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H C Flemming

Publications and source records attributed to H C Flemming.

18 recordsLinked to original sources
PubMed2004

Contamination potential of drinking water distribution network biofilms.

Drinking water distribution system biofilms were investigated for the presence of hygienically relevant microorganisms. Early biofilm formation was evaluated in biofilm reactors on stainless steel, copper, polyvinyl chloride (PVC) and polyethylene coupons exposed to unchlorinated drinking water. After 12 to 18 months, a plateau phase of biofilm development was reached. Surface colonization on the materials ranged between 4 x 10(6) and 3 x 10(7) cells/cm2, with heterotrophic plate count (HPC) bacteria between 9 x 10(3) and 7 x 10(5) colony-forming units (cfu)/cm2. Established biofilms were investigated in 18 pipe sections (2 to 99 years old) cut out from distribution pipelines. Materials included cast iron, galvanized steel, cement and PVC. Colonization ranged from 4 x 10(5) to 2 x 10(8) cells/cm2, HPC levels varied between 1 and 2 x 10(5) cfu/cm2. No correlation was found between extent of colonization and age of the pipes. Using cultural detection methods, coliform bacteria were rarely found, while Escherichia coli, Pseudomonas aeruginosa and Legionella spp. were not detected in the biofilms. In regular operation, distribution system biofilms do not seem to be common habitats for pathogens. However, nutrient-leaching materials like rubber-coated valves were observed with massive biofilms which harboured coliform bacteria contaminating drinking water.

Biofilms↗
PubMed2003

Role and levels of real-time monitoring for successful anti-fouling strategies--an overview.

Biofouling is a biofilm problem and any anti-fouling strategy will be greatly improved if the site and extent of biofilm growth can be monitored. A suitable monitoring system will provide early warning capacity and allow for specific optimization of countermeasures. As water samples do not give reliable information about biofilms, surface sampling is mandatory. Conventional biofilm monitoring techniques rely on removal of material from representative sites or on analysis of test surfaces which have been exposed. This procedure is time consuming and, depending on the parameters to be measured, requires skilled laboratory personnel. There is a strong demand for direct, on-line, in situ, continuous, non-destructive real-time information about biofilms in a system. Such demands can only be fulfilled by physical or physico-chemical methods, a number of which have already been successfully applied for biofilm monitoring. It is important, however, to be aware of the actual parameter they refer to in order to interpret the data properly. Three levels of information can be identified: (i) systems which detect increase and decrease of material accumulating on a surface but cannot differentiate between biomass and other components of a deposit, (ii) systems which provide biological information and distinguish between biotic and abiotic material, and (iii) systems which provide detailed chemical information. Examples for all three levels are presented and discussed.

Bacteria↗
PubMed2003

On-line monitoring of biofilm formation in a brewery water pipeline system with a fibre optical device.

Any advanced anti-fouling strategy must be based on early warning systems which allow for timely, precisely directed and optimized countermeasures. Such systems must be able to detect biofilm growth on representative surfaces. In order to meet this requirement, a fibre optical device (FOS) has been developed. It is based on light which is scattered by objects deposited on the tip of an optical fibre. A receiving fibre collects the signal and forwards it to a detection and quantification unit. Both the sending and the receiving fibre are mounted in a measuring head which is integrated evenly on the inner surface of a water pipeline at representative sites. This device was applied to a water system of a brewery in order toput its reliability to test under practical conditions. The FOS detected the build-up of a deposit which was identified independently as consisting of microorganisms, i.e., a biofilm. A stable, well detectable and reproducible signal could be obtained above a colonization of 10(5) cells cm-2. Adjustment of the sensitivity of the amplifier allowed for detection of biofilms up to 10(10) cells cm-2. Cleaning countermeasures could be detected clearly by a decrease of backscattered light intensity. The system proved to be suitable for on-line, non-destructive, real-time and automatic monitoring for a period of almost two years, and thus, provides an important constituent for an advanced anti-fouling strategy.

Automation↗
PubMed2001

Uniaxial compression measurement device for investigation of the mechanical stability of biofilms.

The mechanical stability of biofilms is important for biotechnology, as sloughing of the biomass due to mechanical failure of the biofilm matrix can lead to severe interferences with biofilm processes. In cases of biofouling, biofilms have to be removed, in which case their mechanical stability must be overcome. The apparent modulus of elasticity and the yield strength as obtained from uniaxial compression experiments can be taken as parameters indicative for the mechanical stability of a biofilm. A film rheometer is presented which allows for the determination of these quantities, using model biofilms of Pseudomonas aeruginosa grown on membrane filters. The compressive stress-strain behaviour up to the point of failure is recorded at a compression speed of 1 microm s(-1). In accordance with the stress-strain curve, the investigated biofilm can be described as viscoelastic material, which demonstrates plastic flow properties. The extracellular polymeric substances (EPS), which keep biofilms together, form a temporary network of fluctuating junction points. Above the yield point, the gel structure fails and the system behaves as a highly viscous fluid. The apparent modulus of elasticity and the yield point are considered to be useful parameters for characterizing the mechanical properties of biofilms.

