Erik Arvin

Long-Term Succession of Structure and Diversity of a Biofilm Formed in a Model Drinking Water Distribution System

ABSTRACT In this study, we examined the long-term development of the overall structural morphology and community composition of a biofilm formed in a model drinking water distribution system with biofilms from 1 day to 3 years old. Visualization and subsequent quantification showed how the biofilm developed from an initial attachment of single cells through the formation of independent microcolonies reaching 30 μm in thickness to a final looser structure with an average thickness of 14.1 μm and covering 76% of the surface. An analysis of the community composition by use of terminal restriction fragment length polymorphisms showed a correlation between the population profile and the age of the sample, separating the samples into young (1 to 94 days) and old (571 to 1,093 days) biofilms, whereas a limited spatial variation in the biofilm was observed. A more detailed analysis with cloning and sequencing of 16S rRNA fragments illustrated how a wide variety of cells recruited from the bulk water initially attached and resulted in a species richness comparable to that in the water phase. This step was followed by the growth of a bacterium which was related to Nitrospira, which constituted 78% of the community by day 256, and which resulted in a reduction in the overall richness. After 500 days, the biofilm entered a stable population state, which was characterized by a greater richness of bacteria, including Nitrospira, Planctomyces, Acidobacterium, and Pseudomonas. The combination of different techniques illustrated the successional formation of a biofilm during a 3-year period in this model drinking water distribution system.

Identification of Bacteria in Biofilm and Bulk Water Samples from a Nonchlorinated Model Drinking Water Distribution System: Detection of a Large Nitrite-Oxidizing Population Associated with Nitrospira spp.

ABSTRACT In a model drinking water distribution system characterized by a low assimilable organic carbon content (<10 μg/liter) and no disinfection, the bacterial community was identified by a phylogenetic analysis of rRNA genes amplified from directly extracted DNA and colonies formed on R2A plates. Biofilms of defined periods of age (14 days to 3 years) and bulk water samples were investigated. Culturable bacteria were associated with Proteobacteria and Bacteriodetes, whereas independently of cultivation, bacteria from 12 phyla were detected in this system. These included Acidobacteria, Nitrospirae, Planctomycetes, and Verrucomicrobia, some of which have never been identified in drinking water previously. A cluster analysis of the population profiles from the individual samples divided biofilms and bulk water samples into separate clusters (P = 0.027). Bacteria associated with Nitrospira moscoviensis were found in all samples and encompassed 39% of the sequenced clones in the bulk water and 25% of the biofilm community. The close association with Nitrospira suggested that a large part of the population had an autotrophic metabolism using nitrite as an electron donor. To test this hypothesis, nitrite was added to biofilm and bulk water samples, and the utilization was monitored during 15 days. A first-order decrease in nitrite concentration was observed for all samples with a rate corresponding to 0.5 × 105 to 2 × 105 nitrifying cells/ml in the bulk water and 3 × 105 cells/cm2 on the pipe surface. The finding of an abundant nitrite-oxidizing microbial population suggests that nitrite is an important substrate in this system, potentially as a result of the low assimilable organic carbon concentration. This finding implies that microbial communities in water distribution systems may control against elevated nitrite concentrations but also contain large indigenous populations that are capable of assisting the depletion of disinfection agents like chloramines.

Identification of organic compounds migrating from polyethylene pipelines into drinking water

A study of the diffusion of organic additives from four polyethylene (PE) materials into drinking water was conducted. Various structures of organic chemicals were identified in the water extracts by means of gas chromatography-mass spectrometry analysis. Most of them presented a basic common structure characterised by a phenolic ring typically substituted with hindered alkyl groups in positions 2 and 6 on the aromatic ring. The structures attributed to some of the chemicals have been confirmed using commercial or purposely synthesised standards. Unprocessed granules of raw PE were also analysed, in order to investigate the origin of the chemicals detected in the water samples. Consequently, the presence of some of the compounds was attributed to impurities or by-products of typical phenolic additives used as antioxidants in pipeline production. Finally, the occurrence of the identified chemicals was tested under field conditions, i.e. in water samples from newly installed pipelines in a distribution system. Here, the presence of three of the compounds identified in vitro was detected.

Bulk water phase and biofilm growth in drinking water at low nutrient conditions.

