Evaluation of production of recombinant human interleukin-2 in fluidized bed bioreactor

The production of recombinant human interleukin-2 in a fluidized bed bioreactor containing porous glass carriers is described. Cultivations were carried out with different medium formulations over 80 days. Maximal cell densities and product yield could be maintained even when protein free medium was perfused, with less than 10% cell washout. Due to this effective immobilization of the cells in the reactor, continuous operation was easy to perform. Final cell densities on the order of 3.8 x 10(8) mL(-1) intrasphere volume were reached while the interleukin-2 production rate was 0.75 mg L(-1) d(-1). The production rate showed a maximum of a 1.9 fold decrease compared with a homogeneous stirred bubble-free aerated system. This result was in contrast to that achieved with hybridoma cell lines, where better performance was obtained with the fluidized bed bioreactor. The situation may reflect the problems caused by the dense cell culture with adherent cells, as previously shown in a hollow-fiber bioreactor with the same cell line.     

Detachment of multi species biofilm in circulating fluidized bed bioreactor

In this study, the detachment rates of various microbial species from the aerobic and anoxic biofilms in a circulating fluidized bed bioreactor (CFBB) with two entirely separate aerobic and anoxic beds were investigated. Overall detachment rate coefficients for biomass, determined on the basis of volatile suspended solids (VSS), glucose and protein as well as for specific microbial groups, i.e., for nitrifiers, denitrifiers, and phosphorous accumulating organisms (PAOs), were established. Biomass detachment rates were found to increase with biomass attachment on carrier media in both beds. The detachment rate coefficients based on VSS were significantly affected by shear stress, whereas for protein, glucose and specific microbial groups, no significant effect of shear stress was observed. High detachment rates were observed for the more porous biofilm structure. The presence of nitrifiers in the anoxic biofilm and denitrifiers in the aerobic biofilm was established by the specific activity measurements. Detachment rates of PAOs in aerobic and anoxic biofilms were evaluated.     

Bioartificial liver system based on choanoid fluidized bed bioreactor improve the survival time of fulminant hepatic failure pigs

Bioartificial liver (BAL) support system has been proposed as potential treatment method for end-stage liver diseases. We described an improved BAL system based on a choanoid fluidized bed bioreactor containing alginate-chitosan encapsulated primary porcine hepatocytes. The feasibility, safety, and efficiency of this device were estimated using an allogeneic fulminant hepatic failure (FHF) model. FHF was induced with intravenous administration of D-galactosamine. Thirty FHF pigs were divided into three groups: (1) an FHF group which was only given intensive care; (2) a sham BAL group which was treated with the BAL system with empty encapsulation, and (3) a BAL group which was treated with the BAL system containing encapsulated freshly isolated primary porcine hepatocytes. The survival times and biochemical parameters of these animals were measured, and properties of the encapsulations and hepatocytes before and after perfusion were also evaluated. Compared to the two control groups, the BAL-treated group had prolonged the survival time and decreased the blood lactate levels, blood glucose, and amino acids remained stable. No obvious ruptured beads or statistical decline in viability or function of encapsulated hepatocytes were observed. This new fluidized bed BAL system is safe and efficient. It may represent a feasible alternative in the treatment of liver failure.     

Biotechnological production of xylitol in a three-phase fluidized bed bioreactor with immobilized yeast cells in Ca-alginate beads

Cells of Candida guilliermondii entrapped in Ca-alginate beads were used for xylitol production, from concentrated hemicellulose hydrolyzate of sugarcane bagasse, in a fluidized bed bioreactor (FBR). The maximum xylitol concentration 28.9 g xylitol/L was obtained at a high aeration rate of 600 mL/min after 70 h of fermentation, indicating that the use of high aeration rate in this system is favored for better oxygen transfer into the immobilized cells. The specific xylitol productivity and the xylitol yield were of 0.4 g xylitol/L.h and 0.58 g xylitol/g xylose respectively. The immobilization efficiency at the end of the fermentation was of 65 %. After 90 h of fermentation xylitol productivity and yield decreased to 0.25 g xylitol/L.h and 0.47 g xylitol/g xylose respectively, indicating the beginning of xylitol consumption by the yeast. The use of FBR system with immobilized cells presented high xylitol yield and productivity.     

