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.     

Enhancement of Anaerobic Digestion Using Particulate Growth Systems

Attached media reactors are used for enhancement of wastewater treatment processes including anaerobic condition. Selection of a suitable biofilm carrier is a compelling method to improve anaerobic digestion systems. This study investigates the performance of four fibrous biofilms installed in batch biogas reactors for treatment of cow manure. BioCords HS1, HS2, LS1, and LS2 are manufactured by Bishop Water Technologies, ON, Canada. Effluents and attached growth media were analyzed after batch experiment; methane production, methane yield, transfer efficiencies, organic and solid removal efficiencies, pH, and attached volatile suspended solid (VSS) were measured; VSS attached to biofilms mainly correlated with the specific surface area of each biofilm. Additionally, SEM (scanning electron microscopy) was used for further understanding of biofilm formation process for BioCords and the dissimilarity in their performance. The results indicated that BioCord LS2 had positive impact on achieving higher methane production and removal efficiencies compared to other support media utilized in batch reactors. It was also demonstrated from the experiment that BioCord LS2 potentially could generate higher methane production than conventional batch bioreactor.     

Comparative analysis of hydrogen-producing bacterial biofilms and granular sludge formed in continuous cultures of fermentative bacteria

A system for biohydrogen production was developed based on long-term continuous cultures grown on sugar beet molasses in packed bed reactors. In two separate cultures, consortia of fermentative bacteria developed as biofilms on granitic stones. In one of the cultures, a granular sludge was also formed. Metagenomic analysis of the microbial communities by 454-pyrosequencing of amplified 16S rDNA fragments revealed that the overall biodiversity of the hydrogen-producing cultures was quite small. The stone biofilm from the culture without granular sludge was dominated by Clostridiaceae and heterolactic fermentation bacteria, mainly Leuconostocaeae. Representatives of the Leuconostocaeae and Enterobacteriaceae were dominant in both the granules and the stone biofilm formed in the granular sludge culture. The culture containing granular sludge produced hydrogen significantly more effectively than that containing only the stone biofilm: 5.43 vs. 2.8 mol H₂/mol sucrose from molasses, respectively. The speculations that lactic acid bacteria may favor hydrogen production are discussed.     

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.     

Treatment of heavy metal-polluted wastewaters using the biofilms of a multistage rotating biological contactor

The ability of the biofilms of a three-stage rotating biological contactor (RBC) to treat wastewater contaminated with cadmium, copper and zinc was investigated. The system successfully removed the metals, in the order Cu > Zn > Cd with removal capacities of approximately 73, 42 and 33% respectively. Analysis of the contribution of each reactor indicated that metal removal was not uniform, with Reactor 1 showing a much higher removal capacity than Reactors 2 and 3. Energy dispersive X-ray spectroscopy (EDS) revealed the presence of all three metals on the surface of the biofilms in all three reactors. Closer inspection of the biofilms, in terms of biomass and biofilm thickness, revealed that the low metal removal in Reactors 2 and 3 was probably attributable to poor biofilm development in these two reactors compared to that in Reactor 1. The poor biofilm development was substantiated by low chemical oxygen demand (COD) removal in the latter two reactors.     

Assessment of anaerobic wastewater treatment failure using terminal restriction fragment length polymorphism analysis

AIMS: The suitability of genetic fingerprinting to study the microbiological basis of anaerobic bioreactor failure is investigated. METHODS AND RESULTS: Two laboratory-scale anaerobic expanded granular sludge bed bioreactors, R1 and R2, were used for the mesophilic (37 degrees C) treatment of high-strength [10 g chemical oxygen demand (COD) l(-1)] synthetic industrial-like wastewater over a 100-day trial period. A successful start up was achieved by both bioreactors with COD removal over 90%. Both reactors were operated under identical parameters; however, increased organic loading during the trial induced a reduction in the COD removal of R1, while R2 maintained satisfactory performance (COD removal >90%) throughout the experiment. Specific methanogenic activity measurements of biomass from both reactors indicated that the main route of methane production was hydrogenotrophic methanogenesis. Terminal restriction fragment length polymorphism (TRFLP) analysis was applied to the characterization of microbial community dynamics within the system during the trial. The principal differences between the two consortia analysed included an increased abundance of Thiovulum- and Methanococcus-like organisms and uncultured Crenarchaeota in R1. CONCLUSIONS: The results indicated that there was a microbiological basis for the deviation, in terms of operational performance, of R1 and R2. SIGNIFICANCE AND IMPACT OF THE STUDY: High-throughput fingerprinting techniques, such as TRFLP, have been demonstrated as practically relevant for biomonitoring of anaerobic reactor communities.     

