Comparison of polyhydroxyalkanoates production by activated sludges from anaerobic and oxic zones of an enhanced biological phosphorus removal system: effect of sludge retention time

This study compared the PHAs production behavior of sludges from the anaerobic and oxic phases of an enhanced biological phosphorus removal (EBPR) system. This was accomplished by using the kinetics and stoichiometric coefficients obtained from aerobic batch tests to evaluate the performance of these two sludges. Experimental results indicated that the metabolic behavior of the sludges for PHAs production depend significantly on the operating sludge retention time (SRT) of the EBPR system. The oxic sludge with 5 days of SRT exhibited better PHAs production performance than anaerobic sludge. Conversely, the anaerobic sludge with 15 days of SRT had superior PHAs production capability compared to oxic sludge. These comparisons suggest that whether anaerobic or oxic sludge should be employed for PHAs production depends mainly on the operating SRT of the EBPR system.     

Scanning electron microscopy of biofilm development in anaerobic fixed-bed reactors: Influence of the inoculum

Summary Scanning electron microscopy was applied to evaluate the influence of inoculum on efficiency of initial biofilm formation and reactor performance. Five anaerobic fixed-bed reactors were inoculated with anaerobic sludges from different sources and operated in parallel under identical conditions with defined wastewater and acetate, propionate and butyrate as constituents In all sludges Methanothrix sp. was the predominant acetotroph. The reactors inoculated with anaerobic sludge adapted to the wastewater achieved the highest space loading with 21.0 g COD/l·d after 58 days. The inoculation with granular sludge from an upflow anaerobic sludge blanket (UASB) reactor resulted in significantly less reactor efficiency. Time course of biofilm formation and biofilm thickness (ranging from 20–200 m) depended on the type of inoculum.     

Feasibility studies on the treatment of dairy wastewaters with upflow anaerobic sludge blanket reactors

The feasibility of using upflow anaerobic sludge blanket (UASB) reactors for the treatment of dairy wastewaters was explored. Two types of UASBs were used--one operating on anaerobic sludge granules developed by us from digested cowdung slurry (DCDS) and the other on the granules obtained from the reactors of M/s EID Parry treating sugar industry wastewaters. The reactors were operated at HRT of 3 and 12 h and on COD loading rates ranging from 2.4 kg per m3 of digester volume, per day to 13.5 kg m(-3) d(-1). At the 3 h HRT, the maximum COD reduction in the DCDS-seeded and the industrial sludge-seeded reactors was 95.6% and 96.3%, respectively, better than at 12 h HRT (90% and 92%, respectively). In both the reactors, the maximum, the second best, and the third best COD reduction occurred at the loading rates of 10.8, 8.6 and 7.2 kg m3 d(-1), respectively. At loading rates higher than 10.8 kg, the reactor performance dropped precipitously. Whereas in the first few months the reactors operating on sludge from EID Parry achieved better biodegradation of the waste, compared to the reactors operated on DCDS, the performance of the latter gradually improved and matched with the performance of the former.     

Long-term effect of weak nitrogen limitation on polyhydroxyalkanoates production of propionate-fed activated sludge operated at long sludge retention time

A sequencing batch reactor (SBR) submitted to aerobic dynamic feeding condition was fed with weakly nitrogen-limited substrate (COD/N ratio of 60?mg/mg, propionate as the sole carbon source), and operated at 15?days sludge retention time (SRT). Then, the nitrogen-limited sludges at different cultivating periods (45th, 120th, 195th and 240th days) were harvested to conduct aerobic batch experiments of PHAs production under various nitrogen concentrations. Experimental results indicated that PHAs accumulating behavior of the propionate-fed sludge declined significantly after long-term cultivation, implying that weak nitrogen limitation and long SRT operation cannot achieve a stable PHAs producing performance of the sludge. Supplementing enough nitrogen was consequently a requisite to guarantee the stability of the propionate-fed PHAs accumulating culture. Besides, the sludge with higher PHAs content in SBR did not ensure a better PHAs producing capability in the batch PHAs production phase. Batch experiments also indicated that the propionate-fed sludge with 0.7?N?mmol/L exhibited a better PHAs producing capability than with other nitrogen concentrations.     

