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Low-temperature inhibition of the activated sludge process by an industrial discharge containing the azo dye acid black 1.

A municipal wastewater treatment plant (WWTP) receiving industrial dyeing discharge containing acid black 1 (AB1) failed to meet NH(3) and BOD(5) discharge limits, especially for NH(3) during the winter. Dyeing discharge was combined with domestic sewage in volumetric ratios reflecting the range received by the WWTP and fed to sequencing batch reactors at 22 and 7 degrees C. Analysis of the various nitrogen species revealed complete nitrification failure at 7 degrees C with more rapid nitrification failure as the dye concentration increased. Slight nitrification inhibition occurred at 22 degrees C: NH(3) removal decreased from 99.9% for the control compared to only 97.0% removal with dye addition. Dye-bearing wastewater also reduced COD removal by half at 7 degrees C and by one-fifth at 22 degrees C, and increased effluent TSS nearly three-fold at 7 degrees C. Activated sludge quality at 7 degrees C deteriorated after exposure to AB1, as indicated by excessive foaming and the presence of filamentous bacteria and by a decrease in endogenous and exogenous oxygen uptake. Decreasing AB1 loading resulted in partial activated sludge recovery. Eliminating the dye-bearing discharge to the full-scale WWTP led to improved performance bringing the WWTP into compliance with discharge limits.

Amido Black↗

Nitrobacter and Nitrospira genera as representatives of nitrite-oxidizing bacteria: detection, quantification and growth along the lower Seine River (France).

Pollution from agriculture and urban effluents influences the ecology and biochemical functioning of the Seine River. Nitrification dominates nitrogen transformations downstream of the effluents of the Paris wastewater treatment plant (WWTP) at Achères, treating, by activated sludge the wastewater of 6.5 million inhabitant equivalents from Paris and its suburbs, without nitrification and denitrification treatment. It discharges effluents containing large amounts of nitrogen, ammonium mostly (approximately 30 mg L(-1) N-NH(4+) L(-1)), on average 45 mg L(-1) of suspended particulate matter, high quantities of total organic carbon (approximately 30 mg C L(-1)) largely biodegradable (40%), and high concentration in total phosphorus ( approximately 3 mg Tot P L(-1)), as well as microorganisms. Ammonium, brought into the river system, is slowly nitrified in the lower Seine River and especially in the freshwater estuary. The nitrifying activities can be observed by measuring inorganic nitrogen compound concentrations and potential activities. To understand the contributions of the WWTP effluents, the upstream agricultural runoff water and the Seine tributaries, it is useful to investigate the bacterial community. Whereas ammonia oxidation has been widely studied, the second step, i.e. nitrite oxidation, is less well understood. We have previously analysed the ammonium-oxidizing bacterial (AOB) community in the Seine (Cébron, A., Berthe, T., Garnier, J., 2003. Nitrification and nitrifying bacteria in the lower Seine River and estuary (France). Appl. Environ. Microbiol. 69, 7091-7100; Cébron, A., Coci, M., Garnier, J., Laanbroek, H.J., 2004. DGGE analysis of the ammonia oxidizing bacterial community structure in the lower Seine River: impact of the Paris wastewater effluents. Appl. Environ. Microbiol. 70, 6726-6737), and focus here on the composition of the nitrite-oxidizing bacterial (NOB) community. As no general molecular probe targeting all known NOBs is currently available, we chose to target and quantify (by competitive PCR) the two genera Nitrobacter and Nitrospira assumed to be the major players in nitrite oxidation in freshwater environments. Nitrobacter species were dominant in the upstream Seine River basin but Nitrospira was the dominant NOB downstream of the WWTP. These two genera were equally represented in WWTP effluents. In the Seine River estuary, especially in the salinity gradient, the Nitrobacter proportion increases and that of Nitrospira disappears, possibly due dilution by seawater.

Bacteria↗

Estimating the combined effects of copper and phenol to nitrifying bacteria in wastewater treatment plants.