In this study, the bacterial growth dynamics of a drinking water distribution system at low nutrient conditions was studied in order to determine bacterial growth rates by a range of methods, and to compare growth rates in the bulk water phase and the biofilm. A model distribution system was used to quantify the effect of retention times at hydraulic conditions similar to those in drinking water distribution networks. Water and pipe wall samples were taken and examined during the experiment. The pipes had been exposed to drinking water at approximately 13 degrees C, for at least 385 days to allow the formation of a mature quasi-stationary biofilm. At retention times of 12 h, total bacterial counts increased equivalent to a net bacterial growth rate of 0.048 day(-1). The bulk water phase bacteria exhibited a higher activity than the biofilm bacteria in terms of culturability, cell-specific ATP content, and cell-specific leucine incorporation rate. Bacteria in the bulk water phase incubated without the presence of biofilm exhibited a bacterial growth rate of 0.30 day(-1). The biofilm was radioactively labelled by the addition of 14C-benzoic acid. Subsequently, a biofilm detachment rate of 0.013 day(-1) was determined by measuring the release of 14C-labelled bacteria of the biofilm. For the quasi-stationary phase biofilm, the detachment rate was equivalent to the net growth rate. The growth rates determined in this study by different independent experimental approaches were comparable and within the range of values reported in the literature.

A potential approach for monitoring drinking water quality from groundwater systems using organic matter fluorescence as an early warning for contamination events

The fluorescence characteristics of natural organic matter in a groundwater based drinking water supply plant were studied with the aim of applying it as a technique to identify contamination of the water supply. Excitation-emission matrices were measured and modeled using parallel factor analysis (PARAFAC) and used to identify which wavelengths provide the optimal signal for monitoring contamination events. The fluorescence was characterized by four components: three humic-like and one amino acid-like. The results revealed that the relative amounts of two of the humic-like components were very stable within the supply plant and distribution net and changed in a predictable fashion depending on which wells were supplying the water. A third humic-like component and an amino acid-like component did not differ between wells. Laboratory contamination experiments with wastewater revealed that combined they could be used as an indicator of microbial contamination. Their fluorescence spectra did not overlap with the other components and therefore the raw broadband fluorescence at the wavelengths specific to their fluorescence could be used to detect contamination. Contamination could be detected at levels equivalent to the addition of 60 μg C/L in drinking water with a TOC concentration of 3.3 mg C/L. The results of this study suggest that these types of drinking water systems, which are vulnerable to microbial contamination due to the lack of disinfectant treatment, can be easily monitored using online organic matter fluorescence as an early warning system to prompt further intensive sampling and appropriate corrective measures.

Activity of toluene-degrading pseudomonas putida in the early growth phase of a biofilm for waste gas treatment

A biological trickling filter for treatment of toluene-containing waste gas was studied. The overall kinetics of the biofilm growth was followed in the early growth phase. A rapid initial colonization took place during the first three days. The biofilm thickness increased exponentially, whereas the incease of active biomass and polymers was linear. In order to investigate the toluene degradation, various toluene degraders from the multispecies biofilm were isolated, and a Pseudomonas putida was chosen as a representative of the toluene-degrading population. A specific rRNA oligonucleotide probe was used to follow the toluene-degrading P. putida in the multispecies biofilm in the filter by means of number and cellular rRNA content. P. putida appeared to detach from the biofilm during the first three days of growth, after which P. putida was found at a constant level of 10% of the active biomass in the biofilm. Based on the rRNA content, the in situ activity was estimated to be reduced to 20% of cells grown at maximum conditions in batch culture. The toluene degraded by P. putida was estimated to be a minor part (11%) of the overall toluene degradation.

Microbial degradation of phenols and aromatic hydrocarbons in creosote-contaminated groundwater under nitrate-reducing conditions

Batch experiments were carried out to investigate the biodegradation of phenols and aromatic hydrocarbons under anaerobic, nitrate-reducing conditions in groundwater from a creosote-contaminated site at Fredensborg, Denmark. The bacteria in the creosote-contaminated groundwater degraded a mixture of toluene, phenol, the cresols (o-, m- and p-cresol) and the dimethylphenols 2,4-DMP and 3,4-DMP at both 10° and 20°C. Benzene, the xylenes, napthalene, 2,3-DMP, 2,5-DMP, 2,6-DMP and 3,5-DMP were resistant to biodegradation during 7–12 months of incubation. It was demonstrated that the degradation of toluene, 2,4-DMP, 3,4-DMP and p-cresol depended on nitrate or nitrite as electron acceptors. 40–80% of the nitrate consumed during degradation of the aromatic compounds was recovered as nitrite, and the consumption of nitrate was accompanied by a production of ATP. Stoichiometric calculations indicated that in addition to the phenols are toluene other carbon sources present in the groundwater contributed to the consumption of nitrate. If the groundwater was incubated under anaerobic conditions without nitrate, sulphate-reducing conditions evolved after ∼ 1 month at 20°C and ∼2 months at 10°C. In the sulphate-reducing batches disappearance of toluene, phenol, o-cresol and o-cresol was observed, whereas no removal of benzene, the xylenes, naphthalane, 2,3-DMP, 2,4-DMP, 2,5-DMP and 3,5-DMP was detected during 7 months of incubation.

