Influence of waste water composition on biofilm development in laboratory methanogenic fluidized bed reactors

Summary The influence of the volatile fatty acid composition of waste waters on biofilm development and on the time course of reactor start-up was investigated in laboratory scale fluidized bed reactors. It was found that biofilm development proceeded in a similar way with either acetate, butyrate, propionate or a mixture of these compounds as carbon source in the waste water. Startup was retarded, however, with propionate as sole carbon source. Scanning electron microscopic examination revealed that immobilization of bacteria on the sand used as adhesive support initially occurred in crevices and that thereupon the surface of the sand particles became colonized. The composition of the newly developed biomass was determined when reactors reached steady state. Significant differences in the relative substrate spectra and in the amounts of hydrogenotrophic and acetotrophic methanogenic bacteria were measured. The differences reflected the differences in the composition of the waste waters. The results obtained emphasize the role of the structure of the carrier surface in start-up of methanogenic fluidized bed reactors.Abbreviations used Aw ash weight - COD chemical oxygen demand - EB fluidized bed - hbi vitamin B₁₂-HBI - spt sarcinapterin - UASB upflow anaerobic sludge blanket - VFA volatile fatty acid - VSS volatile suspended solids - Ww wet weight     

Inhibition of propionate degradation by acetate in methanogenic fluidized bed reactors

Summary The degradation of acetate, propionate and butyrate was monitored during start-up of five lab-scale methanogenic fluidized bed reactors on an artificially prepared waste water. The acetate concentration in the reactor content was found to influence the degradation of propionate but not of butyrate. In general, at acetate levels over 200 mg/l the degradation of propionate was below 60%, whereas the degradation was complete at acetate levels under 100 mg/l. The rationale of the inhibition of propionate degradation by acetate is discussed.     

Variation of parameters affecting the start-up of methanogenic fluidized bed reactors

Summary The influence of the medium composition, the inoculum and the inoculation procedure on initial biofilm development in methanogenic fluidized bed reactors was studied on laboratory scale. Trace minerals but not vitamins were found to be essential for biofilm development. Inoculation with heterogeneous bacterial cultures of potentially sand-colonizing microorganisms and/or with pure cultures ofMethanothrix soehngenii did not accelerate biofilm development significantly as compared to inoculation with effluent from a fully operative fluidized bed reactor.     

Early stages in biofilm development in methanogenic fluidized-bed reactors

Summary Biofilm development in methanogenic fluidized-bed reactors with sand as the carrier was studied on a laboratory scale. The microorganisms present in consecutive layers of the biofilm of mature sludge granules were preliminarily characterized on the basis of their morphology, element composition and adhesion capacity and were compared to bacteria which take part in the initial colonization of sand. The early phase of biofilm development was monitored with reactors receiving waste-waters containing different mixtures of volatile fatty acids and inoculated with fluidized-bed reactor effluent for different lengths of time. The results obtained indicate that facultative anaerobic bacteria abundantly present in the outermost biofilm layers of mature sludge granules are probably the main primary colonizers of the sand. Methanothrix spp. or other methanogens were rarely observed among the primary colonizers. The course of biofilm formation was comparable under the various start-up conditions employed including variations in waste-water composition, inoculation and anaerobicity. However, omission of waste-water and thus of substrate resulted in rapid wash-out of the attached biomass. Offprint requests to: W. Heinen     

Brewery wastewater treatment using anaerobic inverse fluidized bed reactors

Two anaerobic inverse fluidized bed reactors were utilized to evaluate organic matter removal from brewery wastewater, applying different OLR and testing two support materials. Hydrodynamic tests varying liquid flow and solid concentration were developed on the supports in order to establish operational conditions. A batch colonization stage was applied using 25% active volume of extendosphere and triturated polyethylene as support materials. The reactors were subsequently operated continuously with stepwise increments in organic loading rate until limiting conditions was reached. For the supports studied, IFBR technology was suitable for organic matter removal present in brewery wastewater with COD removal efficiencies greater than 90%. The reactor with triturated polyethylene support showed an excellent COD removal with OLR values up to 10 g COD/Ld, whereas the reactor with extendosphere support had an excellent hydrodynamic and biologic behavior working with OLR values up to 70 g COD/Ld.     

