Effect of starch addition on the biological conversion and microbial community in a methanol-fed UASB reactor during long-term continuous operation

The effect of starch addition on the microbial composition and the biological conversion was investigated using two upflow anaerobic sludge bracket (UASB) reactors treating methanolic wastewater: one reactor was operated with starch addition, and another reactor was operated without starch addition. Approximately 300 days of operation were performed at 30 kg COD/m³/d, and then, the organic load of the reactors was gradually increased to 120 kg COD/m³/d. Successful operation was achieved at 30 kg COD/m³/d in both reactors; however, the methanol-fed reactor did not perform well at 120 kg COD/m³/d while the methanol-starch-fed reactor did. The granule analysis revealed the granule developed further only in the methanol-starch-fed reactor. The results of the microbial community analysis revealed more Methanosaeta cells were present in the methanol-starch-fed reactor, suggesting the degradation of starch produced acetate as an intermediate, which stimulated the growth of Methanosaeta cells responsible for the extension of granules.     

Anaerobic digestion and wastewater treatment systems

Upflow Anaerobic Sludge Bed (UASB) wastewater (pre-)treatment systems represent a proven sustainable technology for a wide range of very different industrial effluents, including those containing toxic/inhibitory compounds. The process is also feasible for treatment of domestic wastewater with temperatures as low as 14–16° C and likely even lower. Compared to conventional aerobic treatment systems the anaerobic treatment process merely offers advantages. This especially is true for the rate of start-up. The available insight in anaerobic sludge immobilization (i.e. granulation) and growth of granular anaerobic sludge in many respects suffices for practice. In anaerobic treatment the immobilization of balanced microbial communities is essential, because the concentration of intermediates then can be kept sufficiently low.So far ignored factors like the death and decay rate of organisms are of eminent importance for the quality of immobilized anaerobic sludge. Taking these factors into account, it can be shown that there does not exist any need for phase separation when treating non- or slightly acidified wastewaters. Phase separation even is detrimental in case the acidogenic organisms are not removed from the effluent of the acidogenic reactor, because they deteriorate the settleability of granular sludge and also negatively affect the formation and growth of granular sludge. The growing insight in the role of factors like nutrients and trace elements, the effect of metabolic intermediates and end products opens excellent prospects for process control, e.g. for the anaerobic treatment of wastewaters containing mainly methanol.Anaerobic wastewater treatment can also profitably be applied in the thermophilic and psychrophilic temperature range. Moreover, thermophilic anaerobic sludge can be used under mesophilic conditions.The Expanded Granular Sludge Bed (EGSB) system particularly offers big practical potentials, e.g. for very low strength wastewaters (COD 1 g/l) and at temperatures as low as 10° C. In EGSB-systems virtually all the retained sludge is employed, while compared to UASB-systems also a substantially bigger fraction of the immobilized organisms (inside the granules) participates in the process, because an extraordinary high substrate affinity prevails in these systems. It looks necessary to reconsider theories for mass transfer in immobilized anaerobic biomass.Instead of phasing the digestion process, staging of the anaerobic reactors should be applied. In this way mixing up of the sludge can be significantly reduced and a plug flow is promoted. A staged process will provide a higher treatment efficiency and a higher process stability. This especially applies for thermophilic systems.     

Pentachlorophenol (PCP) dechlorination in horizontal-flow anaerobic immobilized biomass (HAIB) reactors

