WorldWideScience

Sample records for bioreactor cultivation conditions

  1. High cell density cultivation of recombinant yeasts and bacteria under non-pressurized and pressurized conditions in stirred tank bioreactors.

    Science.gov (United States)

    Knoll, Arnd; Bartsch, Stefan; Husemann, Bernward; Engel, Philip; Schroer, Kirsten; Ribeiro, Betina; Stöckmann, Christoph; Seletzky, Juri; Büchs, Jochen

    2007-10-31

    This study demonstrates the applicability of pressurized stirred tank bioreactors for oxygen transfer enhancement in aerobic cultivation processes. The specific power input and the reactor pressure was employed as process variable. As model organism Escherichia coli, Arxula adeninivorans, Saccharomyces cerevisiae and Corynebacterium glutamicum were cultivated to high cell densities. By applying specific power inputs of approx. 48kWm(-3) the oxygen transfer rate of a E. coli culture in the non-pressurized stirred tank bioreactor was lifted up to values of 0.51moll(-1)h(-1). When a reactor pressure up to 10bar was applied, the oxygen transfer rate of a pressurized stirred tank bioreactor was lifted up to values of 0.89moll(-1)h(-1). The non-pressurized stirred tank bioreactor was able to support non-oxygen limited growth of cell densities of more than 40gl(-1) cell dry weight (CDW) of E. coli, whereas the pressurized stirred tank bioreactor was able to support non-oxygen limited growth of cell densities up to 225gl(-1) CDW of A. adeninivorans, 89gl(-1) CDW of S. cerevisiae, 226gl(-1) CDW of C. glutamicum and 110gl(-1) CDW of E. coli. Compared to literature data, some of these cell densities are the highest values ever achieved in high cell density cultivation of microorganisms in stirred tank bioreactors. By comparing the specific power inputs as well as the k(L)a values of both systems, it is demonstrated that only the pressure is a scaleable tool for oxygen transfer enhancement in industrial stirred tank bioreactors. Furthermore, it was shown that increased carbon dioxide partial pressures did not remarkably inhibit the growth of the investigated model organisms.

  2. Lipase production by solid-state fermentation: cultivation conditions and operation of tray and packed-bed bioreactors.

    Science.gov (United States)

    Gutarra, Melissa L E; Cavalcanti, Elisa D C; Castilho, Leda R; Freire, Denise M G; Sant'Anna, Geraldo L

    2005-01-01

    The production of lipase by Penicillium simplicissimum in solid-state fermentation was studied using babassu cake as the basal medium. Tray-type and packed-bed bioreactors were employed. In the former, the influence of temperature; content of the medium, and medium supplementation with olive oil, sugarcane molasses, corn steep liquor, and yeast hydrolysate was studied. For all combinations of supplements, a temperature of 30 degrees C, a moisture content of 70%, and a concentration of carbon source of 6.25% (m/m, dry basis) provided optimum conditions for lipase production. When used as single supplements olive oil and molasses also were able to provide high lipase activities (20 U/g). Using packed-bed bioreactors and molasses-supplemented medium, optimum conditions for enzyme production were air superficial velocities above 55 cm/min and temperatures below 28 degrees C. The lower temperature optimum found for these reactors is probably related to radial heat gradient formation inside the packed bed. Maximum lipase activities obtained in these bioreactors (26.4 U/g) were 30% higher than in tray-type reactors.

  3. LTCC based bioreactors for cell cultivation

    Science.gov (United States)

    Bartsch, H.; Welker, T.; Welker, K.; Witte, H.; Müller, J.

    2016-01-01

    LTCC multilayers offer a wide range of structural options and flexibility of connections not available in standard thin film technology. Therefore they are considered as material base for cell culture reactors. The integration of microfluidic handling systems and features for optical and electrical capturing of indicators for cell culture growth offers the platform for an open system concept. The present paper assesses different approaches for the creation of microfluidic channels in LTCC multilayers. Basic functions required for the fluid management in bioreactors include temperature and flow control. Both features can be realized with integrated heaters and temperature sensors in LTCC multilayers. Technological conditions for the integration of such elements into bioreactors are analysed. The temperature regulation for the system makes use of NTC thermistor sensors which serve as real value input for the control of the heater. It allows the adjustment of the fluid temperature with an accuracy of 0.2 K. The tempered fluid flows through the cell culture chamber. Inside of this chamber a thick film electrode array monitors the impedance as an indicator for the growth process of 3-dimensional cell cultures. At the system output a flow sensor is arranged to monitor the continual flow. For this purpose a calorimetric sensor is implemented, and its crucial design parameters are discussed. Thus, the work presented gives an overview on the current status of LTCC based fluid management for cell culture reactors, which provides a promising base for the automation of cell culture processes.

  4. Cultivation of mammalian cells using a single-use pneumatic bioreactor system.

    Science.gov (United States)

    Obom, Kristina M; Cummings, Patrick J; Ciafardoni, Janelle A; Hashimura, Yasunori; Giroux, Daniel

    2014-10-10

    Recent advances in mammalian, insect, and stem cell cultivation and scale-up have created tremendous opportunities for new therapeutics and personalized medicine innovations. However, translating these advances into therapeutic applications will require in vitro systems that allow for robust, flexible, and cost effective bioreactor systems. There are several bioreactor systems currently utilized in research and commercial settings; however, many of these systems are not optimal for establishing, expanding, and monitoring the growth of different cell types. The culture parameters most challenging to control in these systems include, minimizing hydrodynamic shear, preventing nutrient gradient formation, establishing uniform culture medium aeration, preventing microbial contamination, and monitoring and adjusting culture conditions in real-time. Using a pneumatic single-use bioreactor system, we demonstrate the assembly and operation of this novel bioreactor for mammalian cells grown on micro-carriers. This bioreactor system eliminates many of the challenges associated with currently available systems by minimizing hydrodynamic shear and nutrient gradient formation, and allowing for uniform culture medium aeration. Moreover, the bioreactor's software allows for remote real-time monitoring and adjusting of the bioreactor run parameters. This bioreactor system also has tremendous potential for scale-up of adherent and suspension mammalian cells for production of a variety therapeutic proteins, monoclonal antibodies, stem cells, biosimilars, and vaccines.

  5. Bioreactor cultivation of anatomically shaped human bone grafts.

    Science.gov (United States)

    Temple, Joshua P; Yeager, Keith; Bhumiratana, Sarindr; Vunjak-Novakovic, Gordana; Grayson, Warren L

    2014-01-01

    In this chapter, we describe a method for engineering bone grafts in vitro with the specific geometry of the temporomandibular joint (TMJ) condyle. The anatomical geometry of the bone grafts was segmented from computed tomography (CT) scans, converted to G-code, and used to machine decellularized trabecular bone scaffolds into the identical shape of the condyle. These scaffolds were seeded with human bone marrow-derived mesenchymal stem cells (MSCs) using spinner flasks and cultivated for up to 5 weeks in vitro using a custom-designed perfusion bioreactor system. The flow patterns through the complex geometry were modeled using the FloWorks module of SolidWorks to optimize bioreactor design. The perfused scaffolds exhibited significantly higher cellular content, better matrix production, and increased bone mineral deposition relative to non-perfused (static) controls after 5 weeks of in vitro cultivation. This technology is broadly applicable for creating patient-specific bone grafts of varying shapes and sizes.

  6. Disposable orbitally shaken TubeSpin bioreactor 600 for Sf9 cell cultivation in suspension.

    Science.gov (United States)

    Monteil, Dominique T; Shen, Xiao; Tontodonati, Giulia; Baldi, Lucia; Hacker, David L; Wurm, Florian M

    2016-07-15

    Disposable orbitally shaken TubeSpin bioreactor 600 tubes (TS600s) were recently developed for the bench-scale cultivation of animal cells in suspension. Here we compared batch cultures of Sf9 insect cells in TS600s, spinner flasks, and shake flasks. Superior cell growth was observed in TS600s and shake flasks as compared with spinner flasks, and more favorable oxygen-enriched cell culture conditions were observed in TS600s as compared with either spinner or shake flasks. The results demonstrated the suitability of TS600s as a disposable vessel for the cultivation of Sf9 cells in suspension.

  7. Lactose autoinduction with enzymatic glucose release: characterization of the cultivation system in bioreactor.

    Science.gov (United States)

    Mayer, Sonja; Junne, Stefan; Ukkonen, Kaisa; Glazyrina, Julia; Glauche, Florian; Neubauer, Peter; Vasala, Antti

    2014-02-01

    The lactose autoinduction system for recombinant protein production was combined with enzymatic glucose release as a method to provide a constant feed of glucose instead of using glycerol as a carbon substrate. Bioreactor cultivation confirmed that the slow glucose feed does not prevent the induction by lactose. HPLC studies showed that with successful recombinant protein production only a very low amount of lactose was metabolized during glucose-limited fed-batch conditions by the Escherichia coli strain BL21(DE3)pLysS in well-aerated conditions, which are problematic for glycerol-based autoinduction systems. We propose that slow enzymatic glucose feed does not cause a full activation of the lactose operon. However recombinant PDI-A protein (A-domain of human disulfide isomerase) was steadily produced until the end of the cultivation. The results of the cultivations confirmed our earlier observations with shaken cultures showing that lactose autoinduction cultures based on enzymatic glucose feed have good scalability, and that this system can be applied also to bioreactor cultivations.

  8. Bioreactor Cultivation of Zeltnera beyrichii (Torr. & A. Gray Mans.: A Novel Source of Biologically Active Compounds

    Directory of Open Access Journals (Sweden)

    Miloš Radović

    2013-08-01

    Full Text Available With regard to world’s increasing demand for biologically active compounds, a novel source of xanthones and secoiridoid glycosides has been studied . Zeltnera beyrichii (Torr. & A. Gray Mans., an insufficiently acknowledged North American medicinal plant species, may be considered a pharmacological substitute for commercial C. erythraea Rafn, since it accumulates in aerial parts nearly the same amount of secoiridoid glycosides: swertiamarin, gentiopicrin, and sweroside (13.76, 7.56, and 0.17 mmol per 100 g dry weight, respectively in plants grown under greenhouse condition, and a considerable amount of xanthones: decussatin and eustomin. Additionally, Z. beyrichii produced as much biomass during cultivation in RITA ® temporary immersion bioreactors as greenhouse-grown plants, in a third of the time. Plants grown in bioreactors contained moderate levels of total phenolics and total flavonoids, and possessed modest antioxidant activity and antimicrobial potential against eight bacterial and eight fungal species. Therefore, this species may be highly recommended for cultivation either in natural environment, or in bioreactors under in vitro conditions, for producing compounds of interest of modern pharmacology and food industry.

  9. Anaerobic membrane bioreactor under extreme conditions (poster)

    NARCIS (Netherlands)

    Munoz Sierra, J.D.; De Kreuk, M.K.; Spanjers, H.; Van Lier, J.B.

    2013-01-01

    Membrane bioreactors ensure biomass retention by the application of micro or ultrafiltration processes. This allows operation at high sludge concentrations. Previous studies have shown that anaerobic membrane bioreactors is an efficient way to retain specialist microorganisms for treating wastewater

  10. Features of development of Stevia rebaudiana shoots cultivated in the roller bioreactor and their production of steviol glycosides.

    Science.gov (United States)

    Bondarev, Nikolai; Reshetnyak, Oxana; Nosov, Alexander

    2002-08-01

    Growth and development of Stevia rebaudiana shoots cultivated in the roller bioreactor and their production of steviol glycosides (SGs) were investigated. It was found that, owing to the highly favorable conditions of shoot cultivation created in such an apparatus, the intensity of shoot growth and SG production appeared to be 1.5 - 2.0 times higher than those of the shoots grown in tubes. These results indicate the existence of a positive correlation between these two processes. The data obtained suggest that the enhanced SG production is due to the differentiation of chlorenchyma cells and formation of specific subcellular structures for the glycoside to be accumulated.

  11. Shear conditions in clavulanic acid production by Streptomyces clavuligerus in stirred tank and airlift bioreactors.

    Science.gov (United States)

    Cerri, M O; Badino, A C

    2012-08-01

    In biochemical processes involving filamentous microorganisms, the high shear rate may damage suspended cells leading to viability loss and cell disruption. In this work, the influence of the shear conditions in clavulanic acid (CA) production by Streptomyces clavuligerus was evaluated in a 4-dm(3) conventional stirred tank (STB) and in 6-dm(3) concentric-tube airlift (ALB) bioreactors. Batch cultivations were performed in a STB at 600 and 800 rpm and 0.5 vvm (cultivations B1 and B2) and in ALB at 3.0 and 4.1 vvm (cultivations A1 and A2) to define two initial oxygen transfer conditions in both bioreactors. The average shear rate ([Formula: see text]) of the cultivations was estimated using correlations of recent literature based on experimental data of rheological properties of the broth (consistency index, K, and flow index, n) and operating conditions, impeller speed (N) for STB and superficial gas velocity in the riser (UGR) for ALB. In the same oxygen transfer condition, the [Formula: see text] values for ALB were higher than those obtained in STB. The maximum [Formula: see text] presented a strong correlation with a maximum consistency index (K (max)) of the broth. Close values of maximum CA production were obtained in cultivations A1 and A2 (454 and 442 mg L(-1)) with similar maximum [Formula: see text] values of 4,247 and 4,225 s(-1). In cultivations B1 and B2, the maximum CA production of 269 and 402 mg L(-1) were reached with a maximum [Formula: see text] of 904 and 1,786 s(-1). The results show that high values of average shear rate increase the CA production regardless of the oxygen transfer condition and bioreactor model.

  12. Modeling of hydrodynamics in hollow fiber membrane bioreactor for mammalian cells cultivation

    Directory of Open Access Journals (Sweden)

    N. V. Menshutina

    2016-01-01

    Full Text Available The mathematical modelling in CFD-packages are powerfull instrument for design and calculation of any engineering tasks. CFD-package contains the set of programs that allow to model the different objects behavior based on the mathematical lows. ANSYS Fluent are widely used for modelling of biotechnological and chemical-technological processes. This package is convenient to describe their hydrodynamics. As cell cultivation is one of the actual scientific direction in modern biotechnology ANSYS Fluent was used to create the model of hollow fiber membrane bioreactor. The fibers are hollow cylindrical membrane to be used for cell cultivation. The criterion of process effectiveness for cell growth is full filling of the membrane surface by cells in the bioreactor. While the cell growth the fiber permeability is decreased which effects to feed flow through membrane pores. The specific feature of this process is to ensure such feed flow to deliver the optimal nutrition for the cells on the external membrane surface. The velocity distribution inside the fiber and in all bioreactor as a whole has been calculated based on mass an impulse conservation equations taking into account the mathematical model assumptions. The hydrodynamics analysis in hollow fiber membrane bioreactor is described by the three-dimensional model created in ANSYS Fluent. The specific features of one membrane model are considered and for whole bioreactor too.

  13. Carbon dioxide fixation by microalgae cultivated in open bioreactors

    Energy Technology Data Exchange (ETDEWEB)

    Centeno da Rosa, Ana Priscila; Fernandes Carvalho, Lisiane; Goldbeck, Luzia [Laboratory of Biochemical Engineering, College of Chemistry and Food, Federal University of Rio Grande (FURG), P.O. Box 474, Rio Grande, RS 96201-900 (Brazil); Vieira Costa, Jorge Alberto, E-mail: dqmjorge@furg.br [Laboratory of Biochemical Engineering, College of Chemistry and Food, Federal University of Rio Grande (FURG), P.O. Box 474, Rio Grande, RS 96201-900 (Brazil)

    2011-08-15

    Highlights: {yields} We studied the growth and CO{sub 2} fixation by Spirulina LEB18 and Chlorella kessleri. {yields} The maximum dailyfixation was obtained for Spirulina with an injection of 6% of CO{sub 2}. {yields} The microalgae presented growth during the 20 d of culture with up to 18% of CO{sub 2}. {yields} The use of CO{sub 2} from industrial generation decreases the cost of producing biomass. - Abstract: The biofixation of carbon dioxide (CO{sub 2}) by microalgae has been proven to be an efficient and economical method, mainly due to the photosynthetic ability of these microorganisms to use this gas as a source of nutrients for their development. The aim of this work was to study the growth of Spirulina LEB18 and Chlorella kessleri microalgae, exposed to controlled and non-controlled conditions, with the injection of different concentrations of CO{sub 2}. The cultures was carried out in 6 L open raceway ponds, under controlled conditions at 30 {sup o}C and 39 {mu}E m{sup -2} s{sup -1} and under non-controlled conditions, protected by a tunnel of transparent film. The experiments were subjected to CO{sub 2} injections at concentrations of 0.038, 6, 12 and 18% (v/v). The highest concentration of biomass (4.95 g L{sup -1}) and maximum daily fixation (0.21 g g{sup -1} d{sup -1}) were obtained for Spirulina LEB18 in culture that was prepared in non-controlled conditions with an injection of 6% (v/v) of CO{sub 2}. C. kessleri had maximum (p < 0.0008) specific growth rate (0.84 d{sup -1}) when grown with 18% (v/v) of CO{sub 2} in non-controlled conditions of cultivation.

  14. Bioreactor

    Science.gov (United States)

    1996-01-01

    The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues currently being cultured in rotating bioreactors by investigators

  15. A novel milliliter-scale chemostat system for parallel cultivation of microorganisms in stirred-tank bioreactors.

    Science.gov (United States)

    Schmideder, Andreas; Severin, Timm Steffen; Cremer, Johannes Heinrich; Weuster-Botz, Dirk

    2015-09-20

    A pH-controlled parallel stirred-tank bioreactor system was modified for parallel continuous cultivation on a 10 mL-scale by connecting multichannel peristaltic pumps for feeding and medium removal with micro-pipes (250 μm inner diameter). Parallel chemostat processes with Escherichia coli as an example showed high reproducibility with regard to culture volume and flow rates as well as dry cell weight, dissolved oxygen concentration and pH control at steady states (n=8, coefficient of variation bioreactor on a liter-scale. Thus, parallel and continuously operated stirred-tank bioreactors on a milliliter-scale facilitate timesaving and cost reducing steady state studies with microorganisms. The applied continuous bioreactor system overcomes the drawbacks of existing miniaturized bioreactors, like poor mass transfer and insufficient process control.

  16. Transfer and consumption of oxygen during the cultivation of the ectomycorrhizal fungus Rhizopogon nigrescens in an airlift bioreactor.

    Science.gov (United States)

    Rossi, Márcio José; Nascimento, Francisco Xavier; Giachini, Admir José; Oliveira, Vetúria Lopes; Furigo, Agenor

    2017-02-01

    The study had the objective of examining the aspects involved in the cultivation of ectomycorrhizal fungi for the production of commercially sustainable inoculant to attend the demands of the seedling nursery industry. It focused on certain parameters, such as the oxygen consumption levels, during the cultivation of the ectomycorrhizal fungus Rhizopogon nigrescens CBMAI 1472, which was performed in a 5-L airlift bioreactor. The dynamic method was employed to determine the volumetric coefficient for the oxygen transfer (k L a) and the specific oxygen uptake rate (Q O2 ). The results indicate that specific growth rates (μ X ) and oxygen consumption decline rapidly with time, affected mainly by increases in biomass concentration (X). Increases in X are obtained primarily by increases in the size of pellets that are formed, altering, consequently, the cultivation dynamics. This is the result of natural increases in transferring resistance that are observed in these environments. Therefore, to avoid critical conditions that affect viability and the productivity of the process, particular settings are discussed.

  17. A Novel Seeding and Conditioning Bioreactor for Vascular Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Julia Schulte

    2014-07-01

    Full Text Available Multiple efforts have been made to develop small-diameter tissue engineered vascular grafts using a great variety of bioreactor systems at different steps of processing. Nevertheless, there is still an extensive need for a compact all-in-one system providing multiple and simultaneous processing. The aim of this project was to develop a new device to fulfill the major requirements of an ideal system that allows simultaneous seeding, conditioning, and perfusion. The newly developed system can be actuated in a common incubator and consists of six components: a rotating cylinder, a pump, a pulse generator, a control unit, a mixer, and a reservoir. Components that are in direct contact with cell media, cells, and/or tissue allow sterile processing. Proof-of-concept experiments were performed with polyurethane tubes and collagen tubes. The scaffolds were seeded with fibroblasts and endothelial cells that were isolated from human saphenous vein segments. Scanning electron microscopy and immunohistochemistry showed better seeding success of polyurethane scaffolds in comparison to collagen. Conditioning of polyurethane tubes with 100 dyn/cm2 resulted in cell detachments, whereas a moderate conditioning program with stepwise increase of shear stress from 10 to 40 dyn/cm2 induced a stable and confluent cell layer. The new bioreactor is a powerful tool for quick and easy testing of various scaffold materials for the development of tissue engineered vascular grafts. The combination of this bioreactor with native tissue allows testing of medical devices and medicinal substances under physiological conditions that is a good step towards reduction of animal testing. In the long run, the bioreactor could turn out to produce tissue engineered vascular grafts for human applications “at the bedside”.

  18. Cultivation of methanogenic community from 2-km deep subseafloor coalbeds using a continuous-flow bioreactor

    Science.gov (United States)

    Imachi, H.; Tasumi, E.; Morono, Y.; Ito, M.; Takai, K.; Inagaki, F.

