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).     

Degradation of phthalate esters in an activated sludge wastewater treatment plant

Efficient removal of phthalate esters (PE) in wastewater treatment plants (WWTP) is becoming an increasing priority in many countries. In this study, we examined the fate of dimethyl phthalate (DMP), dibutyl phthalate (DBP), butylbenzyl phthalate (BBP), and di-(2-ethylhexyl) phthalate (DEHP) in a full scale activated sludge WWTP with biological removal of nitrogen and phosphorus. The mean concentrations of DMP, DBP, BBP, and DEHP at the WWTP inlet were 1.9, 20.5, 37.9, and 71.9 μg/L, respectively. Less than 0.1%, 42%, 35%, and 96% of DMP, DBP, BBP, and DEHP was associated with suspended solids, respectively. The overall microbial degradation of DMP, DBP, BBP, and DEHP in the WWTP was estimated to be 93%, 91%, 90%, and 81%, respectively. Seven to nine percent of the incoming PE were recovered in the WWTP effluent. Factors affecting microbial degradation of DEHP in activated sludge were studied using [U-¹⁴C-ring] DEHP as tracer. First order rate coefficients for aerobic DEHP degradation were 1.0×10⁻², 1.4×10⁻², and 1.3×10⁻³ at 20, 32, and 43 °C, respectively. Aerobic degradation rates decreased dramatically under aerobic thermophilic conditions (<0.1×10⁻² h⁻¹ at 60 °C). The degradation rate under anoxic denitrifying conditions was 0.3×10⁻² h⁻¹, whereas the rate under alternating conditions (aerobic-anoxic) was 0.8×10⁻² h⁻¹. Aerobic DEHP degradation in activated sludge samples was stimulated 5-9 times by addition of a phthalate degrading bacterium. The phthalate degrading bacterium was isolated from activated sludge, and maintained a capacity for DEHP degradation while growing on vegetable oil. Collectively, the results of the study identified several controls of microbial PE degradation in activated sludge. These controls may be considered to enhance PE degradation in activated sludge WWTP with biological removal of nitrogen and phosphorus.     

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.     

Diurnal variations in the occurrence and the fate of hormones and antibiotics in activated sludge wastewater treatment in Oslo, Norway

We present an assessment of the dynamics in the influent concentration of hormones (estrone, estriol) and antibiotics (trimethoprim, sulfamethoxazole, tetracycline, ciprofloxacin) in the liquid phase including the efficiency of biological municipal wastewater treatment. The concentration of estradiol, 17-α−ethinylestradiol, doxycycline, oxytetracycline, demeclocycline, chlortetracycline, cefuroxime, cyclophosphamide, and ifosfamide were below the limit of detection in all of the sewage samples collected within this study. Two different types of diurnal variation pattern were identified in the influent mass loads of selected antibiotics and hormones that effectively correlate with daily drug administration patterns and with the expected maximum human hormone release, respectively. The occurrence of natural hormones and antimicrobials, administered every 12 hours, shows a daily trend of decreasing contaminant mass load, having the maximum values in the morning hours. The occurrence of antibiotics, typically administered every 8 hours, indicates a daily peak value in samples collected under the highest hydraulic loading. The efficiency of biological removal of both hormones and antibiotics is shown to be limited. Compared to the values obtained in the influent samples, increased concentrations are observed in the biologically treated effluent for trimethoprim, sulfamethoxazole and ciprofloxacin, mainly as a result of deconjugation processes. Ciprofloxacin is shown as the predominant antimicrobial compound in the effluent, and it is present at quantities approximately 10 fold greater than the total mass of the other of the compounds due to poor removal efficiency and alternating solid-liquid partitioning behaviour. Our results suggest that, to increase the micro-pollutant removal and the chemical dosing efficiency in enhanced tertiary treatment, significant benefits can be derived from the optimisation of reactor design and the development of control schemes that accounts for diurnal secondary effluent micro-pollutant and hydraulic loading patterns.     