This study verifies the potential applicability of horizontal-flow anaerobic immobilized biomass (HAIB) reactors to pentachlorophenol (PCP) dechlorination. Two bench-scale HAIB reactors (R1 and R2) were filled with cubic polyurethane foam matrices containing immobilized anaerobic sludge. The reactors were then continuously fed with synthetic wastewater consisting of PCP, glucose, acetic acid, and formic acid as co-substrates for PCP anaerobic degradation. Before being immobilized in polyurethane foam matrices, the biomass was exposed to wastewater containing PCP in reactors fed at a semi-continuous rate of 2.0 μg PCP g⁻¹ VS. The applied PCP loading rate was increased from 0.05 to 2.59 mg PCP l⁻¹ day⁻¹ for R1, and from 0.06 to 4.15 mg PCP l⁻¹ day⁻¹ for R2. The organic loading rates (OLR) were 1.1 and 1.7 kg COD m⁻³ day⁻¹ at hydraulic retention times (HRT) of 24 h for R1 and 18 h for R2. Under such conditions, chemical oxygen demand (COD) removal efficiencies of up to 98% were achieved in the HAIB reactors. Both reactors exhibited the ability to remove 97% of the loaded PCP. Dichlorophenol (DCP) was the primary chlorophenol detected in the effluent. The adsorption of PCP and metabolites formed during PCP degradation in the packed bed was negligible for PCP removal efficiency.     

Influence of carbon source and inoculum type on anaerobic biomass adhesion on polyurethane foam in reactors fed with acid mine drainage

This paper analyzes the influence of carbon source and inoculum origin on the dynamics of biomass adhesion to an inert support in anaerobic reactors fed with acid mine drainage. Formic acid, lactic acid and ethanol were used as carbon sources. Two different inocula were evaluated: one taken from an UASB reactor and other from the sediment of a uranium mine. The values of average colonization rates and the maximum biomass concentration (C_max) were inversely proportional to the number of carbon atoms in each substrate. The highest C_max value (0.35 g TVS g⁻¹ foam) was observed with formic acid and anaerobic sludge as inoculum. Maximum colonization rates (v_max) were strongly influenced by the type of inoculum when ethanol and lactic acid were used. For both carbon sources, the use of mine sediment as inoculum resulted in a v_max of 0.013 g TVS g⁻¹ foam day⁻¹, whereas 0.024 g TVS g⁻¹ foam day⁻¹ was achieved with anaerobic sludge.     

Cultivation of shear stress sensitive microorganisms in disposable bag reactor systems

Technical scale (≥5 l) cultivations of shear stress sensitive microorganisms are often difficult to perform, as common bioreactors are usually designed to maximize the oxygen input into the culture medium. This is achieved by mechanical stirrers, causing high shear stress. Examples for shear stress sensitive microorganisms, for which no specific cultivation systems exist, are many anaerobic bacteria and fungi, such as basidiomycetes. In this work a disposable bag bioreactor developed for cultivation of mammalian cells was investigated to evaluate its potential to cultivate shear stress sensitive anaerobic Eubacterium ramulus and shear stress sensitive basidiomycetes Flammulina velutipes and Pleurotus sapidus. All cultivations were compared with conventional stainless steel stirred tank reactors (STR) cultivations. Good growth of all investigated microorganisms cultivated in the bag reactor was found. E. ramulus showed growth rates of μ = 0.56 h⁻¹ (bag) and μ = 0.53 h⁻¹ (STR). Differences concerning morphology, enzymatic activities and growth in fungal cultivations were observed. In the bag reactor growth in form of small, independent pellets was observed while STR cultivations showed intense aggregation. F. velutipes reached higher biomass concentrations (21.2 g l⁻¹ DCW vs. 16.8 g l⁻¹ DCW) and up to 2-fold higher peptidolytic activities in comparison to cell cultivation in stirred tank reactors.     

Comparison of different pilot scale bioreactors for the treatment of a real wastewater from the textile industry

