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.     

Nitrous oxide emissions for early warning of biological nitrification failure in activated sludge

Experiments were carried out to establish whether nitrous oxide (N₂O) could be used as a non-invasive early warning indicator for nitrification failure. Eight experiments were undertaken; duplicate shocks DO depletion, influent ammonia increases, allylthiourea (ATU) shocks and sodium azide (NaN₃) shocks were conducted on a pilot-scale activated sludge plant which consisted of a 315 L completely mixed aeration tank and 100 L clarifier. The process performed well during pre-shock stable operation; ammonia removals were up to 97.8% and N₂O emissions were of low variability (<0.5 ppm). However, toxic shock loads produced an N₂O response of a rise in off-gas concentrations ranging from 16.5 to 186.3 ppm, followed by a lag-time ranging from 3 to 5 h ((0.43-0.71) × HRT) of increased NH₃-N and/or NO₂⁻ in the effluent ranging from 3.4 to 41.2 mg L⁻¹. It is this lag-time that provides the early warning for process failure, thus mitigating action can be taken to avoid nitrogen contamination of receiving waters.     

Nitrous oxide emission during wastewater treatment

Nitrous oxide (N₂O), a potent greenhouse gas, can be emitted during wastewater treatment, significantly contributing to the greenhouse gas footprint. Measurements at lab-scale and full-scale wastewater treatment plants (WWTPs) have demonstrated that N₂O can be emitted in substantial amounts during nitrogen removal in WWTPs, however, a large variation in reported emission values exists. Analysis of literature data enabled the identification of the most important operational parameters leading to N₂O emission in WWTPs: (i) low dissolved oxygen concentration in the nitrification and denitrification stages, (ii) increased nitrite concentrations in both nitrification and denitrification stages, and (iii) low COD/N ratio in the denitrification stage. From the literature it remains unclear whether nitrifying or denitrifying microorganisms are the main source of N₂O emissions. Operational strategies to prevent N₂O emission from WWTPs are discussed and areas in which further research is urgently required are identified.     

Effect of heat recovery from raw wastewater on nitrification and nitrogen removal in activated sludge plants

By recovery of heat from the raw wastewater in the sewer system, the influent temperature of a wastewater treatment plant (WWTP) is reduced. This can have a negative effect on nitrification in the WWTP, since this process strongly depends on temperature. The analysis of the temperature regime in the WWTP of Zurich, Switzerland, revealed that in the cold season, the effluent temperature is about 0.7 degrees C higher than the influent temperature and that nitrification is not affected by a decrease of the influent wastewater temperature lasting for a couple of hours only, but is significantly affected by a longer lasting temperature decrease. Three diagrams were developed with a steady-state model, from which the consequences of a permanent temperature decrease on the nitrification safety factor, aerobic sludge retention time and total nitrogen removal can be evaluated. Using simulations with a dynamic model, calibrated for the Zurich WWTP, a quantitative relationship between the wastewater temperature and the ammonium effluent concentration was established. This relationship can, in combination with measured effluent concentrations of an existing WWTP, be used to predict the increase of the ammonium effluent concentration in this plant resulting from a permanent decrease of the wastewater influent temperature.     

Effects of chromium on activated sludge and on the performance of wastewater treatment plants: A review