Evaluation of methanogenic kinetics in an anaerobic fluidized bed reactor (AFBR)

A kinetic study of the methanogenic phase was carried out on a pilot lab scale anaerobic fluidized bed reactor (AFBR) in batch mode. An examination of the effect of initial acetate concentration, bed expansion and bed segregation is presented.Experimental data observed for the acetate removal against time were adjusted to a zero-order kinetic equation, over the chemical oxygen demand (COD) range studied (1430–5340 mg litre⁻¹), independently of the bed expansion (11–37%). The kinetic constant was calculated using robust regression analysis. The zero-order kinetic constant, K₀ was between 1180–1380 mg COD litre⁻¹ h⁻¹ on the fixed bed volume basis, and the maximum specific substrate utilization rate, k, was between 145–198 mg COD g VS⁻¹ h⁻¹.The kinetic behaviour was found to be different throughout the reactor, on the fixed bed volume basis and the activity at the bottom of the bed was lower than the activity in the upper region. However, on an attached volatile solids basis, the activity at the bottom level was the greatest.     

Growth and productivity of Beta vulgaris cell culture in fluidized bed reactors

Beta vulgaris cells were cultured in specially constructed fluidized bed reactors on Murashige and Skoog medium with 3% sucrose as a carbon source. Biomass density levels of approximately 17–18 g dry cell weight liter^–1 for 5 liter fermenters and 9–10 g dry cell weight liter^–1 for 50 liter reactor were observed. Depending on particular cell line, cell content of betalains reached levels ranging from 12 to 25 mg g dry cell weight^–1, representing the productivities of 14 to 17 mg of pigments per liter and day.The authors would like to thank the Chemap Tetra Laval group and the Kommission zur Förderung der wissenschaftlichen Forschung for the financial support of this project (KWF No. 2057.1).     

Real evidence about zeolite as microorganisms immobilizer in anaerobic fluidized bed reactors

Using the scanning electronic microscopy, it was observed that natural zeolite possesses excellent physical characteristics as a support medium in anaerobic fluidized bed reactors (AFBR). Samples for biomass analysis were taken from two identical laboratory-scale AFBR (R-1 and R-2), which were operated with 25% of fluidization. These reactors treated distillery wastewaters (vinasses) at mesophilic temperature (30 ± 2 °C). The experiments were carried out with 0.25–0.50 and 0.50–0.80 mm zeolite particle diameter in reactors R-1 and R-2, respectively. The biomass concentration attached to zeolite in both reactors was found to be in the range of 40–45 g volatile solids/l. COD removal efficiencies as high as 90% were achieved at organic loading rate (OLRs) of up to 20 g COD/l day. The volatile fatty acid (VFA) levels were always lower that the suggested limits for digester failure. The yield coefficient of methane production was 0.29 l CH₄(at STP)/g COD consumed and was virtually independent of the OLR applied. A hybridization technique (fluorescence in situ hybridization, FISH) helped determine the predominant anaerobic microorganisms that colonized the natural zeolite, which were found to be Methanosaeta and Methanosarcinaceae, observing a reduced number of sulphate reducing bacteria. The results obtained for reactors R-1 and R-2 were very similar, showing that the particle size did not significantly influence the microbial community immobilized on zeolite.     

A simple model describing nitrate and nitrite reduction in fluidized bed biological reactors

A uniquely simple model is developed to describe the NO(3) (-) and NO(2) (-) concentration profiles within a dentrification fluidized bed biological reactor. This simple model is compared to experimental data, and to a more complex model similar to those previously proposed in the literature. The simple model fits the experimental data at least as well as the more complex model. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 543-548, 1997.     