The inhibition of nitrification by phenol and copper mixtures was studied to investigate the differences between individual and mixture inhibition as the change of phenol and copper speciation due to their reactions. This study showed a decrease in the ammonium removal rate (k, first-order rate coefficient), indicating an increase in the relative % inhibition on nitrification, as the phenol and mixed liquor suspended solids (MLSS) concentrations were increased. In the case of copper, the ammonium removal rate decreased as the copper concentration increased and that of the MLSS decreased. This was attributed to the enhanced sorption of copper to heterotrophs at high MLSS concentrations. The relative % inhibition was plotted against free Cu2+ ions, which showed a close relationship. In the tests on copper and phenol mixtures, the mixture effects turned out overestimated by the calculation based on the additional chemical concentrations. In most Cu-phenol mixture tests, the presence of phenol as a complexation ligand caused a reduction of the inhibition rate to nitrifiers over the entire exposure range. Consequently, the prediction of nitrification inhibition in mixture environments, such as wastewater treatment plants, is influenced by the presence of a number of parameters that affect the activity of nitrifiers and, therefore, chemical speciation should be taken into consideration.

Adsorption↗

Effect of soluble microbial products on microbial metabolisms related to nutrient removal.

In this study, the effect of soluble microbial products (SMP) on the metabolisms related to phosphate or nitrogen removal of activated sludge was investigated. Two anaerobic-aerobic activated sludge processes were operated, one with a hydraulic retention time (HRT) of 48 h (RunL) and the other 6.4 h (RunS). The longer HRT of RunL was intended to promote the accumulation of SMPs in the supernatant. With the sludge from RunS and the supernatant from both of the runs, supernatant exchange batch experiments (SEBEs) were conducted, in which the acetate uptake rate and phosphate release rates under anaerobic conditions and the phosphate uptake rate under aerobic conditions were measured as these metabolisms are related to enhanced biological phosphorus removal. The nitrification rate was also measured. The statistical analyses of the results from the SEBEs showed that the supernatant from RunL had an inhibitory effect on the anaerobic acetate uptake and nitrification of the sludge from RunS. The cause of which was attributed to SMPs in the supernatant from RunL. As a result, the inhibitory effect of SMPs on nitrification and anaerobic acetate uptake was confirmed.

Sewage↗

Treatment of anaerobically pre-treated domestic sewage by a rotating biological contactor.

The performance of a rotating biological contactor (RBC) for the post-treatment of the effluent of an up-flow anaerobic sludge blanket (UASB) was the subject of this study. Different hydraulic and organic loading rates have been investigated. The removal efficiencies of COD(total), COD(suspended), COD(colloidal) and COD(soluble) increased at a higher hydraulic retention time (HRT) and a lower influent organic loading rate. The results obtained indicated that a two-stage RBC reactor at an HRT of 10 h and an organic loading rate of 6.4g COD m(-2) d(-1) represents an effective post-treatment process. Most COD(suspended) and COD(colloidal) were removed in the first stage while nitrification proceeded in the second stage. The overall nitrification efficiency was 92% at an ammonia loading rate of 1.1 gm(-2) d(-1). Total E. coli removal at HRTs of 10, 5 and 2.5h were 99.5%, 99.0% and 89.0%. respectively. The major part of suspended E. coli ( >4.4 microm) was removed by sedimentation or by adsorption in the biofilm of the first stage of RBC (99.66%). However, E. coli in the colloidal fraction (<4.4 to >0.45 microm) was eliminated in the second stage of RBC (99.78%). A comparison of the performance of a one-stage versus two-stage RBC system, operated at the same total loading rate, revealed an improvement in the effluent quality of the two-stage effluent as compared to the one-stage effluent. The two stages RBC were used to examine the effect of hydraulic shock loads on reactor performance in terms of COD, nitrification and E. coli removal.

Adsorption↗

Determination of the bacterial processes which are sources of nitrous oxide production in marine samples.

Partial denitrification and the initial step of nitrification are the main biological processes which produce nitrous oxide. In order to determine the contribution that these processes have in nitrous oxide production, the efficiency of different inhibitors on nitrifying activity has been tested, and the effect on denitrifying activity has been investigated, using culture strains and natural marine samples. A good nitrification inhibitor should not affect denitrification. A low partial pressure of C2H2 provided the best conditions, inhibiting 75%, Nitrosococcus oceanus (culture sample) and 100% (natural sample) of the nitrifying activity and having only a small inhibitory effect (12%) on denitrifying activity. These conditions have been applied on samples from the dilution plume of the Rhĵne River, an area characterized as a source of nitrous oxide. Using these inhibitors, it has been shown that in this area, incomplete denitrification is the main process producing nitrous oxide in the surface layers at the mouth of the river and in the bottom nepheloid layer, whereas in the marine surface layer the dominant process is nitrification.