    2013-12-01

    Deep subseafloor environments associated with hydrocarbon reservoirs have been least explored by previous scientific drilling and hence the nature of deep subseafloor life and its ecological roles in the carbon cycle remain largely unknown. In this study, we performed cultivation of subseafloor methanogenic communities using a continuous-flow bioreactor with polyurethane sponges, called down-flow hanging sponge (DHS) reactor. The sample used for the reactor cultivation was obtained from 2 km-deep coalbeds off the Shimokita Peninsula of Japan, the northwestern Pacific, during the Integrated Ocean Drilling Program (IODP) Expedition 337 using a riser drilling technology of the drilling vessel Chikyu. The coalbed samples were incubated anaerobically in the DHS reactor at the in-situ temperature of 40°C. Synthetic seawater supplemented with a tiny amount of yeast extract, acetate, propionate and butyrate was provided into the DHS reactor. After 34 days of the bioreactor operation, a small production of methane was observed. The methane concentration was gradually increased and the stable carbon isotopic composition of methane was consistency 13C-depleted during the bioreactor operation, indicating the occurrence of microbial methanogenesis. Microscopic observation showed that the enrichment culture contained a variety of microorganisms, including methanogen-like rod-shaped cells with F420 auto-fluorescence. Interestingly, many spore-like particles were observed in the bioreactor enrichment. Phylogenetic analysis of 16S rRNA genes showed the growth of phylogenetically diverse bacteria and archaea in the DHS reactor. Predominant archaeal components were closely related to hydrogenotrophic methanogens within the genus Methanobacterium. Some predominant bacteria were related to the spore-formers within the class Clostridia, which are overall in good agreement with microscopic observations. By analyzing ion images using a nano-scale secondary ion mass spectrometry (Nano

  19. Fully automated single-use stirred-tank bioreactors for parallel microbial cultivations.

    Science.gov (United States)

    Kusterer, Andreas; Krause, Christian; Kaufmann, Klaus; Arnold, Matthias; Weuster-Botz, Dirk

    2008-04-01

    Single-use stirred tank bioreactors on a 10-mL scale operated in a magnetic-inductive bioreaction block for 48 bioreactors were equipped with individual stirrer-speed tracing, as well as individual DO- and pH-monitoring and control. A Hall-effect sensor system was integrated into the bioreaction block to measure individually the changes in magnetic field density caused by the rotating permanent magnets. A restart of the magnetic inductive drive was initiated automatically each time a Hall-effect sensor indicates one non-rotating gas-inducing stirrer. Individual DO and pH were monitored online by measuring the fluorescence decay time of two chemical sensors immobilized at the bottom of each single-use bioreactor. Parallel DO measurements were shown to be very reliable and independently from the fermentation media applied in this study for the cultivation of Escherichia coli and Saccharomyces cerevisiae. The standard deviation of parallel pH measurements was pH 0.1 at pH 7.0 at the minimum and increased to a standard deviation of pH 0.2 at pH 6.0 or at pH 8.5 with the complex medium applied for fermentations with S. cerevisiae. Parallel pH-control was thus shown to be meaningful with a tolerance band around the pH set-point of +/- pH 0.2 if the set-point is pH 6.0 or lower.

  20. Feeding strategies enhance high cell density cultivation and protein expression in milliliter scale bioreactors.

    Science.gov (United States)

    Faust, Georg; Janzen, Nils H; Bendig, Christoph; Römer, Lin; Kaufmann, Klaus; Weuster-Botz, Dirk

    2014-10-01

    Miniature bioreactors under parallel fed-batch operations are not only useful screening tools for bioprocess development but also provide a suitable basis for eventual scale-up. In this study, three feeding strategies were investigated: besides the established intermittent feeding by a liquid handler, an optimized microfluidic device and a new enzymatic release system were applied for parallel fed-batch cultivation of Escherichia coli HMS174(DE3) and BL21(DE3) strains in stirred-tank bioreactors on a 10 mL scale. Lower fluctuation in dissolved oxygen (DO) and higher optical densities were measured in fed-batch processes applying the microfluidic device or the enzymatic glucose/fructose release system (conversion of intermittently added sucrose by an invertase), but no difference in dry cell weights (DCW) were observed. With all three feeding strategies high cell densities were realized on a milliliter scale with final optical density measured at 600 nm (OD600 ) of 114-133 and final DCW concentrations of 69-70 g L(-1) . The effect of feeding strategies on the expression of two heterologous proteins was investigated. Whereas no impact was observed on the expression of the spider silk protein eADF4(C16), the fluorescence of enhanced green fluorescence protein (eGFP) was reproducibly lower, if an intermittent glucose feed was applied. Thus, the impact of feeding strategy on expression is strongly dependent on the E. coli strain and/or expressed protein. As a completely continuous feed supply is difficult to realize in miniature bioreactors, the enzymatic release approach from this study can be easily applied in all microfluidic system to reduce fluctuations of glucose supply and DO concentrations.

  1. High-EPA Biomass from Nannochloropsis salina Cultivated in a Flat-Panel Photo-Bioreactor on a Process Water-Enriched Growth Medium

    DEFF Research Database (Denmark)

    Safafar, Hamed; Hass, Michael Z.; Møller, Per

    2016-01-01

    Nannochloropsis salina was grown on a mixture of standard growth media and pre-gasified industrial process water representing effluent from a local biogas plant. The study aimed to investigate the effects of enriched growth media and cultivation time on nutritional composition of Nannochloropsis...... salina biomass, with a focus on eicosapentaenoic acid (EPA). Variations in fatty acid composition, lipids, protein, amino acids, tocopherols and pigments were studied and results compared to algae cultivated on F/2 media as reference. Mixed growth media and process water enhanced the nutritional quality...... of Nannochloropsis salina in laboratory scale when compared to algae cultivated in standard F/2 medium. Data from laboratory scale translated to the large scaleusing a 4000 L flat panel photo-bioreactor system. The algae growth rate in winter conditions in Denmark was slow, but results revealed that large...

  2. Optimizing of Culture Condition in Horizontal Rotating Bioreactor

    Institute of Scientific and Technical Information of China (English)

    Yan-Fang ZHANG; Huai-Qing CHEN; Hua HUANG

    2005-01-01

    @@ 1 Introduction Bioreactor is the most important equipment in tissue engineering. It can mimic the micro-environment of cell growth in vitro. At present, horizontal rotating bioreactor is the most advanced equipment for cell culture in the world.

  3. Cultivation of diploid and tetraploid hairy roots of Datura stramonium L. in stirred tank bioreactor for tropane alkaloids production

    Directory of Open Access Journals (Sweden)

    ATANAS PAVLOV

    2012-01-01

    Full Text Available Biomass accumulation and tropane alkaloids production by diploid and tetraploid hairy root cultures of Datura stramonium L. cultivated in stirred tank bioreactor at different aeration rates were investigated. The maximal growth for both hairy root cultures (ADB = 8.3 g/L and 6.8 g/L for diploid and tetraploid line, respectively was achieved at aeration rate of 15.0 L/(L.h. The corresponding growth indexes were remarkably high (GIDW = 9.0 and 7.8 for diploid and tetraploid line, respectively compared to the values, usually reported for other hairy root cultures. The optimal aeration rate for biomass accumulation was also optimal for alkaloids biosynthesis. According to our survey, the achieved maximal amounts of accumulated hyoscyamine (35.0 mg/L and 27.0 mg/L for diploid and tetraploid line were the highest reported in the scientific literature for D. stramonium L. hairy roots. During the cultivation in stirred tank bioreactor, the hairy roots biosynthesized pharmaceutically important alkaloid scopolamine in minor concentrations. This is an important observation since scopolamine was not detected during submerged cultivation of these hairy root lines in other bioreactor types. However, the ploidy level was found to be the most important factor concerning scopolamine production by D. stramonium L. hairy roots cultures. The present work demonstrated the effect of ploidity levels on biomass accumulation and tropane alkaloids production by D. stramonium L. hairy roots cultivated in stirred tank bioreactor. This investigation show that the stirred tank bioreactor could be successfully applied for both maximal biomass accumulations, as well as for manipulation of tropane alkaloids production by diploid and tetraploid D. stramonium L. hairy root cultures.

  4. Effect of operating conditions in production of diagnostic Salmonella Enteritidis O-antigen-specific monoclonal antibody in different bioreactor systems.

    Science.gov (United States)

    Ayyildiz-Tamis, Duygu; Nalbantsoy, Ayse; Elibol, Murat; Deliloglu-Gurhan, Saime Ismet

    2014-01-01

    In this study, different cultivation systems such as roller bottles (RB), 5-L stirred-tank bioreactor (STR), and disposable bioreactors were used to cultivate hybridoma for lab-scale production of Salmonella Enteritidis O-antigen-specific monoclonal antibody (MAb). Hybridoma cell line was cultivated in either serum-containing or serum-free medium (SFM) culture conditions. In STR, MAb production scaled up to 4 L, and production capabilities of the cells were also evaluated in different featured production systems. Moreover, the growth parameters of the cells in all production systems such as glucose consumption, lactate and ammonia production, and also MAb productivities were determined. Collected supernatants from the reactors were concentrated by a cross-flow filtration system. In conclusion, cells were not adapted to SFM in RB and STR. Therefore, less MAb titer in both STR and RB systems with SFM was observed compared to the cultures containing fetal bovine serum-supplemented medium. A higher MAb titer was gained in the membrane-aerated system compared to those in STR and RB. Although the highest MAb titer was obtained in the static membrane bioreactor system, the highest productivity was obtained in STR operated in semicontinuous mode with overlay aeration.

  5. Cultivation of oleaginous Rhodotorula mucilaginosa in airlift bioreactor by using seawater.

    Science.gov (United States)

    Yen, Hong-Wei; Liao, Yu-Ting; Liu, Yi Xian

    2016-02-01

    The enormous water resource consumption is a concern to the scale-up fermentation process, especially for those cheap fermentation commodities, such as microbial oils as the feedstock for biodiesel production. The direct cultivation of oleaginous Rhodotorula mucilaginosa in a 5-L airlift bioreactor using seawater instead of pure water led to a slightly lower biomass being achieved, at 17.2 compared to 18.1 g/L, respectively. Nevertheless, a higher lipid content of 65 ± 5% was measured in the batch using seawater as compared to the pure water batch. Both the salinity and osmotic pressure decreased as the cultivation time increased in the seawater batch, and these effects may contribute to the high tolerance for salinity. No effects were observed for the seawater on the fatty acid profiles. The major components for both batches using seawater and pure water were C16:0 (palmitic acid), C18:1 (oleic acid) and C18:2 (linoleic acid), which together accounted for over 85% of total lipids. The results of this study indicated that seawater could be a suitable option for scaling up the growth of oleaginous R. mucilaginosa, especially from the perspective of water resource utilization.

  6. Comparison between controlled landfill reactor and conditioned landfill bioreactor

    Institute of Scientific and Technical Information of China (English)

    LUO Feng; CHEN Wan-zhi; SONG Fu-zhong; LI Xiao-peng; ZHANG Guo-qing

    2004-01-01

    Bioreactor landfills allow a more active landfill management that recognizes the biological, chemical and physical processes involved in a landfill environment. The results of laboratory-scale simulators of landfill reactors treating municipal solid wastes were studied, the effect of solid waste size, leachate recirculation, nutrient balance, pH value, moisture content and temperature on the rate of municipal solid waste(MSW) biodegradation were determined, and it indicated the optimum pH value, moisture content and temperature can used to decompose MSW. The results of waste biodegradation were compared with that of the simulators of the leachate-recirculated landfill and conservative sanitary landfill. In the control experiment the antitheses of a decreasing trend of the organic load, measured as biological oxygen demand and chemical oxygen demand, was shown, and heavy metals concentration was observed. An obvious enhancement of effective disposal from simulator of conservative sanitary landfill(CSL), to that of leachate-recirculated landfill(LRL) and to that of conditioned bioreactor landfill(CBL) would be noted, through displaying the compared results of solid waste settlement, heavy metal concentration in leachate, methane production rate, biogas composition, BOD and COD as well as their ratio.

  7. High-EPA Biomass from Nannochloropsis salina Cultivated in a Flat-Panel Photo-Bioreactor on a Process Water-Enriched Growth Medium

    Directory of Open Access Journals (Sweden)

    Hamed Safafar

    2016-07-01

    Full Text Available Nannochloropsis salina was grown on a mixture of standard growth media and pre-gasified industrial process water representing effluent from a local biogas plant. The study aimed to investigate the effects of enriched growth media and cultivation time on nutritional composition of Nannochloropsis salina biomass, with a focus on eicosapentaenoic acid (EPA. Variations in fatty acid composition, lipids, protein, amino acids, tocopherols and pigments were studied and results compared to algae cultivated on F/2 media as reference. Mixed growth media and process water enhanced the nutritional quality of Nannochloropsis salina in laboratory scale when compared to algae cultivated in standard F/2 medium. Data from laboratory scale translated to the large scale using a 4000 L flat panel photo-bioreactor system. The algae growth rate in winter conditions in Denmark was slow, but results revealed that large-scale cultivation of Nannochloropsis salina at these conditions could improve the nutritional properties such as EPA, tocopherol, protein and carotenoids compared to laboratory-scale cultivated microalgae. EPA reached 44.2% ± 2.30% of total fatty acids, and α-tocopherol reached 431 ± 28 µg/g of biomass dry weight after 21 days of cultivation. Variations in chemical compositions of Nannochloropsis salina were studied during the course of cultivation. Nannochloropsis salina can be presented as a good candidate for winter time cultivation in Denmark. The resulting biomass is a rich source of EPA and also a good source of protein (amino acids, tocopherols and carotenoids for potential use in aquaculture feed industry.

  8. Modelling the damage potential of fluid flows for animal cells undergoing cultivation in bioreactors.

    Science.gov (United States)

    Stanford Keen, Giles

    1996-11-01

    Mechanical disruption and injury sustained by animal cells undergoing cultivation in bioreactors is an important problem in biotechnology. Damage to cells is thought to be caused primarily by bubbles bursting at the free surface of the culture medium. Here we present computational studies applying a mathematical model for the cell damage rates experienced by cells in laminar flow. Two fluid dynamical systems are considered - namely a converging channel and a single bursting bubble. The flows are calculated using a fourth-order finite difference technique on a stretched grid, and a boundary integral method respectively. It is possible to obtain an estimate for the number of cells in a particular population which are likely to be disrupted by the forces they experience in the flow. This is done by calculating the maximum rate of strain experienced by fluid particles, and combining this with experimental data on the strength and size of cells, obtained by micromanipulation techniques. The resulting information is then used together with the cell damage model to produce a cell damage prediction. The computational results are compared with experimental measurements of cell death, to validate the model for cell damage.

  9. High cell density cultivation and recombinant protein production with Escherichia coli in a rocking-motion-type bioreactor

    Directory of Open Access Journals (Sweden)

    Adams Thorsten

    2010-05-01

    Full Text Available Abstract Background Single-use rocking-motion-type bag bioreactors provide advantages compared to standard stirred tank bioreactors by decreased contamination risks, reduction of cleaning and sterilization time, lower investment costs, and simple and cheaper validation. Currently, they are widely used for cell cultures although their use for small and medium scale production of recombinant proteins with microbial hosts might be very attractive. However, the utilization of rocking- or wave-induced motion-type bioreactors for fast growing aerobic microbes is limited because of their lower oxygen mass transfer rate. A conventional approach to reduce the oxygen demand of a culture is the fed-batch technology. New developments, such as the BIOSTAT® CultiBag RM system pave the way for applying advanced fed-batch control strategies also in rocking-motion-type bioreactors. Alternatively, internal substrate delivery systems such as EnBase® Flo provide an opportunity for adopting simple to use fed-batch-type strategies to shaken cultures. Here, we investigate the possibilities which both strategies offer in view of high cell density cultivation of E. coli and recombinant protein production. Results Cultivation of E. coli in the BIOSTAT® CultiBag RM system in a conventional batch mode without control yielded an optical density (OD600 of 3 to 4 which is comparable to shake flasks. The culture runs into oxygen limitation. In a glucose limited fed-batch culture with an exponential feed and oxygen pulsing, the culture grew fully aerobically to an OD600 of 60 (20 g L-1 cell dry weight. By the use of an internal controlled glucose delivery system, EnBase® Flo, OD600 of 30 (10 g L-1 cell dry weight is obtained without the demand of computer controlled external nutrient supply. EnBase® Flo also worked well in the CultiBag RM system with a recombinant E. coli RB791 strain expressing a heterologous alcohol dehydrogenase (ADH to very high levels, indicating that

  10. Design of photo-bioreactor and the application for cultivating algae%藻类光生物反应器的设计及应用研究

    Institute of Scientific and Technical Information of China (English)

    董汝晶; 谯顺彬; 田辉; 张义明; 罗芳; 陶希芹

    2012-01-01

    A new photo-bioreactor was designed basing on the growth trait of the algae.The volume of the reactor was 10.0L,the length,width and height was 320,80,390mm,respectively.Used the reactor to culture Spirulina platensis,and employed the response surface methodology to optimize the fermentation conditions.Then,researched the optimization concentration levels and the relations between these factors and building up a quadratic regression equation with dry weight as the dependent,light intensity,air flow,time of cultivation and volume of medium as independent.Under the optimistically conditions,the final dry weight was 1.298g/L.According to the experiment results,the photo-bioreactor designed was fit to cultivate the algae.%根据藻类的生长特点设计了一个容积为10.0L的光生物反应器,其长×宽×高分别为320mm×80mm×390mm。利用该反应器进行螺旋藻培养实验,采用响应面法对其培养条件进行优化研究,建立以藻体干重为响应值,以光照强度、通气量、培养时间和装液量为自变量的二次多项式数学模型。培养条件优化后螺旋藻最终干重为1.298g/L。实验结果表明,所设计的反应器能很好地满足藻类生长,其培养产率也明显提高。

  11. Performance of a membrane-dialysis bioreactor with a radial-flow fixed bed for the cultivation of a hybridoma cell line.

    Science.gov (United States)

    Bohmann, A; Pörtner, R; Märkl, H

    1995-10-01

    A bioreactor system for the continuous cultivation of animal cells with a high potential for scale-up is presented. This reactor system consists of radial-flow fixed-bed units coupled with a dialysis module The dialysis membrane enables the supply of low-molecular-weight nutrients and removal of toxic metabolites, while high-molecular-weight nutrients and products (e.g., monoclonal antibodies) are retained and accumulated. This concept was investigated on the laboratory scale in a bioreactor with an integrated dialysis membrane. The efficiency of the reactor system and the reproducibility of the cell activity (hybridoma cells) under certain process conditions could be demonstrated in fermentations up to 77 days. Based on model calculations, an optimized fermentation strategy was formulated and experimentally confirmed. Compared to chemostat cultures with suspended cells, a ten-times higher mAb concentration (383 mg1(-1)) could be obtained. The highest volumetric specific mAb production rate determined was 6.1 mg mAb (1 fixed bed)-1h-1.

  12. On-line near infrared bioreactor monitoring of cell density and concentrations of glucose and lactate during insect cell cultivation.

    Science.gov (United States)

    Qiu, Jiang; Arnold, Mark A; Murhammer, David W

    2014-03-10

    Near infrared spectroscopy is demonstrated as a suitable method for monitoring real time cell density and concentrations of glucose and lactate during insect cell cultivation. The utility of this approach is illustrated during the cultivation of Trichoplusia ni BTI-Tn-5B1-4 insect cells in a stirred-tank bioreactor. On-line near infrared measurements are made by passing unaltered culture medium through an autoclavable near infrared flow-through sample cell during the cultivation process. Single-beam near infrared spectra were collected over the combination spectral range (5000-4000cm(-1)) through a 1.5mm path length sample. Cell density calibration model was established by uni-variable linear regressions with measured mean absorbance values of on-line spectra collected during a cultivation run. Calibration models are generated for glucose and lactate by regression analysis of both off line and on line spectra collected during a series of pre-measurement cultivation runs. Analyte-specific calibration models are generated by using a combination of spectra from both natural, unaltered samples and samples spiked with known levels of glucose and lactate. Spiked samples are used to destroy concentration correlations between solutes, thereby enhancing the selectivity of the calibration models. Absorbance spectra are used to build partial least squares calibration models for glucose and lactate. The calibration model for cell density corresponds to a univariate linear regression calibration model based on the mean absorbance between 4750 and 4250cm(-1). The standard errors of prediction are 1.54mM, 0.83mM, and 0.38×10(6)cells/mL for the glucose, lactate, and cell density models, respectively.

  13. Production of Forskolin by Axenic Coleus forskohlii Roots Cultivated in Shake Flasks and 20-l Glass Jar Bioreactors*.

    Science.gov (United States)

    Krombholz, R; Mersinger, R; Kreis, W; Reinhard, E

    1992-08-01

    Root cultures of COLEUS FORSKOHLII Briq. were initiated from primary callus or IBA-treated suspension cultures and maintained on Gamborg's B5 medium containing 1 mg/l IBA. Transformed root cultures were established by infecting surface-sterilized leaves with AGROBACTERIUM RHIZOGENES strain 15834. Transformation was confirmed by mannopine detection. These cultures displayed the typical characteristics of hairy root cultures, with the sole exceptions of slow growth in hormone-free medium and accelerated growth on medium containing phytohormones. All root cultures examined formed forskolin and its derivatives in amounts ranging from 500 to 1300 mg/kg dry weight, corresponding to about 4 to 5 mg/l. During cultivation roots could be cut into small pieces without affecting growth and forskolin production. Scale-ups of the cultivation procedure were performed in 20-l glass jars with a working volume of 10 to 13l. Forskolin production in bioreactors was better than in shake flasks. Levels of almost 14 mg/l could be reached after 21 d of cultivation. As in the shake flask experiments cutting the roots did not affect growth or productivity in a negative way.

  14. Production of the biopesticide azadirachtin by hairy root cultivation of Azadirachta indica in liquid-phase bioreactors.

    Science.gov (United States)

    Srivastava, Smita; Srivastava, Ashok K

    2013-11-01

    Batch cultivation of Azadirachta indica hairy roots was carried out in different liquid-phase bioreactor configurations (stirred-tank, bubble column, bubble column with polypropylene basket, and polyurethane foam disc as root supports) to investigate possible scale-up of the A. indica hairy root culture for in vitro production of the biopesticide azadirachtin. The hairy roots failed to grow in the conventional bioreactor designs (stirred tank and bubble column). However, modified bubble column reactor (with polyurethane foam as root support) configuration facilitated high-density culture of A. indica hairy roots with a biomass production of 9.2 g l(-1)dry weight and azadirachtin yield of 3.2 mg g(-1) leading to a volumetric productivity of azadirachtin as 1.14 mg l(-1) day(-1). The antifeedant activity in the hairy roots was also evaluated by no choice feeding tests with known concentrations of the hairy root powder and its solvent extract separately on the desert locust Schistocerca gregaria. The hairy root powder and its solvent extract demonstrated a high level of antifeedant activity (with an antifeedant index of 97 % at a concentration of 2 % w/v and 83 % at a concentration of 0.05 % (w/v), respectively, in ethanol).

