Showing papers on "Bioreactor published in 2003"

Design of a flow perfusion bioreactor system for bone tissue-engineering applications.

Abstract: Several different bioreactors have been investigated for tissue-engineering applications. Among these bioreactors are the spinner flask and the rotating wall vessel reactor. In addition, a new type of culture system has been developed and investigated, the flow perfusion culture bioreactor. Flow perfusion culture offers several advantages, notably the ability to mitigate both external and internal diffusional limitations as well as to apply mechanical stress to the cultured cells. For such investigation, a flow perfusion culture system was designed and built. This design is the outgrowth of important design requirements and incorporates features crucial to successful experimentation with such a system.

Metabolic model for glycogen-accumulating organisms in anaerobic/aerobic activated sludge systems.

Abstract: Glycogen-accumulating organisms (GAO) have the potential to directly compete with polyphosphate-accumulating organisms (PAO) in EBPR systems as both are able to take up VFA anaerobically and grow on the intracellular storage products aerobically. Under anaerobic conditions GAO hydrolyse glycogen to gain energy and reducing equivalents to take up VFA and to synthesise polyhydroxyalkanoate (PHA). In the subsequent aerobic stage, PHA is being oxidised to gain energy for glycogen replenishment (from PHA) and for cell growth. This article describes a complete anaerobic and aerobic model for GAO based on the understanding of their metabolic pathways. The anaerobic model has been developed and reported previously, while the aerobic metabolic model was developed in this study. It is based on the assumption that acetyl-CoA and propionyl-CoA go through the catabolic and anabolic processes independently. Experimental validation shows that the integrated model can predict the anaerobic and aerobic results very well. It was found in this study that at pH 7 the maximum acetate uptake rate of GAO was slower than that reported for PAO in the anaerobic stage. On the other hand, the net biomass production per C-mol acetate added is about 9% higher for GAO than for PAO. This would indicate that PAO and GAO each have certain competitive advantages during different parts of the anaerobic/aerobic process cycle.

New hepatocyte in vitro systems for drug metabolism: metabolic capacity and recommendations for application in basic research and drug development, standard operation procedures.

Abstract: Primary hepatocytes represent a well-accepted in vitro cell culture system for studies of drug metabolism, enzyme induction, transplantation, viral hepatitis, and hepatocyte regeneration. Recently, a multicentric research program has been initiated to optimize and standardize new in vitro systems with hepatocytes. In this article, we discuss five of these in vitro systems: hepatocytes in suspension, perifusion culture systems, liver slices, co-culture systems of hepatocytes with intestinal bacteria, and 96-well plate bioreactors. From a technical point of view, freshly isolated or cryopreserved hepatocytes in suspension represent a readily available and easy-to-handle in vitro system that can be used to characterize the metabolism of test substances. Hepatocytes in suspension correctly predict interspecies differences in drug metabolism, which is demonstrated with pantoprazole and propafenone. A limitation of the hepatocyte suspensions is the length of the incubation period, which should not exceed 4hr. This incubation period is sufficiently long to determine the metabolic stability and to allow identification of the main metabolites of a test substance, but may be too short to allow generation of some minor, particularly phase II metabolites, that contribute less than 3% to total metabolism. To achieve longer incubation periods, hepatocyte culture systems or bioreactors are used. In this research program, two bioreactor systems have been optimized: the perifusion culture system and 96-well plate bioreactors. The perifusion culture system consists of collagen-coated slides allowing the continuous superfusion of a hepatocyte monolayer with culture medium as well as establishment of a constant atmosphere of 13% oxygen, 82% nitrogen, and 5% CO2. This system is stable for at least 2 weeks and guarantees a remarkable sensitivity to enzyme induction, even if weak inducers are tested. A particular advantage of this systemis that the same bioreactor can be perfused with different concentrations of a test substance in a sequential manner. The 96-well plate bioreactor runs 96 modules in parallel for pharmacokinetic testing under aerobic culture conditions. This system combines the advantages of a three-dimensional culture system in collagen gel, controlled oxygen supply, and constant culture medium conditions, with the possibility of high throughput and automatization. A newly developed co-culture system of hepatocytes with intestinal bacteria offers the possibility to study the metabolic interaction between liver and intestinal microflora. It consists of two chambers separated by a permeable polycarbonate membrane, where hepatocytes are cultured under aerobic and intestinal bacteria in anaerobic conditions. Test substances are added to the aerobic side to allow their initial metabolism by the hepatocytes, followed by the metabolism by intestinal bacteria at the anaerobic side. Precision-cut slices represent an alternative to isolated hepatocytes and have been used fo the investigation of hepatic metabolism, hepatotoxicity, and enzyme induction. A specific advantage of liver slices is the possibility to study toxic effects on hepatocytes that are mediated or modified by nonparenchymal cells (e.g., by cytokine release from Kupffer cells) because the physiological liver microarchitecture is maintained in cultured slices. For all these in vitro systems, a prevalidation has been performed using standard assays for phase I and II enzymes. Representative results with test substances and recommendations for application of these in vitro systems, as well as standard operation procedures are given.