Bacteria↗

A simple biofilm model of bacterial competition for attached surface.

A simple biofilm model of competition in bacterial growth for an attached surface is developed. Competition for the attached surface is expressed with the crowded and detachment effects. The developed model is verified by comparing simulated results with data obtained in the experiments of batch culture of nitrifier and continuous treatment of actual sewage with biofilm reactor. This model can favorably simulate the growth competition between autotrophic and heterotrophic bacteria for the attached surface. Then some parameters for nitrification process are discussed with this model. It is clarified that the effective removal of organic matter before nitrification tank is required for effective nitrification in the biofilm reactor.

Biodegradation, Environmental↗

Influence of aeration and sludge retention time on ammonium oxidation to nitrite and nitrate.

Partial nitrification to nitrite was reported to be technically feasible and economically favourable, especially when wastewater with high ammonium concentrations or low C/N ratios are treated. Nitritation can be obtained by selectively inhibiting nitrite oxidizing microrganisms through appropriate regulation of the system's pH, temperature, and sludge retention time. In addition to already known methods, the work showed that aeration patterns may play a relevant role too. Nitrification tests were performed in two lab-scale reactors operated under continuous and intermittent aeration, respectively. In both plants, temperature was maintained at 32 degrees C and pH was regulated at 7.2 by providing external buffer capacity when needed. The results showed that partial nitrification to nitrite was steadily obtained under oxygen limitation, independent of the sludge age. Therefore, the aeration pattern is proposed as an alternative parameter to the sludge retention time for controlling ammonium oxidation to nitrite.

Bacteria↗

Congruence in the performance of model nitrifying activated sludge plants located in Germany, Scotland and Spain.

A laboratory model nitrifying activated sludge plant treating OECD synthetic sewage was designed and constructed by each of three laboratories in Germany, Scotland and Spain in order to produce a sludge inoculum for 5 rapid toxicity bioassays. The plants were run for 3 years and produced sludge for the microbially based bioassays Vibrio fischeri bioluminescence, ATP luminescence and respiration, and, nitrification and enzyme inhibition. Although the initial sludge inoculum for the plants differed, as did some of the running conditions such as temperature regime, the sludge produced within the different countries had similar characteristics with respect to sludge age, total suspended solids and volatile suspended solids. Nitrification was generally maintained over the 3-year period although there were occasions when the process was inconsistent. Nitrification recovery was afforded by reseeding with a nitrifying sludge from a local wastewater treatment works (WWTW) or imposition of starvation conditions for a period of time. The sludge produced was used to carry out toxicity testing and results compared well with those using sludge from a WWTW. Overall, the use of sludge generated in the laboratory could be used for toxicity testing negating the need to resort to the use of natural WWTW sludge, which may contain a range of toxic substances due to uncontrolled industrial and domestic inputs and an unbalanced microbial consortium.

Adenosine Triphosphate↗

Effect of temperature and disinfection strategies on ammonia-oxidizing bacteria in a bench-scale drinking water distribution system.

The establishment of ammonia-oxidizing bacteria (AOB), a group of autotrophic microorganisms responsible for nitrification in chloraminated distribution systems, was studied in a bench-scale distribution system. The potential significance of temperature and disinfectant residual associated with chloramination in full-scale drinking water distribution systems was assessed. Biofilm development was primarily monitored using AOB abundance and nitrite concentrations. The bench-scale system was initially operated under typical North American summer (22 degrees C) and fall (12 degrees C) temperatures, representing optimal and less optimal growth ranges for these microorganisms. Additional experimentation investigated AOB establishment at a suboptimal winter distribution system temperature of 6 degrees C. The effect of chloramine residual on AOB establishment was studied at higher (0.2-0.6mg/L) and lower (0.05-0.1mg/L) ranges, using a 3:1 (w/w) chlorine:ammonia dosing ratio. Conditions were selected to represent those typically found in a North American distribution system, in areas of low flow and longer retention times, respectively. Finally, the effect of a free chlorine residual on an established nitrifying biofilm was briefly examined. Results clearly indicate that AOB development occurs at all examined temperatures, as well as at selected monochloramine residuals. The maintenance of a disinfectant residual was difficult at times, but was more inhibitory to the nitrifying biofilm than the lower temperature. It can be concluded from the data that nitrification may not be adequately inhibited during the winter months, which may result in more advanced stages of nitrification the following season. Free chlorination can be effective in controlling AOB activity in the short term, but may not prevent reestablishment of a nitrifying biofilm upon return to chloramination.

