in increased shear sensitivity estimates. The shear sensitivity may be used for the estimation of en equivalent change in the Gibb's energy of cell adhesion (delta (delta Gad/RT)).

Bacterial Adhesion↗