Characterising organic matter in recirculating aquaculture systems with fluorescence EEM spectroscopy.

The potential of recirculating aquaculture systems (RAS) in the aquaculture industry is increasingly being acknowledged. Along with intensified application, the need to better characterise and understand the accumulated dissolved organic matter (DOM) within these systems increases. Mature RASs, stocked with rainbow trout and operated at steady state at four feed loadings, were analysed by dissolved organic carbon (DOC) analysis and fluorescence excitation-emission matrix (EEM) spectroscopy. The fluorescence dataset was then decomposed by PARAFAC analysis using the drEEM toolbox. This revealed that the fluorescence character of the RAS water could be represented by five components, of which four have previously been identified in fresh water, coastal marine water, wetlands and drinking water. The fluorescence components as well as the DOC showed positive correlations with feed loading, however there was considerable variation between the five fluorescence components with respect to the degree of accumulation with feed loading. The five components were found to originate from three sources: the feed; the influent tap water (groundwater); and processes related to the fish and the water treatment system. This paper details the first application of fluorescence EEM spectroscopy to assess DOM in RAS, and highlights the potential applications of this technique within future RAS management strategies.

Designing water supplies: Optimizing drinking water composition for maximum economic benefit.

It is possible to optimize drinking water composition based on a valuation of the impacts of changed water quality. This paper introduces a method for assessing the potential for designing an optimum drinking water composition by the use of membrane desalination and remineralization. The method includes modeling of possible water quality blends and an evaluation of corrosion indices. Based on concentration-response relationships a range of impacts on public health, material lifetimes and consumption of soap have been valued for Perth, Western Australia and Copenhagen, Denmark. In addition to water quality aspects, costs of water production, fresh water abstraction and CO(2)-emissions are integrated into a holistic economic assessment of the optimum share of desalinated water in water supplies. Results show that carefully designed desalination post-treatment can have net benefits up to €0.3 ± 0.2 per delivered m(3) for Perth and €0.4(±0.2) for Copenhagen. Costs of remineralization and green house gas emission mitigation are minor when compared to the potential benefits of an optimum water composition. Finally, a set of optimum water quality criteria is proposed for the guidance of water supply planning and management.

Optimal drinking water composition for caries control in populations.

Apart from the well-documented effect of fluoride in drinking water on dental caries, little is known about other chemical effects. Since other ions in drinking water may also theoretically influence caries, as well as binding of fluoride in the oral environment, we hypothesized that the effect of drinking water on caries may not be limited to fluoride only. Among 22 standard chemical variables, including 15 ions and trace elements as well as gases, organic compounds, and physical measures, iterative search and testing identified that calcium and fluoride together explained 45% of the variations in the numbers of decayed, filled, and missing tooth surfaces (DMF-S) among 52,057 15-year-old schoolchildren in 249 Danish municipalities. Both ions had reducing effects on DMF-S independently of each other, and could be used in combination for the design of optimal drinking water for caries control in populations.

Monitoring toxicity of industrial wastewater and specific chemicals to a green alga, nitrifying bacteria and an aquatic bacterium.

Treatment plants may be exposed to a whole range of toxic organic and inorganic compounds that may inhibit the removal of organic matter and nitrogen. In order to secure maximum treatment efficiency, the plant manager has to monitor the toxicity of the influent sewage. With regard to the receiving water the manager also has to make sure that toxicity in the influent is significantly reduced during treatment. Because a whole range of chemicals may be present, chemical analysis may be insufficient and expensive as a control instrument. Instead, direct toxicity measurements are preferable to capture the complexity of the wastewater. The monitoring methods have to be relevant and sensitive for the processes in the treatment plant, i.e. removal of organic matter and nutrients. The methods also have to be simple and inexpensive. The paper reports on recent results from the application of nitrification, algae and Biotox tests, and summarises the experience with monitoring of toxicity. Although the sensitivity of the tests varies with respect to individual chemicals or group of chemicals, the application of a combination of the tests gives a high likelihood of detecting toxic impacts on treatment plants and receiving waters.

Methanotrophic biodegradation of cis-1,2-dichloroethylene in a continuously fed fixed-film bioreactor.

Co-metabolic biodegradation of cis-dichloroethylene (cis-DCE) was investigated in a bench-scale fixed-film bioreactor inoculated with a mixed culture of methane oxidising bacteria. The aim of this work was to identify factors that affect the cis-DCE biodegradation. It was observed that the presence of methane was necessary to enhance the biodegradation of cis-DCE, but an excess of methane inhibited the cis-DCE removal. cis-DCE did not inhibit the methane biodegradation at concentrations up to 300 microg/L. Maximum cis-DCE removal was observed with a methane bulk concentration ranging from 0.2 to 0.7 mg/L. It was found that the activity of the biofilm was located in the upper 100 microm of the biofilm. On the basis of this study it is concluded that careful control of the oxygen and methane concentrations as well as of the biofilm thickness is necessary in order to optimise the biodegradation of cis-DCE in fixed film bioreactors.

Monitoring biofilm formation and activity in drinking water distribution networks under oligotrophic conditions.