Chromium is a heavy metal of commercial importance, thus significant amounts are released in wastewaters. Chromium in wastewaters and in the aquatic environment is primarily encountered in oxidation stages +3 (Cr^(III)) and +6 (Cr^(VI)). Recent publications suggest that Cr^(VI) compounds are more toxic than Cr^(III) ones, while Cr^(III) has been identified as trace element, at least for complex organisms. With respect to chromium species mobility, Cr^(VI) can cross cellular membranes, which then may be oxidized to Cr^(III) and react with intracellular biomolecules. Clear conclusions cannot be derived about the critical chromium concentrations that affect activated sludge growth, as the latter is a function of a number of factors. Broadly, may be supported that activated sludge growth is stimulated at Cr^(III) concentrations up to 15 mg L⁻¹, above which is inhibited, with lethal doses lying above 160 mg Cr^(III) L⁻¹. On the other hand, literature data on Cr^(VI) effects on activated sludge are even more controversial. A number of reports support that Cr^(VI) is toxic to activated sludge at concentrations above 5 mg L⁻¹, while others report growth stimulation at concentrations up to 25 mg L⁻¹. However, all reports agree that Cr^(VI) is definitely an activated sludge growth inhibitor at higher concentrations, while 80 mg Cr^(VI) L⁻¹ have been identified as lethal dose. A number of factors have been identified to influence chromium toxicity on activated sludge, such as, pH, biomass concentration, presence of organic substances or other heavy metals, acclimation process, exposure time, etc. Naturally, the presence of chromium species in wastewaters may affect the performance of wastewater treatment plants often causing malfunctions, particularly for industrial wastewaters containing relatively high chromium concentrations. The present work reviews in a critical way the published literature on chromium effects on activated sludge, and on the operation of wastewater treatment plants.     

Tetracycline resistance genes in activated sludge wastewater treatment plants

The development and proliferation of antibiotic resistance in pathogenic, commensal, and environmental microorganisms is a major public health concern. The extent to which human activities contribute to the maintenance of environmental reservoirs of antibiotic resistance is poorly understood. In the current study, wastewater treatment plants (WWTPs) were investigated as possible sources of tetracycline resistance via qualitative PCR and quantitative PCR (qPCR). Various WWTPs and two freshwater lakes were surveyed for the presence of an array of 10 tetracycline resistance determinants (tet(R)): tet(A)-(E), tet(G), tet(M), tet(O), tet(Q), tet(S). All WWTP samples contained more different types of tet(R) genes, as compared to the lake water samples. Gene copy numbers of tet(G) and tet(Q) in these samples were quantified via qPCR and normalized to both the volume of original sample and to the amount of DNA extracted per sample (a proxy for bacterial abundance). Concentrations of tet(Q) were found to be highest in wastewater influent while tet(G) concentrations were highest in activated sludge. Investigation of the effects of UV disinfection on wastewater effluent showed no reduction in the number of detectable tet(R) gene types.     

Nitrous oxide emissions from the oxidation tank of a pilot activated sludge plant

This study discusses the results of the continuous monitoring of nitrous oxide emissions from the oxidation tank of a pilot conventional wastewater treatment plant. Nitrous oxide emissions from biological processes for nitrogen removal in wastewater treatment plants have drawn great attention over the last years, due to the high greenhouse effect. However, even if several studies have been carried out to quantify nitrous oxide emission rates from different types of treatment, quite wide ranges have been reported. Only grab samples or continuous measurements over limited periods were considered in previous studies, which can account for the wide variability of the obtained results. Through continuous monitoring over several months, our work tries to fill this gap of knowledge and get a deeper insight into nitrous oxide daily and weekly emission dynamics. Moreover, the influence of some operating conditions (sludge age, dissolved oxygen concentration in the oxidation tank, nitrogen load) was studied to determine good practices for wastewater treatment plant operation aiming at the reduction of nitrous oxide emissions. The dissolved oxygen set-point is shown to play a major role in nitrous oxide emissions. Low sludge ages and high nitrogen loads are responsible for higher emissions as well. An interesting pattern has been observed, with quite negligible emissions during most of the day and a peak with a bell-like shape in the morning in the hours of maximum nitrogen load in the plant, correlated to the ammonia and nitrite peaks in the tank.     