Pilot study on the upgrading configuration of UASB-MBBR with two carriers: Treatment effect,sludge reduction and functional microbial identification

Two kinds of biocarriers were adopted and a combined process of “AMC (Anaerobic microorganism carrier)-UASB and PBG (Porous bio-gel)-MBBR” was operated at the pilot scale for the treatment of real textile wastewater. The influence mechanism of the two carriers on the start-up, pollutant removal and sludge reduction were investigated within 118 days of operation. The dominant functional bacteria in anaerobic and aerobic systems were identified by high-throughput sequencing, and the possible ways and related mechanisms of nutrient removal and sludge reduction were analyzed based on the data. 37.0 ± 7.5 % and 53 ± 12.7 % of COD removal efficiencies were achieved in anaerobic system and aerobic system, respectively. Ammonia nitrogen concentration decreased from 20 to 45 to 3.49 ± 0.54 mg/L after treatment. An anaerobe was found to be closely related to color removal, which existed in both anaerobic and aerobic systems, achieving 84.0 % of color removal. With the operation of the system, the sludge yield decreased gradually. The sludge yields of anaerobic and aerobic systems were calculated individually and compared with similar studies. Aging biofilms were characterized to explore the factors associated with biofilm renewal.     

A novel UASB–MFC–BAF integrated system for high strength molasses wastewater treatment and bioelectricity generation

An up-flow anaerobic sludge blanket reactor–microbial fuel cell–biological aerated filter (UASB–MFC–BAF) system was developed for simultaneous bioelectricity generation and molasses wastewater treatment in this study. The maximum power density of 1410.2 mW/m² was obtained with a current density of 4947.9 mA/m² when the high strength molasses wastewater with chemical oxygen demand (COD) of 127,500 mg/l was employed as the influent. The total COD, sulfate and color removal efficiencies of the proposed system were achieved of 53.2%, 52.7% and 41.1%, respectively. Each unit of this system had respective function and performed well when integrated together. The UASB reactor unit was mainly responsible for COD removal and sulfate reduction, while the MFC unit was used for the oxidation of generated sulfide with electricity generation. The BAF unit dominated color removal and phenol derivatives degradation. This study is a beneficial attempt to combine MFC technology with conventional anaerobic–aerobic processes for actual wastewater treatment.     

Treatment of winery effluent with upflow anaerobic sludge blanket (UASB) – granular sludges enriched with Enterobacter sakazakii

Three upflow anaerobic sludge blankets (UASBs) were evaluated for the treatment of winery wastewater: the first was seeded with granular sludge enriched with Enterobacter sakazakii and reached a 90% COD removal within 17 d at hydraulic retention time of 24 h; the second was seeded with brewery granules and achieved 85% COD removal within 50 d, the third was seeded with just sludge and showed the typical problems encountered with conventional sludge seeding and had continuously to be re-seeded. A PCR-based technique was developed for the rapid detection of E. sakazakii in the granular sludge.     

Acetate Degradation at 70 degrees C in Upflow Anaerobic Sludge Blanket Reactors and Temperature Response of Granules Grown at 70 degrees C

Anaerobic acetate degradation at 70 degrees C and at 55 degrees C (as a reference) was studied by running laboratory upflow anaerobic sludge blanket (UASB) reactors inoculated with mesophilic granular sludge. In UASB reactors fed with acetate-containing media (3 g of chemical oxygen demand [COD] per liter, corresponding to 47 mM acetate) approximately 50 days was needed at 70 degrees C and less than 15 days was needed at 55 degrees C to achieve an effluent COD of 500 to 700 mg/liter. In the UASB reactors at both 70 and 55 degrees C up to 90% of the COD was removed. Batch assays showed that sludges from two 70 degrees C UASB reactors, one run at a low effluent acetate concentration and the other run at a high effluent acetate concentration, exhibited slightly different responses to temperatures in the range from 37 to 70 degrees C. Both 70 degrees C sludges, as well as the 55 degrees C sludge, produced methane at temperatures of 37 to 73 degrees C. The 55 degrees C sludge exhibited shorter lag phases than the 70 degrees C sludges and higher specific methane production rates between 37 and 65 degrees C.     

