Dynamics of biofilm detachment in biofilm airlift suspension reactors

The dynamic change in the overall detachment rate of spherical biofilms in a biofilm airlift suspension reactor was measured after a downshift of the substrate loading rate to zero while all other conditions remained constant. In contrast to the expectations, the overall detachment rate decreased rapidly to a nearly stable level. Correlations available from literature were not able to describe this phenomenon. Concepts were formulated which can describe the observations from this study. Research under dynamic conditions and careful monitoring of the biofilm surface area and biofilm morphology are necessary to elucidate and discriminate biofilm detachment mechanisms. (c) 1995 John Wiley & Sons, Inc.     

Influence of dissolved oxygen concentration on nitrite accumulation in a biofilm airlift suspension reactor

The biofilm airlift suspension (BAS) reactor can treat wastewater at a high volumetric loading rate combined with a low sludge loading. Two BAS reactors were operated, with an ammonium load of 5 kg N/(m(3) d), in order to study the influence of biomass and oxygen concentration on the nitrification process. After start-up the nitrifying biomass in the reactors gradually increased up to 30 g VSS/L. Due to this increased biomass concentration the gas-liquid mass transfer coefficient was negatively influenced. The resulting gradual decrease in dissolved oxygen concentration (over a 2-month period) was associated with a concomitantly nitrite build-up. Short term experiments showed a similar relation between dissolved oxygen concentration (DO) and nitrite accumulation. It was possible to obtain full ammonium conversion with approximately 50% nitrate and 50% nitrite in the effluent. The facts that (i) nitrite build up occurred only when DO dropped, (ii) the nitrite formation was stable over long periods, and (iii) fully depending on DO levels in short term experiments, led to the conclusion that it was not affected by microbial adaptations but associated with intrinsic characteristics of the microbial growth system. A simple biofilm model based on the often reported difference of oxygen affinity between ammonium and nitrite oxydizers was capable of adequately describing the phenomena.Measurements of biomass density and concentration are critical for the interpretation of the results, but highly sensitive to sampling procedures. Therefore we have developed an independent method, based on the residence time of Dextran Blue, to check the experimental methods. There was a good agreement between procedures.The relation between biomass concentration, oxygen mass transfer rate and nitrification in a BAS reactor is discussed. (c) 1997 John Wiley & Sons, Inc.     

Hydrodynamic characteristics and gas-liquid mass transfer in a biofilm airlift suspension reactor

The hydrodynamics and mass transfer, specifically the effects of gas velocity and the presence and type of solids on the gas hold-up and volumetric mass transfer coefficient, were studied on a lab-scale airlift reactor with internal draft tube. Basalt particles and biofilm-coated particles were used as solid phase. Three distinct flow regimes were observed with increasing gas flow rate. The influence of the solid phase on the hydrodynamics was a peculiar characteristic of the regimes. The volumetric mass transfer coefficient was found to decrease with increasing solid loading and particle size. This could be predominantly related to the influence that the solid has on gas hold-up. The ratio between gas hold-up and volumetric mass transfer coefficient was found to be independent of solid loading, size, or density, and it was proven that the presence of solids in airlift reactors lowers the number of gas bubbles without changing their size. To evaluate scale effects, experimental results were compared with theoretical and empirical models proposed for similar systems.     

Control of heterotrophic layer formation on nitrifying biofilms in a biofilm airlift suspension reactor

A Biofilm Airlift Suspension (BAS) reactor was operated with nitrifying biofilm growth and heterotrophic suspended growth, simultaneously converting ammonium and acetate. Growth of heterotrophs in suspension decreases the diffusion limitation for the nitrifiers, and enlarges the nitrifying capacity of a biofilm reactor. Neither nitrifiers nor heterotrophs suffer from additional oxygen diffusion limitation when the heterotrophs grow in suspension. Control of the location of heterotrophic growth, either in suspension or in biofilms over the nitrifying biofilms, was possible by manipulation of the hydraulic retention time. A time delay for formation and disappearance of the heterotrophic biofilms of 10 to 15 days was observed. Surprisingly, it was found that in the presence of the heterotrophic layers the maximum specific activity on ammonia of the nitrifying biofilms increased. The reason for the increase in activity is unknown. The effect of heterotrophic biofilm formation on oxygen diffusion limitation for the nitrifiers is discussed. Some phenomena compensating the increased mass transfer resistance due to the growth of a heterotrophic layer are also presented. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 397-405, 1997.     