  15. Application of airlift bioreactor for the cultivation of aerobic oleaginous yeast Rhodotorula glutinis with different aeration rates.

    Science.gov (United States)

    Yen, Hong-Wei; Liu, Yi Xian

    2014-08-01

    The high cost of microbial oils produced from oleaginous microorganisms is the major obstacle to commercial production. In this study, the operation of an airlift bioreactor is examined for the cultivation of oleaginous yeast-Rhodotorula glutinis, due to the low process cost. The results suggest that the use of a high aeration rate could enhance cell growth. The maximum biomass concentration of 25.40 g/L was observed in the batch with a 2.0 vvm aeration rate. In addition, a higher aeration rate of 2.5 vvm could achieve the maximum growth rate of 0.46 g/L h, about twice the 0.22 g/L h obtained in an agitation tank. However, an increase in tank pressure instead of the aeration rate did not enhance cell growth. The operation of airlift bioreactor described in this work has the advantages of simple operation and low energy consumption, thus making it suitable for the accumulation of microbial oils.

  16. Influence of stationary and bioreactor cultivation on Lentinula edodes (berk pegler lignocellulolitic activity

    Directory of Open Access Journals (Sweden)

    Magali Regina

    2008-04-01

    Full Text Available This work aimed to study the stationary and periodically mixed culture of L. edodes to the production of lignocellulolitic enzymes activity. LE 95/17, LE 96/22 and Leax strains were incubated in 25 g of eucalyptus sawdust substrate in Erlenmeyer flasks in stationary culture at 25º C and in a bioreactor with four complete rotations daily at 25º C and 3% CO2. The samples were collected at 8, 11, 14, 17 and 20 days after the incubation. Oxidative and hydrolytic enzymes analyses were performed. Lignin peroxidase enzyme was not found in the lignolytic system for LE 95/17, LE 96/22 and Leax strains in the different incubation methods. The use of bioreactor could be a practicable system to induce the laccase activity for L22 and Leax and MnP activity for L17 and L22. The activity of the hydrolytic enzymes was higher in the stationary system in comparison to periodically mixed system in the bioreactor.Lentinula edodes pertence ao grupo de fungos causadores da podridão branca, conhecidos pela capacidade efetiva de degradar compostos lignocelulósicos. No cultivo deste fungo o método de incubação, substratos e características genéticas determinam o comportamento enzimático. Este trabalho teve como objetivo verificar a influência do cultivo estacionário e em bioreator na atividade de enzimas lignocelulolíticas do L. edodes. As linhagens LE 95/17, LE 96/22 e Leax foram cultivadas em 25 g de substrato composto de serragem de eucalipto em frascos de Erlenmeyer, em cultura estacionária, sob de temperatura controlada a 25° C e em bioreator com quatro rotações diárias e completas, a 25° C e 3% de CO2. Foram coletados dados aos 8, 11, 14, 17 e 20 dias depois da incubação para atividade de enzimas oxidativas e hidrolíticas. A enzima lignina peroxidase não foi encontrada em nenhum dos tratamentos. O uso de bioreator pode ser visto como um sistema praticável para induzir atividade de laccase para L22 e Leax e atividade de MnP para L17 e L22. A

  17. Membrane photobioreactors for integrated microalgae cultivation and nutrient remediation of membrane bioreactors effluent.

    Science.gov (United States)

    Marbelia, L; Bilad, M R; Passaris, I; Discart, V; Vandamme, D; Beuckels, A; Muylaert, K; Vankelecom, Ivo F J

    2014-07-01

    The feasibility of a new concept of wastewater treatment by combining a membrane bioreactor (MBR) and a microalgae membrane photobioreactor (MPBR) is assessed in this study. In this system, the organic carbon present in wastewater is expected to be fully oxidized in the MBR, while the nutrients are removed via the subsequent MPBR treatment. The effluent of a lab-scale MBR was fed into a PBR and a MPBR which served as growing medium for Chlorella vulgaris. The MPBRs demonstrated their superiority by limiting the algae wash-out, thus increasing the allowable optimum dilution rate (Dopt). At these corresponding Dopt values, 3.5 and 2 times higher biomass concentrations and volumetric productivities respectively were achieved by the MPBR. It is also possible to run the MPBR at still higher biomass concentration, thus enabling a smaller footprint and higher nutrient removal efficiency. However, reduced nutrient removal efficiencies were found to be one possible drawback.

  18. Bioreactor Conditioning for Accelerated Remodeling of Fibrin-Based Tissue Engineered Heart Valves

    Science.gov (United States)

    Schmidt, Jillian Beth

    Fibrin is a promising scaffold material for tissue engineered heart valves, as it is completely biological, allows for engineered matrix alignment, and is able to be degraded and replaced with collagen by entrapped cells. However, the initial fibrin matrix is mechanically weak, and extensive in vitro culture is required to create valves with sufficient mechanical strength and stiffness for in vivo function. Culture in bioreactor systems, which provide cyclic stretching and enhance nutrient transport, has been shown to increase collagen production by cells entrapped in a fibrin scaffold, accelerating strengthening of the tissue and reducing the required culture time. In the present work, steps were taken to improve bioreactor culture conditions with the goal of accelerating collagen production in fibrin-based tissue engineered heart valves using two approaches: (i) optimizing the cyclic stretching protocol and (ii) developing a novel bioreactor system that permits transmural and lumenal flow of culture medium for improved nutrient transport. The results indicated that incrementally increasing strain amplitude cyclic stretching with small, frequent increments in strain amplitude was optimal for collagen production in our system. In addition, proof of concept studies were performed in the novel bioreactor system and increased cellularity and collagen deposition near the lumenal surface of the tissue were observed.

  19. A Pulsatile Bioreactor for Conditioning of Tissue-Engineered Cardiovascular Constructs under Endoscopic Visualization

    Directory of Open Access Journals (Sweden)

    Bassil Akra

    2012-07-01

    Full Text Available Heart valve disease (HVD is a globally increasing problem and accounts for thousands of deaths yearly. Currently end-stage HVD can only be treated by total valve replacement, however with major drawbacks. To overcome the limitations of conventional substitutes, a new clinical approach based on cell colonization of artificially manufactured heart valves has been developed. Even though this attempt seems promising, a confluent and stable cell layer has not yet been achieved due to the high stresses present in this area of the human heart. This study describes a bioreactor with a new approach to cell conditioning of tissue engineered heart valves. The bioreactor provides a low pulsatile flow that grants the correct opening and closing of the valve without high shear stresses. The flow rate can be regulated allowing a steady and sensitive conditioning process. Furthermore, the correct functioning of the valve can be monitored by endoscope surveillance in real-time. The tubeless and modular design allows an accurate, simple and faultless assembly of the reactor in a laminar flow chamber. It can be concluded that the bioreactor provides a strong tool for dynamic pre-conditioning and monitoring of colonized heart valve prostheses physiologically exposed to shear stress.

  20. A Pulsatile Bioreactor for Conditioning of Tissue-Engineered Cardiovascular Constructs under Endoscopic Visualization.

    Science.gov (United States)

    König, Fabian; Hollweck, Trixi; Pfeifer, Stefan; Reichart, Bruno; Wintermantel, Erich; Hagl, Christian; Akra, Bassil

    2012-07-19

    Heart valve disease (HVD) is a globally increasing problem and accounts for thousands of deaths yearly. Currently end-stage HVD can only be treated by total valve replacement, however with major drawbacks. To overcome the limitations of conventional substitutes, a new clinical approach based on cell colonization of artificially manufactured heart valves has been developed. Even though this attempt seems promising, a confluent and stable cell layer has not yet been achieved due to the high stresses present in this area of the human heart. This study describes a bioreactor with a new approach to cell conditioning of tissue engineered heart valves. The bioreactor provides a low pulsatile flow that grants the correct opening and closing of the valve without high shear stresses. The flow rate can be regulated allowing a steady and sensitive conditioning process. Furthermore, the correct functioning of the valve can be monitored by endoscope surveillance in real-time. The tubeless and modular design allows an accurate, simple and faultless assembly of the reactor in a laminar flow chamber. It can be concluded that the bioreactor provides a strong tool for dynamic pre-conditioning and monitoring of colonized heart valve prostheses physiologically exposed to shear stress.

  1. Development of thin-film photo-bioreactor and its application to outdoor culture of microalgae.

    Science.gov (United States)

    Yoo, Jae Jun; Choi, Seung Phill; Kim, Jaoon Y H; Chang, Won Seok; Sim, Sang Jun

    2013-06-01

    Photosynthetic microalgae have received much attention as a microbial source of diverse useful biomaterials through CO(2) fixation and various types of photo-bioreactors have been developed for efficient microalgal cultivation. Herein, we developed a novel thin-film photo-bioreactor, which was made of cast polypropylene film, considering outdoor mass cultivation. To develop optimal design of photo-bioreactor, we tested performance of three shapes of thin-film photo-bioreactors (flat, horizontal and vertical tubular shapes) and various parts in the bioreactor. Collectively, vertical tubular bioreactor with H/D ratio 6:1 and cylindrical stainless steel spargers showed the most outstanding performance. Furthermore, the photo-bioreactor was successfully applied to the cultivation of other microalgae such as Chlamydomonas reinhardtii and Chlorella vulgaris. The scalability of photo-bioreactor was confirmed by gradually increasing culture volume from 4 to 25 L and the biomass productivity of each reactor was quite consistent (0.05-0.07 g/L/day) during the cultivation of H. pluvialis under indoor and outdoor conditions. Especially, we also achieved dry cell weight of 4.64 g/L and astaxanthin yield of 218.16 mg/L through long-term cultivation (100 days) under outdoor condition in 15 L photo-bioreactor using Haematococcus pluvialis, which means that the astaxanthin yield from outdoor cultivation is equal or superior to that obtained from controlled indoor condition. Therefore, these results indicate that we can apply this approach to development of optimal photo-bioreactor for the large-scale culture of microalgae and production of useful biomaterials under outdoor condition.

  2. Experimental mixture design as a tool to enhance glycosyl hydrolases production by a new Trichoderma harzianum P49P11 strain cultivated under controlled bioreactor submerged fermentation.

    Science.gov (United States)

    Delabona, Priscila da Silva; Farinas, Cristiane Sanchez; Lima, Deise Juliana da Silva; Pradella, José Geraldo da Cruz

    2013-03-01

    This work investigates the glycosyl hydrolase (GH) profile of a new Trichoderma harzianum strain cultivated under controlled bioreactor submerged fermentation. The influence of different medium components (delignified steam-exploded sugarcane bagasse, sucrose, and soybean flour) on GH biosynthesis was assessed using experimental mixture design (EMD). Additionally, the effect of increased component concentrations in culture media selected from the EMD was studied. It was found that that a mixed culture medium could significantly maximize GH biosynthesis rate, especially for xylanase enzymes which achieved a 2-fold increment. Overall, it was demonstrated that T. harzianumP49P11 enzymes have a great potential to be used in the deconstruction of biomass.

  3. Modelling of a Batch Whey Cultivation of Kluyveromyces marxianus var. lactis MC 5 with Investigation of Mass Transfer Processes in the Bioreactor

    OpenAIRE

    Mitko Petrov; Tatiana Ilkova; Juris Vanags

    2015-01-01

    This study presents a mathematical model of a batch fermentation of lactose oxidation from a natural substratum in a cultivation by the strain Kluyweromyces marxianus var. lactis MC 5. In the model of the process, the mass transfer in the bioreactor for oxygen concentration in the gas phase (GP) and in the liquid phase (LP) is based on the dispersion model of the GP. In addition, perfect mixing in LP is included. Nine models were investigated for specific growth rate and specific oxygen consu...

  4. Screening of phenylpyruvic acid producers and optimization of culture conditions in bench scale bioreactors.

    Science.gov (United States)

    Coban, Hasan B; Demirci, Ali; Patterson, Paul H; Elias, Ryan J

    2014-11-01

    Alpha keto acids are deaminated forms of amino acids that have received significant attention as feed and food additives in the agriculture and medical industries. To date, their production has been commonly performed at shake-flask scale with low product concentrations. In this study, production of phenylpyruvic acid (PPA), which is the alpha keto acid of phenylalanine was investigated. First, various microorganisms were screened to select the most efficient producer. Thereafter, growth parameters (temperature, pH, and aeration) were optimized in bench scale bioreactors to maximize both PPA and biomass concentration in bench scale bioreactors, using response surface methodology. Among the four different microorganisms evaluated, Proteus vulgaris was the most productive strain for PPA production. Optimum temperature, pH, and aeration conditions were determined as 34.5 °C, 5.12, and 0.5 vvm for PPA production, whereas 36.9 °C, pH 6.87, and 0.96 vvm for the biomass production. Under these optimum conditions, PPA concentration was enhanced to 1,054 mg/L, which was almost three times higher than shake-flask fermentation concentrations. Moreover, P. vulgaris biomass was produced at 3.25 g/L under optimum conditions. Overall, this study demonstrated that optimization of growth parameters improved PPA production in 1-L working volume bench-scale bioreactors compared to previous studies in the literature and was a first step to scale up the production to industrial production.

  5. Iron and phosphorus speciation in Fe-conditioned membrane bioreactor activated sludge.

    Science.gov (United States)

    Wu, Hao; Ikeda-Ohno, Atsushi; Wang, Yuan; Waite, T David

    2015-06-01

    Iron dosing of membrane bioreactors (MBRs) is widely used as a means of meeting effluent phosphorus targets but there is limited understanding of the nature of iron and phosphorus-containing solids that are formed within the bioreactor (an important issue in view of the increasing interest in recovering phosphorus from wastewaters). Of particular challenge is the complexity of the MBR system and the variety of reactions that can occur on addition of iron salts to a membrane bioreactor. In this study, the performances of bench scale MBRs with dosing of either ferrous or ferric salts were monitored for a period of four months. The distributions of Fe and P-species in the Fe-conditioned sludges were determined using X-ray absorption spectroscopy (XAS) at the Fe K-edge and the P K-edge. Regardless of whether iron was dosed to the anoxic or aerobic chambers and regardless of whether ferrous (Fe(II)) or ferric (Fe(III)) iron was dosed, iron present in the minerals in the conditioned sludges was consistently in the +III oxidation state. Fitting of the Fe K-edge EXAFS spectra revealed that an Fe(III)-phosphate species was the main Fe species present in all cases with the remaining fraction dominated by lepidocrocite (γ-FeOOH) in the Fe(II)-dosed case and ferrihydrite (am-FeOOH) in the Fe(III)-dosed case. Approximately half the phosphorus in the activated sludge samples was present as a distinct Fe-PO4 mineral (such as strengite or an amorphous ferric hydroxyl phosphate analogue of strengite) and half as phosphorus adsorbed to an iron oxyhydroxide mineral phase indicating that both co-precipitation and adsorption of phosphorus by iron contribute to removal of phosphorus from the MBR supernatant.

  6. NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Biotechnology Specimen Temperature Controller (BSTC) will cultivate cells until their turn in the bioreactor; it can also be used in culturing experiments that do not require the bioreactor. The BSTC comprises four incubation/refrigeration chambers individually set at 4 to 50 deg. C (near-freezing to above body temperature). Each chamber holds three rugged tissue chamber modules (12 total), clear Teflon bags holding 30 ml of growth media, all positioned by a metal frame. Every 7 to 21 days (depending on growth rates), an astronaut uses a shrouded syringe and the bags' needleless injection ports to transfer a few cells to a fresh media bag, and to introduce a fixative so that the cells may be studied after flight. The design also lets the crew sample the media to measure glucose, gas, and pH levels, and to inspect cells with a microscope. The controller is monitored by the flight crew through a 23-cm (9-inch) color computer display on the face of the BSTC. This view shows the BTSC with the front panel open. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  7. In Vitro Model for Hepatotoxicity Studies Based on Primary Human Hepatocyte Cultivation in a Perfused 3D Bioreactor System

    Directory of Open Access Journals (Sweden)

    Fanny Knöspel

    2016-04-01

    Full Text Available Accurate prediction of the potential hepatotoxic nature of new pharmaceuticals remains highly challenging. Therefore, novel in vitro models with improved external validity are needed to investigate hepatic metabolism and timely identify any toxicity of drugs in humans. In this study, we examined the effects of diclofenac, as a model substance with a known risk of hepatotoxicity in vivo, in a dynamic multi-compartment bioreactor using primary human liver cells. Biotransformation pathways of the drug and possible effects on metabolic activities, morphology and cell transcriptome were evaluated. Formation rates of diclofenac metabolites were relatively stable over the application period of seven days in bioreactors exposed to 300 µM diclofenac (300 µM bioreactors (300 µM BR, while in bioreactors exposed to 1000 µM diclofenac (1000 µM BR metabolite concentrations declined drastically. The biochemical data showed a significant decrease in lactate production and for the higher dose a significant increase in ammonia secretion, indicating a dose-dependent effect of diclofenac application. The microarray analyses performed revealed a stable hepatic phenotype of the cells over time and the observed transcriptional changes were in line with functional readouts of the system. In conclusion, the data highlight the suitability of the bioreactor technology for studying the hepatotoxicity of drugs in vitro.

  8. In Vitro Model for Hepatotoxicity Studies Based on Primary Human Hepatocyte Cultivation in a Perfused 3D Bioreactor System.

    Science.gov (United States)

    Knöspel, Fanny; Jacobs, Frank; Freyer, Nora; Damm, Georg; De Bondt, An; van den Wyngaert, Ilse; Snoeys, Jan; Monshouwer, Mario; Richter, Marco; Strahl, Nadja; Seehofer, Daniel; Zeilinger, Katrin

    2016-04-16

    Accurate prediction of the potential hepatotoxic nature of new pharmaceuticals remains highly challenging. Therefore, novel in vitro models with improved external validity are needed to investigate hepatic metabolism and timely identify any toxicity of drugs in humans. In this study, we examined the effects of diclofenac, as a model substance with a known risk of hepatotoxicity in vivo, in a dynamic multi-compartment bioreactor using primary human liver cells. Biotransformation pathways of the drug and possible effects on metabolic activities, morphology and cell transcriptome were evaluated. Formation rates of diclofenac metabolites were relatively stable over the application period of seven days in bioreactors exposed to 300 µM diclofenac (300 µM bioreactors (300 µM BR)), while in bioreactors exposed to 1000 µM diclofenac (1000 µM BR) metabolite concentrations declined drastically. The biochemical data showed a significant decrease in lactate production and for the higher dose a significant increase in ammonia secretion, indicating a dose-dependent effect of diclofenac application. The microarray analyses performed revealed a stable hepatic phenotype of the cells over time and the observed transcriptional changes were in line with functional readouts of the system. In conclusion, the data highlight the suitability of the bioreactor technology for studying the hepatotoxicity of drugs in vitro.

  9. Optimization of Cultivating Conditions for Triterpenoids Production from Antrodia cinnmomea.

    Science.gov (United States)

    He, Yuan-Chuan; He, Kai-Ze; Pu, Qiang; Li, Jing; Zhao, Zong-Jie

    2012-12-01

    The submerged cultivating conditions for triterpenoids production from Antrodia cinnamomea were optimized using uniform design method and the one-factor-at-a-time method was adopted to investigate the effect of plants oils and glucose supply on triterpenoids production and mycelia growth. Corn starch and culturing time were identified as more significant variables for triterpenoids production. The optimal conditions for triterpenoids production was 20.0 g/L corn starch, 20.0 g/L wheat bran, 1.85 g/L MgSO4, initial pH 3 and 16 days of cultivation. In addition, investigation of plant oils and glucose supply showed that 0.3 % (v/v) olive oil supply at the beginning of fermentation stimulated mycelia growth and significantly increased triterpenoids production; 0.2 % (w/v) glucose supplement at 10th day enhanced production of triterpenoids with slight effect on biomass, which is reported for the first time. The triterpenoids production experimentally obtained under the optimal conditions was 7.23 % (w/w). The uniform design method may be used to optimize many environmental and genetic factors such as temperature and agitation that can also affect the triterpenoids production from A. cinnamomea.

  10. Development of an in situ detachment protocol of Vero cells grown on Cytodex1 microcarriers under animal component-free conditions in stirred bioreactor.

    Science.gov (United States)

    Rourou, Samia; Riahi, Nesrine; Majoul, Samy; Trabelsi, Khaled; Kallel, Héla

    2013-08-01

    Subcultivation of Vero cells grown in a proprietary animal component-free medium named IPT-AFM, on microcarriers, was studied. TrypLE Select, a non-animal-derived protease, was used as an alternative to trypsin for cell passaging. We first studied the effect of increasing concentrations of TrypLE Select toward cell growth and then studied the inactivation of the protease using either soybean trypsin inhibitor (STI) or the soy hydrolysate Hypep 1510, in six-well plates. Data showed that cell growth was impaired by residual level of TrypLE Select; STI was identified as an efficient agent to neutralize this effect. To restore cell growth and inactivate TrypLE Select, STI should be added to the medium at least at 0.2 g L(-1). Cells were also grown in spinner flask on 2 g L(-1) Cytodex1 in IPT-AFM. In these conditions, the cell detachment yield was equal to 78 ± 8 %. Furthermore, cells exhibited a typical growth profile when using the dislodged cells to seed a new culture. A cell detachment yield of 70 ± 19 % was also achieved when the cells were grown in a 2-L stirred bioreactor in IPT-AFM, on 3 g L(-1) Cytodex1. This protocol can be of great interest to scale-up the process of Vero cells cultivation in IPT-AFM on Cytodex1 from one stirred bioreactor culture to another.

  11. Bioreactors for high cell density and continuous multi-stage cultivations: options for process intensification in cell culture-based viral vaccine production.