Bulking in activated sludge plants treating paper mill wastewaters

As part of a larger project to examine the causes of bulking in activated sludge plants treating wastewaters from paper mills, two laboratory-scale activated sludge plants were run in parallel. This was to evaluate the impact of variable environmental and operational parameters on the sludge ecology, and in particular, on the filamentous bacteria in the sludge. A survey of paper mills in the UK showed that there was a significant difference between the activated sludge plants at the paper mills using virgin fibre and those using recycled fibre. An examination of samples from activated sludge plants at the paper mills showed that there did not appear to be any significant correlation between filament abundance and the settlement characteristics as measured by stirred specific volume index (SSVI). The surface charge carried by the sludge particles was also measured and it was found that this parameter was better related to the SSVI. Similar results were found for samples from the laboratory plants. Seven different sets of operational conditions were applied to the laboratory-scale plants. These were aimed at simulating the conditions noted for the full-scale plants during the mill survey. The effects of low dissolved oxygen and low organic loading rates were examined. The effect of inorganic sulphur compounds and volatile fatty acids was studied. A change in raw material from virgin fibre to recycled fibre (corrugated paper) caused an immediate and continuous deterioration in settlement. Divalent cations, calcium and magnesium, were found to be successful in controlling settlement in the final clarifier.     

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.     

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.     

Nitrous oxide emissions from secondary activated sludge in nitrifying conditions of urban wastewater treatment plants: effect of oxygenation level

In order to better understand the mechanisms of N(2)O emissions from nitrifying activated sludge of urban WWTPs, sludge from the Valenton plant (Paris conurbation) are subjected to lab-scale batch experiments under various conditions of oxygenation. The results show that the highest N(2)O emissions (7.1 microgN-N(2)OgSS(-1) h(-1) in average) occur at a dissolved oxygen (DO) concentration of around 1mgO(2)L(-1). These high emissions at low oxygenation (from 0.1 to 2 mg O(2)L(-1)) are due to two processes: autotrophic nitrifier denitrification and heterotrophic denitrification. Nitrifier denitrification always dominates, representing from 58% to 83% of the N(2)O production. This N(2)O production originating from nitrifying activated sludge becomes 8 times higher when nitrite is added at a DO of 1 mg O(2)L(-1); a decrease is observed both at higher and lower oxygenation. Heterotrophic denitrification represents less than 50% of the N(2)O production, decreasing from 42% to 17% when oxygenation increases from 0.1 to 2 mg O(2) L(-1). We show that ammonium oxidizing bacteria (AOB) can shift to nitrifier denitrification when oxygen is depleted in the environments including in the WWTPs, nitrite then plays the role of oxygen as the final electron acceptor. As opposed to what happens in nitrification, the end products of nitrifier denitrification are gaseous forms of nitrogen, where N(2)O is not negligible compared to N(2). Overall, N(2)O emissions represent 0.1-0.4% of oxidized NH(4)(+), depending on the oxygenation level. N(2)O emissions would range from 0.11 to 0.42 TN-N(2)O day(-1) for a tertiary treatment of the Paris wastewater effluents, consisting exclusively of activated sludge nitrification.     

Influence of pharmaceutical residues on the structure of activated sludge bacterial communities in wastewater treatment bioreactors

Concern is growing over contamination of the environment with pharmaceuticals because of their widespread use and incomplete removal during wastewater treatment, where microorganisms drive the key processes. The influence of pharmaceuticals on bacterial community structure in activated sludge was assessed in small-scale wastewater treatment bioreactors containing different concentrations (5, 50, 200 and 500 μg L⁻¹) of several commonly used pharmaceuticals (ibuprofen, naproxen, ketoprofen, diclofenac and clofibric acid). T-RFLP analyses of the bacterial 16S rRNA genes indicated a minor but consistent shift in the bacterial community structure in the bioreactor R50 supplied with pharmaceuticals at a concentration of 50 μg L⁻¹, compared to the control reactor R0, which was operated without addition of pharmaceuticals. In the reactors operated with higher concentrations of pharmaceuticals, a greater structural divergence was observed. Bacterial community composition was further investigated by preparation of two clone libraries of bacterial 16S rRNA genes from reactors R0 and R50. Most clones in both libraries belonged to the Betaproteobacteria, among which Thauera, Sphaerotilus, Ideonella and Acidovorax-related spp. dominated. Nitrite-oxidizing bacteria of the genus Nitrospira sp., which are key organisms for the second stage of nitrification in wastewater treatment plants, were found only in the clone library of the reactor without pharmaceuticals. In addition, diversity indices were calculated for the two clone libraries, indicating a reduced diversity of activated sludge bacterial community in the reactor supplied with 50 μg L⁻¹ of each of selected pharmaceuticals.     