Startup of anaerobic fluidized bed reactors with acetic acid as the substrate

The startup of anaerobic fluidized bed reactors, which use Manville R-633 beads as the growth support media, acetate enriched bacterial culture as the inoculum, and acetic acid as the sole substrate, is studied. Tow startup strategies are evaluated: one based on maximum and stable substrate utilization and another based on maximum substrate loading controlled by reactor pH. The startup process is characterized using a number of operational parameters.The reactors again excellent total organic carbon (TOC) removal (i.e., > 97% at a feed concentration of 5000 mg TOC/L) and stable methane production (i.e., 0.90 L CH(4)/g TOC, where TOC(r) is TOC removed) at a early stage of the startup process, regardless of the strategies applied. The loading can be increased rapidly without the danger of being overloaded. Significant losses of growth support media and biomass caused by gas effervescence at higher loadings limits the maximum loading that can be safely applied during startup process.A high reactor immobilized biomass inventory is achievable using the porous growth support media (e.g., Manville 633 beads). A rapid increase in loading creates a substrate rich environment that yields more viable reactor biomass. Both substrate utilization rate (batch and continuous) and immobilized biomass inventory stabilize concomitantly at the late stage of the startup process, indicating the attainment of steady-state conditions in reactors. Therefore, they are better parameters that TOC removal and methane production for characterizing the entire startup process of aerobic fluidized bed reactor.The strategy based on maximum substrate loading controlled by reactor pH significantly shortens the startup time. In this case, the reactor attains steady-state conditions approximately 140 days after startup. On the other hand, a startup time of 200 days is required when the strategy based maximum substrate utilization is adopted. (c) 1993 John Wiley & Sons, Inc.     

Comprehensive modeling of methanogenic biofilms in fluidized bed systems: Mass transfer limitations and multisubstrate aspects

A cognitive model for anaerobic digestion in fluidized bed reactors is developed. The general pathway of the process is divided into five main reactions performed by different bacterial groups. Molecular diffusion of each substrate involved in the reaction scheme is described. Effectiveness factor calculations are performed in steady state for each bacterial group taken into account in the process. The case of a single substrate removal is discussed, and optimal biofilm sizes are found. Sequential substrate removal is investigated, and different kinetic regimes are characterized. The influence of biofilm size and primary substrate removal is discussed in the case of standard concentrations in the liquid phase. This study shows that, according to the theoretical model the limiting step of the process may be different and depends in a large way on mass transfer effects. Finally, importance of biofilm size is compared for acidogenic and methano-genic steps: each reaction is found to be optimized for different biofilm thicknesses. This result may be of interest for design purposes and further dynamic modeling. Concluding remarks concerning the validation of the model are made, and a comparison to experimental data from the literature is presented. (c) 1995 John Wiley & Sons, Inc.     

Biological cyanide degradation in aerobic fluidized bed reactors: treatment of almond seed wastewater

The continuous aerobic transformation of synthetic cyanide waste-water, amygdalin solutions and almond seed extract containing cyanide was investigated in several fluidized bed reactors. Various inocula consisting of activated sludge or soil slurry were used. Successful inoculation was achieved with simple soil slurry. No significant influence was found between the performance of the systems inoculated with a cyanide contaminated soil and a garden soil. The performance and stability of the reactors with respect to degradation rate were tested for a range of cyanide loading conditions, with feed containing only cyanide, and with different additional carbon sources, as well as various CN ratios at a hydraulic retention time of 24 h. No growth with cyanide as the sole source of carbon and nitrogen was observed. The system with lactate as the organic C-source was capable of operating at cyanide concentrations of 160 ppm cyanide with a conversion rate of 0.125 kg cyanide/m³ d. Ammonia was the end product and the effluent concentration was 0.5 ppm CN^–. The systems with ethanol as the organic C-source could degrade only 0.05 kg cyanide/m³ d, whose feed concentration was 60 ppm cyanide. Amygdalin, an organic cyanide-containing compound present in stone fruit seeds, was fed as a model substrate. Degradation rates up to 1.2 kg COD/m³ d could be measured with no free or organically bound cyanide in the effluent. These rates were limited by oxygen transfer, owing to the large amount of degradable COD. The further investigations with almond seed extracts, confirmed the applicability of the aerobic process to treat food-processing waste streams having low concentrations of cyanide with high COD content.The authors with to thank Dr. Ö.M. Kurt for useful discussions.     

An evaluation of single- and separated-phase anaerobic industrial wastewater treatment in fluidized bed reactors

Four fluidized bed reactors were used to evaluate single-and separated-phase anaerobic treatments of a high strength wastewater. Two reactors were fed with a synthetic wastewater, containing glucose as the primary carbon source, with a COD of 1.2 x 10(4) mg/L while the remaining pair were fed with a wastewater with a COD of 6000 mg/L. AT each influent strength, one fluidized bed reactor was operated as a single-phase system while the other was operated as a methanogenic reactor which was preceded by an acidification reactor in a separatedphase system. The reactors were operated under steady-state and variable process conditions. The separated-phase system consistently gave a better quality effluent with lower effluent suspended solids and total COD, and the methane yield was also improved. Under variable process conditions, the separated-phase system was inherently more stable and recovered more rapidly following a shock loading. Propionate and acetate degradation studies indicated that the biomass in the methanogenic fluidized beds of the two-phase systems was more adapted to volatile acid degradation than the biomass in the single-phase fluidized beds.     