Wastewater from textile industry usually undergoes activated sludge biotreatment ahead of refining treatments, final discharge or reuse. To identify the most effective bioreactor typology for the secondary treatment of a wastewater resulting from a textile industry of the Biella district (Italy), four pilot units characterized by a different configuration and fluid dynamics (i.e., Bioflotation^®, Fixed Bed Biofilm Reactor (FBBR), flow-jet aeration and standard aerobic sludge reactors) were operated in parallel, inoculated with the same microbial consortium and fed with identical streams of wastewater discharged from wet textile processes of the industy. COD, TC and non-ionic surfactants were monitored in effluents of the compared bioreactors working under continuous mode and the cultivable heterotrophic microorganisms prevailing in each of them were isolated and characterized as the end of the study. The results demonstrated that the air supply system greatly influenced the treatment efficiency which reached the highest value in the case of Bioflotation^® and FBBR technology. A highly specialized bacterial biomass mostly composed by strains of the Pseudomonas, Stenotrophomonas and Ochrobactrum genera was isolated in such reactors, thus suggesting that a direct correlation between reactor configuration, decontamination performances and microbial biomass composition exist.     

Effect of seed sludge and operation conditions on performance and archaeal community structure of low-temperature anaerobic solvent-degrading bioreactors

Two laboratory-scale expanded granular sludge bed (EGSB) anaerobic bioreactors (R1 and R2) were inoculated with biomass from different mesophilic (37 °C) treatment plants, and used for the treatment of an organic solvent-based wastewater at 9–14 °C at applied organic loading rates (OLRs) of 1.2–3.6 kg chemical oxygen demand (COD) m⁻³ d⁻¹. Replicated treatment performance was observed at 10–14 °C, which suggested the feasibility of the process at pilot-scale. Stable and efficient COD removal, along with high methane productivity, was demonstrated at 9 °C at an applied OLR of 2.4 kg COD m⁻³ d⁻¹. Clonal libraries and fluorescence in situ hybridization (FISH) indicated that the seed sludges were dominated (>60%) by acetoclastic Methanosaeta-like organisms. Specific methanogenic activity (SMA) profiles indicated shifts in the physiological profiles of R1 and R2 biomass, including the development of psychrotolerant methanogenic activity. Acetoclastic methanogenesis represented the primary route of methane production in R1 and R2, which is in contrast with several previous reports from low-temperature bioreactor trials. A reduction in the abundance of Methanosaeta-like clones (R2), along with the detection of hydrogenotrophic methanogenic species, coincided with altered granule (sludge) morphology and the development of hydrogenotrophic SMA after prolonged operation at 9 °C.     

Examination of microbial fuel cell start-up times with domestic wastewater and additional amendments

Rapid startup of microbial fuel cells (MFCs) and other bioreactors is desirable when treating wastewaters. The startup time with unamended wastewater (118 h) was similar to that obtained by adding acetate or fumarate (110-115 h), and less than that with glucose (181 h) or Fe(III) (353 h). Initial current production took longer when phosphate buffer was added, with startup times increasing with concentration from 149 h (25 mM) to 251 h (50 mM) and 526 h (100 mM). Microbial communities that developed in the reactors contained Betaproteobacteria, Acetoanaerobium noterae, and Chlorobium sp. Anode biomass densities ranged from 200 to 600 μg/cm² for all amendments except Fe(Ш) (1650 μg/cm²). Wastewater produced 91 mW/m², with the other MFCs producing 50 mW/m² (fumarate) to 103 mW/m² (Fe(III)) when amendments were removed. These experiments show that wastewater alone is sufficient to acclimate the reactor without the need for additional chemical amendments.     

Pathogen removal in farm-scale psychrophilic anaerobic digesters processing swine manure

This study assessed the efficiency of commercial-scale psychrophilic anaerobic digestion in sequencing batch reactors (PADSBRs) for pathogen removal from pig manure. The impact of treatment cycle length and of hydraulic flow regimes on pathogen removal efficiency was investigated. Two conventionally operated SBRs (BR1 and BR2) and two SBRs simultaneously fed during the draw step (BR3 and BR4) were monitored over a two-year period. PADSBRs significantly decreased the concentration of coliforms, Salmonella, Campylobacter spp., and Y. enterocolitica, respectively from about 10⁶, 10³ CFU g⁻¹, 10³, and 10⁴ CFU g⁻¹ to undetectable levels in most samples. Densities of the gram-positive Clostridium perfringens and Enterococcus spp. remained high (10⁵ CFU g⁻¹) in the digesters throughout treatment. The PADSBRs maintained the same level of pathogen removal when the treatment cycle length was reduced from 2 to 1 week. Mass balances on volatile fatty acids (VFAs) revealed short-circuits of inlet flow respectively averaging 6.3% and 6.4% for BR3 and BR4, significantly reducing the overall performance of these reactors regarding pathogens removal. The results obtained in this study show the ability of low temperature anaerobic digestion to remove or significantly reduce indicator and pathogen concentration from raw pig manure.     