    Science.gov (United States)

    Tapia, Felipe; Vázquez-Ramírez, Daniel; Genzel, Yvonne; Reichl, Udo

    2016-03-01

    With an increasing demand for efficacious, safe, and affordable vaccines for human and animal use, process intensification in cell culture-based viral vaccine production demands advanced process strategies to overcome the limitations of conventional batch cultivations. However, the use of fed-batch, perfusion, or continuous modes to drive processes at high cell density (HCD) and overextended operating times has so far been little explored in large-scale viral vaccine manufacturing. Also, possible reductions in cell-specific virus yields for HCD cultivations have been reported frequently. Taking into account that vaccine production is one of the most heavily regulated industries in the pharmaceutical sector with tough margins to meet, it is understandable that process intensification is being considered by both academia and industry as a next step toward more efficient viral vaccine production processes only recently. Compared to conventional batch processes, fed-batch and perfusion strategies could result in ten to a hundred times higher product yields. Both cultivation strategies can be implemented to achieve cell concentrations exceeding 10(7) cells/mL or even 10(8) cells/mL, while keeping low levels of metabolites that potentially inhibit cell growth and virus replication. The trend towards HCD processes is supported by development of GMP-compliant cultivation platforms, i.e., acoustic settlers, hollow fiber bioreactors, and hollow fiber-based perfusion systems including tangential flow filtration (TFF) or alternating tangential flow (ATF) technologies. In this review, these process modes are discussed in detail and compared with conventional batch processes based on productivity indicators such as space-time yield, cell concentration, and product titers. In addition, options for the production of viral vaccines in continuous multi-stage bioreactors such as two- and three-stage systems are addressed. While such systems have shown similar virus titers compared to

  12. Design and Validation of a Cyclic Strain Bioreactor to Condition Spatially-Selective Scaffolds in Dual Strain Regimes

    Directory of Open Access Journals (Sweden)

    J. Matthew Goodhart

    2014-03-01

    Full Text Available The objective of this study was to design and validate a unique bioreactor design for applying spatially selective, linear, cyclic strain to degradable and non-degradable polymeric fabric scaffolds. This system uses a novel three-clamp design to apply cyclic strain via a computer controlled linear actuator to a specified zone of a scaffold while isolating the remainder of the scaffold from strain. Image analysis of polyethylene terephthalate (PET woven scaffolds subjected to a 3% mechanical stretch demonstrated that the stretched portion of the scaffold experienced 2.97% ± 0.13% strain (mean ± standard deviation while the unstretched portion experienced 0.02% ± 0.18% strain. NIH-3T3 fibroblast cells were cultured on the PET scaffolds and half of each scaffold was stretched 5% at 0.5 Hz for one hour per day for 14 days in the bioreactor. Cells were checked for viability and proliferation at the end of the 14 day period and levels of glycosaminoglycan (GAG and collagen (hydroxyproline were measured as indicators of extracellular matrix production. Scaffolds in the bioreactor showed a seven-fold increase in cell number over scaffolds cultured statically in tissue culture plastic petri dishes (control. Bioreactor scaffolds showed a lower concentration of GAG deposition per cell as compared to the control scaffolds largely due to the great increase in cell number. A 75% increase in hydroxyproline concentration per cell was seen in the bioreactor stretched scaffolds as compared to the control scaffolds. Surprisingly, little differences were experienced between the stretched and unstretched portions of the scaffolds for this study. This was largely attributed to the conditioned and shared media effect. Results indicate that the bioreactor system is capable of applying spatially-selective, linear, cyclic strain to cells growing on polymeric fabric scaffolds and evaluating the cellular and matrix responses to the applied strains.

  13. Pseudomonas putida response in membrane bioreactors under salicylic acid-induced stress conditions.

    Science.gov (United States)

    Collado, Sergio; Rosas, Irene; González, Elena; Gutierrez-Lavin, Antonio; Diaz, Mario

    2014-02-28

    Starvation and changing feeding conditions are frequently characteristics of wastewater treatment plants. They are typical causes of unsteady-state operation of biological systems and provoke cellular stress. The response of a membrane bioreactor functioning under feed-induced stress conditions is studied here. In order to simplify and considerably amplify the response to stress and to obtain a reference model, a pure culture of Pseudomonas putida was selected instead of an activated sludge and a sole substrate (salicylic acid) was employed. The system degraded salicylic acid at 100-1100mg/L with a high level of efficiency, showed rapid acclimation without substrate or product inhibition phenomena and good stability in response to unsteady states caused by feed variations. Under starvation conditions, specific degradation rates of around 15mg/gh were achieved during the adaptation of the biomass to the new conditions and no biofilm formation was observed during the first days of experimentation using an initial substrate to microorganisms ratio lower than 0.1. When substrate was added to the reactor as pulses resulting in rapidly changing concentrations, P. putida growth was observed only for substrate to microorganism ratios higher than 0.6, with a maximum YX/S of 0.5g/g. Biofilm development under changing feeding conditions was fast, biomass detachment only being significant for biomass concentrations on the membrane surface that were higher than 16g/m(2).

  14. Production of bacterial cellulose membranes in a modified airlift bioreactor by Gluconacetobacter xylinus.

    Science.gov (United States)

    Wu, Sheng-Chi; Li, Meng-Hsun

    2015-10-01

    In this study, a novel bioreactor for producing bacterial cellulose (BC) is proposed. Traditional BC production uses static culture conditions and produces a gelatinous membrane. The potential for using various types of bioreactor, including a stirred tank, conventional airlift, and modified airlift with a rectangular wire-mesh draft tube, in large-scale production has been investigated. The BC obtained from these bioreactors is fibrous or in pellet form. Our proposed airlift bioreactor produces a membrane-type BC from Gluconacetobacter xylinus, the water-holding capacity of which is greater than that of cellulose types produced using static cultivation methods. The Young's modulus of the product can be manipulated by varying the number of net plates in the modified airlift bioreactor. The BC membrane produced using the proposed bioreactor exhibits potential for practical application.

  15. Pseudomonas putida response in membrane bioreactors under salicylic acid-induced stress conditions

    Energy Technology Data Exchange (ETDEWEB)

    Collado, Sergio; Rosas, Irene; González, Elena; Gutierrez-Lavin, Antonio; Diaz, Mario, E-mail: mariodiaz@uniovi.es

    2014-02-01

    Highlights: • MBR under feed-induced stress conditions: starvation and changing feeding conditions. • High capacity of MBR to withstand high variations in feed loads. • Slow biofilm formation under starvation conditions during the first days. • Observed growth of P. putida for substrate to microorganism ratio higher than 0.6 g/g. • Maximum specific growth rate and growth yield values of around 37.5 h{sup −1} and 0.5 g/g. - Abstract: Starvation and changing feeding conditions are frequently characteristics of wastewater treatment plants. They are typical causes of unsteady-state operation of biological systems and provoke cellular stress. The response of a membrane bioreactor functioning under feed-induced stress conditions is studied here. In order to simplify and considerably amplify the response to stress and to obtain a reference model, a pure culture of Pseudomonas putida was selected instead of an activated sludge and a sole substrate (salicylic acid) was employed. The system degraded salicylic acid at 100–1100 mg/L with a high level of efficiency, showed rapid acclimation without substrate or product inhibition phenomena and good stability in response to unsteady states caused by feed variations. Under starvation conditions, specific degradation rates of around 15 mg/g h were achieved during the adaptation of the biomass to the new conditions and no biofilm formation was observed during the first days of experimentation using an initial substrate to microorganisms ratio lower than 0.1. When substrate was added to the reactor as pulses resulting in rapidly changing concentrations, P. putida growth was observed only for substrate to microorganism ratios higher than 0.6, with a maximum Y{sub X/S} of 0.5 g/g. Biofilm development under changing feeding conditions was fast, biomass detachment only being significant for biomass concentrations on the membrane surface that were higher than 16 g/m{sup 2}.

  16. NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101825 for a version with major elements labeled, and No. 0103180 for an operational schematic. 0101816

  17. Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors.

    Science.gov (United States)

    Rafiq, Qasim A; Coopman, Karen; Nienow, Alvin W; Hewitt, Christopher J

    2016-03-01

    Production of human mesenchymal stem cells for allogeneic cell therapies requires scalable, cost-effective manufacturing processes. Microcarriers enable the culture of anchorage-dependent cells in stirred-tank bioreactors. However, no robust, transferable methodology for microcarrier selection exists, with studies providing little or no reason explaining why a microcarrier was employed. We systematically evaluated 13 microcarriers for human bone marrow-derived MSC (hBM-MSCs) expansion from three donors to establish a reproducible and transferable methodology for microcarrier selection. Monolayer studies demonstrated input cell line variability with respect to growth kinetics and metabolite flux. HBM-MSC1 underwent more cumulative population doublings over three passages in comparison to hBM-MSC2 and hBM-MSC3. In 100 mL spinner flasks, agitated conditions were significantly better than static conditions, irrespective of donor, and relative microcarrier performance was identical where the same microcarriers outperformed others with respect to growth kinetics and metabolite flux. Relative growth kinetics between donor cells on the microcarriers were the same as the monolayer study. Plastic microcarriers were selected as the optimal microcarrier for hBM-MSC expansion. HBM-MSCs were successfully harvested and characterised, demonstrating hBM-MSC immunophenotype and differentiation capacity. This approach provides a systematic method for microcarrier selection, and the findings identify potentially significant bioprocessing implications for microcarrier-based allogeneic cell therapy manufacture.

  18. Following an Optimal Batch Bioreactor Operations Model

    DEFF Research Database (Denmark)

    Ibarra-Junquera, V.; Jørgensen, Sten Bay; Virgen-Ortíz, J.J.;

    2012-01-01

    The problem of following an optimal batch operation model for a bioreactor in the presence of uncertainties is studied. The optimal batch bioreactor operation model (OBBOM) refers to the bioreactor trajectory for nominal cultivation to be optimal. A multiple-variable dynamic optimization of fed-b...

  19. Degradation of municipal solid waste in simulated landfill bioreactors under aerobic conditions.

    Science.gov (United States)

    Slezak, Radoslaw; Krzystek, Liliana; Ledakowicz, Stanislaw

    2015-09-01

    In this study the municipal solid waste degradation processes in simulated landfill bioreactors under aerobic and anaerobic conditions is investigated. The effect of waste aeration on the dynamics of the aerobic degradation processes in lysimeters as well as during anaerobic processes after completion of aeration is presented. The results are compared with the anaerobic degradation process to determine the stabilization stage of waste in both experimental modes. The experiments in aerobic lysimeters were carried out at small aeration rate (4.41⋅10(-3)lmin(-1)kg(-1)) and for two recirculation rates (24.9 and 1.58lm(-3)d(-1)). The change of leachate and formed gases composition showed that the application of even a small aeration rate favored the degradation of organic matter. The amount of CO2 and CH4 released from anaerobic lysimeter was about 5 times lower than that from the aerobic lysimeters. Better stabilization of the waste was obtained in the aerobic lysimeter with small recirculation, from which the amount of CO2 produced was larger by about 19% in comparison with that from the aerobic lysimeter with large leachate recirculation.

  20. Effect of activated sludge properties and membrane operation conditions on fouling characteristics in membrane bioreactors.

    Science.gov (United States)

    Choi, Hyeok; Zhang, Kai; Dionysiou, Dionysios D; Oerther, Daniel B; Sorial, George A

    2006-06-01

    Biofouling control is considered to be a major challenge in operating membrane bioreactors (MBRs) for the treatment of wastewater. This study examined the impact of biological, chemical, and physical properties of activated sludge on membrane filtration performance in laboratory-scale MBRs. Sludges with different microbial communities were produced using pseudo-continuous stirred-tank reactors and pseudo-plug flow reactors treating a synthetic paper mill wastewater. Various filtration resistances were used to investigate membrane fouling characteristics, and molecular biology tools targeting 16S ribosomal DNA gene sequences were used to identify predominant bacterial populations in the sludges or attached to the fouled membranes. Filtration experiments using axenic cultures of Escherichia coli, Acinetobacter calcoaceticus, and Gordonia amarae were also performed to better understand the initiation and development of biofouling. The results showed that the tendency of membranes to biofoul depended upon membrane operating conditions as well as the properties of the activated sludge in the MBR systems. Specific bacterial populations, which were not dominant in the activated sludges, were selectively accumulated on the membrane surface leading to the development of irreversible biofouling.

  1. Microbial sulfate reduction under sequentially acidic conditions in an upflow anaerobic packed bed bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Jong, T.; Parry, D.L. [Charles Darwin University, Darwin, NT (Australia). Faculty for Educational Health & Science

    2006-07-15

    The aim of this study was to operate an upflow anaerobic packed bed reactor (UAPB) containing sulfate reducing bacteria (SRB) under acidic conditions similar to those found in acid mine drainage (AMD). The UAPB was filled with sand and operated under continuous flow at progressively lower pH and was shown to be capable of supporting sulfate reduction at pH values of 6.0, 5.0, 4.5, 4.0 and 3.5 in a synthetic medium containing 53.5 mmol l{sup -1} lactate. Sulfate reduction rates of 553-1052 mmol m{sup -3} d{sup -1} were obtained when the influent solution pH was progressively lowered from pH 6.0 to 4.0, under an optimal flow rate of 2.61 ml min{sup -1}. When the influent pH was further lowered to pH 3.5, sulfate reduction was substantially reduced with only about 1% sulfate removed at a rate of 3.35 mmol m{sup -3} d{sup -1} after 20 days of operation. However, viable SRB were recovered from the column, indicating that the SRB population was capable of surviving and metabolizing at low levels even at pH 3.5 conditions for at least 20 days. The changes in conductivity in the SRB column did not always occur with changes in pH and redox potential, suggesting that conductivity measurements may be more sensitive to SRB activity and could be used as an additional tool for monitoring SRB activity. The bioreactor containing SRB was able to reduce sulfate and generate alkalinity even when challenged with influent as low as pH 3.5, indicating that such treatment systems have potential for bioremediating highly acidic, sulfate contaminated waste waters.

  2. Bioreactor landfill

    Institute of Scientific and Technical Information of China (English)

    WANG Hao; XING Kai; Anthony Adzomani

    2004-01-01

    Following the population expansion, there is a growing threat brought by municipal solid waste (MSW) against environment and human health. Sanitary landfill is the most important method of MSW disposal in China. In contrast to the conventional landfill, this paper introduces a new technique named bioreactor landfill (BL). Mechanisms, operation conditions as well as the advantages and disadvantages of BL are also discussed in this paper.

  3. Modelling of a Batch Whey Cultivation of Kluyveromyces marxianus var. lactis MC 5 with Investigation of Mass Transfer Processes in the Bioreactor

    Directory of Open Access Journals (Sweden)

    Mitko Petrov

    2015-04-01

    Full Text Available This study presents a mathematical model of a batch fermentation of lactose oxidation from a natural substratum in a cultivation by the strain Kluyweromyces marxianus var. lactis MC 5. In the model of the process, the mass transfer in the bioreactor for oxygen concentration in the gas phase (GP and in the liquid phase (LP is based on the dispersion model of the GP. In addition, perfect mixing in LP is included. Nine models were investigated for specific growth rate and specific oxygen consumptions rate: Monod, Mink, Tessier, Aiba, Andrews, Haldane, Luong, Edward and Han-Levenspiel. In regard to the parameter estimation, the worst observed error was used for all experiments as an objective function. This approach is a special case of multi objective parameter estimation problems allowing the parameter estimation problem to become a min-max problem. The results obtained (values of criteria, relative error and statistics λ for the specific growth rate showed that the best fit to experimental data is achieved when applying the Mink model. In a combination a Mink, and Monod, Mink, Luong, Haldane, and Han-Levenspiel are used for specific oxygen consumptions rate. Based on the investigation, it was discovered that the best fit belonged to the models of Mink and Haldane, Mink and Luong and Mink and Han-Levenspiel. Therefore, these particular models are used for modeling the batch processes.

  4. Evaluation of a new mist-chamber bioreactor for biotechnological applications.

    Science.gov (United States)

    Tscheschke, Bernd; Dreimann, Janis; von der Ruhr, Jürgen W; Schmidt, Timo; Stahl, Frank; Just, Lothar; Scheper, Thomas

    2015-06-01

    In this article we describe the development, the characterization and the evaluation of a novel bioreactor type for the cultivation of different pro- and eukaryotic cell-systems: the mist-chamber bioreactor. This innovative bioreactor meets the demand of cultivation systems for shear stress sensitive cells with high requirements for gas supply. Within the mist-chamber bioreactor the cells are cultivated inside an aerosol of vaporized medium generated by ultrasonic vaporization. In contrast to many established bioreactor systems the mist-chamber bioreactor offers an environment with an excellent gas supply without any impeller or gas bubble induced shear stress. A mist-chamber bioreactor prototype has been manufactured and characterized during this work. In the technical and chemical characterization we evaluated the vaporization process, resulting in a vaporization performance of 32 mL/h at working conditions. On this basis we calculated a biomass of 1.4 g (S. cerevisiae, qs  = 3.45 × 10-3 mol/g/h) and 3.4 g (Aspergillus niger, qs  = 1.33 × 10-3 mol/g/h) where the growth rate becomes limited by transport processes. Additionally, we determined a homogenous cultivation area to a height of 3 cm giving a total volume of 0.45 L for the cultivation. Medium components were examined according to their stability during vaporization with the result that all components are stable for at least 5 days. After the technical characterization we demonstrated the feasibility to cultivate S. cerevisiae and F. velupites in the mist-chamber bioreactor. The results demonstrated that the mist-chamber bioreactor is able to transport a sufficient amount of nutrients consistently to the cell samples and offers an excellent oxygen supply without any shear stress inducing aeration. Furthermore we successfully cultivated F. velupites in a solid state cultivation in a long term experiment. The data indicate that the new bioreactor concept can contribute to

  5. High cell density cultivation of Escherichia coli K4 in a microfiltration bioreactor: a step towards improvement of chondroitin precursor production

    Directory of Open Access Journals (Sweden)

    De Rosa Mario

    2011-02-01

    Full Text Available Abstract Background The bacteria Escherichia coli K4 produces a capsular polysaccharide (K4 CPS whose backbone is similar to the non sulphated chondroitin chain. The chondroitin sulphate is one of the major components of the extra-cellular matrix of the vertebrate connective tissues and a high value molecule, widely employed as active principle in the treatment of osteoarthritis. It is usually obtained by extraction from animal tissues, but the risk of virus contaminations, as well as the scarceness of raw material, makes this productive process unsafe and unable to satisfy the growing market demand. In previous studies a new biotechnological process to produce chondroitin from Escherichia coli K4 capsular polysaccharide was investigated and a 1.4 g·L-1 K4 CPS concentration was reached using fed-batch fermentation techniques. In this work, on the trail of these results, we exploited new fermentation strategies to further improve the capsular polysaccharide production. Results The inhibitory effect of acetate on the bacterial cells growth and K4 CPS production was studied in shake flask conditions, while a new approach, that combined the optimization of the feeding profiles, the improvement of aeration conditions and the use of a microfiltration bioreactor, was investigated in three different types of fermentation processes. High polysaccharide concentrations (4.73 ± 0.2 g·L-1, with corresponding average yields (0.13 ± 0.006 gK4 CPS·gcdw-1, were obtained; the increase of K4 CPS titre, compared to batch and fed-batch results, was of 16-fold and 3.3-fold respectively, while average yield was almost 3.5 and 1.4 fold higher. Conclusion The increase of capsular polysaccharide titre confirmed the validity of the proposed fermentation strategy and opened the way to the use of the microfiltration bioreactor for the biotechnological production of chondroitin.

  6. Over-pressurized bioreactors: application to microbial cell cultures.

    Science.gov (United States)

    Lopes, Marlene; Belo, Isabel; Mota, Manuel

    2014-01-01

    In industrial biotechnology, microbial cultures are exposed to different local pressures inside bioreactors. Depending on the microbial species and strains, the increased pressure may have detrimental or beneficial effects on cellular growth and product formation. In this review, the effects of increased air pressure on various microbial cultures growing in bioreactors under moderate total pressure conditions (maximum, 15 bar) will be discussed. Recent data illustrating the diversity of increased air pressure effects at different levels in microbial cells cultivation will be presented, with particular attention to the effects of oxygen and carbon dioxide partial pressures on cellular growth and product formation, and the concomitant effect of oxygen pressure on antioxidant cellular defense mechanisms.

  7. Production of alginate by Azotobacter vinelandii grown at two bioreactor scales under oxygen-limited conditions.

    Science.gov (United States)

    Díaz-Barrera, Alvaro; Gutierrez, Javiera; Martínez, Fabiola; Altamirano, Claudia

    2014-06-01

    The oxygen transfer rate (OTR) was evaluated as a scale-up criterion for alginate production in 3- and 14-L stirred fermentors. Batch cultures were performed at different agitation rates (200, 300, and 600 rpm) and airflow rates (0.25, 0.5, and 1 vvm), resulting in different maximum OTR levels (OTRmax). Although the two reactors had a similar OTRmax (19 mmol L(-1) h(-1)) and produced the same alginate concentration (3.8 g L(-1)), during the cell growth period the maximum molecular weight of the alginate was 1,250 kDa in the 3-L stirred fermentor and 590 kDa in 14-L stirred fermentor. The results showed for the first time the evolution of the molecular weight of alginate and OTR profiles for two different scales of stirred fermentors. There was a different maximum specific oxygen uptake rate between the two fermenters, reaching 8.3 mmol g(-1) h(-1) in 3-L bioreactor and 10.6 mmol g(-1) h(-1) in 14-L bioreactor, which could explain the different molecular weights observed. These findings open the possibility of using [Formula: see text] instead of OTRmax as a scaling criterion to produce polymers with similar molecular weights during fermentation.

  8. Bioreactors and bioseparation.

    Science.gov (United States)

    Zhang, Siliang; Cao, Xuejun; Chu, Ju; Qian, Jiangchao; Zhuang, Yingping

    2010-01-01

    Along with the rapid development of life science, great attention has been increasingly given to the biotechnological products of cell cultivation technology. In the course of industrialization, bioreactor and bioproduct separation techniques are the two essential technical platforms. In this chapter, the current situation and development prospects of bioreactor techniques in China are systematically discussed, starting with the elucidation of bioreactor processes and the principle of process optimization. Separation technology for biological products is also briefly introduced.At present, a series of bioreactors made by Chinese enterprises have been widely used for laboratory microbial cultivation, process optimization studies, and large-scale production. In the course of bioprocess optimization studies, the complicated bioprocesses in a bioreactor could be resolved into different reaction processes on three scales, namely genetic, cellular, and bioreactor scales. The structural varieties and nonlinear features of various scales of bioprocess systems was discussed through considering the mutual effects of different scale events, namely material flux, energy flux, and information flux, and the optimization approach for bioprocesses was proposed by taking the analysis of metabolic flux and multiscale consideration as a core strategy.In order to realize such an optimization approach, a bioreactor system based on association analysis of multiscale parameters was elaborated, and process optimization of many biological products were materialized, which resulted in great improvement in production efficiency. In designing and manufacturing large-scale bioreactors, the principle of scaling up a process incorporated with flow field study and physiological features in a bioreactor was suggested according to the criterion for the scale-up of cellular physiological and metabolic traits. The flow field features of a bioreactor were investigated through computational fluid

  9. Bioreactors for Plant Embryogenesis and Beyond.

    Science.gov (United States)

    Fei, Liwen; Weathers, Pamela

    2016-01-01

    A variety of different bioreactors have been developed for use in initiating and cultivating somatic embryos. The various designs for embryogenesis and culture are critically evaluated here. Bioreactor optimization and operation methods are also described along with recommendations for use based on desired outcome.