Improvement of activated sludge dewaterability by mild thermal treatment in CaCl2 solution

Activated sludge dewatering is of great importance in sludge treatment and disposal. To enhance the dewaterability, a novel method was performed by treating the sludge under mild temperature (50-90 °C) in CaCl₂ solution (3.7-1110.0 mg/g dry sludge). The capillary suction time, zeta potential, Fourier-transformed infrared spectra, concentration of soluble protein and carbohydrates were employed to characterize the dewaterability and influencing mechanism. The sludge dewaterability was deteriorated with single thermal treatment, but significantly promoted in CaCl₂ solution and advanced further together with thermal treatment. An increasing CaCl₂ dosage reduced the surface charge remarkably, and a higher temperature could strengthen this impact. The spectra indicate that Ca²⁺ could interact with the protein, phenols and O-H functional group in the flocs. The thermal treatment could cause the solubilization of protein and carbohydrates, providing more binding sites for Ca²⁺ to establish a strong bridging among the flocs. As CaCl₂ dosage elevated, the soluble carbohydrates showed a reduction trend, while the soluble protein lowered firstly and then bounced back except that remained unchanged at room temperature. A bridging equilibrium is presumed to exist between Ca²⁺ and the soluble protein. And the bridging between Ca²⁺ and the soluble carbohydrates plays a more important role in the dewatering. The sludge dewaterability was successfully and economically improved by thermal treatment in CaCl₂ solution.     

Biodegradability of activated sludge organics under anaerobic conditions

From an experimental and theoretical investigation of the continuity of activated sludge organic (COD) compounds along the link between the fully aerobic or N removal activated sludge and anaerobic digestion unit operations, it was found that the unbiodegradable particulate organics (i) originating from the influent wastewater and (ii) generated by the activated sludge endogenous process, as determined from response of the activated sludge system, are also unbiodegradable under anaerobic digestion conditions. This means that the activated sludge biodegradable organics that can be anaerobically digested can be calculated from the active fraction of the waste activated sludge based on the widely accepted ordinary heterotrophic organism (OHO) endogenous respiration/death regeneration rates and unbiodegradable fraction. This research shows that the mass balances based steady state and dynamic simulation activated sludge, aerobic digestion and anaerobic digestion models provide internally consistent and externally compatible elements that can be coupled to produce plant wide steady state and dynamic simulation WWTP models.     

Effects of sonication on bacteria viability in wastewater treatment plants evaluated by flow cytometry--fecal indicators, wastewater and activated sludge

The application of sonication to wastewater or sludge contributes to the dispersion of aggregates, the solubilisation of particulate matter with an increase in its biodegradability, the damage of microorganisms due to the loss of cellular membrane integrity. This research is aimed at investigating the effects of sonication at 20kHz frequency on viability of microorganisms present in raw wastewater and activated sludge taken from a municipal wastewater treatment plant, as well as pure strains of Escherichia coli and E. faecalis. Flow cytometry was applied for the identification and quantification of viable and dead bacteria free in the bulk liquid, after the fluorescent staining of cellular nucleic acids. The main results showed that: (i) cells of E. coli were highly sensitive to sonication, even at low specific ultrasonic energy (E(s)), and disintegration of a large amount of cells was observed; (ii) on the contrary E. faecalis were more resistant than E. coli, even if high levels of E(s) were applied; (iii) bacteria in raw wastewater exhibited a dynamic of viable and dead bacteria similar to E. coli; (iv) in activated sludge samples, low levels of E(s) produced a prevalent disaggregation of flocs releasing single cells in the bulk liquid, while disruption of bacteria was induced only by very high levels of E(s).     