In this study, the construction a model distribution system suitable for studies of attached and suspended microbial consisted of two loops connected in series with a total of 140 biofilm sampling points. The biofilm from the system was studied using 11 different microbial methods and the results were compared and discussed. The methods were used for biomass quantification (AODC, HPC and ATP determination), visualisation of structure (CLSM), activity measurement (leucine incorporation, AOC removal rate, respiration of benzoic acid, CTC and live/dead stains), and microbial diversity profiling (clone libraries and DGGE).

Biofilm formation in a hot water system.

The biofilm formation rate was measured in situ in a hot water system in an apartment building by specially designed sampling equipment, and the net growth of the suspended bacteria was measured by incubation of water samples with the indigeneous bacteria. The biofilm formation rate reached a higher level in the hot water distribution system (2.1 d(-1) to 2.3 d(-1)) than in the hot water tank (1.4 d(-1) to 2.2 d(-1)) indicating an important area for surface associated growth. The net growth rate of the suspended bacteria measured in hot water from the top, middle and bottom of the hot water tank, in the sludge, or in the water from the distribution system was negligible. This indicated that bacterial growth took place on the inner surfaces in the hot water system and biofilm formation and detachment of bacteria could account for most of the suspended bacteria actually measured in hot water. Therefore, attempts to reduce the number of bacteria in a hot water system have to include the distribution system as well as the hot water tank.

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.

The use of mathematical models in teaching wastewater treatment engineering.

Mathematical modeling of wastewater treatment processes has become increasingly popular in recent years. To prepare students for their future careers, environmental engineering education should provide students with sufficient background and experiences to understand and apply mathematical models efficiently and responsibly. Approaches for introducing mathematical modeling into courses on wastewater treatment engineering are discussed depending on the learning objectives, level of the course and the time available.

Modelling the growth of a methanotrophic biofilm: estimation of parameters and variability.

This article discusses the growth of methanotrophic biofilms. Several independent biofilm growths scenarios involving different inocula were examined. Biofilm growth, substrate removal and product formation were monitored throughout the experiments. Based on the oxygen consumption it was concluded that heterotrophs and nitrifiers co-existed with methanotrophs in the biofilm. Heterotrophic biomass grew on soluble polymers formed by the hydrolysis of dead biomass entrapped in the biofilm. Nitrifier populations developed because of the presence of ammonia in the mineral medium. Based on these experimental results, the computer program AQUASIM was used to develop a biological model involving methanotrophs, heterotrophs and nitrifiers. The modelling of six independent growth experiments showed that stoichiometric and kinetic parameters were within the same order of magnitude. Parameter estimation yielded an average maximum growth rate for methanotrophs, micron, of 1.5 +/- 0.5 d-1, at 20 degrees C, a decay rate, bm, of 0.24 +/- 0.1 d-1, a half saturation constant, KS(CH4), of 0.06 +/- 0.05 mg CH4/L, and a yield coefficient, YCH4, of 0.57 +/- 0.04 g X/g CH4. In addition, a sensitivity analysis was performed on this model. It indicated that the most influential parameters were those related to the biofilm (i.e. density; solid-volume fraction; thickness). This suggests that in order to improve the model, further research regarding the biofilm structure and composition is needed.

Examination of reproducibility in microbiological degradation experiments.

Experimental data indicate that certain microbiological degradation experiments have a limited reproducibility. Nine identical batch experiments were carried out on 3 different days to examine reproducibility. A pure culture, isolated from soil, grew with toluene as the only carbon and energy source. Toluene was degraded under aerobic conditions at a constant temperature of 28 degrees C. The experiments were modelled by a Monod model--extended to meet the air/liquid system, and the parameter values were estimated using a statistical nonlinear estimation procedure. Model reduction analysis resulted in a simpler model without the biomass decay term. In order to test for model reduction and reproducibility of parameter estimates, a likelihood ratio test was employed. The limited reproducibility for these experiments implied that all 9 batch experiments could not be described by the same set of parameter values. However, experiments carried out the same day (within the same run) were more uniform than experiments carried out on different days (between runs), and a common set of parameter estimates could be accepted for experiments within runs, but not for experiments from different runs. The limited reproducibility may be caused by variability in the preculture, or more precisely, variations in the physiological state of the bacteria in the precultures just before used as inoculum.

Improved Dechlorinating Performance of Upflow Anaerobic Sludge Blanket Reactors by Incorporation of Dehalospirillum multivorans into Granular Sludge