  10. Cultivation Conditions for Phytase Production from Recombinant Escherichia coli DH5α

    OpenAIRE

    Rafidah Mohd Ariff; Anwar Fitrianto; Mohd Yazid Abd. Manap; Aini Ideris; Azhar Kassim; Afinah Suhairin; Anis Shobirin Meor Hussin

    2013-01-01

    Response surface methodology (RSM) was used to optimize the cultivation conditions for the production of phytase by recombinant Escherichia coli DH5α. The optimum predicted cultivation conditions for phytase production were at 3 hours seed age, a 2.5% inoculum level, an L-arabinose concentration of 0.20%, a cell concentration of 0.3 (as measured at 600 nm) and 17 hours post-induction time with a predicted phytase activity of 4194.45 U/mL. The model was validated and the results showed no sign...

  11. Removal of Cr(VI) from aqueous solutions by a bacterial biofilm supported on zeolite: optimisation of the operational conditions and Scale-Up of the bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Pazos, M. [IBB - Instituto de Biotecnologia e Bioengenharia, Centro de Engenharia Biologica, Universidade do Minho, Braga (Portugal); Departamento de Ingenieria Quimica, Universidade de Vigo, Vigo (Spain); Branco, M.; Tavares, T. [IBB - Instituto de Biotecnologia e Bioengenharia, Centro de Engenharia Biologica, Universidade do Minho, Braga (Portugal); Neves, I.C. [Departamento de Quimica, Centro de Quimica, Universidade do Minho, Braga (Portugal); Sanroman, M.A. [Departamento de Ingenieria Quimica, Universidade de Vigo, Vigo (Spain)

    2010-12-15

    The aim of this study was to investigate the feasibility of a bioreactor system and its scale-up to remove Cr(VI) from solution. The bioreactor is based on an innovative process that combines bioreduction of Cr(VI) to Cr(III) by the bacterium Arthrobacter viscosus and Cr(III) sorption by a specific zeolite. Batch studies were conducted in a laboratory-scale bioreactor, taking into account different operating conditions. Several variables, such as biomass concentration, pH and zeolite pre-treatment, were evaluated to increase removal efficiency. The obtained results suggest that the Cr removal efficiency is improved when the initial biomass concentration is approximately 5 g L{sup -1} and the pH in the system is maintained at an acidic level. Under the optimised conditions, approximately 100 % of the Cr(VI) was removed. The scale-up of the developed biofilm process operating under the optimised conditions was satisfactorily tested in a 150-L bioreactor. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  12. Cultivation of Isochrysis galbana in phototrophic, heterotrophic, and mixotrophic conditions.

    Science.gov (United States)

    Alkhamis, Yousef; Qin, Jian G

    2013-01-01

    This study compared the growth and biomass production of Isochrysis galbana under hetero-, mixo-, and phototrophic conditions using different organic carbon sources. The growth of I. galbana was inhibited in heterotrophy but was enhanced in mixotrophy compared to that in phototrophy. Subsequently, the influences of organic carbon and environmental factors (light and salinity) on the growth of I. galbana were further investigated. Algal dry weight increased as glycerol concentrations increased from 0 to 200 mmol and the highest algal production occurred at 50 mmol glycerol. At a range of light intensities of 25-200 μ mol photons m(-2) s(-2), the highest algal growth rate occurred at 100 photons μ mol m(-2) s(-2). The growth of I. galbana was significantly affected by photoperiod, and the maximal dry weight was obtained at 12 h light and 12 h dark. In the salinity test, I. galbana could grow in a wide range of salinities from 10 to 65 ‰, but the 35 ‰ salinity was optimal. This study suggests that the growth and production of I. galbana can be improved using mixotrophic culture at 50 mmol glycerol in 35 ‰ salinity.

  13. Alleviating liver failure conditions using an integrated hybrid cryogel based cellular bioreactor as a bioartificial liver support

    Science.gov (United States)

    Damania, Apeksha; Hassan, Mohsin; Shirakigawa, Nana; Mizumoto, Hiroshi; Kumar, Anupam; Sarin, Shiv K.; Ijima, Hiroyuki; Kamihira, Masamichi; Kumar, Ashok

    2017-01-01

    Conventionally, some bioartificial liver devices are used with separate plasmapheresis unit to separate out plasma from whole blood and adsorbent column to detoxify plasma before it passes through a hepatocytes-laden bioreactor. We aim to develop a hybrid bioreactor that integrates the separate modules in one compact design improving the efficacy of the cryogel based bioreactor as a bioartificial liver support. A plasma separation membrane and an activated carbon cloth are placed over a HepG2-loaded cryogel scaffold in a three-chambered bioreactor design. This bioreactor is consequently connected extracorporeally to a rat model of acute liver failure for 3 h and major biochemical parameters studied. Bilirubin and aspartate transaminase showed a percentage decrease of 20–60% in the integrated bioreactor as opposed to 5–15% in the conventional setup. Urea and ammonia levels which showed negligible change in the conventional setup increase (40%) and decrease (18%), respectively in the integrated system. Also, an overall increase of 5% in human albumin in rat plasma indicated bioreactor functionality in terms of synthetic functions. These results were corroborated by offline evaluation of patient plasma. Hence, integrating the plasmapheresis and adsorbent units with the bioreactor module in one compact design improves the efficacy of the bioartificial liver device. PMID:28079174

  14. Development of Cultivation Technology for the Escherichia coli Recombinant Strain Producing Argininedeiminase of Mycoplasma hominis

    OpenAIRE

    Fayura, L R; Boretsky, Yu.R.; Pynyaha, Yu.V.; Martynyuk, N.B.; Skorohod, V.V.; Sybyrny, А.А.

    2014-01-01

    The recombinant Escherichia coli strain producing arginine deiminase of Mycoplasma hominis has been constructed. Storage conditions that provide stabilization of most productive clones of the producer were found. Terms for cultivation of the arginine deiminase producer using bioreactors of different volume were optimized. Highly purified samples of arginine deiminase were obtained and their longterm storage conditions were selected.

  15. Development of Cultivation Technology for the Escherichia coli Recombinant Strain Producing Argininedeiminase of Mycoplasma hominis

    Directory of Open Access Journals (Sweden)

    Fayura, L.R.

    2014-07-01

    Full Text Available The recombinant Escherichia coli strain producing arginine deiminase of Mycoplasma hominis has been constructed. Storage conditions that provide stabilization of most productive clones of the producer were found. Terms for cultivation of the arginine deiminase producer using bioreactors of different volume were optimized. Highly purified samples of arginine deiminase were obtained and their longterm storage conditions were selected.

  16. Diversity and interactions of microbial functional genes under differing environmental conditions: insights from a membrane bioreactor and an oxidation ditch

    Science.gov (United States)

    Xia, Yu; Hu, Man; Wen, Xianghua; Wang, Xiaohui; Yang, Yunfeng; Zhou, Jizhong

    2016-01-01

    The effect of environmental conditions on the diversity and interactions of microbial communities has caused tremendous interest in microbial ecology. Here, we found that with identical influents but differing operational parameters (mainly mixed liquor suspended solid (MLSS) concentrations, solid retention time (SRT) and dissolved oxygen (DO) concentrations), two full-scale municipal wastewater treatment systems applying oxidation ditch (OD) and membrane bioreactor (MBR) processes harbored a majority of shared genes (87.2%) but had different overall functional gene structures as revealed by two datasets of 12-day time-series generated by a functional gene array-GeoChip 4.2. Association networks of core carbon, nitrogen and phosphorus cycling genes in each system based on random matrix theory (RMT) showed different topological properties and the MBR nodes showed an indication of higher connectivity. MLSS and DO were shown to be effective in shaping functional gene structures of the systems by statistical analyses. Higher MLSS concentrations resulting in decreased resource availability of the MBR system were thought to promote positive interactions of important functional genes. Together, these findings show the differences of functional potentials of some bioprocesses caused by differing environmental conditions and suggest that higher stress of resource limitation increased positive gene interactions in the MBR system.

  17. Diversity and interactions of microbial functional genes under differing environmental conditions: insights from a membrane bioreactor and an oxidation ditch

    Science.gov (United States)

    Xia, Yu; Hu, Man; Wen, Xianghua; Wang, Xiaohui; Yang, Yunfeng; Zhou, Jizhong

    2016-01-01

    The effect of environmental conditions on the diversity and interactions of microbial communities has caused tremendous interest in microbial ecology. Here, we found that with identical influents but differing operational parameters (mainly mixed liquor suspended solid (MLSS) concentrations, solid retention time (SRT) and dissolved oxygen (DO) concentrations), two full-scale municipal wastewater treatment systems applying oxidation ditch (OD) and membrane bioreactor (MBR) processes harbored a majority of shared genes (87.2%) but had different overall functional gene structures as revealed by two datasets of 12-day time-series generated by a functional gene array-GeoChip 4.2. Association networks of core carbon, nitrogen and phosphorus cycling genes in each system based on random matrix theory (RMT) showed different topological properties and the MBR nodes showed an indication of higher connectivity. MLSS and DO were shown to be effective in shaping functional gene structures of the systems by statistical analyses. Higher MLSS concentrations resulting in decreased resource availability of the MBR system were thought to promote positive interactions of important functional genes. Together, these findings show the differences of functional potentials of some bioprocesses caused by differing environmental conditions and suggest that higher stress of resource limitation increased positive gene interactions in the MBR system. PMID:26743465

  18. Model-based analysis of the effect of different operating conditions on fouling mechanisms in a membrane bioreactor.

    Science.gov (United States)

    Sabia, Gianpaolo; Ferraris, Marco; Spagni, Alessandro

    2016-01-01

    This study proposes a model-based evaluation of the effect of different operating conditions with and without pre-denitrification treatment and applying three different solids retention times on the fouling mechanisms involved in membrane bioreactors (MBRs). A total of 11 fouling models obtained from literature were used to fit the transmembrane pressure variations measured in a pilot-scale MBR treating real wastewater for more than 1 year. The results showed that all the models represent reasonable descriptions of the fouling processes in the MBR tested. The model-based analysis confirmed that membrane fouling started by pore blocking (complete blocking model) and by a reduction of the pore diameter (standard blocking) while cake filtration became the dominant fouling mechanism over long-term operation. However, the different fouling mechanisms occurred almost simultaneously making it rather difficult to identify each one. The membrane "history" (i.e. age, lifespan, etc.) seems the most important factor affecting the fouling mechanism more than the applied operating conditions. Nonlinear regression of the most complex models (combined models) evaluated in this study sometimes demonstrated unreliable parameter estimates suggesting that the four basic fouling models (complete, standard, intermediate blocking and cake filtration) contain enough details to represent a reasonable description of the main fouling processes occurring in MBRs.

  19. Anaerobic submerged membrane bioreactor (AnSMBR) for municipal wastewater treatment under mesophilic and psychrophilic temperature conditions.

    Science.gov (United States)

    Martinez-Sosa, David; Helmreich, Brigitte; Netter, Thomas; Paris, Stefania; Bischof, Franz; Horn, Harald

    2011-11-01

    A pilot scale anaerobic submerged membrane bioreactor (AnSMBR) with an external filtration unit for municipal wastewater treatment was operated for 100 days. Besides gas sparging, additional shear was created by circulating sludge to control membrane fouling. During the first 69 days, the reactor was operated under mesophilic temperature conditions. Afterwards, the temperature was gradually reduced to 20 °C. A slow and linear increase in the filtration resistance was observed under critical flux conditions (7 L/(m2 h)) at 35 °C. However, an increase in the fouling rate probably linked to an accumulation of solids, a higher viscosity and soluble COD concentrations in the reactor was observed at 20 °C. The COD removal efficiency was close to 90% under both temperature ranges. Effluent COD and BOD5 concentrations were lower than 80 and 25 mg/L, respectively. Pathogen indicator microorganisms (fecal coliforms bacteria) were reduced by log(10)5. Hence, the effluent could be used for irrigation purposes in agriculture.

  20. Two new disposable bioreactors for plant cell culture: The wave and undertow bioreactor and the slug bubble bioreactor.

    Science.gov (United States)

    Terrier, Bénédicte; Courtois, Didier; Hénault, Nicolas; Cuvier, Arnaud; Bastin, Maryse; Aknin, Aziz; Dubreuil, Julien; Pétiard, Vincent

    2007-04-01

    The present article describes two novel flexible plastic-based disposable bioreactors. The first one, the WU bioreactor, is based on the principle of a wave and undertow mechanism that provides agitation while offering convenient mixing and aeration to the plant cell culture contained within the bioreactor. The second one is a high aspect ratio bubble column bioreactor, where agitation and aeration are achieved through the intermittent generation of large diameter bubbles, "Taylor-like" or "slug bubbles" (SB bioreactor). It allows an easy volume increase from a few liters to larger volumes up to several hundred liters with the use of multiple units. The cultivation of tobacco and soya cells producing isoflavones is described up to 70 and 100 L working volume for the SB bioreactor and WU bioreactor, respectively. The bioreactors being disposable and pre-sterilized before use, cleaning, sterilization, and maintenance operations are strongly reduced or eliminated. Both bioreactors represent efficient and low cost cell culture systems, applicable to various cell cultures at small and medium scale, complementary to traditional stainless-steel bioreactors.

  1. Bioreactor principles

    Science.gov (United States)

    2001-01-01

    Cells cultured on Earth (left) typically settle quickly on the bottom of culture vessels due to gravity. In microgravity (right), cells remain suspended and aggregate to form three-dimensional tissue. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  2. Cells growing in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. Shown here, clusters of cells slowly spin inside a bioreactor. On Earth, the cells continually fall through the buffer medium and never hit bottom. In space, they are naturally suspended. Rotation ensures gentle stirring so waste is removed and fresh nutrient and oxygen are supplied. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  3. Optimization of medium and cultivation conditions for enhanced exopolysaccharide yield by marine Cyanothece sp. 113

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Cyanothece sp. 113 is a unicellular, aerobic, diazotrophic and photosynthetic marine cyanobacterium. The optimal medium for exopolysaccharide yield by the strain was 70.0 g/L of NaCl,and 0.9 g/L of MgSO4 based on the modified F/2 medium for cultivation of marine algae. The optimal cultivation condition for exopolysaccharide yield by this cyanobacterial strain was 29℃, aeration, and continuous illumination at 86.0 μE/M2/S. Under the optimal conditions, over 18.4 g/L of exopolysaccharide was produced within 12 days. This was so far the highest exopolysaccharide yield produced with strains of Cyanothece sp. obtained.

  4. [Is it possible to "cancel" aging process of cell cultures under optimal conditions for cultivation?].

    Science.gov (United States)

    Bozhkov, A I; Kovaleva, M K; Menzianova, N G

    2011-01-01

    The characteristics of the cells epigenotypes Dunaliella viridis Teod. in the process of chronological and replicative aging were investigated. By 40th day of accumulative cultivation (which coincided with the stationary growth phase) DNA content in the cells of Dunaliella viridis increased 2 times, triacylglycerides 3 times, beta-carotene and carbonyl proteins 2 times, RNA content decreased in comparison with cells in exponential growth phase, i. e., the 40th day of growth of culture forms the age-related epigenotype. 4 received subcultures were being transplanted during 2 years in mid-logarithmic growth phase (subculture-10), early stationary phase of growth (subculture-20), in the mid-stationary growth phase (subculture-30), and late stationary growth phase (subculture-40). It is shown that epigenotype of subculture-10 remained unchanged over 2 years of cultivation, i. e., it does not manifest replicative aging. At the same time, the subculture-20, although long enough (at least 40 passages), maintained epigenotype characteristic of young cultures, and showed age-related changes. Pronounced age-dependent changes of epigenotype in the course of cultivation were identified for subculture-30, and subculture-40 was characterized by unstable epigenotype. Thus, cultivation conditions determine the intensity of replicative aging in Dunaliella viridis.

  5. Photo-mixotrophic Cultivation of Algae Euglena gracilis for Lipid Production

    Directory of Open Access Journals (Sweden)

    Tonči Rezić

    2013-03-01

    Full Text Available In the future, due to limited resources, a crisis of energy storing molecules (fuels, which are currently produced from crude mineral oil, is expected. One strategy to compensate a part of the oil deficiency is the production of biodiesel from microalgal lipids. As model microorganism for lipid production microalgae Euglena gracilis was selected and photo mixotrophic cultivation was performed in the stirred tank photobioreactor. During this research, medium composition and operational conditions of photo-bioreactor were optimized in order to define adequate cultivation conditions for algae biomass and lipid production. As low-cost and available complex carbon/ nitrogen source, corn steep liquor (CSL was used to promote E. gracilis growth and lipid production. Due to the optimization of medium composition and cultivation conditions, lipid production was increased up to 29% of biomass dry weight in a two stage cultivation process inside one photo-bioreactor. Promising results obtained in this research encouraged us for further investigation.

  6. Removal of nitrogen and organic matter in a submerged-membrane bioreactor operating in a condition of simultaneous nitrification and denitrification

    Directory of Open Access Journals (Sweden)

    Izabela Major Barbosa

    2016-04-01

    Full Text Available This study evaluated the removal of nitrogen and organic matter in a membrane bioreactor system operating in a condition of simultaneous nitrification and denitrification controlled by intermittent aeration. A submerged-membrane system in a bioreactor was used in a pilot scale to treat domestic wastewater. The dissolved oxygen concentration was maintained between 0.5 and 0.8 mg L-1. The concentration of the mixed liquor suspended solids (MLSS in the system ranged from 1 to 6 g L-1. The system efficiency was evaluated by the removal efficiency of organic matter, quantified by Chemical Oxygen Demand (COD, Biochemical Oxygen Demand (BOD5 and Total Organic Carbon (TOC. Nitrogen removal was assessed by quantifying Total Kjeldahl Nitrogen (TKN and ammonia nitrogen. During the system start-up, the removal efficiencies of COD and NTK were around 90% and 80%, respectively. After the simultaneous nitrification and denitrification (SND conditions were established, the removal efficiencies of COD and NTK were 70% and 99%, respectively. These results showed that sewage treatment with the membrane bioreactor (MBR system, operating with simultaneous nitrification and denitrification conditions, was able to remove organic matter and promote nitrification and denitrification in a single reactor, producing a high-quality permeate.

  7. Design challenges for space bioreactors

    Science.gov (United States)

    Seshan, P. K.; Petersen, G. R.

    1989-01-01

    The design of bioreactors for operation under conditions of microgravity presents problems and challenges. Absence of a significant body force such as gravity can have profound consequences for interfacial phenomena. Marangoni convection can no longer be overlooked. Many speculations on the advantages and benefits of microgravity can be found in the literature. Initial bioreactor research considerations for space applications had little regard for the suitability of the designs for conditions of microgravity. Bioreactors can be classified in terms of their function and type of operation. The complex interaction of parameters leading to optimal design and operation of a bioreactor is illustrated by the JSC mammalian cell culture system. The design of a bioreactor is strongly dependent upon its intended use as a production unit for cell mass and/or biologicals or as a research reactor for the study of cell growth and function. Therefore a variety of bioreactor configurations are presented in rapid summary. Following this, a rationale is presented for not attempting to derive key design parameters such as the oxygen transfer coefficient from ground-based data. A set of themes/objectives for flight experiments to develop the expertise for design of space bioreactors is then proposed for discussion. These experiments, carried out systematically, will provide a database from which engineering tools for space bioreactor design will be derived.

  8. Water resource recovery by means of microalgae cultivation in outdoor photobioreactors using the effluent from an anaerobic membrane bioreactor fed with pre-treated sewage.

    Science.gov (United States)

    Viruela, Alexandre; Murgui, Mónica; Gómez-Gil, Tao; Durán, Freddy; Robles, Ángel; Ruano, María Victoria; Ferrer, José; Seco, Aurora

    2016-10-01

    With the aim of assessing the potential of microalgae cultivation for water resource recovery (WRR), the performance of three 0.55m(3) flat-plate photobioreactors (PBRs) was evaluated in terms of nutrient removal rate (NRR) and biomass production. The PBRs were operated outdoor (at ambient temperature and light intensity) using as growth media the nutrient-rich effluent from an AnMBR fed with pre-treated sewage. Solar irradiance was the most determining factor affecting NRR. Biomass productivity was significantly affected by temperatures below 20°C. The maximum biomass productivity (52.3mgVSS·L(-1)·d(-1)) and NRR (5.84mgNH4-N·L(-1)·d(-1) and 0.85mgPO4-P·L(-1)·d(-1)) were achieved at solar irradiance of 395μE·m(-2)·s(-1), temperature of 25.5°C, and HRT of 8days. Under these conditions, it was possible to comply with effluent nutrient standards (European Directive 91/271/CEE) when the nutrient content in the influent was in the range of 40-50mgN·L(-1) and 6-7mg P·L(-1).