Role of extracellular polymeric substances in bioflocculation of activated sludge microorganisms under glucose-controlled conditions

Extracellular polymeric substances (EPS) secreted by suspended cultures of microorganisms from an activated sludge plant in the presence of glucose were characterized in detail using colorimetry, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy. EPS produced by the multi-species community were similar to literature reports of pure cultures in terms of functionalities with respect to C and O but differed subtly in terms of N and P. Hence, it appears that EPS produced by different microorganisms maybe homologous in major chemical constituents but may differ in minor components such as lipids and phosphodiesters. The role of specific EPS constituents on microbial aggregation was also determined. The weak tendency of microorganisms to bioflocculate during the exponential growth phase was attributed to electrostatic repulsion when EPS concentration was low and acidic in nature (higher fraction of uronic acids to total EPS) as well as reduced polymer bridging. However, during the stationary phase, polymeric interactions overwhelmed electrostatic interactions (lower fraction of uronic acids to total EPS) resulting in improved bioflocculation. More specifically, microorganisms appeared to aggregate in the presence of protein secondary structures including aggregated strands, β-sheets, α- and 3-turn helical structures. Bioflocculation was also favored by increasing O-acetylated carbohydrates and overall C-(O,N) and functionalities.     

Decay processes of nitrifying bacteria in biological wastewater treatment systems

A knowledge of the decay rates of autotrophic bacteria is important for reliably modeling nitrification in activated sludge plants. The introduction of nitrite to activated sludge models also requires the separate determination of the kinetics of ammonia- and nitrite-oxidizing bacteria. Batch experiments were carried out in order to study the effects of different oxidiation-reduction potential conditions and membrane separation on the separate decay of these bacteria. It was found that decay is negligible in both cases under anoxic conditions. No significant differences were detected between the membrane and conventional activated sludge. The aerobic decay of these two types of bacteria did not diverge significantly either. However, the measured loss of autotrophic activity was only partly explained by the endogenous respiration concept as incorporated in activated sludge model no. 3 (ASM3). In contrast to nitrite-oxidizing bacteria, ammonia-oxidizing bacteria needed 1-2 h after substrate addition to reach their maximum growth rate measured as a maximum OUR. This pattern could be successfully modeled using the ASM3 extended by enzyme kinetics. The significance of these findings on wastewater treatment is discussed on the basis of the extended ASM3.     

Predicting odour emissions from wastewater treatment plants by means of odour emission factors

In this study, the results of odour concentration measurements on different wastewater treatment plants are presented and used in order to estimate the odour emission factors relevant to single odour sources. An odour emission factor is a representative value that relates the quantity of odour released to the atmosphere to a specific activity index, which in this case was the plant treatment capacity, resulting in an odour emission factor expressed in odour units per cubic metre of treated sewage. The results show that the major odour source of a wastewater treatment plant is represented by the primary sedimentation (with an OEF equal to 1.9 × 10⁵ ou_E m⁻³). In general, the highest OEFs are observed in correspondence of the first steps of the wastewater depuration cycle (OEF between 1.1 × 10⁴ ou_E m⁻³ and 1.9 × 10⁵ ou_E m⁻³) and tend to decrease along the depuration process (OEF between 7.4 × 10³ ou_E m⁻³ and 4.3 × 10⁴ ou_E m⁻³). In general, the OEFs calculated according to this approach represent a model for a rough prediction of odour emissions independently from the specific characteristics of the different plants.     

DNA microarray detection of nitrifying bacterial 16S rRNA in wastewater treatment plant samples

A small scale DNA microarray containing a set of oligonucleotide probes targeting the 16S rRNAs of several groups of nitrifying bacteria was developed for the monitoring of wastewater treatment plant samples. The microarray was tested using reference rRNAs from pure cultures of nitrifying bacteria. Characterization of samples collected from an industrial wastewater treatment facility demonstrated that nitrifying bacteria could be detected directly by microarray hybridization without the need for PCR amplification. Specifically, the microarray detected Nitrosomonas spp. but did not detect Nitrobacter. The specificity and sensitivity of direct detection was evaluated using on-chip dissociation analysis, and by two independent analyses--an established membrane hybridization format and terminal restriction fragment length polymorphism fingerprinting (T-RFLP). The latter two analyses also revealed Nitrospira and Nitrobacter to be contributing populations in the treatment plant samples. The application of DNA microarrays to wastewater treatment systems, which has been demonstrated in the current work, should offer improved monitoring capabilities and process control for treatment systems, which are susceptible to periodic failures.     