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.     

Biomass retention and performance of anaerobic fixed-film reactors treating acetic acid wastewater

An acetic-acid-based synthetic wastewater of different organic concentrations was successfully treated at 35 degrees C in anaerobic downflow fixed-film reactors operated at high organic loading rates and short hydraulic retention times (HRTs). Substrate removal and methane production rates close to theoretical values of complete volumetric chemical oxygen demand (COD) removal and maximum methane conversion were obtained. A high concentration of biofilm biomass was retained in the reactor. Steady-state biofilm concentration increased with increased organic loading rate and decreased HRTs, reaching a maximum of 8.3 kg VFS/m(3) at a loading rate of 17 kg COD/m(3) day. Biofilm substrate utilization rates of up to 1.6 kg COD/kg VFS day were achieved. Soluble COD utilization rates at various COD concentrations can be described by half-order reaction kinetics.     

Thermophilic anaerobic treatment of sulphur rich forest industry wastewater

Thermophilic anaerobic treatment of sulphur-rich paper mill wastewater (0.8-3.1 gCOD/l, 340–850 mgSO₄/l; COD:SO₄ 3.4-5.3) was studied in three laboratory-scale, upflow anaerobic sludge blanket (UASB) reactors and in bioassays. The reactors were inoculated with non-adapted thermophilic granular sludge. In the bioassays, no inhibition of the inoculum was detected and about 62% COD removal (sulphide stripped) was obtained. About 70 to 80% of the removed COD was methanised. In the reactors, up to 60–74% COD removal (effluent sulphide stripped) was obtained at loading rates up to 10–30 kgCOD/m³d and hydraulic retention times down to 6 to 2 hours. The effluent total sulphide was up to 150–250 mg/l. Sulphide inhibition could not be confirmed from the reactor performances. The results from bioassays suggested that both the inoculum and sludge from the UASB reactor used acetate mainly for methane production, while sulphide was produced from hydrogen or its precursors.     

A novel nonwoven hybrid bioreactor (NWHBR) for enhancing simultaneous nitrification and denitrification

This study proposed a nonwoven hybrid bioreactor (NWHBR) in which the nonwoven fabric played dual roles as a biofilm carrier and membrane‐like separation of the flocculent sludge in the reactor. The results of long‐term monitoring demonstrated that the NWHBR could achieve simultaneous nitrification and denitrification (SND), with nearly complete ammonium removal and 80% removal of total nitrogen. The biofilm attached to the nonwoven fabric removed 27% of the chemical oxygen demand (COD) and 36% of the nitrate in the reactor, an enhanced elimination of nutrients that was attributed to the increased mass transfer within the biofilm due to permeate drag. The results of batch experiments showed that the flocculent sludge played a more dominant role in nitrification and denitrification (79% and 61%, respectively) than the biofilm (21% and 36%, respectively). The batch experiments also revealed that the enforced mass transfer, with an effluent recirculation rate of 4.3 L/m²h (which was the same as the flux during the reactor's long‐term operation), improved the denitrification rate by 58% (i.e., from 9.0 to 14.2 mg‐NO‐N/h). Pyrosequencing of the 16S rRNA gene amplification revealed a high microbial diversity in both the flocculent sludge and biofilm, with Proteobacteria, Bacteroidetes and Chloroflexi as the dominant groups. A phylogenetic (P) test indicated that the NWHBR contained phylogenetically distinct microbial communities: those in the biofilm differed from those in the flocculent sludge. However, the communities on the exterior and interior of the biofilm were more similar to each other. Due to its good SND performance, low physical back‐washing frequency and low air‐to‐water ratio, the NWHBR represents an attractive alternative for the wider application of either low‐cost membrane bioreactors or biofilm reactors. Biotechnol. Bioeng. 2013; 110: 1903–1912. © 2013 Wiley Periodicals, Inc.     