Assessment of activated sludge microbial community analysis in full-scale biological wastewater treatment plants using patterns of fatty acid isopropyl esters (FAPEs)

This investigation introduces the application of a relatively rapid technique to obtain information about the dynamic nature of microbial communities in activated sludge. The objective has been to consider variability due to measurement errors and protocol changes within the same quantitative framework as the analysis of systematic differences in microbial communities in large-scale aerobic activated sludge secondary wastewater treatment systems. Adjustments to the methodology were considered due to their potential for simplifying and shortening the analysis procedure. All modifications to the protocols used to assay the composition of microbial fatty acids (MFAs) of activated sludge imposed some bias to the chromatographic data. This methodological bias was similar in magnitude to the level of discrimination between activated sludge microbial community structures that were considered as part of the present study. MFA analysis supported the expectations of subtle but systematic community structure differences and shifts in activated sludge based on the current understanding of these wastewater treatment systems. A standardized MFA methodology was shown to be sensitive to minor systematic changes in activated sludge communities due the anticipated underlying factors of selective pressures from the process configuration, history, operational conditions and/or nutrient status. The chemometric approach of fatty acid isopropyl ester analysis of activated sludge can provide a routine tool for meaningful and quantitative information of changes in activated sludge quality in full-scale treatment systems.     

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.     

Heterotroph anoxic yield in anoxic aerobic activated sludge systems treating municipal wastewater

As input to the steady state design and kinetic simulation models for the activated sludge system, the correct value for the heterotroph anoxic yield is essential to provide reliable estimates for the system denitrification potential. This paper examines activated sludge anoxic yield values in the literature, and presents experimental data quantifying the value. In the literature, in terms of the structure of ASM1 and similar models, theoretically it has been shown that the anoxic yield should be reduced to approximately 0.79 the value of the aerobic yield. This theoretical value is validated with data from corresponding aerobic OUR and anoxic nitrate time profiles in a batch fed laboratory scale long sludge age activated sludge system treating municipal wastewater. The value also is in close agreement with values in the literature measured with both artificial substrates and municipal wastewater. Thus, it is concluded that, in ASM1 and similar models, for an aerobic yield of 0.67mg COD/mg COD, the anoxic yield should be about 0.53 mg COD/mg COD. Including such a lower anoxic yield in ASM1 and similar models will result in a significant increase in denitrification potential, due to increased denitrification with wastewater RBCOD as substrate. In terms of the structure of ASM3, for the proposed substrate storage yields and the aerobic yield of 0.63 mg COD/mg COD, experimental data indicate that the corresponding anoxic yield should be about 0.42 mg COD/mg COD. This is significantly lower than the proposed value of 0.54 mg COD/mg COD, and requires further investigation.     

Zinc-induced antibiotic resistance in activated sludge bioreactors

Increased levels of bacterial resistance to antibiotics noted in recent decades poses a significant obstacle to the effective treatment and prevention of disease. Although overuse of antibiotics in agriculture and medicine is partially responsible, environmental exposure to heavy metals may also contribute to antibiotic resistance, even in the absence of antibiotics themselves. In this study, a series of eight lab-scale activated-sludge reactors were amended with Zn and/or a suite of three antibiotics (oxytetracycline, ciprofloxacin, and tylosin), in parallel with unamended controls. Classical spread-plating methods were used to assess resistance to these compounds in culturable bacteria over 21 weeks. After seven weeks of general acclimation and development of baseline resistance levels (phase 1), 5.0 mg/L Zn was added to half of the reactors, which were then operated for an additional 7 weeks (phase 2). For the final seven weeks (phase 3), two of the Zn-amended reactors and two of the control reactors were amended with all three antibiotics, each at 0.2 mg/L. Zn amendment alone did not significantly change resistance levels at the 95% confidence level in phase 2. However, tylosin resistance increased significantly during phase 3 in the Zn-only reactors and resistance to all three antibiotics significantly increased as a consequence of combined Zn+antibiotic amendments. Ambient dissolved Zn levels in the reactors were only 12% of added levels, indicating substantial Zn removal by adsorption and/or precipitation. These results show that sub-toxic levels of Zn can cause increased antibiotic resistance in waste treatment microbial communities at comparatively low antibiotic levels, probably due to developed cross-resistance resulting from pre-exposure to Zn.     

Characterization and treatability of aerobic bacterial thermophilically treated wastewater by a conventional activated sludge and granular activated carbon

An industrial wastewater that was pretreated by an aerobic thermophilic bacterial consortium (THE) was subjected to additional treatability studies by granular activated carbon (GAC) and a conventional activated sludge (CAS). The removal of dissolved organic carbon (DOC) in both systems was generally found to be similar. While GAC was able to attain better effluent concentrations of toluene and methyl isobutyl ketone (MIBK), the CAS was much more efficient at removing acetone. Furthermore, unlike the GAC, the performance of the CAS was not influenced by the high degree of variability in the influent wastewater. Characterization of the influent thermophilic wastewater using gas chromatography-mass spectroscopy (GC/MS) was performed to quantify the micropollutants as well as to evaluate removal efficiencies from the GAC and CAS systems.     