Dechlorination of tetrachloroethene, also known as perchloroethylene (PCE), was investigated in an upflow anaerobic sludge blanket (UASB) reactor after incorporation of the strictly anaerobic, reductively dechlorinating bacterium Dehalospirillum multivorans into granular sludge. This reactor was compared to the reference 1 (R1) reactor, where the granules were autoclaved to remove all dechlorinating abilities before inoculation, and to the reference 2 (R2) reactor, containing only living granular sludge. All three reactors were fed mineral medium containing 3 to 57 µM PCE, 2 mM formate, and 0.5 mM acetate and were operated under sterile conditions. In the test reactor, an average of 93% (mole/mole) of the effluent chloroethenes was dichloroethene (DCE), compared to 99% (mole/mole) in the R1 reactor. The R2 reactor, with no inoculation, produced only trichloroethene (TCE), averaging 43% (mole/mole) of the effluent chloroethenes. No dechlorination of PCE was observed in an abiotic control consisting of sterile granules without inoculum. During continuous operation with stepwise-reduced hydraulic retention times (HRTs), both the test reactor and the R1 reactor showed conversion of PCE to DCE, even at HRTs much lower than the reciprocal maximum specific growth rate of D. multivorans, indicating that this bacterium was immobilized in the living and autoclaved granular sludge. In contrast, the R2 reactor, with no inoculation of D. multivorans, only converted PCE to TCE under the same conditions. Immobilization could be confirmed by using fluorescein-labeled antibody probes raised against D. multivorans. In granules obtained from the R1 reactor, D. multivorans grew mainly in microcolonies located in the centers of the granules, while in the test reactor, the bacterium mainly covered the surfaces of granules.

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.

Activity and three-dimensional distribution of toluene-degrading Pseudomonas putida in a multispecies biofilm assessed by quantitative in situ hybridization and scanning confocal laser microscopy.

As a representative member of the toluene-degrading population in a biofilter for waste gas treatment, Pseudomonas putida was investigated with a 16S rRNA targeting probe. The three-dimensional distribution of P. putida was visualized in the biofilm matrix by scanning confocal laser microscopy, demonstrating that P. putida was present throughout the biofilm. Acridine orange staining revealed a very heterogeneous structure of the fully hydrated biofilm, with cell-free channels extending from the surface into the biofilm. This indicated that toluene may penetrate to deeper layers of the biofilm, and consequently P. putida may be actively degrading toluene in all regions of the biofilm. Furthermore, measurements of growth rate-related parameters for P. putida showed reduced rRNA content and cell size (relative to that in a batch culture), indicating that the P. putida population was not degrading toluene at a maximal rate in the biofilm environment. Assuming that the rRNA content reflected the cellular activity, a lower toluene degradation rate for P. putida present in the biofilm could be estimated. This calculation indicated that P. putida was responsible for a significant part (65%) of the toluene degraded by the entire community.

Method development for trace analysis of heteroaromatic compounds in contaminated groundwater.

An analytical method providing high sensitivity (limit of quantitation of 50 ng/l) with acceptable reproducibility (mean R.S.D. 19%) has been developed for determining heteroaromatic compounds in creosote-contaminated groundwater. The best technique (highest recovery and reproducibility) found between liquid-liquid extraction using either dichloromethane, diethyl ether or pentane and solid-phase extraction with reversed-phase bonded columns, was the classical liquid extraction with dichloromethane from weak basic solutions and GC-MS (selective ion monitoring) analysis of concentrated extracts.

Effects of creosote compounds on the aerobic bio-degradation of benzene.

The inhibitory effect of creosote compounds on the aerobic degradation of benzene was studied in microcosm experiments. A total removal of benzene was observed after twelve days of incubation in microcosms where no inhibition was observed. Thiophene and benzothiophene, two heterocyclic aromatic compounds containing sulfur (S-compounds), had a significant inhibitory effect on the degradation of benzene, but also an inhibitory effect of benzofuran (an O-compound) and 1-methylpyrrole (a N-compound) could be observed, although the effect was weaker. The NSO-compounds also had an inhibitory effect on the degradation of p-xylene, o-xylene, and naphthalene, while they only had a weak influence on the degradation of 1-methylnaphthalene, o-cresol and 2,4-dimethylphenol. The phenolic compounds seemed to have a weak stimulating effect on the degradation of benzene whereas the monoaromatic hydrocarbons and the naphthalenes had no significant influence on the benzene degradation. The inhibitory effect of the NSO-compounds on the aerobic degradation of benzene could be identified as three different phenomena. The lag phase increased, the degradation rate decreased, and a residual concentration of benzene was observed in microcosms when NSO-compounds were present. The results show that NSO-compounds can have a potential inhibitory effect on the degradation of many creosote compounds, and that inhibitory effects in mixtures can be important for the degradation of different compounds.

The influence of creosote compounds on the aerobic degradation of toluene.

The inhibiting effect of 14 typical creosote compounds on the aerobic degradation of toluene was studied in batch experiments. Four NSO-compounds (pyrrole, 1-methylpyrrole, thiophene, and benzofuran) strongly inhibited the degradation of toluene. When the NSO-compounds were present together with toluene, little or no degradation of toluene was observed during 16 days of incubation, compared with a total removal of toluene within 4 days when the four compounds were absent. Indole (an N-compound) and three phenolic compounds (phenol, o-cresol, and 2,4-dimethylphenol) also inhibited the degradation of toluene, though the effect was much weaker that of the four NSO-compounds. O-xylene, p-xylene, naphthalene and 1-methylnaphthalene seemed to stimulate the degradation even though the influence was very weak. No effects of benzothiophene (an S-compound) and quinoline (an N-compound) were observed. Benzofuran (an O-compound) was identified as the compound that most inhibited the degradation of toluene. An effect could be detected even at low concentrations (40 micrograms/l).