Performances and microbial features of an aerobic packed-bed biofilm reactor developed to post-treat an olive mill effluent from an anaerobic GAC reactor

Background  

Olive mill wastewater (OMW) is the aqueous effluent of olive oil producing processes. Given its high COD and content of phenols, it has to be decontaminated before being discharged. Anaerobic digestion is one of the most promising treatment process for such an effluent, as it combines high decontamination efficiency with methane production. The large scale anaerobic digestion of OMWs is normally conducted in dispersed-growth reactors, where however are generally achieved unsatisfactory COD removal and methane production yields. The possibility of intensifying the performance of the process using a packed bed biofilm reactor, as anaerobic treatment alternative, was demonstrated. Even in this case, however, a post-treatment step is required to further reduce the COD. In this work, a biological post-treatment, consisting of an aerobic biological "Manville" silica bead-packed bed aerobic reactor, was developed, tested for its ability to complete COD removal from the anaerobic digestion effluents, and characterized biologically through molecular tools.     

Anaerobic treatment of highly concentrated aniline wastewater using packed-bed biofilm reactor

The performance of packed-bed biofilm reactor (PBBR) with self-floating bio-carriers was investigated to treat highly concentrated organic nitrogenous aniline wastewater with a COD value as high as 24,000 mg/L. With 45 vol% of carrier charge inside the reactor, the aniline wastewater can be effectively treated with 94% of COD removal efficiency at a low organic loading rate (OLR) of 0.9 kg COD/(m³ d). The removal efficiency decreased gradually down to 75% when OLR increased to 12.27 kg COD/(m³ d) that corresponded to 1 day of HRT. Separate tests with biofilm alone showed that the conversion contribution of the biofilm was about half of the overall COD conversion by the biofilm plus sludge system at the same OLRs of 3–4 kg COD/(m³ d), and that the biofilm had higher activity than suspended sludge. Ammonium released from decomposed aniline was increased gradually from 500 to 1700 mg/L with the OLR increase from 0.9 to 12.27 kg COD/(m³ d), which resulted in inhibitory effect to the microorganism due to the toxicity of free ammonia. Batch anaerobic toxicity tests showed that the biofilm was less sensitive to toxic compounds than suspended sludge and could tolerate higher concentration of free ammonia.     

Microflora from leaf debris is suitable for treatment of starch industry wastewater

Biological treatment of industrial waste is a widely practiced technique that generates comparatively less environmentally hazardous waste than other chemical treatment processes. Wet milling of maize generates huge amount of wastewater (5 m³/ton) of low pH with organic matter and nutrients. Anaerobic methanogenic and aerobic bacteria are mostly highly sensitive to low pH. The treatment of wastewater causes huge cost of chemical neutralization or hydraulic recirculation for maintaining neutral pH. In the present study, different microbial consortia isolated from cow dung, active sludge from an anaerobic reactor for treatment of industrial wastewater, and leaf debris from benthic soil were screened for tolerance against low pH and for potential of chemical oxygen demand (COD) removal in order to find out an alternative microbial population for industrial water treatment at low pH. The most effective consortia found from leaf debris were further investigated for optimal operation. The microscopic analysis of leaf debris sludge showed abundance of Gram‐negative methanococci, which was found tolerant to low pH in plate culture method. On further investigation for COD removal from starch industry effluent, they were found to be most effective at pH 5 with highest COD removal rate of 70% and lowest biomass generation of 81%. Hence, it was concluded that the low pH‐tolerant methanogen bacteria, enriched from leaf debris sludge, is highly beneficial for anaerobic treatment of wastewater from several industries including corn starch industry by reducing cost of operation for neutralization to neutral pH and through reducing excess waste sludge production by the treatment system.     

Metagenomic and metaproteomic analyses of Accumulibacter phosphatis‐enriched floccular and granular biofilm