Application of two component biodegradable carriers in a particle-fixed biofilm airlift suspension reactor: development and structure of biofilms

Two component biodegradable carriers for biofilm airlift suspension (BAS) reactors were investigated with respect to development of biofilm structure and oxygen transport inside the biofilm. The carriers were composed of PHB (polyhydroxybutyrate), which is easily degradable and PCL (caprolactone), which is less easily degradable by heterotrophic microorganisms. Cryosectioning combined with classical light microscopy and CLSM was used to identify the surface structure of the carrier material over a period of 250 days of biofilm cultivation in an airlift reactor. Pores of 50 to several hundred micrometers depth are formed due to the preferred degradation of PHB. Furthermore, microelectrode studies show the transport mechanism for different types of biofilm structures, which were generated under different substrate conditions. At high loading rates, the growth of a rather loosely structured biofilm with high penetration depths of oxygen was found. Strong changes of substrate concentration during fed-batch mode operation of the reactor enhance the growth of filamentous biofilms on the carriers. Mass transport in the outer regions of such biofilms was mainly driven by advection.     

Influence of detachment,substrate loading and reactor scale on the formation of biofilms in airlift reactors

 For a stable and reliable operation of the biofilm airlift suspension reactor (BAS reactor) means to control biomass concentration, biofilm thickness and biofilm morphology are required. For this reason, the influence of applied detachment forces and surface substrate loading on the formation of heterotrophic biofilms in laboratory-scale BAS reactors was studied. Detachment forces were altered by variation of the initial bare carrier concentration or the superficial air velocity. In addition, the dynamics of biofilm formation during start-up of a full scale BAS reactor (300 m³) was monitored and compared with the laboratory-scale start-up (3 l). This study shows that the biofilm morphology and strength were influenced to a large extent by the surface substrate loading and applied detachment forces. A moderate surface substrate loading and a high detachment force yielded smooth and strong biofilms. The combination of a high surface substrate loading and low detachment forces did lead to rough biofilms, but did not lead to the expected high amount of biomass on the carrier, apparently because of the formation of weaker biofilms. The strength of the bio-films appeared to be related to the detachment forces applied during biofilm formation, in combination with the surface substrate loading. The biofilm morphology and biomass on carrier in the BAS reactor can be controlled using the carrier concentration, substrate loading rate and the superficial air velocity as parameters. The dynamics of biofilm formation during the start-up of a full-scale BAS reactor proved to be similar to heterotrophic biofilm formation in laboratory-scale reactors. This indicates that a model system on the laboratory scale can successfully be applied to predict dynamic phenomena in the full-scale reactor. Received: 31 March 1995/Received revision: 11 August 1995/Accepted: 22 August 1995     

Effect of the nonilluminated part of suspension on biomass production in an algal reactor

The production of algal biomass in the illuminated and nonilluminated part of the suspension in an algal reactor was analyzed with regard to the biological inertia of the algae. The calculations indicate a considerable effect of nonilluminated part of the reactor on algal biomass production. The intensity of suspension stirring affects biomass production only slightly.     

Nitrifying activity monitoring and kinetic parameters determination in a biofilm airlift reactor by respirometry

The applicability of batch respirometry, as a simple technique for monitoring off-line nitrifying activity and kinetic parameters, was evaluated using two sets of ammonia and nitrite concentrations. The O₂ uptake rate (OUR) profiles obtained from the assays were adjusted to a substrate inhibition model. The maximum specific ammonia-oxidizing biomass activity (r_Smax) was 0.079 g N-NH₄ ⁺ g VSS^–1 d^–1 with a half saturation coefficient (K_S) of 11 mg N-NH₄ ⁺ l^–1 and an inhibition coefficient (K_i) of 3300 mg N-NH₄ ⁺ l^–1. Besides, the maximum specific value of nitrite-oxidizing activity was 0.082 g N-NO₂ ^– g VSS^–1 d^–1 with a K_S of 4.1 mg N-NO₂ ^– l^–1 and K_i of 1400 mg N-NO₂ ^– l^–1.     