Biofilms↗
PubMed2001

Capability of mucoid Pseudomonas aeruginosa to survive in chlorinated water.

Mucoid strains of Pseudomonas aeruginosa are characterized by an overproduction of the extracellular polysaccharide alginate. When suspended into chlorinated swimming-pool water or drinking water samples, mucoid bacteria revealed enhanced survival compared with isogenic nonmucoid cells. Removal of slime from mucoid bacteria abolished chlorine resistance, addition of purified alginate to washed bacteria again enhanced survival. Thus, alginate-containing slime confers protection on P. aeruginosa against chlorine and may contribute to survival of these bacteria in chlorinated water systems.

Chlorine Compounds↗
PubMed2001

Relevance of microbial extracellular polymeric substances (EPSs)--Part I: Structural and ecological aspects.

Extracellular polymeric substances are the construction materials for microbial aggregates such as biofilms, flocs ("planktonic biofilms") and sludge. Their major components are not only polysaccharides but also proteins and in some cases lipids, with minor contents of nucleic acids and other biopolymers. In the EPS, biofilm organisms can establish stable arrangements and function multicellularly as synergistic microconsortia. The matrix facilitates the retention of exoenzymes, cellular debris and genetic material; it can be considered as a microbial recycling yard. Gradients can develop due to the physiological activity and the fact that diffusive mass transport prevails over convective transport in the matrix. Biofilm cells tolerate higher concentrations of many biocides. The EPS matrix sequesters nutrients from the water phase. In photosynthetic communities, EPS molecules can function as light transmitters and provide photons to organisms located deeper in a microbial mat. The EPS matrix is a dynamic system, constructed by the organisms and responding to environmental changes. It enables the cells to function in a manner similar to multicellular organisms.

Biofilms↗
PubMed2001

Agarose hydrogels as EPS models.

For investigating the influence of extracellular polymeric substances (EPSs) on biofilm properties, artificial models of defined composition and structure can be very helpful. Different immobilised cell systems can be useful in the fitting of experimental results. Two different types of of artificial EPS matrix model were developed earlier. Homogeneous agarose beads (50-500 microns diameter) and porous beads (260 microns mean diameter) containing pores with diameters from 10 to 80 microns (28 microns on average), allowed the embedding of cells, particles and typical EPS matrix components such as proteins and polysaccharides. In this paper, some physico-chemical properties of the artificial EPS matrices were described and compared with results known from natural EPS. The stability of the artificial matrix polymer against solution in the aqueous medium was studied in batch experiments. The water binding and water retaining capabilities of the EPS models were investigated by drying and re-swelling experiments. The simulation of protective effects of the artificial EPS matrix against toxic substances like biocides in comparison to such known protective effects of the EPS of native biofilms were proved by the application of sodium hypochlorite (0.5 mg/l, 30 min) and subsequent microscopic investigation of the cell population after LIVE/DEAD staining (Molecular Probes).

Biofilms↗
PubMed2001

A new method for extraction of extracellular polymeric substances from biofilms and activated sludge suitable for direct quantification of sorbed metals.

A method for extraction of extracellular polymeric substances (EPSs) with a dicyclohexyl-18-crown-6 ether was developed to determine levels of organic and inorganic contaminants sorbed to EPS. The crown ether selectively binds alkaline and alkaline earth metals but not heavy metals. The effectiveness of the extraction procedure was higher than that of 2 other methods tested and comparable with that of a method based on a cation exchange resin. On average it was possible to extract 20% of the TOC, 12% of the total protein content, and 4% of the total carbohydrate content of sludge or biofilm biomass. Metal sorption studies in activated sludge showed no influence of exposure time on the fractionation of metals within the biomass. Metals sorbed mostly to cellular material. In biofilms 12.2% of the cadmium and 9.1% of the zinc added was found in the EPS. In activated sludge EPS contained only 2.9% zinc. The distribution of metals within the biomass was dose dependent. The percentage of metals found in EPS decreased with increasing metal concentration. This indicates a higher affinity of metals for cellular binding sites. Time course experiments in a rotating biofilm annular reactor, which consisted of an external cylinder with removable slides and an internal solid drum, revealed a gradual change in zinc concentration associated with EPS, although the total zinc concentration in the biomass remained constant. Concurrently, the amount of extractable EPS decreased. This was a consequence of a microbial population shift, with bacterial counts decreasing and algal and fungal biomass increasing. Using confocal laser scanning microscopy and the fluorescent metal complexing agent Newport Green for in situ detection of zinc it was shown that metals were bound to algae and fungi in the latter part of the experiment. The biofilm became more and more heterogeneous coinciding with a decrease in EPS. To summarize, the observed sorption behavior of metals cannot be explained with the conventional paradigm of EPS as hydrophilic gel. Obviously, different binding mechanisms must be invoked to explain the role of EPS in the sorption and removal of toxic substances in activated sludge and biofilm systems. It is important to consider the microbial population to understand differences in sorption in different matrices.