Development of a simultaneous partial nitrification and anaerobic ammonia oxidation process in a single reactor

Up-flow oxygen-controlled biofilm reactors equipped with a non-woven fabric support were used as a single reactor system for autotrophic nitrogen removal based on a combined partial nitrification and anaerobic ammonium oxidation (anammox) reaction. The up-flow biofilm reactors were initiated as either a partial nitrifying reactor or an anammox reactor, respectively, and simultaneous partial nitrification and anammox was established by careful control of the aeration rate. The combined partial nitrification and anammox reaction was successfully developed in both biofilm reactors without additional biomass inoculation. The reactor initiated as the anammox reactor gave a slightly higher and more stable mean nitrogen removal rate of 0.35 (± 0.19) kg-N m⁻³ d⁻¹ than the reactor initiated as the partial nitrifying reactor (0.23 (± 0.16) kg-N m⁻³ d⁻¹). FISH analysis revealed that the biofilm in the reactor started as the anammox reactor were composed of anammox bacteria located in inner anoxic layers that were surrounded by surface aerobic AOB layers, whereas AOB and anammox bacteria were mixed without a distinguishable niche in the biofilm in the reactor started as the partial nitrifying reactor. However, it was difficult to efficiently maintain the stable partial nitrification owing to inefficient aeration in the reactor, which is a key to development of the combined partial nitrification and anammox reaction in a single biofilm reactor.     

Effect of chitosan on UASB treating POME during a transition from mesophilic to thermophilic conditions

The effects of chitosan addition on treatment of palm oil mill effluent were investigated using two lab-scale upflow anaerobic sludge bed (UASB) reactors: (1) with chitosan addition at the dosage of 2 mg chitosan per g volatile suspended solids on the first day of the operation (R1), (2) without chitosan addition (the control, R2). The reactors were inoculated with mesophilic anaerobic sludge which was acclimatized to a thermophilic condition with a stepwise temperature increase of 5 °C from 37 to 57 °C. The OLR ranged from 2.23 to 9.47 kg COD m⁻³ day⁻¹. The difference in biogas production rate increased from non-significant to 18% different. The effluent volatile suspended solids of R1 was 65 mg l⁻¹ lower than that of R2 on Day 123. 16S rRNA targeted denaturing gradient gel electrophoresis (DGGE) fingerprints of microbial community indicated that some methanogens in the genus Methanosaeta can be detected in R1 but not in R2.     

Limitations of thermophilic anaerobic wastewater treatment and the consequences for process design

Thermophilic anaerobic digestion offers an attractive alternative for the treatment of medium- and high-strength wastewaters. However, literature reports reveal that thermophilic wastewater treatment systems are often more sensitive to environmental changes than the well-defined high-rate reactors at the mesophilic temperature range. Also, in many cases a poorer effluent quality is experienced while the carry over of suspended solids in the effluent is relatively high. In this paper recent achievements are discussed regarding the process stability of thermophilic anaerobic wastewater treatment systems. Laboratory experiments reveal a relatively low sensitivity to temperature changes if high-rate reactors with immobilized biomass are used. Other results show that if a staged process is applied, thermophilic reactors can be operated for prolonged periods of time under extreme loading conditions (80–100 kg chemical oxygen demand.m^-3.day^-1), while the concentrations of volatile fatty acids in the effluent remain at a low level.     