Continuous ethanol production by flocculating yeast in the fluidized bed bioreactor

Abstract: Continuous fermentation by a highly flocculant strain of the yeast Saccharomyces cerevisiae was carried out in a tower fluidized-bed bioreactor. The synthetic and molasses media with a total sugar concentration of 17% (w/v) were used for fermentation. Different dilution rates were tested. Stable cell densities of 50 kg m^-3(dry weight) were maintained for all dilution rates. The ethanol productivity was increasing linearly with dilution rates up to 15—20 kg m^-3 h^-1. Aeration of the culture stabilized flocculating activity and viability of yeast and also permitted long-term operation of the bioreactor.     

Biodegradation of 3,4-dichloroaniline in a fluidized bed bioreactor and a steady-state biofilm Kinetic model

Mixed culture of microorganisms immobilized onto Celite diatomaceous earth particles were used to degrade 3,4-dichloroaniline (34DCA) in a three-phase draft tube fluidized bed bioreactor. Biodegradation was confirmed as the dominant removal mechanism by measurements of the concomitant chloride ion evolution. Degradation efficiencies of 95% were obtained at a reactor retention time of 1.25 h. A mathematical model was used to describe the simultaneous diffusion and reaction of 34DCA and oxygen in the biofilms on the particles in the reactor. The parameters describing freely suspended cell growth on 34DCA were obtained in batch experiments. The model was found to describe the system well for three out of four steady states and to predict qualitatively the experimentally observed transition in the biofilm kinetics from 34DCA to oxygen limitation.     

Mathematical description of bikaverin production in a fluidized bed bioreactor

Summary  Growth of Gibberella fujikuroi in submerged cultures occurs as micelles or filamentous hyphae dispersed in fluid and pellets or stable, spherical agglomerations. Gibberella fujikuroi growth, substrate consumption and bikaverin production kinetics obtained from submerged batch fermentation were fitted to three different sigmoid models: two and three-parameter Gompertz models and one Logistic model. Growth fitting was used to compare between models and select the best one by means of an F test. The best model for describing growth was the two-parameter Gompertz model and was used for glucose consumption and bikaverin production fitting. Data from eight different schemes of fermentations were analysed and parameter estimation was carried out by means of minimization of residual sum of squares. Some characteristic values obtained with the two-parameter Gompertz model fit are: μ=0.028 h⁻¹, Y_x/s=0.1089 g substrate/g biomass, α =0.1384 g product/g biomass.     

Feasibility study of degradation of phenol in a fluidized bed bioreactor with a cyclodextrin polymer as biofilm carrier

This work is focused on the evaluation of a beta-cyclodextrin polymer as a carrier medium in a fluidized bed bioreactor treating aqueous phenol as a model pollutant. The insoluble polymer support was obtained in the shape of spherical beads by crosslinking beta-cyclodextrin with epichlorohydrin. A batch of swollen polymer particles was loaded into the reactor and inoculated with a mixed bacterial culture. Bacterial growth on the polymer beads was initially stimulated by glucose addition to the medium, and then gradually replaced with phenol. The operational variables studied after the acclimation period included phenol load, hydraulic residence time and recirculation flow rate. Low hydraulic residence times and moderate phenol loads were applied. The elimination capacity was usually about 1.0 kg-phenol/m(3)d, although a maximum of 2.8 kg-phenol/m(3)d was achieved with a retention time of only 0.55 h. The depuration efficiency was not affected by the recirculation flow rate in the range studied. Neither operational nor support stability problems were detected during the operation. A high degree of expansion was achieved in the bioreactor due to the hydrogel nature of the cyclodextrin polymer and, consequently, a low energy requirement was necessary to fluidize the bed.     