  9. Effect of Cultivation Time and Medium Condition in Production of Bacterial Cellulose Nanofiber for Urease Immobilization

    Directory of Open Access Journals (Sweden)

    M. Pesaran

    2015-01-01

    Full Text Available A new nanoporous biomatrix originated from bacterial resources has been chosen for urease immobilization. Urease has been immobilized on synthesized bacterial cellulose nanofiber since this enzyme has a key role in nitrogen metabolism. Gluconacetobacter xylinum ATCC 10245 has been cultivated for synthesis of a nanofiber with the diameter of 30–70 nm. Different cultivation processes in the aspect of time and cultivation medium conditions were chosen to study the performance of immobilized enzyme on four types of bacterial cellulose nanofibers (BCNs. Urease immobilization into the nanofiber has been done in two steps: enzyme adsorption and glutaraldehyde cross-linking. The results showed that the immobilized enzymes were relatively active and highly stable compared to the control samples of free enzymes. Optimum pH was obtained 6.5 and 7 for different synthesized BCNs, while the optimum temperature for immobilized urease was 50°C. Finding of the current experiment illustrated that the immobilized enzyme in optimum condition lost its initial activity by 41% after 15 weeks.

  10. Cultivation and characters of aerobic granules for pentachlorophenol (PCP) degradation under microaerobic condition

    Institute of Scientific and Technical Information of China (English)

    LAN Hui-xia; CHEN Yuan-cai; CHEN Zhong-hao; CHEN Rong

    2005-01-01

    Cultivation of aerobic granular sludge for pentachlorophenol (PCP) degradation under microaerobic condition ( DO concentration was controlled at 0.2-0.7 mg/L) was studied in this paper. Anaerobic granules were selected as inoculum. The changes of appearance were observed and the variations of SVI, VSS/TSS, PN/PS and the size of sludge were measured during cultivating. The capabilities for degradation of PCP, AOX and CODcr were also studied. Observations on mature granules were carried out by scanning electron microscope, and the results indicated bacillus was dominant on the surface of granules while in the inner of granules both bacillus and coccus were the dominant microorganisms. K, Na, Fe, Ca, Mg, Ni, Co, Mn, Cu and Zn were detected in the granules by element analysis.

  11. Oxygen transfer in a pressurized airlift bioreactor.

    Science.gov (United States)

    Campani, Gilson; Ribeiro, Marcelo Perencin Arruda; Horta, Antônio Carlos Luperni; Giordano, Roberto Campos; Badino, Alberto Colli; Zangirolami, Teresa Cristina

    2015-08-01

    Airlift bioreactors (ALBs) offer advantages over conventional systems, such as simplicity of construction, reduced risk of contamination, and efficient gas-liquid dispersion with low power consumption. ALBs are usually operated under atmospheric pressure. However, in bioprocesses with high oxygen demand, such as high cell density cultures, oxygen limitation may occur even when operating with high superficial gas velocity and air enriched with oxygen. One way of overcoming this drawback is to pressurize the reactor. In this configuration, it is important to assess the influence of bioreactor internal pressure on the gas hold-up, volumetric oxygen transfer coefficient (k(L)a), and volumetric oxygen transfer rate (OTR). Experiments were carried out in a concentric-tube airlift bioreactor with a 5 dm(3) working volume, equipped with a system for automatic monitoring and control of the pressure, temperature, and inlet gas flow rate. The results showed that, in disagreement with previous published results for bubble column and external loop airlift reactors, overpressure did not significantly affect k(L)a within the studied ranges of pressure (0.1-0.4 MPa) and superficial gas velocity in the riser (0.032-0.065 m s(-1)). Nevertheless, a positive effect on OTR was observed: it increased up to 5.4 times, surpassing by 2.3 times the oxygen transfer in a 4 dm(3) stirred tank reactor operated under standard cultivation conditions. These results contribute to the development of non-conventional reactors, especially pneumatic bioreactors operated using novel strategies for oxygen control.

  12. Crossing Methods and Cultivation Conditions for Rapid Production of Segregating Populations in Three Grain Amaranth Species.

    Science.gov (United States)

    Stetter, Markus G; Zeitler, Leo; Steinhaus, Adrian; Kroener, Karoline; Biljecki, Michelle; Schmid, Karl J

    2016-01-01

    Grain amaranths (Amaranthus spp.) have been cultivated for thousands of years in Central and South America. Their grains are of high nutritional value, but the low yield needs to be increased by selection of superior genotypes from genetically diverse breeding populations. Amaranths are adapted to harsh conditions and can be cultivated on marginal lands although little is known about their physiology. The development of controlled growing conditions and efficient crossing methods is important for research on and improvement of this ancient crop. Grain amaranth was domesticated in the Americas and is highly self-fertilizing with a large inflorescence consisting of thousands of very small flowers. We evaluated three different crossing methods (open pollination, hot water emasculation and hand emasculation) for their efficiency in amaranth and validated them with genetic markers. We identified cultivation conditions that allow an easy control of flowering time by day length manipulation and achieved flowering times of 4 weeks and generation times of 2 months. All three different crossing methods successfully produced hybrid F1 offspring, but with different success rates. Open pollination had the lowest (10%) and hand emasculation the highest success rate (74%). Hot water emasculation showed an intermediate success rate (26%) with a maximum of 94% success. It is simple to perform and suitable for a more large-scale production of hybrids. We further evaluated 11 single nucleotide polymorphism (SNP) markers and found that they were sufficient to validate all crosses of the genotypes used in this study for intra- and interspecific hybridizations. Despite its very small flowers, crosses in amaranth can be carried out efficiently and evaluated with inexpensive SNP markers. Suitable growth conditions strongly reduce the generation time and allow the control of plant height, flowering time, and seed production. In combination, this enables the rapid production of segregating

  13. Crossing methods and cultivation conditions for rapid production of segregating populations in three grain amaranth species

    Directory of Open Access Journals (Sweden)

    Markus G Stetter

    2016-06-01

    Full Text Available Grain amaranths (Amaranthus spp. have been cultivated for thousands of years in Central and South America. Their grains are of high nutritional value, but the low yield needs to be increased by selection of superior genotypes from genetically diverse breeding populations. Amaranths are adapted to harsh conditions and can be cultivated on marginal lands although little is known about their physiology. The development of controlled growing conditions and efficient crossing methods is important for research on and improvement of this ancient crop. Grain amaranth was domesticated in the Americas and is highly self-fertilizing with a large inflorescence consisting of thousands of very small flowers. We evaluated three different crossing methods (open pollination, hot water emasculation and hand emasculation for their efficiency in amaranth and validated them with genetic markers. We identified cultivation conditions that allow an easy control of flowering time by manipulating day length and achieved flowering times of four weeks and generation times of two months. All three different crossing methods successfully produced hybrid F1 offspring, but with different success rates. Open pollination had the lowest (10% and hand emasculation the highest success rate (74%. Hot water emasculation showed an intermediate success rate (26% with a maximum of 94% success. It is simple to perform and suitable for a more large-scale production of hybrids. We further evaluated 11 single nucleotide polymorphism (SNP markers and found that they were sufficient to validate all crosses of the genotypes used in this study for intra- and interspecific hybridisations. Despite its very small flowers, crosses in amaranth can be carried out efficiently and evaluated with inexpensive SNP markers. Suitable growth conditions strongly reduce the generation time and allow the control of plant height, flowering time and seed production. In combination, this enables the rapid

  14. Fate of aniline and sulfanilic acid in UASB bioreactors under denitrifying conditions.

    Science.gov (United States)

    Pereira, Raquel; Pereira, Luciana; van der Zee, Frank P; Madalena Alves, M

    2011-01-01

    Two upflow anaerobic sludge blanket (UASB) reactors were operated to investigate the fate of aromatic amines under denitrifying conditions. The feed consisted of synthetic wastewater containing aniline and/or sulfanilic acid and a mixture of volatile fatty acids (VFA) as the primary electron donors. Reactor 1 (R1) contained a stoichiometric concentration of nitrate and Reactor 2 (R2) a stoichiometric nitrate and nitrite mixture as terminal electron acceptors. The R1 results demonstrated that aniline could be degraded under denitrifying conditions while sulfanilic acid remains. The presence of nitrite in the influent of R2, caused a chemical reaction that led to immediate disappearance of both aromatic amines and the formation of an intense yellow coloured solution. HPLC analysis of the influent solution, revealed the emergence of three product peaks: the major one at retention time (R(t)) 14.3 min and two minor at R(t) 17.2 and 21.5 min. In the effluent, the intensity of the peaks at R(t) 14.3 and 17.2 min was very low and of that at R(t) 21.5 min increased (∼3-fold). Based on the mass spectrometry analysis, we propose the structures of some possible products, mainly azo compounds. Denitrification activity tests suggest that biomass needed to adapt to the new coloured compounds, but after a 3 days lag phase, activity is recovered and the final (N(2) + N(2)O) is even higher than that of the control.

  15. Oscillating Cell Culture Bioreactor

    Science.gov (United States)

    Freed, Lisa E.; Cheng, Mingyu; Moretti, Matteo G.

    2010-01-01

    To better exploit the principles of gas transport and mass transport during the processes of cell seeding of 3D scaffolds and in vitro culture of 3D tissue engineered constructs, the oscillatory cell culture bioreactor provides a flow of cell suspensions and culture media directly through a porous 3D scaffold (during cell seeding) and a 3D construct (during subsequent cultivation) within a highly gas-permeable closed-loop tube. This design is simple, modular, and flexible, and its component parts are easy to assemble and operate, and are inexpensive. Chamber volume can be very low, but can be easily scaled up. This innovation is well suited to work with different biological specimens, particularly with cells having high oxygen requirements and/or shear sensitivity, and different scaffold structures and dimensions. The closed-loop changer is highly gas permeable to allow efficient gas exchange during the cell seeding/culturing process. A porous scaffold, which may be seeded with cells, is fixed by means of a scaffold holder to the chamber wall with scaffold/construct orientation with respect to the chamber determined by the geometry of the scaffold holder. A fluid, with/without biological specimens, is added to the chamber such that all, or most, of the air is displaced (i.e., with or without an enclosed air bubble). Motion is applied to the chamber within a controlled environment (e.g., oscillatory motion within a humidified 37 C incubator). Movement of the chamber induces relative motion of the scaffold/construct with respect to the fluid. In case the fluid is a cell suspension, cells will come into contact with the scaffold and eventually adhere to it. Alternatively, cells can be seeded on scaffolds by gel entrapment prior to bioreactor cultivation. Subsequently, the oscillatory cell culture bioreactor will provide efficient gas exchange (i.e., of oxygen and carbon dioxide, as required for viability of metabolically active cells) and controlled levels of fluid

  16. Environmental impacts of barley cultivation under current and future climatic conditions

    DEFF Research Database (Denmark)

    Dijkman, Teunis Johannes; Birkved, Morten; Saxe, Henrik

    2017-01-01

    The purpose of this work is to compare the environmental impacts of spring barley cultivation in Denmark under current (year 2010) and future (year 2050) climatic conditions. Therefore, a Life Cycle Assessment was carried out for the production of 1 kg of spring barley in Denmark, at farm gate....... Both under 2010 and 2050 climatic conditions, four subscenarios were modelled, based on a combination of two soil types and two climates. Included in the assessment were seed production, soil preparation, fertilization, pesticide application, and harvest. When processes in the life cycle resulted in co...... categories, except human and freshwater eco-toxicity, are higher when the barley is produced under climatic circumstances representative for 2050. Comparison of the 2010 and 2050 climatic scenarios indicates that a predicted decrease in barley yields under the 2050 climatic conditions is the main driver...

  17. Cultivation of Thalictrum rugosum cell suspension in an improved airlift bioreactor: stimulatory effect of carbon dioxide and ethylene on alkaloid production.

    Science.gov (United States)

    Kim, D I; Pedersen, H; Chin, C K

    1991-08-05

    Airlift bioreactor operations have been studied for the growth-associated production of secondary metabolites from plant cell suspension cultures. The model system used in this work was Thalictrum rugosum producing berberine, an isoquinoline alkaloid. The airlift system was well suited for growth of Thalictrum cell suspension cultures unless the cell density was high. At high cell density, the airlift system with a draught tube was not adequate due to large aggregates clogging the recirculation paths. This was overcome by use of a cell scraper in the reactor. For berberine production, gas-stripping also played a significant role and it was discovered that CO(2) and ethylene were important for product formation. By supplying a mixture of CO(2) and ethylene into the airlift system, the specific berberine content was increased twofold. It is evident that continuous gas sparging was harmful for the production of berberine without supplementation with other gases.

  18. Influence of sludge retention time and temperature on the sludge removal in a submerged membrane bioreactor: comparative study between pure oxygen and air to supply aerobic conditions.

    Science.gov (United States)

    Rodríguez, F A; Leyva-Díaz, J C; Reboleiro-Rivas, P; González-López, J; Hontoria, E; Poyatos, J M

    2014-01-01

    Performance of a bench-scale wastewater treatment plant, which consisted of a membrane bioreactor, was monitored daily using pure oxygen and air to supply aerobic conditions with the aim of studying the increases of the aeration and sludge removal efficiencies and the effect of the temperature. The results showed the capacity of membrane bioreactor systems for removing organic matter. The alpha-factors of the aeration were determined for six different MLSS concentrations in order to understand the system working when pure oxygen and air were used to supply aerobic conditions in the system. Aeration efficiency was increased between 30.7 and 45.9% when pure oxygen was used in the operation conditions (a hydraulic retention time of 12 h and MLSS concentrations between 4,018 and 11,192 mg/L). Sludge removal efficiency increased incrementally, from 0.2 to 1.5% when pure oxygen was used at low sludge retention time and from 1.5% to 15.4% at medium sludge retention time when temperature conditions were lower than 20°C. Moreover, the difference between calculated and experimental sludge retention time was lesser when pure oxygen was used to provide aerobic conditions, so the influence of the temperature decreased when the pure oxygen was used. These results showed the convenience of using pure oxygen due to the improvement in the performance of the system.

  19. Advanced methods for bioreactor characterization.

    Science.gov (United States)

    Lübbert, A

    1992-08-01

    Bioreactors are characterized by the transport capacities they provide to optimally supply the microorganisms during production process. The transport is performed by flows induced in their cultivation media. In order to understand the extremely complex mixing, mass and heat transfer phenomena encountered, and to perceive their influences on bioreactor performance, sophisticated measuring techniques are required. This review compiles the developments currently in progress to surmount today's shortage of reliable measuring techniques. Measuring techniques are distinguished which can be used on different scales and their application spectra are illustrated by recently obtained results. Several new measuring techniques, which can be employed to resolve the flow structures, are discussed in detail. Only those techniques are considered which can be used to advantage during real cultivations in industrial-scale reactors.

  20. Variability of the microbial abundance of a kefir grain starter cultivated in partially controlled conditions

    Directory of Open Access Journals (Sweden)

    Oger R.

    2005-01-01

    Full Text Available The variability of the abundance in lactobacilli, lactic acid streptococci and yeasts of a kefir grain starter cultivated in partially controlled conditions – milk renewal at room temperature and incubation at defined temperature - was quantified. Expressed by the geometric relative standard deviations of counts repeated over time, it was respectively of 28%, 443% and 35% for each of the three microbial groups. The origin of the microbial variability observed was ascribed to the heterogeneous medium developed around the grains during fermentation and, for the lactic acid streptococci, even more to the initial conditions of fermentation, probably to parameters linked to milk renewal (room temperature, waiting time at room temperature before incubation.

  1. Effect of Gluconacetobacter xylinus cultivation conditions on the selected properties of bacterial cellulose

    Directory of Open Access Journals (Sweden)

    Fijałkowski Karol

    2016-12-01

    Full Text Available The aim of the study was to analyze the changes in the parameters of bacterial cultures and bacterial cellulose (BC synthesized by four reference strains of Gluconacetobacter xylinus during 31-day cultivation in stationary conditions. The study showed that the most visible changes in the analyzed parameters of BC, regardless of the bacterial strain used for their synthesis, were observed in the first 10–14 days of the experiment. It was also revealed, that among parameters showing dependence associated with the particular bacterial strain were the rate and period of BC synthesis, the growth rate of bacteria anchored to the cellulose fibrils, the capacity to absorb water and the water release rate. The results presented in this work may be useful in the selection of optimum culturing conditions and period from the point of view of good efficiency of the cellulose synthesis process.

  2. [Mechanism of malate dehydrogenase isoform formation in Sphaerotilus natans D-507 under different cultivation conditions].

    Science.gov (United States)

    Eprintsev, A T; Falaleeva, M I; Arabtseva, M A; Lavrinenko, I A; Parfenova, I V; Grechkina, M V; Abud, F S

    2011-01-01

    Electrophoretically homogenous preparations of malate dehydrogenase (MDH) isoforms of the bacteria Sphaerotilus natans D-507 with specific activity 7.46 U/mg and 5.74 U/mg with respect to protein concentration have been obtained. The dimeric isoform of the enzyme was shown to function under organotrophic growth conditions, whereas the tetrameric isoform was induced under mixotrophic cultivation conditions. PCR-analysis revealed a single gene encoding the malate dehydrogenase molecule. The topography of the MDH isoform surface was studied by atomic-force microscopy, and a 3D-structure of the enzyme was obtained. Spectraphotometric analysis data allowed us to suggest that stabilization of the tetrameric form of MDH is due to additional bounds implicated in the quaternary structure formation.

  3. Characterization of a pilot plant airlift tower loop bioreactor: II. Evaluation of global mixing properties of the gas phase during yeast cultivation.

    Science.gov (United States)

    Fröhlich, S; Lotz, M; Korte, T; Lübbert, A; Schügerl, K; Seekamp, M

    1991-04-25

    Saccharomyces cerevisiae was cultivated in a 4-m(3) pilot plant airlift tower loop reactor with a draft tube in batch and continuous operations and for comparison in a laboratory airlift tower loop reactor of 0.08 m(3) volume. The reactors were characterized during and after the cultivation by measuring the distributions of the residence times of the gas phase with pseudostochastic tracer signals and mass spectrometer and by evaluating the mixing in the liquid phase with a pulse-shaped volatile tracer signal and mass spectrometer as a detector. The mean residence times and the intensities of the axial mixing in the riser and downcomer, the circulation times of the gas phase, and the fraction of the recirculated gas phase were evaluated and compared.

  4. A stable lipase from Candida lipolytica: cultivation conditions and crude enzyme characteristics.

    Science.gov (United States)

    Pereira-Meirelles, F V; Rocha-Leão, M H; Sant Anna, G L

    1997-01-01

    Although lipases have been intensively studied, some aspects of enzyme production like substrate uptake, catabolite repression, and enzyme stability under long storage periods are seldom discussed in the literature. This work deals with the production of lipase by a new selected strain of Candida lipolytica. Concerning nutrition, it was observed that inorganic nitrogen sources were not as effective as peptone, and that oleic acid or triacylglycerides (TAG) were essential carbon sources. Repression by glucose and stimulation by oleic acid and long chain TAG (triolein and olive oil) were observed. Extracellular lipase activity was only observed at high levels at late stationary phase, whereas intracellular lipase levels were constant and almost undetectable during the cultivation period, suggesting that the produced enzyme was attached to the cell wall, mainly at the beginning of cultivation. The crude lipase produced by this yeast strain shows the following optima conditions: pH 8.0-10.0, temperature of 55 degrees C. Moreover, this preparation maintains its full activity for at least 370 d at 5 degrees C.

  5. INFLUENCE OF CULTIVATION CONDITIONS ON ANTIMICROBIAL PROPERTIES OF Nocardia vaccinii ІMV B-7405 SURFACTANTS

    Directory of Open Access Journals (Sweden)

    Т. P.

    2016-02-01

    Full Text Available The aim of the work was investigation of antimicrobial effect of Nocardia vaccinii ІMV B-7405 surfactants, synthesized in various culture conditions, against phytopathogenic bacteria of genera Pseudomonas, Xanthomonas, and Pectobacterium. The antimicrobial properties of surfactant were determined in suspension culture by Koch method and also by index of the minimum inhibitory concentration. Surfactants were extracted from supernatant of cultural liquid using mixture of chloroform and methanol (2: 1. It has been established that antimicrobial properties of surfactants depend on the nature of the carbon source in the medium (refined vegetable oil, as well as waste oil after frying potatoes and meat, glycerol, the duration of the cultivation (5 and 7 days, the degree of purification of the surfactants (the supernatant of cultural liquid, purified surfactants solution and the test culture type. The highest antimicrobial activity was exhibited by purified surfactants solutions synthesized by microorganisms on the waste oil after potato frying (decreased survival of pathogenic bacteria by 50–95%, and surfactants formed within 7 days of strain B-7405 ІMV cultivation on all test substrates (minimum inhibitory concentration 7–40 µg/mL, which is several times lower than the surfactant, synthesized for 5 days. These data are promising for the development of ecologically friendly biopreparations for the regulation of the number of phytopathogenic bacteria.

  6. Antioxidant and enzyme inhibitory activities of Plebeian herba (Salvia plebeia R. Br.) under different cultivation conditions.

    Science.gov (United States)

    Chen, Lei; Kang, Young-Hwa

    2014-03-12

    An adaptation of cultural management to the specific cultural system, as well as crop demand, can further result in the improvement of the quality of horticultural products. Therefore, this study focused on the antioxidant and enzyme inhibitory activities of Plebeian herba (Salvia plebeia R. Br.) grown in hydroponics in comparison with those of the plant grown in soil. The antioxidant activities of Plebeian herba extract were measured as 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging abilities as well as the reducing power by decreasing nitric oxide (NO) and superoxide dismutase activity (SOD) in vitro. Interestingly, by comparison with hydroponics and traditional cultivation, Plebeian herba cultivated in nutrition-based soil improved inhibitory effect on free radicals of DPPH, ABTS, and NO and increased the contents of phenolics such as caffeic acid (1), luteolin-7-glucoside (2), homoplantaginin (3), hispidulin (4), and eupatorin. Free radical scavenging and SOD activity, as well as α-glucosidase inhibitory effect, were higher in Plebeian herba grown in nutrition-based soil than in plants grown in hydroponics and traditional condition.