Biological kinetics evaluation of anaerobic stabilization pond treatment of palm oil mill effluent

Biological kinetic (bio-kinetic) study of the anaerobic stabilization pond treatment of palm oil mill effluent (POME) was carried out in a laboratory anaerobic bench scale reactor (ABSR). The reactor was operated at different feed flow-rates of 0.63, 0.76, 0.95, 1.27, 1.9 and 3.8 l of raw POME for a day. Chemical oxygen demand (COD) as influent substrates was selected for bio-kinetic study. The investigation showed that the growth yield (Y_G), specific biomass decay (b), maximum specific biomass growth rate (μ_max), saturation constant (K_s) and critical retention time (Θ_c) were in the range of 0.990 g VSS/g COD_removed day, 0.024 day⁻¹, 0.524 day⁻¹, 203.433 g COD l⁻¹ and 1.908 day, respectively.     

Saline adaptation of granules in mesophilic UASB reactors

We exposed mesophilic up-flow anaerobic sludge blanket (UASB) reactors to high concentrations of NaCl to elucidate the saline adaptation capacity of their granular sludge. We operated 10 lab-scale UASB reactors at 37 °C and added NaCl to the influent either abruptly or gradually. With abrupt addition, NaCl concentrations were increased from 0 g L⁻¹ to 20, 30, 35, 40, 45, or 50 g L⁻¹. With gradual addition, the NaCl concentrations were gradually increased from 0 to 64 g L⁻¹ or 0 to 40 g L⁻¹. We successfully saline-adapted the granules up to 32 g NaCl L⁻¹, while maintaining high reactor performance, suggesting that 32 g NaCl L⁻¹ is a practical level for system operation. In the UASB reactors gradually exposed to 32 g L⁻¹ NaCl, methane production decreased by only 13%. We also learned that combining abrupt and gradual salinity increases could shorten the adaptation period. Thus we were able to shorten the adaptation period to only 30 days by increasing the salinity abruptly to 20 g L⁻¹, followed by gradual adaptation to 30 g NaCl L⁻¹.     

The treatment of cheese whey wastewater by sequential anaerobic and aerobic steps in a single digester at pilot scale

The treatment of reconstituted whey wastewater was performed in a 400 L digester at 20 °C, with an anaerobic digestion step, followed by a step of aerobic treatment at low oxygen concentration in the same digester. In a first set of 48 cycles, total cycle time (T_C) of 2, 3 and 4 days were tested at varying organic loading rates (OLR). The COD removal reached 89 ± 4, 97 ± 3 and 98 ± 2% at T_C of 2, 3 and 4 days and OLR of 0.56, 1.04 and 0.78 gCOD L⁻¹ d⁻¹, respectively. The activity of the biomass decreased for the methanogenic population, while increasing by 400% for the acidogens, demonstrating a displacement in the predominant trophic group in the biomass bed. A second set of 16 cycles was performed with higher soluble oxygen concentration in the bulk liquid (0.5 mg L⁻¹) during the aerobic treatment at a T_C of 2 days and an OLR of 1.55 gCOD L⁻¹ d⁻¹, with a soluble COD removal of 88 ± 3%. The biomass specific activities showed a compartmentalization of the trophic group with methanogenic activity maintained in the biomass bed and a high acidogenic activity in the suspended flocs.     

Effect of organic loading rate on VFA production, organic matter removal and microbial activity of a two-stage thermophilic anaerobic membrane bioreactor

This study focused on the VFA (volatile fatty acid) profile variation with organic loading rate (OLR) of a two stage thermophilic anaerobic membrane bioreactor (TAnMBR). The two stage TAnMBR treating high strength molasses-based synthetic wastewater was operated under a side-stream partial sedimentation mode at 55 °C. Reactor performances were studied at different OLR ranging from 5 to 12 kg COD m⁻³ d⁻¹. Operational performance of TAnMBR was monitored by assessing biological activity, organic removal efficiency, and VFA. The major intermediate products of anaerobic digestion were identified as acetate, propionate, iso-butyrate, n-butyrate and valerate. Among them acetate and n-butyrate were identified as the most abundant components. Increase of OLR changes the predominant VFA type from acetic acid to n-butyric acid and the total VFA concentration was increased with increased OLR. Moreover, increased OLR increased organic removal efficiency up to second loading rate and dropped in third loading rate while biological activity was increased continuously.     