The effect of calcium on the treatment of fresh leachate in an expanded granular sludge bed bioreactor

This research investigated the calcium effect on the anaerobic treatment of fresh leachate in an expanded granular sludge bed (EGSB) bioreactor under mesophilic conditions. The observations show that the bioreactor, inoculated with anaerobic granular sludge, can be started up only in about 40 days for the treatment of calcium-containing fresh leachate with chemical oxygen demand (COD) removal efficiency above 90% and organic loading rate up to 72.84 kg COD/m³ day. The calcium accumulation onto the granules was monotonically related to the calcium concentration, accounting for 17-18 wt.% of Ca in the suspended solid in the form of calcium carbonate, phosphates/phosphonates and carboxylates. The mineral formation significantly increased the granule settling velocity (by ∼50%) and the suspended solid concentration (by ∼100%). However, the effect of calcium precipitation on the specific methanogenic activity and the CH₄ production rate was complex, first positive during the start-up but later on negative.     

Evaluation of an integrated sponge--granular activated carbon fluidized bed bioreactor for treating primary treated sewage effluent

An integrated fluidized bed bioreactor (iFBBR) was designed to incorporate an aerobic sponge FBBR (ASB-FBBR) into an anoxic granular activated carbon FBBR (GAC-FBBR). This iFBBR was operated with and without adding a new starch based flocculant (NSBF) to treat synthetic primary treated sewage effluent (PTSE). The NSBF contains starch based cationic flocculants and trace nutrients. The results indicate that the iFBBR with NSBF addition could remove more than 93% dissolved organic carbon (DOC), 61% total nitrogen (T-N) and 60% total phosphorus (T-P) at just a very short hydraulic retention time of 50 min. The optimum frequency of adding NSBF to the iFFBR is four times per day. As a pretreatment to microfiltration, the iFFBR could increase 5 L/m² h of critical flux thus reducing the membrane fouling. In addition, better microbial activity was also observed with high DO consumption (>66%) and specific oxygen uptake rate (>35 mg O₂/g VSS h).     

Anaerobic treatment of 2,4,6-trichlorophenol in an expanded granular sludge bed-anaerobic filter (EGSB-AF) bioreactor at 15 degrees C

Expanded granular sludge bed-anaerobic filter (EGSB-AF) bioreactors were operated at 15 degrees C for the treatment of 2,4,6-trichlorophenol (TCP)-containing volatile fatty acid (VFA)-based wastewaters. The seed sludge used as inoculum for the control (no TCP) and test reactor was unexposed to chlorophenols (CPs) prior to the 425-day trial. TCP supplementation to the feed at 50 mg TCPl(-1) partially inhibited the anaerobic degradation of the VFA feed measured as COD removal efficiency. However, the withdrawal and subsequent application of stepwise increments to the TCP loading resulted in steady COD removal. Terminal restriction fragment length polymorphism analysis showed Methanosaeta-like Archaea in the control reactor over the experimental period. Different methanogenic populations were detected in the test reactor and responded to the changes in feed composition. Bacterial community analyses indicated changes in the community structure over time and suggested the presence of Campylobacter-like, Acidimicrobium-like and Heliophilum-like organisms in the samples. TCP mineralisation was by a reductive dechlorination pathway through 2,4-dichlorophenol (DCP) and 4-chlorophenol (4-CP) or 2-chlorophenol (2-CP). CP degradation rates in sludge granules from the lower chamber of the hybrid EGSB-AF reactor was in the order TCP > DCP > 4-CP > 2-CP. However, a biodegradability order of lower CPs > TCP was observed in fixed-film biomass taken from the upper reactor chamber, thus reflecting the role of this reactor section in the metabolism of residual lower CPs from the lower sludge-bed stage of operation.     

Understanding and modeling retention of mammalian cells in fluidized bed centrifuges

Within the last decade, fully disposable centrifuge technologies, fluidized‐bed centrifuges (FBC), have been introduced to the biologics industry. The FBC has found a niche in cell therapy where it is used to collect, concentrate, and then wash mammalian cell product while continuously discarding centrate. The goal of this research was to determine optimum FBC conditions for recovery of live cells, and to develop a mathematical model that can assist with process scaleup. Cell losses can occur during bed formation via flow channels within the bed. Experimental results with the kSep400 centrifuge indicate that, for a given volume processed: the bed height (a bed compactness indicator) is affected by RPM and flowrate, and dead cells are selectively removed during operation. To explain these results, two modeling approaches were used: (i) equating the centrifugal and inertial forces on the cells (i.e., a force balance model or FBM) and (ii) a two‐phase computational fluid dynamics (CFD) model to predict liquid flow patterns and cell retention in the bowl. Both models predicted bed height vs. time reasonably well, though the CFD model proved more accurate. The flow patterns predicted by CFD indicate a Coriolis‐driven flow that enhances uniformity of cells in the bed and may lead to cell losses in the outflow over time. The CFD‐predicted loss of viable cells and selective removal of the dead cells generally agreed with experimental trends, but did over‐predict dead cell loss by up to 3‐fold for some of the conditions. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1520–1530, 2016     