High-rate acidophilic ferrous iron oxidation in a biofilm airlift reactor and the role of the carrier material

In this study, the feasibility and engineering aspects of acidophilic ferrous iron oxidation in a continuous biofilm airlift reactor inoculated with a mixed culture of Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans bacteria were investigated. Specific attention was paid to biofilm formation, competition between both types of bacteria, ferrous iron oxidation rate, and gas liquid mass transfer limitations. The reactor was operated at a constant temperature of 30 degrees C and at pH values of 0-1.8. Startup of the reactor was performed with basalt carrier material. During the experiments the basalt was slowly removed and the ferric iron precipitates formed served as a biofilm carrier. These precipitates have highly suitable characteristics as a carrier material for the immobilization of ferrous iron-oxidizing bacteria and dense conglomerates were observed. Lowering the pH (0.6-1) resulted in dissolution of the ferric precipitates and induced granular sludge formation. The maximum ferrous iron oxidation rate achieved in this study was about 145 molFe(2+)/m(3).h at a hydraulic residence time of 0.25 h. Optimal treatment performance was obtained at a loading rate of 100 mol/m(3).h at a conversion efficiency as high as 98%. Fluorescent in situ hybridization (FISH) studies showed that when the reactor was operated at high ferrous iron conversion (>85%) for 1 month, the desirable L. ferrooxidans species could out-compete A. ferrooxidans due to the low Fe(2+) and high Fe(3+) concentrations.     

Converting corn wet-milling effluent into high-value fungal biomass in a biofilm reactor

Rhizopus microsporus was grown in an attached growth system using corn wet-milling effluent as a growth medium. This strain was chosen due to its ability to effectively degrade organic matter in corn wet-milling effluent and for its properties to produce significant levels of protein, chitin and chitosan. Fungal growth and organic removal efficiency were examined under both aseptic and non-aseptic conditions with and without nutrient supplementation. Plastic composite support (PCS) tubes, composed of 50% (w/w) polypropylene (PP) and 50% (w/w) agricultural products were used as support media. Significantly higher organic removal measured as chemical oxygen demand (COD) and biomass yield were observed in the bioreactor with PCS tubes than in two control bioreactors; that is with PP tubes alone and suspended growth (without support media). This confirmed that the PCS support medium with agricultural components enhanced fungal growth and organic removal. The results showed that supplementation of nutrients (e.g., mineral salts) under aseptic conditions enhanced the COD removal from 50% to 55% and observed biomass yield from 0.11 to 0.16 g (dry-weight)/g COD(removed) (i.e., from 0.10 to 0.14 g volatile solids (VS)/g COD(removed) approximately). Non-aseptic operation without nutrient supplementation resulted in an observed biomass yield of 0.32 g volatile suspended solids (VSS)/g COD(removed) with no significant improvement in COD removal ( approximately 53%); whereas with nutrient supplementation, the observed biomass yield increased to 0.56 g VSS/g COD(removed) and COD removal improved to 85%. The fungal system was able to degrade the organic matter with concomitant production of high-value fungal biomass. This is the first study that examined the conversion of corn milling waste stream into high value fungal protein.     

Microbial biomass production from rice straw hydrolysate in airlift bioreactors

Rice straw is a by-product of rice production, and a great bioresource as raw biomass material for manufacturing value-adding protein for animal feedstock, which has been paid more and more attention. In the present work, utilizing rice straw hydrolysate as a substrate for microbial biomass production in 11.5L external-loop airlift bioreactors was investigated. Rice straw hydrolysate obtained through acid-hydrolyzing rice straw was used for the culture of yeast Candida arborea AS1.257. The influences of gas flow rate, initial liquid volume, hole diameter of gas sparger and numbers of sieve plates on microbial biomass production were examined. The best results in the external-loop airlift bioreactor were obtained under 9.0 L initial liquid volume, 1.1 (v/v)/min gas flow rate during culture time of 0-24 h and 1.4 (v/v)/min gas flow rate of 24-48 h at 29+/-1 degrees C. The addition of the sieve plates in the riser of the external-loop airlift bioreactor increased productivity. After 48 h, under optimized operation conditions, crude protein productivity with one sieve and two sieves were 13.6 mg/mL and 13.7 mg/mL, respectively, comparing 12.7 mg/mL without sieves in the airlift bioreactor and 11.7 mg/mL in the in the 10-L mechanically stirred tank bioreactor. It is feasible to operate the external-loop airlift bioreactors and possible to reduce the production cost for microbial biomass production from the rice straw hydrolysate.     