Direct quantification of bacterial biomass in influent, effluent and activated sludge of wastewater treatment plants by using flow cytometry

A rapid multi-step procedure, potentially amenable to automation, was proposed for quantifying viable and active bacterial cells, estimating their biovolume using flow cytometry (FCM) and to calculate their biomass within the main stages of a wastewater treatment plant: raw wastewater, settled wastewater, activated sludge and effluent. Fluorescent staining of bacteria using SYBR-Green I + Propidium Iodide (to discriminate cell integrity or permeabilisation) and BCECF-AM (to identify enzymatic activity) was applied to count bacterial cells by FCM. A recently developed specific procedure was applied to convert Forward Angle Light Scatter measured by FCM into the corresponding bacterial biovolume. This conversion permits the calculation of the viable and active bacterial biomass in wastewater, activated sludge and effluent, expressed as Volatile Suspended Solids (VSS) or particulate Chemical Oxygen Demand (COD). Viable bacterial biomass represented only a small part of particulate COD in raw wastewater (4.8 ± 2.4%), settled wastewater (10.7 ± 3.1%), activated sludge (11.1 ± 2.1%) and effluent (3.2 ± 2.2%). Active bacterial biomass counted for a percentage of 30-47% of the viable bacterial biomass within the stages of the wastewater treatment plant.     

Diversity study of nitrifying bacteria in full-scale municipal wastewater treatment plants

We hypothesize that activated-sludge processes having stable and complete nitrification have significant and similar diversity and functional redundancy among its ammonia- and nitrite-oxidizing bacteria, despite differences in temperature, solids retention time (SRT), and other operating conditions. To evaluate this hypothesis, we examined the diversity of nitrifying bacterial communities in all seven water-reclamation plants (WRPs) operated by Metropolitan Water Reclamation District of Greater Chicago (MWRDGC). These plants vary in types of influent waste stream, plant size, water temperature, and SRT. We used terminal restriction fragment length polymorphism (T-RFLP) targeting the 16S rRNA gene and group-specific ammonia-monooxygenase functional gene (amoA) to investigate these hard-to-culture nitrifying bacteria in the full-scale WRPs. We demonstrate that nitrifying bacteria carrying out the same metabolism coexist in all WRPs studied. We found ammonia-oxidizing bacteria (AOB) belonging to the Nitrosomonas europaea/eutropha, Nitrosomonas oligotropha, Nitrosomonas communis, and Nitrosospira lineages in all plants. We also observed coexisting Nitrobacter and Nitrospira genera for nitrite-oxidizing bacteria (NOB). Among the factors that varied among the WRPs, only the seasonal temperature variation seemed to change the nitrifying community, especially the balance between Nitrosospira and Nitrosomonas, although both coexisted in winter and summer samples. The coexistence of various nitrifiers in all WRPs is evidence of functional redundancy, a feature that may help maintain the stability of the system for nitrification.     

Pharmaceutical chemicals and endocrine disrupters in municipal wastewater in Tokyo and their removal during activated sludge treatment