Comparative kinetic study between moving bed biofilm reactor-membrane bioreactor and membrane bioreactor systems and their influence on organic matter and nutrients removal

New technologies regarding wastewater treatment have been developed. Among these technologies, the moving bed biofilm reactor combined with membrane bioreactor (MBBR-MBR) is a recent solution alternative to conventional processes. This paper presents the results obtained from three wastewater treatment plants working in parallel. The first wastewater treatment plant consisted of a membrane bioreactor (MBR), the second one was a MBBR-MBR system containing carriers both in anoxic and aerobic zones, and the last one consisted of a MBBR-MBR system which contained carriers only in the aerobic zone. The reactors operated with a hydraulic retention time of 26.47 h. During the study, the difference between the experimental plants was not statistically significant concerning organic matter and nutrients removal. However, different tendencies regarding nutrients removal are shown by the three wastewater treatment plants. In this sense, the performances in terms of nitrogen and phosphorus removal of the MBBR-MBR system which contained carriers only in the aerobic zone (67.34 ± 11.22% and 50.65 ± 11.13%, respectively) were slightly better than those obtained from another experimental plants. As a whole, the pilot plant which consisted of a MBR showed better performance from the point of view of the kinetics of the heterotrophic and autotrophic biomass with values of μ_m,H = 0.00858 h⁻¹, μ_m,A = 0.07646 h⁻¹, K_M = 2.37 mg O₂ L⁻¹ and K_NH = 1.31 mg N L⁻¹.     

Anaerobic biodegradation of diesel fuel-contaminated wastewater in a fluidized bed reactor

Diesel fuel spills have a major impact on the quality of groundwater. In this work, the performance of an Anaerobic Fluidized Bed Reactor (AFBR) treating synthetic wastewater is experimentally evaluated. The wastewater comprises tap water containing 100, 200 and 300 mg/L of diesel fuel and nutrients. Granular, inert, activated carbon particles are employed to provide support for biomass inside the reactor where diesel fuel is the sole source of carbon for anaerobic microorganisms. For different rates of organic loading, the AFBR performance is evaluated in terms of the removal of diesel fuel as well as chemical oxygen demand (COD) from wastewater. For the aforementioned diesel fuel concentrations and a wastewater flow rate of 1,200 L/day, the COD removal ranges between 61.9 and 84.1%. The concentration of diesel fuel in the effluent is less than 50 mg/L, and meets the Level II groundwater standards of the MUST guidelines of Alberta.     

对—氯代苯胺（PCA）在填充床生物反应器中的降解作用

以氯代苯胺（ＰＣＡ）为选择基质,用驯化技术从降解对二氯苯（ｐＤＣＢ）的富集培养物中得到了以同化ＰＣＡ为唯一碳源和氮源的混合微生物．将这种固定在填充床反应器中的微生物用于ＰＣＡ的降解作用研究中．在该反应器里,ＰＣＡ的生物降解遵循Ｌｏｇｉｓｔｉｃ方程ｑ＝ｑｍａｘ／（１＋ｅα－βＵｖ）．由方程求出了主要的动力学常数,Ｋｓ（半速率常数）和ｑｍａｘ（最大比基质降解速率）．于ＰＣＡ降解的同时,释放氯离子到培养基中．在水力停留时间３ｈ,进水ＰＣＡ浓度为３６０ｍｇ·Ｌ－１情况下,基质的体积降解率达到１２５ｍｇ·Ｌ－１·ｈ－１;基质的百分去除率为９１％．     

Study on biomass circulation and gasification performance in a clapboard-type internal circulating fluidized bed gasifier