  7. Use of Orbital Shaken Disposable Bioreactors for Mammalian Cell Cultures from the Milliliter-Scale to the 1,000-Liter Scale

    Science.gov (United States)

    Zhang, Xiaowei; Stettler, Matthieu; de Sanctis, Dario; Perrone, Marco; Parolini, Nicola; Discacciati, Marco; de Jesus, Maria; Hacker, David; Quarteroni, Alfio; Wurm, Florian

    Driven by the commercial success of recombinant biopharmaceuticals, there is an increasing demand for novel mammalian cell culture bioreactor systems for the rapid production of biologicals that require mammalian protein processing. Recently, orbitally shaken bioreactors at scales from 50 mL to 1,000 L have been explored for the cultivation of mammalian cells and are considered to be attractive alternatives to conventional stirred-tank bioreactors because of increased flexibility and reduced costs. Adequate oxygen transfer capacity was maintained during the scale-up, and strategies to increase further oxygen transfer rates (OTR) were explored, while maintaining favorable mixing parameters and low-stress conditions for sensitive lipid membrane-enclosed cells. Investigations from process development to the engineering properties of shaken bioreactors are underway, but the feasibility of establishing a robust, standardized, and transferable technical platform for mammalian cell culture based on orbital shaking and disposable materials has been established with further optimizations and studies ongoing.

  8. NASA Bioreactors Advance Disease Treatments

    Science.gov (United States)

    2009-01-01

    The International Space Station (ISS) is falling. This is no threat to the astronauts onboard, however, because falling is part of the ISS staying in orbit. The absence of gravity beyond the Earth s atmosphere is actually an illusion; at the ISS s orbital altitude of approximately 250 miles above the surface, the planet s gravitational pull is only 12-percent weaker than on the ground. Gravity is constantly pulling the ISS back to Earth, but the space station is also constantly traveling at nearly 18,000 miles per hour. This means that, even though the ISS is falling toward Earth, it is moving sideways fast enough to continually miss impacting the planet. The balance between the force of gravity and the ISS s motion creates a stable orbit, and the fact that the ISS and everything in it including the astronauts are falling at an equal rate creates the condition of weightlessness called microgravity. The constant falling of objects in orbit is not only an important principle in space, but it is also a key element of a revolutionary NASA technology here on Earth that may soon help cure medical ailments from heart disease to diabetes. In the mid-1980s, NASA researchers at Johnson Space Center were investigating the effects of long-term microgravity on human tissues. At the time, the Agency s shuttle fleet was grounded following the 1986 Space Shuttle Challenger disaster, and researchers had no access to the microgravity conditions of space. To provide a method for recreating such conditions on Earth, Johnson s David Wolf, Tinh Trinh, and Ray Schwarz developed that same year a horizontal, rotating device called a rotating wall bioreactor that allowed the growth of human cells in simulated weightlessness. Previously, cell cultures on Earth could only be grown two-dimensionally in Petri dishes, because gravity would cause the multiplying cells to sink within their growth medium. These cells do not look or function like real human cells, which grow three-dimensionally in

  9. Production of biomass and bioactive compounds from adventitious roots by optimization of culturing conditions of Eurycoma longifolia in balloon-type bubble bioreactor system.

    Science.gov (United States)

    Lulu, Tao; Park, So-Young; Ibrahim, Rusli; Paek, Kee-Yoeup

    2015-06-01

    The present study aimed to optimize the conditions for the production of adventitious roots from Eurycoma longifolia Jack, an important medicinal woody plant, in bioreactor culture. The effects of the type and concentration of auxin on root growth were studied, as well as the effects of the NH4(+):NO3(-) ratio on adventitious root growth and the production of phenolics and flavonoids. Approximately 5 g L(-1) fresh weight of adventitious roots was inoculated into a 3 L balloon-type bubble bioreactor, which contained 2 L 3/4 MS medium supplemented with 30 g L(-1) sucrose and cultures were maintained in the dark for 7 weeks at 24 ± 1°C. Higher concentrations of IBA (7.0 and 9.0 mg L(-1)) and NAA (5.0 mg L(-1)) enhanced the biomass and accumulation of total phenolics and flavonoids. The adventitious roots were thin, numerous, and elongated in 3/4 MS medium supplemented with 5.0 and 7.0 mg L(-1) IBA, whereas the lateral roots were shorter and thicker with 5.0 mg L(-1) NAA compared with IBA treatment. The optimum biomasses of 50.22 g L(-1) fresh weight and 4.60 g L(-1) dry weight were obtained with an NH4(+):NO3(-) ratio of 15:30. High phenolic and flavonoid productions (38.59 and 11.27 mg L(-1) medium, respectively) were also obtained with a ratio of 15:30. Analysis of the 2,2-diphenyl-1-picrylhydrazyl (DPPH)-scavenging activity indicated higher antioxidant activity with an NH4(+):NO3(-) ratio of 30:15. These results suggest that balloon-type bubble bioreactor cultures are suitable for the large-scale commercial production of E. longifolia adventitious roots which contain high yield of bioactive compounds.

  10. Study of the rheological properties of a fermentation broth of the fungus Beauveria bassiana in a bioreactor under different hydrodynamic conditions.

    Science.gov (United States)

    Núñez-Ramírez, Diola Marina; Medina-Torres, Luis; Valencia-López, José Javier; Calderas, Fausto; López Miranda, Javier; Medrano-Roldán, Hiram; Solís-Soto, Aquiles

    2012-11-01

    Fermentation with filamentous fungi in a bioreactor is a complex dynamic process that is affected by flow conditions and the evolution of the rheological properties of the medium. These properties are mainly affected by the biomass concentration and the morphology of the fungus. In this work, the rheological properties of a fermentation with the fungus Beauveria bassiana under different hydrodynamic conditions were studied and the rheological behavior of this broth was simulated through a mixture of carboxymethyl cellulose sodium and cellulose fibers (CMCNa-SF). The bioreactor was a 10 L CSTR tank operated at different stir velocities. Rheological results were similar at 100 and 300 rpm for both systems. However, there was a significant increase in the viscosity accompanied by a change in the consistence index, calculated according to the power law model, for both systems at 800 rpm. The systems exhibited shear-thinning behavior at all stir velocities, which was determined with the power law model. The mixing time was observed to increase as the cellulose content in the system increased and, consequently, the efficiency of mixing diminished. These results are thought to be due to the rheological and morphological similarities of the two fungal systems. These results will help in the optimization of scale-up production of these fungi.

  11. Projeto e construção de um bioreator para síntese orgânica assimétrica catalisada por saccharomyces cerevisiae (fermento biológico de padaria Project and construction of a bioreactor for reactions catalyzed by baker's yeast (saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Ricardo de Souza Pereira

    1997-10-01

    Full Text Available A model for the construction of a simple and cheap apparatus to be used as bioreactor for reactions catalyzed by baker's yeast (Saccharomyces cerevisiae is described. The bioconversion and separation of cells from products and residual substrates are obtained at the same time. The reactions carried out in this type of reactor are faster than those catalyzed by immobilized cells. Yeast cells can be cultivated in this bioreactor operating with cell recycling at appropriated conditions using glucose and other nutrients.

  12. A Novel bioreactor with mechanical stimulation for skeletal tissue engineering

    Directory of Open Access Journals (Sweden)

    M. Petrović

    2009-01-01

    Full Text Available The provision of mechanical stimulation is believed to be necessary for the functional assembly of skeletal tissues, which are normally exposed to a variety of biomechanical signals in vivo. In this paper, we present a development and validation of a novel bioreactor aimed for skeletal tissue engineering that provides dynamic compression and perfusion of cultivated tissues. Dynamic compression can be applied at frequencies up to 67.5 Hz and displacements down to 5 m thus suitable for the simulation of physiological conditions in a native cartilage tissue (0.1-1 Hz, 5-10 % strain. The bioreactor also includes a load sensor that was calibrated so to measure average loads imposed on tissue samples. Regimes of the mechanical stimulation and acquisition of load sensor outputs are directed by an automatic control system using applications developed within the LabView platform. In addition, perfusion of tissue samples at physiological velocities (10–100 m/s provides efficient mass transfer, as well as the possibilities to expose the cells to hydrodynamic shear and simulate the conditions in a native bone tissue. Thus, the novel bioreactor is suited for studies of the effects of different biomechanical signals on in vitro regeneration of skeletal tissues, as well as for the studies of newly formulated biomaterials and cell biomaterial interactions under in vivo-like settings.

  13. Mycelial cultivation of Phellinus linteus using cheese-processing waste and optimization of bioconversion conditions.

    Science.gov (United States)

    Lee, Changsoo; Lee, Seungyong; Cho, Kyung-Jin; Hwang, Seokhwan

    2011-02-01

    A medicinal mushroom, Phellinus linteus, was successfully cultivated using a cheese-processing waste, whey, and the optimal bioconversion conditions for the maximum mycelial growth rate was also estimated through solid-state cultivation experiments. Response surface analysis with a face-centered design (center point replication = 5) was applied to statistically approximate the simultaneous effects of the three variables, i.e., substrate concentration (10-30 g lactose l⁻¹), temperature (20-30°C), and pH (4-6), on the mycelial growth rate of P. linteus. The following is a partial cubic model where η is the mycelial growth rate (K(r)) and x(k) is the corresponding variable term (k = substrate concentration, temperature, and pH in order): η = -23.8 + 8.67 × 10⁻² x₁ + 1.48x₂ + 1.77x₃ + 8.00 × 10⁻⁴ x₁x₂ + 7.25 × 10⁻² x₁x₃ + 5.13 × 10⁻² x₂x₃ -1.28 × 10⁻² x₁² -3.18 × 10⁻² x²₂. -2.64 × 10⁻¹ x₃² -3.28 × 10⁻³ x₁x₂ x₃ + 4.68 × 10⁻⁴ x₁²x₂. The produced response surface model proved to be significant (r² > 0.99, P-value linteus mycelia. This may provide another potential for managing this nutrient-rich waste in a cost-effective way.

  14. Optimal control of the process of cultivation in the conditions of infection

    Directory of Open Access Journals (Sweden)

    N. V. Sukhanova

    2016-01-01

    Full Text Available The article presents a way of solving of the optimal control problem of antibiotic feeding under condition of infection, consisting in the selection of the optimal control in the field of admissible control, with the aim of achieving a compromise between the losses in production due to the presence of foreign microflora, and the cost of its suppression due to the application of antibiotic. The presence of other microorganisms in the finished product, in particular of the “wild“ ones, considerably impairs the quality indicators of the final product (in particular, it reduces the storage time. In peculiar conditions of production it is possible to improve the quality of target product due to elimination of infection, including, when used antibiotics in the process of cultivation, but due to the lack of efficient algorithms and control systems of their supply the question is still open. We use the system of Lotka-Volterra adapted for microbiological process as a mathematical model adequately describing the situation of competitive interaction of two populations of microorganisms (useful and “wild“ ones due to the consumption of one resource. The aim is to find a control law U(t belonging to the field of admissible control. The control that affords minimum to the optimization criterion in accordance with the principle of maximum is defined by the condition of the maximum of Hamilton function and the resulting canonical system of equations. The modified conjugated system of equations in matrix form is obtained. The solution of system of differential-different equations in the analytical form is found using the method of coordinate transformation. As a result an optimal control law is found (with regard to the selected criterion. This is the control law of application of the antibiotic, allowing to control the concentration of foreign microflora in the process of cultivation of microorganisms and accounting for the specific

  15. Enhancement of Chlorella vulgaris Biomass Cultivated in POME Medium as Biofuel Feedstock under Mixotrophic Conditions

    Directory of Open Access Journals (Sweden)

    M.M. Azimatun Nur

    2015-10-01

    Full Text Available Microalgae cultivated in mixotrophic conditions have received significant attention as a suitable source of biofuel feedstock, based on their high biomass and lipid productivity. POME is one of the wastewaters generated from palm oil mills, containing important nutrients that could be suitable for mixotrophic microalgae growth. The aim of this research was to identify the growth of Chlorella vulgaris cultured in POME medium under mixotrophic conditions in relation to a variety of organic carbon sources added to the POME mixture. The research was conducted with 3 different carbon sources (D-glucose, crude glycerol and NaHCO3 in 40% POME, monitored over 6 days, under an illumination of 3000 lux, and with pH = 7. The biomass was harvested using an autoflocculation method and dry biomass was extracted using an ultrasound method in order to obtain the lipid content. The results show that C. vulgaris using D-glucose as carbon source gained a lipid productivity of 195 mg/l/d.

  16. Effects of site conditions and methods of cultivation on growth of sawtooth oak plantations

    Institute of Scientific and Technical Information of China (English)

    Luozhong TANG; Mukui YU; Dan ZHAO; Chunfeng YAN; Zhilong LIU; Shengzuo FANG

    2009-01-01

    The effects of site conditions and cultivation on the growth of sawtooth oak {Quercus acutissima Carr.) plantations were evaluated at the Hongyashan forest farm, in Chuzhou City, Anhui Province, China. The results indicate that the position on the slope, the amount of gravel and the thickness of the soil were important factors in the growth of the sawtooth oak. Lower slope positions with small amounts of gravel and a thick soil were better for the growth of this species than middle slope positions with more gravel and a thin soil. Given the site conditions of the hilly and mountainous areas in Chuzhou City, the mixed Chinese fir (Cunninghamia lanceolata Hook.) and sawtooth oak forests did not improve forest productivity compared with pure sawtooth oak forests. Both urea and compound fertilizers promoted the growth of sawtooth oak, as did site preparation and intercropping. Two years after planting, the height growth of ordinary seedlings with a starting height of 0.6 m was higher than that of supper seedlings with a starting height of 1.0 m. Compared with planting, the early growth of the coppices was faster, but the later growth of the coppices was slower.

  17. Evaluation of the effects of various culture condition on Cr (VI)reduction by Shewanella oneidensis MR-1 in a novel high-throughputmini-bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Yinjie J.; Laidlaw, David; Gani, Kishen; Keasling, Jay D.

    2006-03-16

    The growth and Cr(VI) reduction by Shewanella oneidensisMR-1 was examined using a mini-bioreactor system that independentlymonitors and controls pH, dissolved oxygen, and temperature for each ofits 24, 10-mL reactors. Independent monitoring and control of eachreactor in the cassette allows the exploration of a matrix ofenvironmental conditions known to influence S. oneidensis chromiumreduction. S. oneidensis MR-1 grew in minimal medium without amino acidor vitamin supplementation under aerobic conditions but required serineand glycine supplementation under anaerobic conditions. Growth wasinhibited by dissolved oxygen concentrations>80 percent. Lactatetransformation to acetate was enhanced by low concentration of dissolvedoxygen during the logarithmic growth phase. Between 11 and 35oC, thegrowth rate obeyed the Arrhenius reaction rate-temperature relationship,with a maximum growth rate occurring at 35oC. S. oneidensis MR-1 was ableto grow over a wide range of pH (6-9). At neutral pH and temperaturesranging from 30-35oC, S. oneidensis MR-1 reduced 100 mu M Cr(VI) toCr(III) within 20 minutes in the exponential growth phase, and the growthrate was not affected by the addition of chromate; it reduced chromateeven faster at temperatures between 35 and 39oC. At low temperatures(<25oC), acidic (pH<6.5), or alkaline (pH>8.5) conditions, 100mu M Cr(VI) strongly inhibited growth and chromate reduction. Themini-bioreactor system enabled the rapid determination of theseparameters reproducibly and easily by performing very few experiments.Besides its use for examining parameters of interest to environmentalremediation, the device will also allow one to quickly assess parametersfor optimal production of recombinant proteins or secondarymetabolites

  18. Using Probabilistic-Risky Programming Models in Identifying Optimized Pattern of Cultivation under Risk Conditions (Case Study: Shoshtar Region

    Directory of Open Access Journals (Sweden)

    Mohammad Kavoosi Kelashemi

    2011-03-01

    Full Text Available Using Telser and Kataoka models of probabilistic-riskymathematical programming, the present research is to determine the optimized pattern of cultivating the agricultural products of Shoshtar region under risky conditions. In order to consider the risk in the mentioned models, time period of agricultural years 1996-1997 till 2004-2005 was taken into account. Results from Telser and Kataoka models showed that due to accepting the risk amounts, most of the optimized amounts suggest the tomato cultivation during the cultivation period of fall, and watermelon cultivation during the cultivation period of spring. On the basis of results, due to allocation of agricultural lands of Shoshtar to tomato and watermeloncultivation and specializing the farming activity in this province,gross profit of agricultural production system can be increasedto 6116047000 and 727782272 thousand Rials, respectively.The results of understudy models were investigated under different income scenarios and probabilistic levels of risk acceptance. Correct policy making in order to offer the suitable equipments for adjusting the effects of lack of certainty and risks due to the climatic unwanted conditions in production process of agricultural products of Shoshtar region improve the life situation of farmers of the mentioned region.

  19. Effect of Different Water Management and Nitrogen Forms on Characteristics of Iron Nutrition of Rice Cultivated Under Aerobic Condition

    Institute of Scientific and Technical Information of China (English)

    QIAN Xiao-qing; SHEN Qi-rong; WANG Juan-juan; YANG Jian-chang; ZHOU Ming-yao; BAI Yan-chao

    2003-01-01

    Although available iron is usually abundant for the growth of rice cultivated in waterlogged condition, the rice crop may suffer from its deficiency when cultivated in aerobic soil since the soil properties are totally different from waterlogged. Solubility of iron is very low in soils with high Eh and/or high Ph. A field experiment with five different depth (10, 20, 30, 40 and 50 cm) of groundwater, and a pot-experiment with five treatments of ammonium-nitrate ratio (100/0, 75/25, 50/50, 25/75 and 0/100) were conducted to study the characteristics of iron nutrition of rice in non-full irrigation condition. Moreover, the contents of iron extracted by 1 mol L-1 HCl of rice plant samples of 8 cultivars from both aerobic and waterlogged cultivation were analyzed to study the effect of water regimes on iron content of rice plants. The results were as follows: (1) The average content of available Fe (2.70 mg kg-1) of 5 layers of the soil treated with 10-cm depth of groundwater was significantly higher than that (0.83 mg kg-1) with 50-cm depth of groundwater, and the iron concentration of rice plant of the former was much higher than that of the later. (2) Iron deficiency of rice became much severe when high ratio of nitrate (more than 75 percent) in nitrogen fertilizer applied at different intervals in aerobic cultivation. (3) The iron concentrations of 3 cultivars, Wuyujing3 (99 mg kg-1), Yangdao4 (87 mg kg-1) and 9520 (95 mg kg-1), of rice plants cultivated in aerobic condition were significantly less than those(195, 197 and 175 mg kg-1) respectively in waterlogged condition at tillering stage. And even much significant differences existed in the iron concentrations of different cultivars growing in the aerobic and waterlogged condition at maturity.

  20. Quantification of gully erosion in a cultivated area in Southern Spain under high rainfall conditions

    Science.gov (United States)

    Castillo, C.; Pérez, R.; Mora, J.; Gómez, J. A.

    2012-04-01

    Gully erosion is a major process contributing to soil degradation on cultivated areas, specially when is triggered by extreme rainfall conditions (e.g. Øygarden, 2003). This study evaluates the gully erosion rates in a Mediterran agricultural area over a five years, 2010-2011, period. This period includes the maximum annual record of the last 50 years. A qualitative field survey (June 2006, to register present gullies and those refilled by tillage) and two field measurement surveys (June 2010 and 2011) were conducted in a set of cultivated catchments located in Cordoba (Spain) within the area named as "Campiña Media del Guadalquivir". The area of these landscapes ranged from 10 to 100 ha, and they were covered by field crops (mostly bean, sunflower and wheat) on vertic soils. The main gully defining properties (widths and depths) were measured after the rainy season, and before tillage operations covered them again, using global position system (GPS), pole and measurement tape. Soil erosion was estimated calculating the gully volume difference between two periods, by assuming simple geometric forms such as a triangle, triangle or trapezium for gully cross sections. Comparing 2006 and 2010 survey results, a mean erosion rate of 10 t ha-1 yr-1 was calculated, under the hyphotesis of only one removing (gully filling) operation. For the 2010-2011 period, a great enlargement of the gully network was observed, including headward migration, cross sectional widening and sidewall sloughing. In most cross sections the widths increase ratio (width in 2011 compared to that in 2010) exceeded 2, reaching in some cases to 3 o 4 times the original dimensions. Gully erosion estimates at the different catchments ranges from 37 to 250 t ha-1 yr-1, exceeding by far the tolerable soil erosion rates. The lack of vegetation, removed during years of continued tillage at valley bottoms, the elimination of natural features such as reach sinuosity by management practices, in combination

  1. Comparison of a production process in a membrane-aerated stirred tank and up to 1000-L airlift bioreactors using BHK-21 cells and chemically defined protein-free medium.

    Science.gov (United States)

    Hesse, Friedemann; Ebel, Maria; Konisch, Nadine; Sterlinski, Reinhard; Kessler, Wolfgang; Wagner, Roland

    2003-01-01

    The applicability of a protein-free medium for the production of recombinant human interleukin-2 with baby hamster kidney cells in airlift bioreactors was investigated. For this purpose, a BHK-21 cell line, adapted to grow and produce in protein-free SMIF7 medium without forming spheroids in membrane-aerated bubble-free bioreactors, was used as the producer cell line. First, cultivation of the cells was established at a 20-L scale using an internal loop airlift bioreactor system. During the culturing process the medium formulation was optimized according to the specific requirements associated with cultivation of mammalian cells under protein-free conditions in a bubble-aerated system. The effects of the addition of an antifoam agent on growth, viability, productivity, metabolic rates, and release of lactate dehydrogenase were investigated. Although it was possible to establish cultivation and production at a 20-L scale without the use of antifoaming substances, the addition of 0.002% silicon-oil-based antifoaming reagent improved the cultivation system by completely preventing foam formation. This reduced the release of lactate dehydrogenase activity to the level found in bubble-free aerated stirred tank membrane bioreactors and led to a reduction in generation doubling times by about 5 h (17%). Using the optimized medium formulation, cells were cultivated at a 1000-L scale, resulting in a culture performance comparable to the 20-L airlift bioreactor. For comparison, cultivations with protein-containing SMIF7 medium were carried out at 20- and 1000-L scales. The application of protein supplements did not lead to a significant improvement in the cultivation conditions. The results were also compared with experiments performed in a bubble-free aerated stirred tank membrane bioreactor to evaluate the influence of bubbles on the investigated culture parameters. The data implied a higher metabolic activity of the cells in airlift bioreactors with a 150% higher glucose

  2. Bioreactors addressing diabetes mellitus.

    Science.gov (United States)

    Minteer, Danielle M; Gerlach, Jorg C; Marra, Kacey G

    2014-11-01

    The concept of bioreactors in biochemical engineering is a well-established process; however, the idea of applying bioreactor technology to biomedical and tissue engineering issues is relatively novel and has been rapidly accepted as a culture model. Tissue engineers have developed and adapted various types of bioreactors in which to culture many different cell types and therapies addressing several diseases, including diabetes mellitus types 1 and 2. With a rising world of bioreactor development and an ever increasing diagnosis rate of diabetes, this review aims to highlight bioreactor history and emerging bioreactor technologies used for diabetes-related cell culture and therapies.