Concentric cylinder bioreactor for production of tissue engineered cartilage: Effect of seeding density and hydrodynamic loading on construct development

Abstract: A concentric cylinder bioreactor has been developed to culture tissue engineered cartilage constructs under hydrodynamic loading. This bioreactor operates in a low shear stress environment, has a large growth area for construct production, allows for dynamic seeding of constructs, and provides for a uniform loading environment. Porous poly-lactic acid constructs, seeded dynamically in the bioreactor using isolated bovine chondrocytes, were cultured for 4 weeks at three seeding densities (60, 80, 100 x 10(6) cells per bioreactor) and three different shear stresses (imposed at 19, 38, and 76 rpm) to characterize the effect of chondrocyte density and hydrodynamic loading on construct growth. Construct seeding efficiency with chondrocytes is greater than 95% within 24 h. Extensive chondrocyte proliferation and matrix deposition are achieved so that after 28 days in culture, constructs from bioreactors seeded at the highest cell densities contain up to 15 x 10(6) cells, 2 mg GAG, and 3.5 mg collagen per construct and exhibit morphology similar to that of native cartilage. Bioreactors seeded with 60 million chondrocytes do not exhibit robust proliferation or matrix deposition and do not achieve morphology similar to that of native cartilage. In cultures under different steady hydrodynamic loading, the data demonstrate that higher shear stress suppresses matrix GAG deposition and encourages collagen incorporation. In contrast, under dynamic hydrodynamic loading conditions, cartilage constructs exhibit robust matrix collagen and GAG deposition. The data demonstrate that the concentric cylinder bioreactor provides a favorable hydrodynamic environment for cartilage construct growth and differentiation. Notably, construct matrix accumulation can be manipulated by hydrodynamic loading. This bioreactor is useful for fundamental studies of construct growth and to assess the significance of cell density, nutrients, and hydrodynamic loading on cartilage development. In addition, studies of cartilage tissue engineering in the well-characterized, uniform environment of the concentric cylinder bioreactor will develop important knowledge of bioprocessing parameters critical for large-scale production of engineered tissues.

Enrichment of denitrifying glycogen-accumulating organisms in anaerobic/anoxic activated sludge system.

Abstract: Denitrifying glycogen-accumulating organisms (DGAO) were successfully enriched in a lab-scale sequencing batch reactor (SBR) running with anaerobic/anoxic cycles and acetate feeding during the anaerobic period. Acetate was completely taken up anaerobically, which was accompanied by the consumption of glycogen and the production of poly-beta-hydroxy-alkanoates (PHA). In the subsequent anoxic stage, nitrate or nitrite was utilized as electron acceptor for the oxidation of PHA, resulting in glycogen replenishment and cell growth. The above phenotype showed by the enrichment culture demonstrates the existence of DGAO. Further, it was found that the anaerobic behavior of DGAO could be predicted well by the anaerobic GAO model of Filipe et al. (2001) and Zeng et al. (2002a). The final product of denitrification during anoxic stage was mainly nitrous oxide (N2O) rather than N-2. The data strongly suggests that N2O production may be caused by the inhibition of nitrous oxide reductase by an elevated level of nitrite accumulated during denitrification. The existence of these organisms is a concern in biological nutrient removal systems that typically have an anaerobic/anoxic/aerobic reactor sequence since they are potential competitors to the polyphosphate-accumulating organisms. (C) 2003 Wiley Periodicals, Inc.