Ammonia↗

Effects of chromium (VI) addition on the activated sludge process.

The effect of hexavalent chromium, Cr(VI), addition on various operating parameters of activated sludge process was evaluated. To accomplish this, two parallel lab-scale continuous-flow activated sludge plants were operated. One was used as a control plant, while the other received Cr(VI) concentrations equal to 0.5, 1, 3 and 5 mgl(-1). Cr(VI) concentrations of 0.5 mgl(-1) caused significant inhibition of the nitrification process (up to 74% decrease in ammonia removal efficiency). On the contrary, the effect of Cr(VI) on organic substrate removal was minor for concentrations up to 5 mgl(-1), indicating that heterotrophic microorganisms are less sensitive to Cr(VI) than nitrifiers. Activated sludge floc size and structure characterization showed that Cr(VI) concentrations higher than 1 mgl(-1) reduced the filaments abundance, causing the appearance of pin-point flocs and free-dispersed bacteria. Additionally, the variability of protozoa and rotifers was reduced. As a result of disperse growth, effluent quality deteriorated, since significant amounts of suspended solids escaped with the effluent. Termination of Cr(VI) addition led to a partial recovery of the nitrification process (up to 57% recovery). Similar recovery signs were not observed for activated sludge floc size and structure. Finally, shock loading to the control plant with 5 mgl(-1) Cr(VI) for 2 days resulted in a significant inhibition of the nitrification process and a reduction in filamentous microorganisms abundance.

Bacteria↗

Impulse responses of a monoethylamine-fed fluidized bed reactor.

The responses of a steady-state, continuous-flow, completely-mixed fluidized bed reactor (FBR) to a range of monoethylamine (MEA) impulses are analyzed in terms of its combined carbon oxidation and nitrification efficiencies. Immobilized cells are cultivated at a mean cell residence time (MCRT) that exceeds 75 days. Responses due to bacterial activities and physical flows are separately estimated using a methodology based on mass balance calculations. MEA inhibition becomes evident when respective critical impulse loadings are exceeded, i.e., 0.12 mg TOC/mg VS for carbon oxidation and 0.021 mg TOC/mg VS for nitrification (TOC: total organic carbon, VS: volatile solids). Nitrifying cells are shown to be more susceptible to MEA impulses than their heterotrophic counterparts. However, the presence of nitrification activities under the conditions tested demonstrates the advantages of cell immobilization that offer greater flexibility when challenged with suddenly increased MEA loads over a short period of time. Mass balance calculations on nitrogen species confirms that 0.583 mg NH(4)(+)-N is produced per mg MEA-C removed when the assimilatory nitrogen requirements for cell synthesis are negligible.

Bioreactors↗

The effect of residual ammonia concentration under aerobic conditions on the growth of Microthrix parvicella in biological nutrient removal plants.

It is demonstrated with two parallel single reactor intermittently aerated nitrification denitrification systems fed municipal wastewater as influent, that Microthrix parvicella bulking can be stimulated and cured by manipulating the ammonia concentration in the aerobic period (by inhibiting the nitrifiers) to high and low values respectively. The proliferation or not of M. parvicella is hypothesized to be due to their requirement for ammonia as a nitrogen source for growth. In terms of this hypothesis, if nitrification is rapid and complete, ammonia is not freely available and will limit M. parvicella growth. If nitrification is not complete for whatever reason, ammonia is available for the growth of the slow growing M. parvicella, enabling their proliferation to cause a bulking sludge. This hypothesis does not overturn or replace the anoxic-aerobic (AA, or low Food/Microorganism, F/M, ratio) filament bulking hypothesis of Casey et al. (Water SA 25(4) (1999) 425) but appears to be additional to it. Future research will focus on determining how elements of both hypotheses superimpose on the conditions in BNR systems, to produce an AA filament bulking sludge or not.