Influence of abrasion on biofilm detachment: evidence for stratification of the biofilm

The objective of this paper was to understand the detachment of multispecies biofilm caused by abrasion. By submitting a biofilm to different abrasion strengths (collision of particles), stratification of biofilm cohesion could be highlighted and related to stratification of biofilm bacterial communities using the PCR-SSCP fingerprint method. The biofilm comprised a thick top layer, weakly cohesive and composed of one dominant species, and a thin basal layer, strongly cohesive and composed of a more diverse population. These observations suggest that microbial composition of biofilms may be an important parameter in understanding biofilm detachment.     

Bioaugmentation as a tool for improving the start-up and stability of a pilot-scale partial nitrification biofilm airlift reactor

The effectiveness of bioaugmentation in the improvement of the start-up of a biofilm airlift reactor to perform partial nitrification was investigated. Two identical biofilm airlift reactors were inoculated. The non-bioaugmented reactor (NB-reactor) was inoculated with conventional activated sludge, whereas the bioaugmented reactor (B-reactor) was seeded with the same conventional activated sludge but bioaugmented with nitrifying activated sludge from a pilot plant performing full nitritation under stable conditions (100% oxidation of influent ammonium to nitrite). The fraction of specialized nitrifying activated sludge in the inoculum of the B-reactor was only 6% (measured as dry matter). To simplify comparison of the results, operational parameters were equivalent for both reactors. Partial nitrification was achieved significantly faster in the B-reactor, showing a very stable operation. The results obtained by fluorescence in situ hybridization assays showed that the specialized nitrifying biomass added to the B-reactor remained in the biofilm throughout the start-up period.     

Bacterial cellulose production by Acetobacter xylinum in a 50-L internal-loop airlift reactor

Bacterial cellulose (BC) production was realized in a batch cultivation of Acetobacter xylinum subsp. sucrofermentans BPR2001 in a 50-L internal-loop airlift reactor. When the bacterium was cultivated with air supply, 3.8 g/L of BC was produced after 67 hours. When oxygen-enriched gas was supplied, the concentration of BC was doubled and the production rate of BC was 0.116 g/L. h, which was two times higher than that of air-supplied culture and comparable to that in a mechanically agitated stirred-tank fermentor. Bacterial cellulose produced by the airlift reactor formed a unique ellipse pellet (BC pellet), different from the fibrous form which was produced in an agitated stirred-tank fermentor. The BC-pellet suspension was demonstrated to have a higher volumetric oxygen transfer coefficient than the fibrous BC suspension in a 50-L internal-loop airlift reactor. The mixing time of BC-pellet suspension in the airlift reactor was also shorter than that in water.     

Gluconic acid production from sucrose in an airlift reactor using a multi-enzyme system

Sucrose from sugarcane is produced in abundance in Brazil, which provides an opportunity to manufacture other high-value products. Gluconic acid (GA) can be produced by multi-enzyme conversion of sucrose using the enzymes invertase, glucose oxidase, and catalase. In this process, one of the byproducts is fructose, which has many commercial applications. This work concerns the batch mode production of GA in an airlift reactor fed with sucrose as substrate. Evaluation was made of the influence of temperature and pH, as well as the thermal stability of the enzymes. Operational conditions of 40 °C and pH 6.0 were selected, based on the enzymatic activity profiles and the thermal stabilities. Under these conditions, the experimental data could be accurately described by kinetic models. The maximum yield of GA was achieved within 3.8 h, with total conversion of sucrose and glucose and a volumetric productivity of around 7.0 g L^?1 h^?1.     