Optimization of the microbial synthesis of dihydroxyacetone from glycerol with Gluconobacter oxydans

Abstract: An optimized repeated-fed-batch fermentation process for the synthesis of dihydroxyacetone (DHA) from glycerol utilizing Gluconobacter oxydans is presented Cleaning, sterilization, and inoculation procedures could be reduced significantly compared to the conventional fed-batch process A stringent requirement was that the product concentration was kept below a critical threshold level at all times in order to avoid irreversible product inhibition of the cells On the basis of experimentally validated model calculations, a threshold value of about 60 kg m-3 DHA was obtained The innovative bioreactor system consisted of a stirred tank reactor combined with a packed trickle-bed column In the packed column, active cells could be retained by in situ immobilization on a hydrophilized Ralu-ring carrier material Within 17 days, the productivity of the process could be increased by 75% to about 28 kg m-3 h-1 However, it was observed that the maximum achievable productivity had not been reached yet

Integrated closed loop system for industrial water purification

Abstract: The present invention relates to an integrated closed loop system for aquaculture in at least one culturing tank and using continuous bioreactor technology for the biological treatment and removal of organic material, nitrogen and phosphorous, comprising: an integrated, partially or wholly closed loop system for waste water treatment, where the water contains nitrogen containing compounds and/or substances, comprising at least one production unit of such nitrogen containing compounds and/or substances and using continuous bioreactor technology for the biological treatment and removal of organic matter, nitrogen and phosphorous from the said water at continuous flow, comprising: a) at least one suspended carrier bioreactor for bacterial growth under anoxic conditions to cause anaerobic denitrification, with one or several compartments, preceding b) at least one suspended-carrier bioreactor for bacterial growth under oxic conditions to cause aerobic nitrification, c) the denitrification taking place after the production unit, and d) the nitrification taking place prior to the production unit in a by-pass mode as part of the continuous flow

H2 production with anaerobic sludge using activated-carbon supported packed-bed bioreactors.

Abstract: Packed-bed bioreactors containing activated carbon as support carrier were used to produce H2 anaerobically from a sucrose-limiting medium while acclimated sewage sludge was used as the H2 producer The effects of bed porosity (eb) and substrate loading rate on H2 fermentation were examined using packed beds with eb of 70–90% being operated at hydraulic retention times (HRT) of 05–4 h Higher eb and lower HRT favored H2 production With 20 g COD l−1 of sucrose in the feed, the optimal H2 production rate (74 l h−1 l−1) was obtained when the bed with eb=90% was operated at HRT = 05 h Flocculation of cells enhanced the retention of sludge for stable operations of the bioreactor at low HRTs The gas products resulting from fermentative H2 production consisted of 30–40% H2 and 60–70% CO2 Butyric acid was the primary soluble product, followed by propionic acid and valeric acid

Anaerobic Granular Sludge Bioreactor Technology

Abstract: Anaerobic digestion is a mature wastewater treatment technology, with worldwide application. The predominantly applied bioreactor designs, such as the upflow anaerobic sludge blanket and expanded granular sludge bed, are based on the spontaneous formation of granular sludge. Despite the exploitation of granular reactors at full-scale for more than two decades, the mechanisms of granulation are not completely understood and numerous theories have been put forward to describe the process from a biological, ecological and engineering point of view. New technological opportunities are emerging for anaerobic digestion, aided by an improved understanding of microbiological and environmental factors affecting the formation and activity of anaerobic granular sludge.

Substrate concentration-independent aerobic granulation in sequential aerobic sludge blanket reactor.

Abstract: The development of aerobic granules was studied in four column‐type sequential aerobic sludge blanket reactors fed with different substrate concentrations ranging from 500 to 3000 mg l−1 COD. Resul...

Growth and ginsenoside production in hairy root cultures of Panax ginseng using a novel bioreactor.

Abstract: We tested the effect of three variables: the bioreactor system (Wave or Spray reactor), medium exchange and culture period, on the capacity of a selected hairy root line of Panax ginseng to produce ginsenosides. Among the reactors, the Wave bioreactor appeared to be the most efficient in promoting hairy root line growth. Periodic exchanges of the medium and a longer culture period increased the growth rate of cultured hairy root line and, consequently, its capacity to produce ginsenosides. Under established optimum conditions (medium exchange every 14 days over a culture period of 56 days using the Wave bioreactor), the initial root fresh weight was enhanced more than 28-fold, giving a root biomass of 284.9 g L(-1) and a ginsenoside content of 145.6 mg L(-1). It is noteworthy that this ginsenoside production exceeded by almost 3-fold that obtained during the shake flask culture of our hairy root line, although it often happens that the scale-up from shake flask to a bioreactor culture results in reduced productivities. To our knowledge this is the first time that a Wave bioreactor has been used for hairy root culture.