Adsorption↗
PubMed2001

Influence of calcium ions on the mechanical properties of a model biofilm of mucoid Pseudomonas aeruginosa.

The mechanical properties of biofilms and in particular their mechanical strength is of great importance for both biofilm reactors and for the removal of undesired biofilms as in cases of biofouling and biocorrosion. By uniaxial compression measurements, it is possible to determine the apparent elastic or Young's modulus and the yield stress as parameters for mechanical stability. This was performed with a recently developed device, using model biofilms of mucoid strain Pseudomonas aeruginosa SG81. The biofilms were grown on membrane filters placed on nutrient agar medium with different concentrations of calcium ions. The compressive stress-strain behaviour up to failure was recorded at a compression speed of 1 micron s-1. The apparent Young's modulus, representing the stiffness of the biofilm, and the yield stress obtained from the stress--strain diagram were used for the description of mechanical properties of biofilms. A certain critical concentration of calcium ions was found where the Young's modulus of the P. aeruginosa biofilms increases strongly and subsequently remains constant for higher calcium concentrations. This behaviour is explained by the presence of calcium ions crosslinking alginate, which is the major component of the extracellular polymeric substances produced by the mucoid P. aeruginosa strain used in this investigation.

Biofilms↗
PubMed2001

Relevance of microbial extracellular polymeric substances (EPSs)--Part II: Technical aspects.

Extracellular polymeric substances (EPSs) are involved in both detrimental and beneficial consequences of microbial aggregates such as biofilms, flocs and biological sludges. In biofouling, they are responsible for the increase of friction resistance, change of surface properties such as hydrophobicity, roughness, colour, etc. In biocorrosion of metals they are involved by their ability to bind metal ions. In bioweathering, they contribute by their complexing properties to the dissolution of minerals. The EPSs represent a sorption site for pollutants such as heavy metal ions and organic molecules. This can lead to a burden in wastewater sludge; on the other hand, the sorption properties can be used for water purification. Other biotechnological uses of EPS exploit their contribution to viscosity, e.g., in food, paints and oil-drilling 'muds'; their hydrating properties are also used in cosmetics and pharmaceuticals. Furthermore, EPSs may have potential uses as biosurfactants, e.g., in tertiary oil production, and as biological glue. EPSs are an interesting component of all biofilm systems and still hold a large biotechnological potential.

Adsorption↗
PubMed2000

Diffusion in Pseudomonas aeruginosa biofilms: a pulsed field gradient NMR study.

A Pseudomonas aeruginosa biofilm is studied with pulsed field gradient echo nuclear magnetic resonance. Although not all spectral components are assigned yet, the experimental results show that a biofilm consists of components with very different diffusion coefficients. The various biofilm components that give motionally narrowed 1H NMR signals, can be grouped into five classes with diffusion coefficients, ranging from 2 x 10(-9) to 1 x 10(-13) m2 s-1. Investigation of the diffusion behavior of water in the biofilm shows three fractions with different diffusion coefficients. Besides the highly mobile bulk water at least two other fractions with much lower diffusion coefficients are detected. It is shown that one of the fractions with the low diffusion coefficient probably arises from intracellular water. Also for another component of the biofilm, glycerol, three fractions with diffusion coefficients that differ more than a factor ten are detected. Also a group of signals exists which result from practically immobile components.

Alginates↗
PubMed2000

Artificial biofilm model--a useful tool for biofilm research.

For biofilm studies, artificial models can be very helpful in studying processes in hydrogels of defined composition and structure. Two different types of artificial biofilm models were developed. Homogeneous agarose beads (50-500 microm diameter) and porous beads (260 microm mean diameter) containing pores with diameters from 10 to 80 microm (28 microm on average) allowed the embedding of cells, particles and typical biofilm matrix components such as proteins and polysaccharides. The characterisation of the matrix structures and of the distribution of microorganisms was performed by confocal laser scanning microscopy. The physiological condition of the embedded bacteria was examined by redox activity (CTC-assay) and membrane integrity (Molecular Probes LIVE/DEAD-Kit). Approximately 35% of the immobilised cells (Pseudomonas aeruginosa SG81) were damaged due to the elevated temperature required for the embedding process. It was shown that the surviving cells were able to multiply when provided with nutrients. In the case of homogeneous agarose beads, cell growth only occurred near the bead surface, while substrate limitation prevented growth of more deeply embedded cells. In the porous hydrogel, cell division was observed across the entire matrix due to better mass transport. It could be shown that embedding in the artificial gel matrix provided protection of immobilized cells against toxic substances such as sodium hypochlorite (0.5 mg/l, 30 min) in comparison to suspended cells, as observed in other immobilized systems. Thus, the model is suited to simulate important biofilm matrix properties.