Effect of seed sludge microbial community and activity on the performance of anaerobic reactors during the start-up period

In this study, two laboratory-scale anaerobic batch reactors started up with different inoculum sludges and fed with the same synthetic wastewater were monitored in terms of performance and microbial community shift by denaturant gradient gel electrophoresis fingerprinting and subsequent cloning, sequencing analysis in order to reveal importance of initial quality of inoculum sludge for operation of anaerobic reactors. For this purpose, two different seed sludge were evaluated. In Reactor1 seeded with a sludge having less diverse microbial community (19 operational taxonomic unit (OTU’s) for Bacterial and 8 OTU’s for Archaeal community, respectively) and a methanogenic activity of 150 ml CH₄ g TVS⁻¹ day⁻¹, a chemical oxygen demand (COD) removal efficiency of 78.8 ± 4.17% was obtained at a substrate to microorganism (S/X) ratio of 0.38. On the other hand, Reactor2, seeded with a sludge having a much more diverse microbial community (24 OTU’s for Bacterial and 9 OTU’s for Archaeal communities, respectively) and a methanogenic activity, 450 ml CH₄ g TVS⁻¹ day⁻¹, operated in the same conditions showed a better start-up performance; a COD removal efficiency of over 98% at a S/X ratio of 0.53. Sequence analysis of Seed2 revealed the presence of diverse fermentative and syntrophic bacteria, whereas excised bands of Seed1 related to fermentative and sulfate/metal-reducing bacteria. This study revealed that a higher degree of bacterial diversity, especially the presence of syntrophic bacteria besides the abundance of key species such as methanogenic Archaea may play an important role in the performance of anaerobic reactors during the start-up period.     

Metabolic transformations and characterisation of the sludge community in an enhanced biological phosphorus removal system

Enhanced biological phosphorus removal was performed in a continuous laboratory-scale two-reactor system with sludge recirculation over a 75-day period. Influent wastewater was a synthetic medium based on acetate, and the sludge age was kept at 12 days. The adapted sludge stored poly-β-hydroxyalkanoic acids (PHA) in the anaerobic reactor with a conversion ratio of 1.45 PHA/acetic acid (based on chemical O₂ demand: COD/COD) and gave ratio of a phosphate-P release to acetic acid uptake of 0.51 P/CH₃COOH (w/w). Fractionation of anaerobic and aerobic sludges showed that the main part of phosphorus taken up, was eluted in the trichloroacetic acid fraction indicating that it was polyphosphate. A total of 60% of the phosphorus in the aerobic sludge was solubilized in the trichloroacetic acid fraction, whereas this fraction accounted for only 32% of the phosphorus in the anaerobic sludge. Only 4% of the total phosphorus in the aerobic sludge and 2% in the anaerobic sludge was found in the EDTA fraction, indicating low amounts of metal-bound phosphates. Isolation on acetate-based agar medium showed that Acinetobacter strains were present in the sludge. However, a more complete analysis of the bacterial community of the sludge was obtained by creating a clone library based on the 16S rRNA gene. A total of 51 partial clone sequences were phylogenetically evaluated. The predominating group was found in the high-(G+C) (mol%) gram-positive bacterial subphylum (31% of the sequenced clones), while the gamma proteobacteria only constituted 9.8% of the clones. Received: 12 June 1997 / Received revision: 26 September 1997 / Accepted: 28 September 1997     

Bioelectricity production from acidic food waste leachate using microbial fuel cells: Effect of microbial inocula

The effects of three different inocula (domestic wastewater, activated sludge, and anaerobic sludge) on the treatment of acidic food waste leachate in microbial fuel cells (MFCs) were evaluated. A food waste leachate (pH 4.76; 1000 mg chemical oxygen demand (COD)/L) was used as the substrate. The results indicate that the leachate itself can enable electricity production in an MFC, but the co-addition of different inocula significantly reduces the start-up time (approximately 7 days). High COD and volatile fatty acids removal (>87%) were obtained in all MFCs but with only low coulombic efficiencies (CEs) (14–20%). The highest power (432 mW/m³) and CE (20%) were obtained with anaerobic sludge as the co-inoculum. Microbial community analysis (PCR-DGGE) of the established biofilms suggested that the superior performance of the anaerobic sludge-MFC was associated with the enrichment of both fermentative (Clostridium sp. and Bacteroides sp.) and electrogenic bacteria (Magnetospirillum sp. and Geobacter sp.) at the anode.     