Actinobacteria↗

Annual cycle of nitrogen removal by a pilot-scale subsurface horizontal flow in a constructed wetland under moderate climate.

The annual course of nitrogen removal in a stable operating subsurface horizontal flow constructed wetland (SSF) in a moderate climate was evaluated using a large pool of data from 4 years of operation. In spring and autumn removal efficiencies were found to depend on the nitrogen load in a linear mode. The efficiencies in winter and summer differed extremely (mean removal rates of 0.15/0.7 g m(-2) d(-1) (11%/53%) in January/August) and were independent of the nitrogen load (0.7-1.7 g m(-2) d(-1)) in principle. Oscillations of the removal rates in spring, forming several maxima, suggest seasonal specific effects caused by the dynamics of the plant-physiology finally determining the nitrification efficiency, i.e. via O(2)-supply. Nitrification is limited by temperature during all seasons and surprisingly in midsummer additionally restricted by other seasonal aspects forming a clear-cut relative nitrification minimum (mean rate of 0.43 g m(-2) d(-1) (32%)) in July. The importance and the effect of the plants' gas exchange and oxygen input into the rhizosphere are discussed. Denitrification was nearly complete in midsummer and was clearly restricted at seasonal temperatures below 15 degrees C.

Climate↗

Organic matter removal in combined anaerobic-aerobic fixed-film bioreactors.

A combination of two fixed-film bioreactors (FFB) with arranged media, the first anaerobic and the second aerobic, connected in series with recirculation was fed continuously for 133 days with wastewater from a poultry slaughterhouse. Oxidation of the organic carbon compounds and nitrification were carried out in the aerobic FFB and methanogenesis and denitrification were performed in the anaerobic FFB. The average organic loading rate was 0.39 kg COD/m3d and 92% removal efficiencies of organic matter were achieved. COD-removal occurred mainly in the anaerobic FFB, increasing when the recirculation ratio rose from 1 to 6 due to the increase in the anoxic denitrification. The influence of the C/N ratio of the raw wastewater over the proportion in which the COD-removal was carried out by oxidation in the aerobic FFB, methanogenesis or denitrification in the anaerobic FFB was studied. When the volume of the aerobic FFB became smaller than that of the anaerobic one the fraction of organic matter removed in the anaerobic FFB increased, but also the ratio between the respective volumetric rates (rCODan/rCODae) increased. High recirculation and low C/NO-N ratio in the anaerobic FFB feed favoured the denitrification to the detriment of the methanogenic process. Regarding nitrogen removal for nitrogen applied loads around 0.064 kg TKN/m3d the removal efficiency was of 95%, which decreased to 84% for 0.14 kg TKN/m3d. The stability of the nitrification process was the controlling factor of the nitrogen removal. High ammonia concentration caused by high recirculation ratio, specially when the aerobic FFB volume was smaller, caused nitrification inhibition which destabilised the system.

Abattoirs↗

Effects of predation and ORP conditions on the performance of nitrifiers in activated sludge systems.

Effects of changing oxidation-reduction potential (ORP) and grazing of protozoa on nitrifiers in activated sludge systems was investigated. This study used sequencing batch reactors which were acclimated under aerobic and alternating anoxic/aerobic conditions, with and without inhibition of protozoa, the predatory microorganisms. The feed used was a synthetic wastewater containing beef and yeast extracts as a carbon source. It was found that the biomass, determined by mixed liquor volatile suspended solids (MLVSS) in the reactors, was significantly affected by predation while ORP (aerobic and alternating anoxic/aerobic conditions) has no impact on the MLVSS regardless of the presence or absence of predation. However, the nitrification rates in the reactors show completely different trends indicating that the ORP of the system has a significant impact on the rates while predation does not. It was found that nitrification rates in alternating reactors were almost double the rates in the aerobic reactors, both with and without predatory inhibition. The decay rate of autotrophic bacteria (b(A)) in aerobic reactors was determined by tracking the decrease of the maximum nitrification rate under both anoxic and aerobic starvation conditions. The b(A) in alternating anoxic/aerobic reactors was also determined under alternating starvation conditions. It was found that, in any case, the alternating anoxic/aerobic autotrophic biomass b(A) was much smaller that the b(A) of aerobic biomass determined under aerobic and anoxic starvation conditions. The alternating anoxic/aerobic b(A) was 62.1% less than the aerobic biomass b(A) under aerobic starvation and 40.2% less for the aerobic biomass starved under anoxic conditions. No statistically significant differences in b(A) were observed between reactors with or without the inhibition of predators.