Biofilms are ubiquitous in nature, forming diverse adherent microbial communities that perform a plethora of functions. Here we operated two laboratory‐scale sequencing batch reactors enriched with Candidatus Accumulibacter phosphatis (Accumulibacter) performing enhanced biological phosphorus removal. Reactors formed two distinct biofilms, one floccular biofilm, consisting of small, loose, microbial aggregates, and one granular biofilm, forming larger, dense, spherical aggregates. Using metagenomic and metaproteomic methods, we investigated the proteomic differences between these two biofilm communities, identifying a total of 2022 unique proteins. To understand biofilm differences, we compared protein abundances that were statistically enriched in both biofilm states. Floccular biofilms were enriched with pathogenic secretion systems suggesting a highly competitive microbial community. Comparatively, granular biofilms revealed a high‐stress environment with evidence of nutrient starvation, phage predation pressure, and increased extracellular polymeric substance and cell lysis. Granular biofilms were enriched in outer membrane transport proteins to scavenge the extracellular milieu for amino acids and other metabolites, likely released through cell lysis, to supplement metabolic pathways. This study provides the first detailed proteomic comparison between Accumulibacter‐enriched floccular and granular biofilm communities, proposes a conceptual model for the granule biofilm, and offers novel insights into granule biofilm formation and stability.     

Ultrasonic sludge disintegration for enhanced methane production in anaerobic digestion: effects of sludge hydrolysis efficiency and hydraulic retention time

Hydrolysis of waste activated sludge (WAS) has been regarded as the rate limiting step of anaerobic sludge digestion. Therefore, in this study, the effect of ultrasound and hydraulic residence time during sludge hydrolysis was investigated with the goal of enhancing methane production from anaerobic digestion (AD). WAS was ultrasonically disintegrated for hydrolysis, and it was semi-continuously fed to an anaerobic digesters at various hydraulic retention times (HRTs). The results of these experiments showed that the solids and chemical oxygen demand (COD) removal efficiencies when using ultrasonically disintegrated sludge were higher during AD than the control sludge. The longer the HRT, the higher the removal efficiencies of solids and COD, while methane production increased with lower HRT. Sludge with 30% hydrolysis produced 7 × more methane production than the control sludge. The highest methane yields were 0.350 m(3)/kg volatile solids (VS)(add) and 0.301 m(3)/kg COD(con) for 16 and 30% hydrolyzed sludge, respectively. In addition, we found that excess ultrasound irradiation may inhibit AD since the 50% hydrolyzed sludge produced lower methane yields than 16 and 30% hydrolyzed sludge.     

Petroleum coke supplementation for enhanced biogas production and phosphate removal under mesophilic conditions

The use of carbon-based conductive materials has been shown to lead to an increase in biogas and methane yields during anaerobic digestion (AD). The effect of these additives on AD using synthetic substrates has been extensively studied, yet their significance for wastewater sludge digestion has not been adequately investigated. Therefore, the aim of this research was to optimize the concentration of petroleum coke (PC) that is a waste by-product of oil refineries, for the anaerobic digestion of wastewater sludge and investigation of phosphate removal in the AD process in the mesophilic temperature range. According to the results of the experiments, supplementing reactors with PC could significantly improve biogas and methane production. Supplementation of reactors with 1.5 g/L PC led to 23.40 ± 0.26% and 42.55 ± 3.97% increase in biogas production and methane generation, respectively. Moreover, the average volatile solids (VS), phosphate, and chemical oxygen demand (COD) removals were 43.43 ± 0.73, 46.74 ± 0.77%, and 60.40 ± 0.38%, respectively.     

Continuous detoxification, transformation, and degradation of nitrophenols in upflow anaerobic sludge blanket (UASB) reactors

The anaerobic transformation and degradation of nitrophenols by granular sludge was investigated in upflow anaerobic sludge blanket (UASB) reactors continuously fed with a volatile fatty acid (VFA) mixture as the primary substrate. During the start-up, subtoxic concentrations of 2-nitrophenol (2-NP), 4-nitrophenol (4-NP), and 2, 4-dinitrophenol (2, 4-DNP) were utilized. 4-NP and 2, 4-DNP were readily converted to the corresponding aromatic amine; whereas 2-NP was converted to nonaromatic products via intermediate formation of 2-aminophenol (2-AP). These conversions led to a dramatic detoxification of the mononitrophenols because the reactors treated the nitrophenolics at the concentrations which were over 25 times higher than those that caused severe inhibition. VFA removal efficiencies greater than 99% were achieved in both reactors at loading rates greater than 11.4 g COD per liter of reactor volume per day even at volumetric loading of mononitrophenols up to 910 mg/L . d.The sludges obtained from each of the reactors at the end of the continuous experiments were assayed for their specific nitrophenol reducing activity in the presence of different primary substrates. Reduction rates of 45 and 26 mg/g volatile suspended solids per day were observed for 2-NP and 4-NP, respectively, when utilizing the VFA mixture as primary substrate. Hydrogen, an interspecies-reduced compound, and substrates that provide interspecies-reducing equivalents-such as butyrate, propionate, and ethanol stimulated nitrophenol reduction, whereas acetate and methanol did not. Anaerobic batch biodegradability tests with the 2-NP-adapted sludge revealed that its corresponding aromatic amine, 2-AP, was degraded to methane at a specific rate of 14.5 mg/g VSS . d. Acetate was observed to be the major intermediate during 2-AP degradation in the presence of a specific methanogenic inhibitor 2-bromoethanesulfonate. The results of this study indicate that UASB reactors can be applied to rapidly detoxify and, under certain circumstances, degrade nitroaromatic compounds. (c) 1996 John Wiley & Sons, Inc.     