Comparison of the performance of one stage and two stage sequential anaerobic-aerobic biological processes for the treatment of reactive-azo-dye-containing synthetic wastewaters

In the present study the performance of anaerobic-aerobic one and two stage processes for the biological treatment of synthetic wastewaters containing Reactive Black 5 (RB5) were studied and compared with each other. In both processes the majority of colour removal by biodegradation occurred under anaerobic environment. The colour change under aerobic conditions was correlated with extent of anaerobic decolourisation in the preceding phase/stage of the process. Partial mineralisation of the anaerobic dye metabolites, roughly to the same extent, was achieved aerobically in both one stage and two stage processes. The majority of COD was removed in the anaerobic stage for two stage processes and aerobic stage in one stage processes. In one stage processes, the exposure of anaerobic sludge to alternating anaerobic-aerobic environment decreased anaerobic decolourisation efficiency and COD removal; when employing activated sludge, the same exposure enhanced anaerobic substrate utilisation whereas the effect on the anaerobic decolourisation efficiency depended on RB5 concentration. The comparative performance of one and two stage processes in terms of overall dye decolourisation depended on RB5 concentration. Both types of processes brought about similar overall COD removal. Increase in RB5 concentration, in the range studied, resulted in decrease in overall COD removal for both processes.     

Two-stage anaerobic treatment of dairy wastewater using HUASB with PUF and PVC carrier

In the present study, an attempt has been made to treat dairy wastewater entirely via anaerobic treatment over a period of 215 days, using two-stage Hybrid Upflow Anaerobic Sludge Blanket (HUASB) reactors, which offer the advantages associated both with fixed film and upflow sludge blanket treatments. A HUASB with polyurethane foam cubes was used for stage I, and a HUASB utilizing PVC-cut rings was used for stage II. The output from stage I was used as the input for stage II. The two-stage reactor was operated at an organic loading rate that varied from 10.7 to 21.4 kg COD m³/d for a period of 215 days, including the start-up period. The ideal organic loading rate for the two-stage reactor was 19.2 kg COD/m³/d. A further 21.4 kg COD m³/d increase in the organic loading rate resulted in the souring of the reactor function in stage I, which consequently reduced the overall reactor performance. Combined COD removal during the stable operation period (10.7 to 19.2 kg COD m³/d) occurred in a range between 97 and 99%. The methane content in the biogas varied from 65 to 70% in stage I, and from 63 to 66% in stage II. The two-stage anaerobic treatment using HUASB with PUF and PVC described in this work is expected to constitute a better alternative for the complete treatment of dairy wastewater than high-rate anaerobic, anaerobic/aerobic, and two-phase anaerobic treatment methods.     

Biogas production from supernatant of hydrothermally treated municipal sludge by upflow anaerobic sludge blanket reactor

The supernatant of hydrothermally treated sludge was treated by an upflow anaerobic sludge blanket (UASB) reactor for a 550-days running test. The hydrothermal parameter was 170 °C for 60 min. An mesophilic 8.6 L UASB reactor was seeded with floc sludge. The final organic loading rate (OLR) could reach 18 kg COD/m³ d. At the initial stage running for 189 days, the feed supernatant was diluted, and the OLR reached 11 kg COD/m³ d. After 218 days, the reactor achieved a high OLR, and the supernatant was pumped into the reactor without dilution. The influent COD fluctuated from 20,000 to 30,000 mg/L and the COD removal rate remained at approximately 70%. After 150 days, granular sludge was observed. The energy balance calculation show that heating 1.0 kg sludge needs 0.34 MJ of energy, whereas biogas energy from the supernatant of the heated sludge is 0.43 MJ.     