We investigated the solid particle flow characteristics and biomass gasification in a clapboard-type internal circulating fluidized bed reactor. The effect of fluidization velocity on particle circulation rate and pressure distribution in the bed showed that fluidization velocities in the high and low velocity zones were the main operational parameters controlling particle circulation. The maximum internal circulation rates in the low velocity zone came almost within the range of velocities in the high velocity zone, when u_H/u_mf = 2.2–2.4 for rice husk and u_H/u_mf = 3.5–4.5 for quartz sand. In the gasification experiment, the air equvalence ratio (ER) was the main controlling parameter. Rice husk gasification gas had a maximum heating value of around 5000 kJ/m³ when ER = 0.22–0.26, and sawdust gasification gas reached around 6000–6500 kJ/m³ when ER = 0.175–0.24. The gasification efficiency of rice husk reached a maximum of 77% at ER = 0.28, while the gasification efficiency of sawdust reached a maximum of 81% at ER = 0.25.     

Selective production of bikaverin in a fluidized bioreactor with immobilized Gibberella fujikuroi

The best culture medium composition for the production of bikaverin by Gibberella fujikuroi in shake-flasks, i.e. 100 g glucose l^–1; 1 g NH₄Cl l^–1; 2 g rice flour l^–1; 5 g KH₂PO₄ l^–1 and 2.5 g MgSO₄ l^–1, was obtained through a fractional factorial design and then scaled-up to a fluidized bioreactor. The effects of carbon and nitrogen concentrations, inoculum size, aeration, flow rate and bead sizes on batch bikaverin production using immobilized G. fujikuroi in a fluidized bioreactor were determined by an orthogonal experimental design. Concentrations of up to 6.83 g bikaverin l^–1 were obtained when the medium contained 100 g glucose l^–1 and 1 g NH₄Cl l^–1 with an inoculum ratio of 10% v/v, an aeration rate of 3 volumes of air per volume of medium min^–1, and a bead size of 3 mm. Based on dry weight, the bikaverin production was 30–100 times larger than found in submerged culture and approximately three times larger than reported for solid substrate fermentation.     

Effect of nickel deprivation on methanol degradation in a methanogenic granular sludge bioreactor

The effect of omitting nickel from the influent on methanol conversion in an Upflow Anaerobic Sludge Bed (UASB) reactor was investigated. The UASB reactor (30°C, pH 7) was operated for 261 days at a 12-h hydraulic retention time (HRT) and at organic loading rates (OLRs) ranging from 2.6 to 7.8 g COD l reactor⁻¹ day⁻¹. The nickel content of the sludge decreased by 66% during the 261-day reactor run because of washout and doubling of the sludge bed volume. Nickel deprivation initially had a strong impact on the methanogenic activity of the sludge with methanol; e.g., after 89 days of operation, this activity was doubled by adding 2 μM nickel. Upon prolonged UASB reactor operation, methanol and VFA effluent concentrations decreased whereas the sludge lost its response to nickel addition in activity tests. This suggests that a less nickel-dependent methanol-converting sludge had developed in the UASB reactor. Received 09 April 2002/ Accepted in revised form 13 July 2002     

Effects of influent DO/COD ratio on the performance of an anaerobic fluidized bed reactor fed low-strength synthetic wastewater

The effect of influent DO/COD (dissolved oxygen/chemical oxygen demand) ratio on the performance of an anaerobic fluidized bed reactor (AFBR) containing GAC was studied. A high influent DO concentration was found to have adverse impacts on organic removal efficiency, methane production, and effluent suspended solids (SS) concentration. These problems resulted with a DO/COD ratio of 0.12, but not at a lower ratio of 0.05. At first organic removal appeared satisfactory at the higher DO/COD ratio at a hydraulic retention time of 0.30 h, but soon a rapid growth of oxygen-consuming zoogloeal-like organisms resulted, eventually causing high effluent SS concentrations. The influent DO also had an inhibitory effect, resulting in a long recovery time for adequate methanogenic activity to return after influent DO removal began. With the growing interest in anaerobic treatment of low COD wastewaters, the increased possibility of similar adverse DO effects occurring needs consideration.