Method for treating wastewater in a membrane bioreactor to produce a low phosphorus effluent

Abstract: Removal of biological nutrients from a wastewater yielding a low phosphorous (e.g., less than 0.25 mg/L) output includes providing a serial multistage bioreactor containing activated sludge having in hydraulic series an anaerobic zone and a downstream aerobic zone, with each zone having an upstream inlet and a downstream outlet. The wastewater is provided to the anaerobic zone inlet. A quantity of chemical sufficient to precipitate soluble and particulate phosphorous is added to the downstream aerobic zone in an amount sufficient to yield a low phosphorous output. Treated water is separated from the activated sludge and precipitated phosphorous and a return activated sludge separated from the treated water is recycled to the anaerobic zone.

Degradation of xenobiotics in a partitioning bioreactor in which the partitioning phase is a polymer.

Abstract: Two-phase partitioning bioreactors (TPPBs) are characterized by a cell-containing aqueous phase and a second immiscible phase that contains toxic and/or hydro- phobic substrates that partition to the cells at subinhibitory levels in response to the metabolic demand of the organisms. To date, the delivery phase in TPPBs has been a hydrophobic solvent that traditionally needed to possess a variety of important properties including biocompatibil- ity, nonbioavailability, low volatility, and low cost, among others. In the present work we have shown that the organic solvent phase can be replaced by inexpensive polymer beads that function in a similar fashion as organic solvents, delivering a toxic substrate to cells based on equilibrium considerations. Specifically, 3.4 mm diameter beads of poly(ethylene-co-vinyl acetate) (EVA) were used to reduce the aqueous concentration of phenol in a bioreactor from toxic levels (~2,000 mg/L) to subinhibi- tory levels (~750 mg/L), after which Pseudomonas putida ATCC 11172 was added to the system and allowed to consume the total phenol loading. Thus, the beads absorbed the toxic substrate and released it to the cells on demand. The EVA beads, which could be reused, were able to absorb 14 mg phenol/g EVA. This work has opened the possibility of using widely mixed cultures in TPPB systems without concern for degradation of the delivery material and without concern of contamination. B 2003 Wiley Periodicals. Biotechnol Bioeng 84: 399-405, 2003.

Comparison Performances of Membrane Bioreactor and Conventional Activated Sludge Processes on Sludge Reduction Induced by Oligochaete

Abstract: Pilot-scale experiments were carried out to compare sludge reduction induced by Oligochaete in a submerged membrane bioreactor (MBR) and a conventional activated sludge (CAS) reactor for 345 d. Worm growth in the CAS reactor was much better than in the MBR. The average worm density of the aeration tank in the CAS reactor was 71 total worms/mg of volatile suspended solids (VSS), much higher than that in the MBR (10 total worms/mg of VSS). Worms did not naturally produce in the MBR, and the dominant worm type in the MBR depended on sludge inoculation from the CAS reactor. Only two types of worms were found in the MBR, Aeolosoma hemprichicii and Nais elinguis. Worm presence and disappearance in the MBR alternated. Worms in the CAS reactor occurred nearly throughout the operating period and were continuously maintained at over 30 total worms/mg of VSS in the aeration tank for 172 d. Three types of worm were found in the CAS reactor, A. hemprichicii, Pristina aequiseta, and N. elinguis, but P. aequiseta was present only occasionally. The alternating dominance of worm types in both reactors changed between Aeolosoma and Nais, and the time of Aeolosoma dominance was longer than that of Nais dominance. Worm growth in the MBR contributed to neither sludge reduction nor improvement of sludge settling characteristics because of low density. But worm presence and bloom in the CAS reactor greatly decreased sludge yield and improved sludge settling characteristics at high density. Both the average sludge yield (0.17 kg of suspended solids (SS)/kg of chemical oxygen demand removed (CODremoved)) and sludge volume index (60 mL/g) in the CAS reactor were much lower than those in the MBR (0.40 kg of SS/kg of CODremoved and 133 mL/g). Nais had more potential for sludge reduction than Aeolosoma. Worm growth had little impact on effluent quality in the MBR but affected effluent quality very much in the CAS reactor.