A study of the efficacy of ultrasonic waves in removing biofilms

doi:10.1111/j.1741‐2358.2009.00325.x
A study of the efficacy of ultrasonic waves in removing biofilms Objective: The removal of adherent biofilms was assessed using ultrasonic waves in a non‐contact mode. Materials and Methods: In in vitro experiments, Streptococcus mutans (S. mutans) biofilms were exposed to ultrasonic waves at various frequencies (280 kHz, 1 MHz, or 2 MHz), duty ratios (0–90%), and exposure times (1–3 minutes), and the optimal conditions for biofilm removal were identified. Furthermore, the effect of adding a contrast medium, such as micro bubbles (Sonazoid^®), was examined. The spatial distribution and architecture of S. mutans biofilms before and after ultrasonic wave exposure were examined via scanning electron microscopy. The biofilm removal effect was also examined in in vivo experiments, using a custom‐made oral cleaning device. Results: When a 280 kHz probe was used, the biofilm‐removing effect increased significantly compared to 1 and 2 MHz probes; more than 80% of the adherent biofilm was removed with a duty cycle of 50–90% and a 3 minutes exposure time. The maximum biofilm‐removing effect was observed with a duty cycle of 80%. Furthermore, the addition of micro bubbles enhanced this biofilm‐removing effect. In in vivo experiments, moderate biofilm removal was observed when a 280 kHz probe was used for 5 minutes. Conclusions: This study demonstrated that ultrasonic wave exposure in a non‐contact mode effectively removed adherent biofilms composed of S. mutans in vitro.     

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.     

High-rate treatment of terephthalate in anaerobic hybrid reactors.

The anaerobic degradation of terephthalate as sole substrate was studied in three anaerobic upflow reactors. Initially, the reactors were operated as upflow anaerobic sludge bed (UASB) reactors and seeded with suspended methanogenic biomass obtained from a full-scale down-flow fixed film reactor, treating wastewater generated during production of purified terephthalic acid. The reactors were operated at 30, 37, and 55 degrees C. The terephthalate removal capacities remained low in all three reactors (<4 mmolxL-1xday-1, or 1 g of chemical oxygen demand (COD)xL-1xday-1) due to limitations in biomass retention. Batch experiments with biomass from the UASB reactors revealed that, within the mesophilic temperature range, optimal terephthalate degradation is obtained at 37 degrees C. No thermophilic terephthalate-degrading culture could be obtained in either continuous or batch cultures. To enhance biomass retention, the reactors were modified to anaerobic hybrid reactors by introduction of two types of reticulated polyurethane (PUR) foam particles. The hybrid reactors were operated at 37 degrees C and seeded with a mixture of biomass from the UASB reactors operated at 30 and 37 degrees C. After a lag period of approximately 80 days, the terephthalate conversion capacity of the hybrid reactors increased exponentially at a specific rate of approximately 0.06 day-1, and high removal rates were obtained (40-70 mmolxL-1xday-1, or 10-17 g of CODxL-1xday-1) at hydraulic retention times between 5 and 8 h. These high removal capacities could be attributed to enhanced biomass retention by the development of biofilms on the PUR carrier material as well as the formation of granular biomass. Biomass balances over the hybrid reactors suggested that either bacterial decay or selective wash-out of the terephthalate fermenting biomass played an important role in the capacity limitations of the systems. The presented results suggest that terephthalate can be degraded at high volumetric rates if sufficiently long sludge ages can be maintained, and the reactor pH and temperature are close to their optima.