Comparing lactate and glycerol as a single-electron donor for sulfate reduction in fluidized bed reactors

Among the greatest challenges to the full implementation of biological sulfate reduction are the cost and availability of the electron source. With the development of the biofuel industry, new organic substrates have become available. Therefore, this work sought to compare the performance of a sulfidogenic process utilizing either lactate or glycerol as the substrate for sulfate-reducing bacteria (SRB) growth. Although sulfate reduction is energetically more favorable with lactate, glycerol is a less expensive alternative because excess production is forecasted with the worldwide development of the biodiesel industry. Continuous experiments were performed in a fluidized bed (FB) reactor containing activated carbon as a carrier for a mixed bacterial population composed of sulfate-reducing and fermentative bacteria. During the lactate-fed phases, incomplete oxidation of lactate to acetate by SRB was the dominant metabolic pathway resulting in as much as 90 % sulfate reduction and high acetate concentrations (2.7 g L^?1). Conversely, in the glycerol-fed phases, glycerol degradation resulted from syntrophic cooperation between sulfate-reducing and fermentative bacteria that produce butyrate along with acetate (1.0 g L^?1) as oxidation products. To our knowledge, this is the first report of butyrate formation during sulfate reduction in a glycerol-fed continuous-flow reactor. Sulfate concentrations were reduced by about 90 % (from 2,000 to 100–300 mg L^?1) when glycerol was being fed to the reactor. Since the FB reactor was able to stand a change from lactate to glycerol, this reactor is recommended as the preferred option should glycerol be selected as a cost-effective alternative to lactate for continuous sulfate reduction.     

Biofilm reactors in anaerobic wastewater treatment

Attached biofilm reactors provide the means for implementing energy-efficient anaerobic wastewater treatment at full scale. Progress has been made in the development of fixed, expanded and fluidized bed anaerobic processes by addressing fundamental reactor design issues. Several new biofilm reactor concepts have evolved from recent studies.     

Biofilm formation in attached microalgal reactors

The objective of this study was to investigate the fundamental question of biofilm formation. First, a drum biofilm reactor was introduced. The drums were coated with three porous substrates (cotton rope, canvas, and spandex), respectively. The relationships among the substrate, extracellular polymeric substances (EPS), and adhesion ratio were analyzed. Second, a plate biofilm reactor (PBR) was applied by replacing the drum with multiple parallel vertical plates to increase the surface area. The plates were coated with porous substrates on each side, and the nutrients were delivered to the cells by diffusion. The influence of nitrogen source and concentration on compositions of EPS and biofilm formation was analyzed using PBR under sunlight. The results indicated that both substrate and nitrogen were critical on the EPS compositions and biofilm formation. Under the optimal condition (glycine with concentration of 1 g l^?1 and substrate of canvas), the maximum biofilm productivity of 54.46 g m^?2 d^?1 with adhesion ratio of 84.4 % was achieved.     

Startup of anaerobic biofilm fluidized bed reactors on molasses and phenol under various conditions

Fluidized sand bed anaerobic biofilm reactors were operated in parallel to study the effects of inoculum, loading, residence time and carrier type on the startup dynamics for the degradation of molasses and phenol. Degradation rates generally depended most directly on concentrations rather than on other operating variables. Residence times did not appear to directly influence startup. Short residence times and high loadings gave the highest specific activities for both substrates. The type of inoculum was found to be most important for the molasses system, and inoculation on fresh carrier was found to be better than reinoculation. The two times higher specific biomass retention on Siran porous glass gave essentially the same degradation rates on a volume basis.List of Symbols L kg/h loading of reactor - M kg/kg biomass per carrier mass - Red. % reduction of feed concentration due to degradation - R kg/(m³ · h) reaction rate - S kg/m³ substrate concentration in reactor and effluent - S ₀ kg/m³ substrate concentration in feed - t h time