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.     

Metal toxicity in municipal wastewater activated sludge investigated by multivariate analysis and in situ hybridization

The toxicity of three heavy metals (copper, zinc and nickel) on an activated sludge biomass was studied by measuring several parameters of microbiological activity and the dynamics of microbial community composition. The effects on the microflora in the activated-sludge tank were evaluated by dosing the tank with different metal concentrations and a non-inhibiting concentration of copper for a 2-week period. The relationship between biological effects and chemical measurements was elucidated by principal component analysis (PCA). The biochemical data suggested a higher sensitivity of nitrifiers to these metals than heterotrophs. This was confirmed by fluorescent in situ hybridization (FISH) analysis on the microbial community. The impact of copper was displayed by biomass deflocculating phenomena (pin-point) and a decrease in the beta-proteo bacteria component. In addition, the zone settling velocity (ZSV) was shown not to be a good indicator of chronic toxicity phenomena. Finally, the sequence of biomass metal accumulation capability was assessed, and corresponds to Cu>Ni>Zn.     

Performance comparison of a pilot-scale UASB and DHS system and activated sludge process for the treatment of municipal wastewater

This study compares the performance of a pilot-scale combination of UASB and DHS system to that of activated sludge process (ASP) for the treatment of municipal sewage. Both systems were operated in parallel with the same sewage as influent. The study was conducted for more than 300 days, which revealed that organic removal efficiency of UASB+DHS system was comparable to that of ASP. Unfiltered BOD removal by both systems was more than 90%. However, UASB+DHS system outperformed ASP for pathogen removal. In addition, volume of excess sludge production from UASB+DHS was 15 times smaller than that from ASP. Moreover, unlike ASP, there is no requirement of aeration for the operation of UASB+DHS system, which makes it an economical treatment system. Considering the above observations, it was concluded that UASB+DHS system can be a cost-effective and viable option for the treatment of municipal sewage over ASP, especially for low-income countries.     

Ammonium adsorption in aerobic granular sludge, activated sludge and anammox granules

The ammonium adsorption properties of aerobic granular sludge, activated sludge and anammox granules have been investigated. During operation of a pilot-scale aerobic granular sludge reactor, a positive relation between the influent ammonium concentration and the ammonium adsorbed was observed. Aerobic granular sludge exhibited much higher adsorption capacity compared to activated sludge and anammox granules. At an equilibrium ammonium concentration of 30 mg N/L, adsorption obtained with activated sludge and anammox granules was around 0.2 mg NH₄-N/g VSS, while aerobic granular sludge from lab- and pilot-scale exhibited an adsorption of 1.7 and 0.9 mg NH₄-N/g VSS, respectively. No difference in the ammonium adsorption was observed in lab-scale reactors operated at different temperatures (20 and 30 °C). In a lab-scale reactor fed with saline wastewater, we observed that the amount of ammonium adsorbed considerably decreased when the salt concentration increased. The results indicate that adsorption or better ion exchange of ammonium should be incorporated into models for nitrification/denitrification, certainly when aerobic granular sludge is used.     

Effect of low ORP in anoxic sludge zone on excess sludge production in oxic-settling-anoxic activated sludge process

This paper studied the effect of oxidation-reduction potential (ORP) in the anoxic sludge zone on the excess sludge production in the oxic-settling-anoxic process (OSA process), a modified activated sludge process. Two pilot-scale activated sludge systems were employed in this study: (1) an OSA process that was modified from a conventional activated sludge process by inserting a sludge holding tank or namely the "anoxic" tank in the sludge return line; and (2) a conventional process used as the reference system. Each was composed of a membrane bioreactor to serve the aeration tank and solid/liquid separator. Both systems were operated with synthetic wastewater for 9 months. During the operation, the OSA system was operated with different ORP levels (+100 to -250 mV) in its anoxic tank. It has been confirmed that the OSA system produced much less excess sludge than the reference system. A lower ORP level than +100 mV in the anoxic tank is in favor of the excess sludge reduction. When the ORP level decreased from +100 to -250 mV the sludge reduction efficiency was increased from 23% to 58%. It has also been found that the OSA system performed better than the reference system with respect to the chemical oxygen demand removal efficiency and sludge settleability. The OSA process may present a potential low-cost solution to the excess sludge problem in an activated sludge process because addition of a sludge holding tank is only needed.     

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.