Biofilms↗
PubMed2000

Biodegradation of cis-1,4-polyisoprene rubbers by distinct actinomycetes: microbial strategies and detailed surface analysis.

Several actinomycetes isolated from nature were able to use both natural rubber (NR) and synthetic cis-1,4-polyisoprene rubber (IR) as a sole source of carbon. According to their degradation behavior, they were divided into two groups. Representatives of the first group grew only in direct contact to the rubber substrate and led to considerable disintegration of the material during cultivation. The second group consisted of weaker rubber decomposers that did not grow adhesively, as indicated by the formation of clear zones (translucent halos) around bacterial colonies after cultivation on NR dispersed in mineral agar. Taxonomic analysis of four selected strains based on 16S rRNA similarity examinations revealed two Gordonia sp. strains, VH2 and Kb2, and one Mycobacterium fortuitum strain, NF4, belonging to the first group as well as one Micromonospora aurantiaca strain, W2b, belonging to the second group. Schiff's reagent staining tests performed for each of the strains indicated colonization of the rubber surface, formation of a bacterial biofilm, and occurrence of compounds containing aldehyde groups during cultivation with NR latex gloves. Detailed analysis by means of scanning electron microscopy yielded further evidence for the two different microbial strategies and clarified the colonization efficiency. Thereby, strains VH2, Kb2, and NF4 directly adhered to and merged into the rubber material, while strain W2b produced mycelial corridors, especially on the surface of IR. Fourier transform infrared spectroscopy comprising the attenuated total reflectance technique was applied on NR latex gloves overgrown by cells of the Gordonia strains, which were the strongest rubber decomposers. Spectra demonstrated the decrease in number of cis-1,4 double bonds, the formation of carbonyl groups, and the change of the overall chemical environment, indicating that an oxidative attack at the double bond is the first metabolic step of the biodegradation process.

Actinomycetales↗
PubMed1999

The role of intermolecular interactions: studies on model systems for bacterial biofilms.

The mechanical stability of biofilms and other microbial aggregates is of great importance for both the maintenance of biofilm processes and the removal of undesired biofilms. The binding forces are weak interactions such as London dispersion forces, electrostatic interactions and hydrogen bonds. In a first attempt to rank their contribution, the viscosity of solutions of extracellular polymeric substances (EPS) from a mucoid strain of Pseudomonas aeruginosa is measured. In order to distinguish the binding forces, substances are chosen which individually address the different types of bonds. Polyacrylic acid is identified as a suitable model system for EPS when molecular interactions are studied. Electrostatic interactions and hydrogen bonds are found to be the dominating forces among macromolecules within the biofilm.

Acrylic Resins↗
PubMed1993

Use of 5-cyano-2,3-ditolyl tetrazolium chloride for quantifying planktonic and sessile respiring bacteria in drinking water.

Direct microscopic quantification of respiring (i.e., viable) bacteria was performed for drinking water samples and biofilms grown on different opaque substrata. Water samples or biofilms developed in flowing drinking water were incubated with the vital redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and R2A medium. One hour of incubation with 0.5 mM CTC was sufficient to obtain intracellular reduction of CTC to the insoluble fluorescent formazan (CTF) product, which was indicative of cellular respiratory (i.e., electron transport) activity. This result was obtained with both planktonic and biofilm-associated cells. Planktonic bacteria were captured on 0.2-microns-pore-size polycarbonate membrane filters and examined by epifluorescence microscopy. Respiring cells containing CTF deposits were readily detected and quantified as red-fluorescing objects on a dark background. The number of CTC-reducing bacteria was consistently greater than the number of aerobic CFU determined on R2A medium. Approximately 1 to 10% of the total planktonic population (determined by counterstaining with 4,6-diamidino-2-phenylindole) were respirometrically active. The proportion of respiring bacteria in biofilms composed of drinking water microflora was greater, ranging from about 5 to 35%, depending on the substratum. Respiring cells were distributed more or less evenly in biofilms, as demonstrated by counterstaining with 4,6-diamidino-2-phenylindole. The amount of CTF deposited in single cells of Pseudomonas putida that formed monospecies biofilms was quantified by digital image analysis and used to indicate cumulative respiratory activity. These data indicated significant cell-to-cell variation in respiratory activity and reduced electron transport following a brief period of nutrient starvation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