Biological removal of phosphorus from wastewater

This paper reviews the present process technology for biological phosphorus (P) removal, the fundamental biological and chemical P removal mechanisms, analytical characterization methods for biological and inorganic P fractions, and finally conventional and new techniques for process design. The understanding of enhanced biological P removal has improved significantly in the past few years. Polyphosphate (poly‐P) accumulating bacteria appear to be selected by introducing an anaerobic fermentation reactor in which the biomass has access to readily biodegradable organics, like volatile acids, etc. The poly‐P bacteria are able to use poly‐P as an energy reserve to absorb and store the readily biodegradable organics, e.g., as s‐polyhy‐droxybutyrate. This gives them an advantage over nonpoly‐P bacteria in their competition for substrate. In some biological P‐removal plants, phosphate precipitation by cations in the sewage may play a significant role. A few milligrams per liter of iron and/or aluminum in the wa...

pH-decrease in nitrifying biofilms

Abstract Nitrification is an acidity producing process. It has been shown theoretically that the diffusional resistance to the transport in the biofilm of the inorganic carbon species as affected by the acidity production in a nitrifying biofilm gives rise to a decreased pH in the interior of the biofilm. These theoretical results have been verified on biofilms developed on the surfaces of a rotating drum under well controlled laboratory conditions. The results show clearly the drop in pH as predicted by theory. The phenomenon can give rise to unexpected effects on the performance of nitrifying biofilms, when most of the bacteria work under a much lower pH than the pH measured in the bulk water.

Biodegradation of phenols in a sandstone aquifer under aerobic conditions and mixed nitrate and iron reducing conditions

Ammonia liquor with very high concentrations of phenol and alkylated phenols is known to have leaked into the subsurface at a former coal carbonization plant in the UK, giving high concentrations of ammonium in the groundwater. In spite of this, no significant concentrations of phenols were found in the groundwater. The potential for biodegradation of the phenols in the sandstone aquifer at the site has been investigated in laboratory microcosms under aerobic (oxygen amended) and mixed nitrate and iron reducing (nitrate enriched and unamended) anaerobic conditions, at a range of concentrations (low: ∼5 mg l−1, high: ∼60 mg l−1, and very high: ∼600 mg l−1) and in the presence of other organic coal–tar compounds (mono- and polyaromatic hydrocarbons (BTEXs and PAHs) and heterocyclic compounds (NSOs)) and ammonia liquor. Sandstone cores and groundwater for the microcosms were collected from within the anaerobic ammonium plume at the field site. Fast and complete degradation of phenol, o- and p-cresol, 2,5- and 3,4-xylenol with no or very short initial lag-phases was observed under aerobic conditions at low concentrations. 2,6- and 3,5-Xylenol were degraded more slowly and 3,5-xylenol degradation was only just complete after about 1 year. The maximum rates of total phenols degradation in duplicate aerobic microcosms were 1.06 and 1.76 mg l−1 day−1. The degradation of phenols in nitrate enriched and unamended anaerobic microcosms was similar. Fast and complete biodegradation of phenol, cresols, 3,4-xylenol and 3,5-xylenol was observed after short lag-phases in the anaerobic microcosms. 2,5-xylenol was partially degraded after a longer lag-phase and 2,6-xylenol persisted throughout the 3 month long experiments. The maximum rates of total phenols degradation in duplicate nitrate enriched and unamended anaerobic microcosms were 0.30–0.38 and 0.29–0.31 mg l−1 day−1, respectively. The highest phenols concentrations in the anaerobic microcosms apparently required very long adaptation periods or inhibited biodegradation of the phenols. For the intermediate concentration level, degradation occurred after comparable lag-phases and at comparable rates to those observed at low concentration. However, after a while degradation of phenols suddenly decreased drastically and then stopped. Dilution by addition of anaerobic groundwater resulted in continued but slow degradation of phenols in unamended microcosms. The effect of other organic coal–tar compounds (BTEXs, PAHs, NSOs) on the degradation of the phenols under unamended conditions was limited to slightly longer lag-phases for some of the phenols. Other constituents of the ammonia liquor did not appear to significantly affect the degradation of the phenols. Fast and complete degradation of 2,3- and 2,4-xylenol was indicated. These experiments were continued for a longer period of time and revealed complete degradation of 2,5-xylenol and, after an approximately 6-month-long lag-phase partial degradation of 2,6-xylenol. The potential for natural attenuation of phenols from process effluents from coal carbonization under aerobic conditions and mixed nitrate and iron reducing conditions appears promising.