Effects of pH profiles on nisin production in biofilm reactor

Apart from its widely accepted commercial applications as a food preservative, nisin emerges as a promising alternative in medical applications for bacterial infection in both humans and livestock. Improving nisin production through optimization of fermentation parameters would make nisin more cost-effective for various applications. Since nisin production by Lactococcus lactis NIZO 22186 was highly influenced by the pH profile employed during fermentation, three different pH profiles were evaluated in this study: (1) a constant pH profile at 6.8 (profile 1), (2) a constant pH profile with autoacidification at 4 h (profile 2), and (3) a stepwise pH profile with pH adjustment every 2 h (profile 3). The results demonstrated that the low-pH stress exerted during the first 4 h of fermentation in profile 3 detrimentally affected nisin production, resulting in a very low maximum nisin concentration (593 IU ml⁻¹). On the other hand, growth and lactic acid production were only slightly delayed, indicating that the loss in nisin production was not a result of lower growth or shifting of metabolic activity toward lactic acid production. Profile 2, in which pH was allowed to drop freely via autoacidification after 4 h of fermentation, was found to yield almost 1.9 times higher nisin (3,553 IU ml⁻¹) than profile 1 (1,898 IU ml⁻¹), possibly as a result of less adsorption of nisin onto producer cells. Therefore, a combination of constant pH and autoacidification period (profile 2) was recommended as the pH profile during nisin production in a biofilm reactor.     

Investigations of cephalosporin C production in an airlift tower loop reactor

Summary Cephalosporin C was produced by Cephalosporium acremonium in a 60 l airlift loop reactor on complex medium (with 30 kg/m³ peanut flour) in fed-batch operation. A final product concentration of 5 kg/m³ and a maximum productivity of 45 g/m³ h were attained. On-line analysis was used to determine ammonia, methionine, phosphate, reducing sugar and cephalosporin C by an autoanalyser, glucose by a flow injection analyser and cephalosporin C, penicillin N, deacetoxycephalosporin C, deacetylce-phalosporin C and methionine by HPLC. The volumetric productivity of the stirred tank reactor was higher than that of the airlift reactor because of differences in cell concentration. Specific productivities in relative to cell mass were similar in the two reactors. The substrate yield coefficient in the airlift reactor was twice that in the stirred tank reactor.Nomenclature E _o2 efficiency of oxygen transfer with regard to the specific power input - K _La volumetric mass transfer coefficient - OTR oxygen transfer rate - P power input - PR volumetric productivity of CPC - q _a volumetric aeration rate/broth volume (vvm) - SPR specific productivity with regard to RNA - V _L broth volume in reactor - z relative height of the aerated reactor     

Outdoor production of Phaeodactylum tricornutum biomass in a helical reactor

The production of microalga Phaeodactylum tricornutum in an outdoor helical reactor was analysed. The influence of temperature, solar irradiance and air flow rate on the yield of the culture was evaluated. Biomass productivities up to 1.5 g l(-1) per day and photosynthetic efficiency up to 14% were obtained by maintaining the cultures below 30 degrees C, dissolved oxygen levels less than 400% Sat. (with respect to air saturated culture) and controlling the cell density in order to achieve an average irradiance within the culture below 250 microE m(-2) s(-1). Under these conditions, the fluorescence parameter, Fv/Fm, which reflects the maximal efficiency of PSII photochemistry, remained roughly 0.6-0.7 and growth rates up to 0.050 h(-1) were achieved. The average irradiance and the light/dark cycle frequency, were the variables determining the behaviour of the cultures. A hyperbolic relationship between growth rate and biomass productivity with the average irradiance was observed, whereas both biomass productivity and photosynthetic efficiency linearly increased with the light/dark cycle frequencies. Optimum design and operational conditions which maximise the production of P. tricornutum biomass in outdoor helical reactors were determined.     

Effect of addition of water-soluble polysaccharides on bacterial cellulose production in a 50-L airlift reactor

Bacterial cellulose (BC) production was carried out in a batch cultivation of Acetobacter xylinum in a 50-L internal loop airlift reactor by addition of water-soluble polysaccharides into the medium. When 0.1% (w/w) agar was added, BC production reached 8.7 g/L compared with 6.3 g/L in the control, and duration of the cultivation period to reach the maximum concentration of BC was almost half of that without addition of polysaccharides. During cultivation, BC was formed into pellets whose size was smaller when the productivity of BC was higher, indicating that increase in the relative viscosity by addition of polysaccharides hindered formation of large clumps of BC and increase in the volumetric oxygen transfer coefficient at high flow rate led to increase in BC productivity.