We measured six acidic analgesics or anti-inflammatories (aspirin, ibuprofen, naproxen, ketoprofen, fenoprofen, mefenamic acid), two phenolic antiseptics (thymol, triclosan), four amide pharmaceuticals (propyphenazone, crotamiton, carbamazepine, diethyltoluamide), three phenolic endocrine disrupting chemicals (nonylphenol, octylphenol, bisphenol A), and three natural estrogens (17beta-estradiol, estrone, estriol) in 24-h composite samples of influents and secondary effluents collected seasonally from five municipal sewage treatment plants in Tokyo. Aspirin was most abundant in the influent, with an average concentration of 7300 ng/L (n = 16), followed by crotamiton (921 ng/L), ibuprofen (669 ng/L), triclosan (511 ng/L), and diethyltoluamide (503 ng/L). These concentrations were 1 order of magnitude lower than those reported in the USA and Europe. This can be ascribed to lower consumption of the pharmaceuticals in Japan. Aspirin, ibuprofen, and thymol were removed efficiently during primary + secondary treatment (> 90% efficiency). On the other hand, amide-type pharmaceuticals, ketoprofen, and naproxen showed poor removal (< 50% efficiency), which is probably due to their lower hydrophobicity (logKow < 3). Because of the persistence of crotamiton during secondary treatment, crotamiton was most abundant among the target pharmaceuticals in the effluent. This is the first paper to report ubiquitous occurrence of crotamiton, a scabicide, in sewage. Because crotamiton is used worldwide and it is persistent during secondary treatment, it is a promising molecular marker of sewage and secondary effluent.     

Enumeration of protozoan ciliates in activated sludge: Determination of replicate number using probability

A new approach to the enumeration of ciliate protozoa in activated sludge is described. A 25 μL sub-sample volume is optimal for routine analysis using a standard slide and 24 × 24 mm cover slip requiring between 20 and 40 min per sub-sample for full enumeration and identification of species. However, to achieve high probability (≥95%) of recovering all species large numbers of replicates are required (i.e. 23-47). To achieve high probabilities of recovery using less replicates it is necessary to neglect rare species with low densities (<0.5%); based on the assumption that they do not play a significant role in plant performance. The precise number of replicates required for different probabilities of recovering species is determined by conducting an initial pilot survey analysing a minimum of 8 replicates and using a probability equation to determine the optimum replicate number for that particular plant. Six replicate 25 μL sub-samples provided excellent species recovery (90-95% excluding up to 3 rare species), while analysing just two or three replicates, as commonly used in previous wastewater studies, only gave probabilities of 25 and 50% respectively for the same recovery. Ciliate analysis should be completed within 8 h of collection with significant changes in community structure occurring beyond this period.     

Occurrence and removal of N-nitrosamines in wastewater treatment plants

The presence of nitrosamines in wastewater might pose a risk to water resources even in countries where chlorination or chloramination are hardly used for water disinfection. We studied the variation of concentrations and removal efficiencies of eight N-nitrosamines among 21 full-scale sewage treatment plants (STPs) in Switzerland and temporal variations at one of these plants. N-nitrosodimethylamine (NDMA) was the predominant compound in STP primary effluents with median concentrations in the range of 5-20 ng/L, but peak concentrations up to 1 μg/L. N-nitrosomorpholine (NMOR) was abundant in all plants at concentrations of 5-30 ng/L, other nitrosamines occurred at a lower number of plants at similar levels. From concentrations in urine samples and domestic wastewater we estimated that human excretion accounted for levels of <5 ng/L of NDMA and <1 ng/L of the other nitrosamines in municipal wastewater, additional domestic sources for <5 ng/L of NMOR. Levels above this domestic background are probably caused by industrial or commercial discharges, which results in highly variable concentrations in sewage. Aqueous removal efficiencies in activated sludge treatment were in general above 40% for NMOR and above 60% for the other nitrosamines, but could be lower if concentrations were below 8-15 ng/L in primary effluent. We hypothesize that substrate competition in the cometabolic degradation explains the occurrence of such threshold concentrations. An additional sand filtration step resulted in a further removal of nitrosamines from secondary effluents even at low concentrations. Concentrations released to surface waters were largely below 10 ng/L, suggesting a low impact on Swiss water resources and drinking water generation considering the generally high environmental dilution and possible degradation. However, local impacts in case a larger fraction of wastewater is present cannot be ruled out.     

ICEAS工艺启动及活性污泥培养驯化应用实例

结合福州市金山污水处理厂工艺及及活性污泥培养驯化实例,分析总结ICEAS工艺启动及活性污泥培养驯化经验。