Animals↗

Advanced treatment of sewage by pre-coagulation and biological filtration process.

A pre-coagulation and bio-filtration process for advanced treatment of sewage was developed and experimentally discussed with a pilot plant. The bio-filtration unit consists of a denitrification filter, a nitrification filter with side stream to the denitrification filter, and a polishing filter with anoxic and aerobic parts. Concentrations of SS, T-COD(Cr), T-carbonaceous BOD, T-N and T-P in the effluent were stably kept at less than 3, 20, 5mg/L, 2mg N/L and 0.2mg P/L, respectively, and transparency at higher than 100 cm, under total hydraulic retention time of 3.2h in the bio-filtration parts (filter-bed). ORP in an anoxic tank before a nitrification tank should be at a low level of less than -120 mV to keep remaining NO(-)(x) - N less than 1mg N/L, but must be maintained at a level higher than -150 mV. The maximum nitrogen-loading rate under a water temperature of 18 degrees C should be less than 0.25 kg N/(m(3)-filter-bed.d). Concentrations of microorganisms kept in the reactors were as high as 4000-5000 mg COD/L-filter-bed. Denitrification activity of 0.4 or 0.7 kg N/(m(3)-filter-bed.d), and nitrification activity of 0.3 kg N/(m(3)-filter-bed.d) were obtained, respectively, under a water temperature of about 18 degrees C. Backwashing in each tank as well as methanol addition and aeration in the polishing filter were operated successfully by the automatic control systems. These results proved that this process is applicable to advanced treatment of sewage with easy maintenance.

Automation↗

Chemical and microbial hypotheses explaining the effect of wastewater treatment plant discharges on the nitrifying communities in freshwater sediment.

Nitrification is a microbial key step of the nitrogen cycle, which performs the oxidation of ammonium to nitrate, via nitrite. In aquatic environments, it mainly takes place in the sediment or is associated with suspended particles. Wastewater treatment plant (WTP) discharges in rivers may disrupt sediment nitrification: this impact is related to nitrogen inputs (mainly NH(4)(+) and organic nitrogen) but could also depend on the nitrifying bacteria inputs which have been proved to survive downstream WTP discharge points. The aim of the present study was to assess the effect of NH(4)(+) and nitrifying bacteria inputs on the two steps of nitrification in freshwater sediments. To avoid natural sites constraints and to control the main environmental parameters, we used microcosms to simulate a river receiving different types of WTP discharges. These microcosms were composed of five glass dual-walls reactors (6 l) containing sediment and continuously filled (controlled flow) with river water and WTP effluent. Two types of effluents were tested: a non-nitrified one (high NH(4)(+) input, very few nitrifying bacteria) and a nitrified one (low NH(4)(+) input, more nitrifying bacteria), at different effluent/freshwater ratios (0/100, 20/80, 40/60 and 80/20). Changes in the ammonium- and nitrite-oxidizing communities were assessed by the Most Probable Number method, and changes in potential ammonium-oxidizing activity and potential nitrite-oxidizing activity were determined by incubations with specific inhibitors (sodium chlorate and allylthiourea). In most of the cases the presence of effluent induced significant changes of the nitrifying bacteria densities and potential activities in the sediment. This effect indicates generally a loss of specific potential activity and in most of the time is significant for a high effluent/river water ratio (40% to 80%). In our experimental conditions and in the case of a large WTP discharge, the nitrifying potential in freshwater sediments could thus be significantly modified.

Bacteria↗