Maximum removal rate of propionic acid as a sole carbon source in UASB reactors and the importance of the macro- and micro-nutrients stimulation

The maximum propionic acid (HPr) removal rate (R_HPr) was investigated in two lab-scale Upflow Anaerobic Sludge Bed (UASB) reactors. Two feeding strategies were applied by modifying the hydraulic retention time (HRT) in the UASB_HRT and the influent HPr concentration in the UASB_HPr, respectively. The experiment was divided into three main phases: phase 1, influent with only HPr; phase 2, HPr with macro-nutrients supplementation and phase 3, HPr with macro- and micro-nutrients supplementation. During phase 1, the maximum R_HPr achieved was less than 3 g HPr-COD L⁻¹ d⁻¹ in both reactors. However, the subsequent supplementation of macro- and micro-nutrients during phases 2 and 3 allowed to increase the R_HPr up to 18.1 and 32.8 g HPr-COD L⁻¹ d⁻¹, respectively, corresponding with an HRT of 0.5 h in the UASB_HRT and an influent HPr concentration of 10.5 g HPr-COD L⁻¹ in the UASB_HPr. Therefore, the high operational capacity of these reactor systems, specifically converting HPr with high throughput and high influent HPr level, was demonstrated. Moreover, the presence of macro- and micro-nutrients is clearly essential for stable and high HPr removal in anaerobic digestion.     

Modeling of biomass production by Spirulinaplatensis as function of phosphate concentrations and pH regimes

Batch cultivations were performed to evaluate the influence of phosphate concentrations (0.25, 0.5, 0.75, and 1.0 g L⁻¹) for pH regimes (9.5, 10.0, and 10.5) on the biomass production by Spirulina platensis. The best condition for cell growth (3.099 g L⁻¹) was found at 0.5 g L⁻¹ phosphate and pH value of 10.0. Cultivation time, phosphate, and pH caused to increase significantly (p < 0.01) in biomass production by S. platensis. Lag time was observed up to 4 h. After then, biomass production increased sharply (p < 0.01) from 0.020 g L⁻¹ to 2.063, 2.213, 1.532, and 0.797 g L⁻¹ at 0.25, 0.5, 0.75, and 1.0 g L⁻¹ phosphate values, respectively. Modified Gompertz model could be regarded as sufficient to describe the biomass production by S. platensis with high determination coefficients and low sum of square value indicated that. Biological parameters for biomass production were successfully predicted by modified Gompertz model.     

Nitrogen removal performance of a hybrid anammox reactor

The anaerobic ammonium oxidation (anammox) process has attracted considerable attention in recent years as an alternative to conventional nitrogen removal technologies. In this study, an innovative hybrid reactor combining fluidized and fixed beds for anammox treatment was developed. The fluidized bed was mechanically stirred and the gaseous product could be rapidly released from the anammox sludge to prevent washout of the sludge caused by floatation. The fixed bed comprising a non-woven biomass carrier could efficiently catch sludge to reduce washout. During the operation, nitrogen loading rates to the reactor were increased to 27.3 kg N/m³/d, with total nitrogen removal efficiencies of 75%. The biomass concentration in the fluidized bed reached 26-g VSS/L. Anammox granules were observed in the reactors, with settling velocities and sludge volumetric index of 27.3 ± 6.5 m/h and 23 mL/g, respectively. Quantification of extracellular polymeric substances revealed the anammox granules contained a significant amount of extracellular proteins.