Enhanced aerobic granulation and nitrogen removal by the addition of zeolite powder in a sequencing batch reactor

The aim of this study was to evaluate the impact of zeolite powders on feasibility of rapid aerobic granulation in the column-type sequencing batch reactors. After 90 days' operation, aerobic granular sludge was formed in both reactors by altering influent chemical oxygen demand/nitrogen (COD/N) ratios. R1 with zeolite powders had better removal capabilities of COD and total nitrogen than R2, which was without zeolite powders. Mixed liquor volatile suspended solid concentrations of the two reactors were 7.36 and 5.45 g/L, while sludge volume index (SVI₃₀) values were 34.9 and 47.9 mg/L, respectively. The mean diameters of aerobic granular sludge in the above two reactors were 2.5 and 1.5 mm, respectively. Both reactors achieved the largest simultaneous nitrification and denitrification (SND) efficiency at an influent COD/N ratio of 8; however, R1 exhibited more excellent SND efficiency than R2. The obtained results could provide a novel technique for rapid aerobic granulation and N removal simultaneously, especially when treating nitrogen-rich industrial wastewater.     

Acetate Degradation at 70°C in Upflow Anaerobic Sludge Blanket Reactors and Temperature Response of Granules Grown at 70°C

Anaerobic acetate degradation at 70°C and at 55°C (as a reference) was studied by running laboratory upflow anaerobic sludge blanket (UASB) reactors inoculated with mesophilic granular sludge. In UASB reactors fed with acetate-containing media (3 g of chemical oxygen demand [COD] per liter, corresponding to 47 mM acetate) approximately 50 days was needed at 70°C and less than 15 days was needed at 55°C to achieve an effluent COD of 500 to 700 mg/liter. In the UASB reactors at both 70 and 55°C up to 90% of the COD was removed. Batch assays showed that sludges from two 70°C UASB reactors, one run at a low effluent acetate concentration and the other run at a high effluent acetate concentration, exhibited slightly different responses to temperatures in the range from 37 to 70°C. Both 70°C sludges, as well as the 55°C sludge, produced methane at temperatures of 37 to 73°C. The 55°C sludge exhibited shorter lag phases than the 70°C sludges and higher specific methane production rates between 37 and 65°C.     

Radiolabelled proteomics to determine differential functioning of Accumulibacter during the anaerobic and aerobic phases of a bioreactor operating for enhanced biological phosphorus removal

Proteins synthesized by the mixed microbial community of two sequencing batch reactors run for enhanced biological phosphorus removal (EBPR) during aerobic and anaerobic reactor phases were compared, using mass spectrometry‐based proteomics and radiolabelling. Both sludges were dominated by polyphosphate‐accumulating organisms belonging to Candidatis Accumulibacter and the majority of proteins identified matched closest to these bacteria. Enzymes from the Embden–Meyerhof–Parnas pathway were identified, suggesting this is the major glycolytic pathway for these Accumulibacter populations. Enhanced aerobic synthesis of glyoxylate cycle enzymes suggests this cycle is important during the aerobic phase of EBPR. In one sludge, several TCA cycle enzymes showed enhanced aerobic synthesis, suggesting this cycle is unimportant anaerobically. The second sludge showed enhanced synthesis of TCA cycle enzymes under anaerobic conditions, suggesting full or partial TCA cycle operation anaerobically. A phylogenetic analysis of Accumulibacter polyphosphate kinase genes from each sludge demonstrated different Accumulibacter populations dominated the two sludges. Thus, TCA cycle activity differences may be due to Accumulibacter strain differences. The major fatty acids present in Accumulibacter‐dominated sludge include palmitic, hexadecenoic and cis‐vaccenic acid and fatty acid content increased by approximately 20% during the anaerobic phase. We hypothesize that this is associated with increased anaerobic phospholipid membrane biosynthesis, to accommodate intracellular polyhydroxyalkanoate granules.     

Anaerobic granule development for removal of pentachlorophenol in an upflow anaerobic sludge blanket (UASB) reactor

The granulation process using synthetic wastewater containing pentachlorophenol (PCP) in four 1.1 l laboratory scale upflow anaerobic sludge blanket (UASB) reactors was studied, and the anaerobic biotransformation of PCP during the granulation process investigated. After 110 days granular sludge was developed and up to 160 and 180 mg/l of PCP was added into the reactors R1 and R2, respectively, when they were inoculated with acclimated anaerobic sludge from an anaerobic digester of a citric acid plant. The inoculum was predominately composed of bacilli and filamentous bacteria. Granulation did not occur in reactors R3 and R4 which were inoculated with acclimated anaerobic sludge from aerobic sludge of the municipal sewage treatment plant which consisted mainly of cocci. Despite similar bacilli in the granule, the filamentous bacteria from reactor R1 were thicker than those of reactor R2. The granular sludge had a maximum diameter of 2.5 and 2.2 mm, and SMA of 1.44 and 1.32 gCOD/gTVS per day for reactors R1 and R2, respectively. Over 98% chemical oxygen demand (COD) removal rate and 99% of PCP removal rate were achieved when reactors R1 and R2 were operated at PCP and COD loading rates of 150 and 7.5 g/l per day, respectively. H₂-producing acetogens were the dominant anaerobes in the granular sludge.     

Effect of sludge type on poliovirus association with and recovery from sludge solids

Sludge type was found to affect the degree of association between seeded poliovirus type 1 (LSc) and sludge solids. The mean percent of solids-associated viruses for activated sludge mixed liquors, anaerobically digested sludges, and aerobically digested percent of solids-associated viruses for activated sludge mixed liquors, anaerobically digested sludges, and aerobically digested sludges was 57.2, 70.4, and 94.7, respectively. The degree of association between poliovirus and sludge solids was significantly greater for aerobically digested sludges than for the other two sludge types. Sludge solids associated viruses were eluted using 0.05 M glycine buffer, pH 10.5-11.0, and subsequently concentrated by organic flocculation. The effectiveness of the glycine method in the recovery of solids-associated viruses was also found to be affected by sludge type. Significantly lower mean poliovirus recovery was found for aerobically digested sludges (14.5%) than for mixed liquors or anaerobically digested sludges (72.3 and 60.2%, respectively). The eluent used in the method was not as effective in dissociating the virus from aerobic sludge solids as it was for the other two sludge types. All other virus adsorption-elution steps of the method (i.e., virus concentration steps) were equally effective in poliovirus recovery for all three sludge types. It is suggested that future methods developed for the recovery of viruses from sludges be evaluated for the various sludge types likely to be tested.     

Long-term impact of dissolved O(2) on the activity of anaerobic granules

The impact of influent dissolved O(2) on the characteristics of anaerobic granular sludge was investigated at various dissolved O(2) concentrations (0.5-8.1 ppm) in 1- and 5-L laboratory-scale upflow anaerobic sludge bed (UASB)-like anaerobic/aerobic coupled reactors with a synthetic wastewater (carbon sources containing 75% sucrose and 25% acetate). The rate of dissolved O(2) supplied to the coupled reactor was as high as 0.40 g O(2)/L(rx).d, and the anaerobic/aerobic coupled reactors maintained excellent methanogenic performances at a COD loading rate of 3 g COD/L(rx).d even after the reactors had been operated with dissolved O(2) for 3 months. The activities of granular sludge on various substrates (glucose, propionate, and hydrogen) were not impaired, and acetate activity was even improved over a short term. However, after 3 months of operation, slight declines on the acetoclastic activities of granules were observed in the coupled reactor receiving the recirculated fluid containing 8.1 ppm dissolved O(2).Methane yield in the anaerobic control reactor and anaerobic/aerobic coupled reactors revealed that a significant aerobic elimination (up to 30%) of substrate occurred in the coupled reactors, as expected. The presence of dissolved O(2) in the recirculated fluid resulted in the development of fluffy biolayers on the granule surface, which imposed a negative impact on the settleability of granular sludge and caused a slightly higher sludge washout. This research shows that the anaerobic/aerobic coupled reactor can be successfully operated under O(2)-limited conditions and is an ideal engineered ecosystem integrating oxic and anaerobic niches. (c) 1996 John Wiley & Sons, Inc.