  3. Modelling the removal of volatile pollutants under transient conditions in a two-stage bioreactor using artificial neural networks.

    Science.gov (United States)

    López, M Estefanía; Rene, Eldon R; Boger, Zvi; Veiga, María C; Kennes, Christian

    2017-02-15

    A two-stage biological waste gas treatment system consisting of a first stage biotrickling filter (BTF) and second stage biofilter (BF) was tested for the removal of a gas-phase methanol (M), hydrogen sulphide (HS) and α-pinene (P) mixture. The bioreactors were tested with two types of shock loads, i.e., long-term (66h) low to medium concentration loads, and short-term (12h) low to high concentration loads. M and HS were removed in the BTF, reaching maximum elimination capacities (ECmax) of 684 and 33 gm(-3)h(-1), respectively. P was removed better in the second stage BF with an ECmax of 130 gm(-3)h(-1). The performance was modelled using two multi-layer perceptrons (MLPs) that employed the error backpropagation with momentum algorithm, in order to predict the removal efficiencies (RE, %) of methanol (REM), hydrogen sulphide (REHS) and α-pinene (REP), respectively. It was observed that, a MLP with the topology 3-4-2 was able to predict REM and REHS in the BTF, while a topology of 3-3-1 was able to approximate REP in the BF. The results show that artificial neural network (ANN) based models can effectively be used to model the transient-state performance of bioprocesses treating gas-phase pollutants.

  4. Increasing Vero viable cell densities for yellow fever virus production in stirred-tank bioreactors using serum-free medium.

    Science.gov (United States)

    Mattos, Diogo A; Silva, Marlon V; Gaspar, Luciane P; Castilho, Leda R

    2015-08-20

    In this work, changes in Vero cell cultivation methods have been employed in order to improve cell growth conditions to obtain higher viable cell densities and to increase viral titers. The propagation of the 17DD yellow fever virus (YFV) in Vero cells grown on Cytodex I microcarriers was evaluated in 3-L bioreactor vessels. Prior to the current changes, Vero cells were repeatedly displaying insufficient microcarrier colonization. A modified cultivation process with four changes has resulted in higher cell densities and higher virus titers than previously observed for 17DD YFV.

  5. Time efficient way to calculate oxygen transfer areas and power input in cylindrical disposable shaken bioreactors.

    Science.gov (United States)

    Klöckner, Wolf; Lattermann, Clemens; Pursche, Franz; Büchs, Jochen; Werner, Sören; Eibl, Dieter

    2014-01-01

    Disposable orbitally shaken bioreactors are a promising alternative to stirred or wave agitated systems for mammalian and plant cell cultivation, because they provide a homogeneous and well-defined liquid distribution together with a simple and cost-efficient design. Cultivation conditions in the surface-aerated bioreactors are mainly affected by the size of the volumetric oxygen transfer area (a) and the volumetric power input (P∕VL ) that both result from the liquid distribution during shaking. Since Computational Fluid Dynamics (CFD)-commonly applied to simulate the liquid distribution in such bioreactors-needs high computing power, this technique is poorly suited to investigate the influence of many different operating conditions in various scales. Thus, the aim of this paper is to introduce a new mathematical model for calculating the values of a and P∕VL for liquids with water-like viscosities. The model equations were derived from the balance of centrifugal and gravitational forces exerted during shaking. A good agreement was found among calculated values for a and P∕VL , CFD simulation values and empirical results. The newly proposed model enables a time efficient way to calculate the oxygen transfer areas and power input for various shaking frequencies, filling volumes and shaking and reactor diameters. All these parameters can be calculated fast and with little computing power.

  6. Column bioreactor use for optimization of pectinase production in solid substrate cultivation Uso de bioreator de coluna para otimização da produção de pectinases por cultivo semi-sólido

    Directory of Open Access Journals (Sweden)

    Giani Andrea Linde

    2007-09-01

    Full Text Available This study aimed to determine the influence of process variables and production, of polygalacturonase (PG and polymetylgalacturonase (PMG by solid substrate cultivation using a fixed bed column bioreactor. A fractional factorial design (FFD was used to study the effect of the following variables: microorganism (Aspergillus oryzae and Aspergillus niger, substratum (wheat bran and defatted rice bran, aeration (40 and 60 ml h-1g-1, pectin (5 and 10 g g-1 and nitrogen (urea and ammonium sulfate. Microorganism, aeration and initial pectin were identified in FFD as significant variables (pO objetivo deste trabalho foi determinar a influência de variáveis de processo na produção das enzimas poligalacturonase (PG e polimetilgalacturonase (PMG por cultivo semi-sólido utilizando bioreator de coluna. Um planejamento fatorial fracionário foi utilizado para determinar o efeito das variáveis "microrganismo" (Aspergillus oryzae e Aspergillus niger, "substrato" (farelo de trigo e farelo desengordurado de arroz, "aeração" (40 e 60 ml h-1 g-1, "pectina" (5 e 10 g g-1 e "nitrogênio" (uréia e sulfato de amônia para a produção de PG. Microrganismo, aeração e pectina foram significantes (p<0,05 para a produção de PG. Utilizando-se um planejamento composto central com quadruplicata no ponto central concluiu-se que Aspergillus niger apresenta maior produção de PG, o substrato e a fonte de nitrogênio não afetam (p<0,05 a produção de PG, a aeração afeta positivamente a produção de PG e negativamente a de PMG, e a concentração inicial de pectina afeta positivamente a atividade de ambas enzimas. Os pontos ótimos de aeração e de concentração inicial de pectina para a produção de PG são 66,13 ml h-1 g-1 e 12,8 g g-1, respectivamente, e para a produção de PMG são 40 ml h-1 g-1 e 15,0 g g-1, respectivamente.

  7. Evaluation of the oil Produced from lettuce crop cultivated under three irrigation conditions

    Directory of Open Access Journals (Sweden)

    Hassan El-Mallah, M.

    2012-10-01

    Full Text Available Three oil lettuce seed samples (lactuca Sativa LS10, LS20, LS30 were cultivated under three irrigation conditions (well, normal and water deficient conditions, after 10, 20 and 30 days respectively to evaluate their oils and to see to what extent the oil lettuce plant resists draught conditions. The oils extracted from the three seed samples were evaluated by determining eight lipid profiles using HPLC in conjunction with capillary GLC. Lettuce seed oils are characterized by high contents of linoleic and oleic acids. Of the triacyl glycerols determined, those containing linoleyl and oleyl acyles are the major ones. The whole sterol profiles include β-sitosterol (as major component followed by 7-stigmasterol, campesterol and 5-stigmasterol, which were found in all the lettuce seed oil samples but with slight differences. Furthermore, sterol patterns of the free and acylated sterols, free and acylated sterylglycosides were determined. It was found that LS30 oil has the highest tocopherol content and α-tocopherol is the only constituent in all the lettuce seed oil samples. On the other hand, the 2-position in the lettuce seed oil samples is mainly acylated by unsaturated fatty acids (98.6% namely, oleic and linoleic acids. According to these results, it can be concluded that irrigation conditions do not affect the lipid constituents of the oil and the oil lettuce plant resists draught and its lipid profiles are in agreement with those of conventional vegetable oils.

    Tres muestras de semillas de lechuga (Lactuca Sativa LS10, LS20, LS30 se cultivaron bajo tres condiciones de riego (bien regado, normal y con deficiencia de agua, después de 10, 20 y 30 días, respectivamente para evaluar sus aceites y ver hasta qué punto el aceite de la planta de lechuga resiste las condiciones de riego. Los aceites extraídos de las tres muestras de semillas se evaluó mediante la determinación de ocho perfiles de lípidos usando cromatograf

  8. Physiology of Geobacter metallireducens under excess and limitation of electron donors. Part II. Mimicking environmental conditions during cultivation in retentostats.

    Science.gov (United States)

    Marozava, Sviatlana; Röling, Wilfred F M; Seifert, Jana; Küffner, Robert; von Bergen, Martin; Meckenstock, Rainer U

    2014-06-01

    The strict anaerobe Geobacter metallireducens was cultivated in retentostats under acetate and acetate plus benzoate limitation in the presence of Fe(III) citrate in order to investigate its physiology under close to natural conditions. Growth rates below 0.003h(-1) were achieved in the course of cultivation. A nano-liquid chromatography-tandem mass spectrometry-based proteomic approach (nano-LC-MS/MS) with subsequent label-free quantification was performed on proteins extracted from cells sampled at different time points during retentostat cultivation. Proteins detected at low (0.002h(-1)) and high (0.06h(-1)) growth rates were compared between corresponding growth conditions (acetate or acetate plus benzoate). Carbon limitation significantly increased the abundances of several catabolic proteins involved in the degradation of substrates not present in the medium (ethanol, butyrate, fatty acids, and aromatic compounds). Growth rate-specific physiology was reflected in the changed abundances of energy-, chemotaxis-, oxidative stress-, and transport-related proteins. Mimicking natural conditions by extremely slow bacterial growth allowed to show how G. metallireducens optimized its physiology in order to survive in its natural habitats, since it was prepared to consume several carbon sources simultaneously and to withstand various environmental stresses.

  9. From transient response of a compact photobioreactor for microalgae cultivation

    Energy Technology Data Exchange (ETDEWEB)

    Dilay, Emerson; Ribeiro, Robert Luis Lara; Pulliam, Raevon; Mariano, Andre Bellin [Universidade Federal do Parana (UFPR), Curitiba, PR (Brazil). Nucleo de Pesquisa e Desenvolvimento em Energia Auto-Sustentavel; Ordonez, Juan Carlos [Florida State University, Tallahassee, FL (United States). Dept. of Mechanical Engineering and Center for Advanced Power Systems], E-mail: ordonez@caps.fsu.edu; Vargas, Jose Viriato Coelho [Universidade Federal do Parana (UFPR), Curitiba, PR (Brazil). Dept. de Engenharia Mecanica

    2010-07-01

    Biofuels from microalgae are currently the subject of funded scientific research in many countries due to their high productivity of oil when compared with other crops. Microalgae can also be used in many important applications such as to obtain compounds of interest for food, chemicals, and pharmaceuticals. The high productivity of microalgae when compared with other crops is achieved because agricultural land is not mandatory for their cultivation, since they can be grown in open ponds, sea or vertical photo bioreactors. In this paper, a mathematical model is introduced for assessing the transient microalgae growth as a function of variable light intensity, temperature and environmental conditions in the daily cycle. Photo bioreactor geometry is considered as well. Light intensity is obtained from sun position, photo bioreactor geometry, and the installation location in the world. The photo bioreactor was discretized in space by the the volume element method. Balances of energy and species together with thermodynamics, heat transfer and chemistry empirical and theoretical correlations are applied to each volume element. Therefore, a system of ordinary differential equations with respect to time only is capable of delivering temperatures and concentrations as functions of space and time, even with a coarse mesh. The numerical results are capable of predicting the transient and steady state photo bioreactor biomass production with low computational time. Microalgae specific growth rate as a function of average light intensity inside the tubes and time was calculated. As a result, the model is expected to be a useful tool for simulation, design, and optimization of compact photo bioreactors. (author)

  10. Hairy root culture: bioreactor design and process intensification.

    Science.gov (United States)

    Stiles, Amanda R; Liu, Chun-Zhao

    2013-01-01

    The cultivation of hairy roots for the production of secondary metabolites offers numerous advantages; hairy roots have a fast growth rate, are genetically stable, and are relatively simple to maintain in phytohormone free media. Hairy roots provide a continuous source of secondary metabolites, and are useful for the production of chemicals for pharmaceuticals, cosmetics, and food additives. In order for hairy roots to be utilized on a commercial scale, it is necessary to scale-up their production. Over the last several decades, significant research has been conducted on the cultivation of hairy roots in various types of bioreactor systems. In this review, we discuss the advantages and disadvantages of various bioreactor systems, the major factors related to large-scale bioreactor cultures, process intensification technologies and overview the mathematical models and computer-aided methods that have been utilized for bioreactor design and development.

  11. Use of Model-Based Nutrient Feeding for Improved Production of Artemisinin by Hairy Roots of Artemisia Annua in a Modified Stirred Tank Bioreactor.

    Science.gov (United States)

    Patra, Nivedita; Srivastava, Ashok K

    2015-09-01

    Artemisinin has been indicated to be a potent drug for the cure of malaria. Batch growth and artemisinin production kinetics of hairy root cultures of Artemisia annua were studied under shake flask conditions which resulted in accumulation of 12.49 g/L biomass and 0.27 mg/g artemisinin. Using the kinetic data, a mathematical model was identified to understand and optimize the system behavior. The developed model was then extrapolated to design nutrient feeding strategies during fed-batch cultivation for enhanced production of artemisinin. In one of the fed-batch cultivation, sucrose (37 g/L) feeding was done at a constant feed rate of 0.1 L/day during 10-15 days, which led to improved artemisinin accumulation of 0.77 mg/g. The second strategy of fed-batch hairy root cultivation involved maintenance of pseudo-steady state sucrose concentration (20.8 g/L) during 10-15 days which resulted in artemisinin accumulation of 0.99 mg/g. Fed-batch cultivation (with the maintenance of pseudo-steady state of substrate) of Artemisia annua hairy roots was, thereafter, implemented in bioreactor cultivation, which featured artemisinin accumulation of 1.0 mg/g artemisinin in 16 days of cultivation. This is the highest reported artemisinin yield by hairy root cultivation in a bioreactor.

  12. Modeling and CFD simulation of nutrient distribution in picoliter bioreactors for bacterial growth studies on single-cell level.

    Science.gov (United States)

    Westerwalbesloh, Christoph; Grünberger, Alexander; Stute, Birgit; Weber, Sophie; Wiechert, Wolfgang; Kohlheyer, Dietrich; von Lieres, Eric

    2015-11-01

    A microfluidic device for microbial single-cell cultivation of bacteria was modeled and simulated using COMSOL Multiphysics. The liquid velocity field and the mass transfer within the supply channels and cultivation chambers were calculated to gain insight in the distribution of supplied nutrients and metabolic products secreted by the cultivated bacteria. The goal was to identify potential substrate limitations or product accumulations within the cultivation device. The metabolic uptake and production rates, colony size, and growth medium composition were varied covering a wide range of operating conditions. Simulations with glucose as substrate did not show limitations within the typically used concentration range, but for alternative substrates limitations could not be ruled out. This lays the foundation for further studies and the optimization of existing picoliter bioreactor systems.

  13. Effect of cultivating conditions on α-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation

    Institute of Scientific and Technical Information of China (English)

    LIU Cai-qin; CHEN Qi-he; CHENG Qian-jun; WANG Jin-ling; HE Guo-qing

    2007-01-01

    The work is intended to achieve optimum culture conditions of α-galactosidase production by a mutant strain Aspergillusfoetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of α-galactosidase production in SSF were 60%initial moisture of medium, 28 ℃ incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0~6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum α-galactosidase production was 2037.51 U/g dry matter near the 144th hour of fermentation.

  14. Influence of feeding conditions on clavulanic acid production in fed-batch cultivation with medium containing glycerol.

    Science.gov (United States)

    Teodoro, Juliana C; Baptista-Neto, Alvaro; Cruz-Hernández, Isara L; Hokka, Carlos O; Badino, Alberto C

    2006-09-01

    First, the effect of different levels of nitrogen source on clavulanic acid (CA) production was evaluated in batch cultivations utilizing complex culture medium containing glycerol and three different levels of soy protein isolate (SPI). Cellular growth, evaluated in terms of the rheological parameter K, was highest with a SPI concentration of 30 g.L(-1) (4.42 g.L(-1) N total). However, the highest production of CA (380 mg.L(-1)) was obtained when an intermediate concentration of 20 g.L(-1) of SPI (2.95 g.L(-1) total N) was used. To address this, the influences of volumetric flow rate (F) and glycerol concentration in the complex feed medium (Cs(F)) in fed-batch cultivations were investigated. The best experimental condition for CA production was F=0.01 L.h(-1) and Cs(F)=120 g.L(-1), and under these conditions maximum CA production was practically twice that obtained in the batch cultivation. A single empirical equation was proposed to relate maximum CA production with F and Cs(F) in fed-batch experiments.

  15. FEATURES OF VENTILATION CONDITIONS BY MUSHROOM CULTIVATION IN MINING UNDERGROUND WORKINGS

    Directory of Open Access Journals (Sweden)

    Vladimir Rendulić

    1991-12-01

    Full Text Available The trial cultivation of mushrooms (Agaricus bisporus in one of the dead faces of the »Krš« pit of the Dalmatian bauxite mines Obrovac proved, that an optimal yield can be attained with the domestic mycelium. The decision has been brought to go on with investments into equipment for new growing-site locations in underground workings of the mine. In order to cultivate high-quality mushrooms, the ventilation of growing sites has been particularly considered. Compressive separate ventilation of growing fields has been applied using the main and the return ventilating pipeline, with the air current regulation according to the growing stage (the paper is published in Croatian.

  16. Efficient PAHs biodegradation by a bacterial consortium at flask and bioreactor scale.

    Science.gov (United States)

    Moscoso, F; Teijiz, I; Deive, F J; Sanromán, M A

    2012-09-01

    In this work, the biodegradation of three polycyclic aromatic hydrocarbons (PAHs) such as Phenanthrene (PHE), Pyrene (PYR) and Benzo[a]anthracene (BaA) has been investigated. A bacterial consortium consisting of two strains was used for the first time based on preliminary promising biodegradation data. They were tentatively identified as Staphylococcus warneri and Bacillus pumilus. Degradation values higher than 85% were obtained for each single PAH when operating at flask scale, whereas minimum levels of 90% of PAHs removal were obtained after just 3 days of cultivation at bioreactor scale. The operation in cometabolic conditions led to maximum levels about 75% and 100% at flask and bioreactor scale, respectively. All the experimental data were analyzed in the light of logistic and Luedeking and Piret type models, with the purpose to better characterize the biodegradation process by S. warneri and B. pumilus. Finally, the metabolic pathway followed to degrade each PAH was ascertained.

  17. Risk of field cultivation of pickling cucumber caused by unfavorable sunshine duration conditions

    OpenAIRE

    2011-01-01

    Solar energy, accessible to plants during sunshine from the sunrise to the sunset is of key importance in productivity of agrocenoses. The aim of the work was to determine risk of pickling cucumber cultivation caused by an unfavorable course of sunshine duration in Poland on the basis of a 40-year research period 1966-2005. The research into the subject was undertaken due to frequent occurrence of sunshine deficiency in Poland and its high temporal and spatial variability. Effect of solar con...

  18. Tissue grown in space in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens of cartilage tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. Constructs grown on Mir (A) tended to become more spherical, whereas those grown on Earth (B) maintained their initial disc shape. These findings might be related to differences in cultivation conditions, i.e., videotapes showed that constructs floated freely in microgravity but settled and collided with the rotating vessel wall at 1g (Earth's gravity). In particular, on Mir the constructs were exposed to uniform shear and mass transfer at all surfaces such that the tissue grew equally in all directions, whereas on Earth the settling of discoid constructs tended to align their flat circular areas perpendicular to the direction of motion, increasing shear and mass transfer circumferentially such that the tissue grew preferentially in the radial direction. A and B are full cross sections of constructs from Mir and Earth groups shown at 10-power. C and D are representative areas at the construct surfaces enlarged to 200-power. They are stained red with safranin-O. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Photo credit: Proceedings of the National Academy of Sciences.

  19. Development of air conditioning system and labor saving technology for efficient hydroponic cultivation; Konoritsuna suiko saibai no tame no kucho to shoryokuka gijutsu no kaihatsu

    Energy Technology Data Exchange (ETDEWEB)

    Okano, T.; Terazoe, H.; Shoji, K. [Central Research Institute of Electric Power Industry, Tokyo (Japan); Yonezawa, K.; Otani, F. [Chugoku Electric Power Co. Inc., Hiroshima (Japan); Sekiyama, T.; Kosakai, K.; Sato, H.

    1997-06-01

    Equipment which made experiments on air conditioning and hydroponic cultivation possible was set up at the technical research center of the Chugoku Electric Power Co., to study an air conditioning system using night power and energy saving technology for the cultivation. Vegetables suitable to the cultivation were selected. For air conditioning, adopted was a water heat storage air conditioning system using night power. The space between the shade curtain and the greenhouse roof was ventilated to prevent increase in cooling load caused by rise in curtain temperature. Moreover, the cultivation equipment was covered with transparent vinyl film to cool the inside of the equipment. The hydroponic cultivation equipment was trially manufactured which makes the continued production by one worker possible. The cultivation of spinach, leaf lettuce and chingensai throughout the year became possible. The yield of chingensai reached the target, but those of spinach and leaf lettuce were approximately 70% of the targets. Vegetables to be produced in the air-conditioned greenhouse by hydroponic cultivation are thought to be those that can have added values such non-pesticides and ingredients, young plants which were increased by cutting or tissue culturing, etc. 5 refs., 19 figs., 8 tabs.

  20. 盐藻的诱变育种及光合反应器培养的初步研究%PRELIMINARY STUDIES ON THE MUTAGEN BREEDING OF Dunaliella salina AND ITS CULTIVATION IN SIMPLE AXIS PHOTOBIOREACTOR

    Institute of Scientific and Technical Information of China (English)

    姜建国; 周世水; 姚汝华

    2001-01-01

    A optimum started strain collected from 3 strains of Dunaliella salina was mutagenized by UV light. A new strain with content of β  carotene 5% higher than that of original strain was obtained. The strain was cultivated with a simple phorobioreactor in the optimum condition of cell growth. The result showed that the biomass of D. salina cultivated in the bioreactor over the triangular flask had a marked increase and the growth period shortened greatly.

  1. Influence of sludge retention time at constant food to microorganisms ratio on membrane bioreactor performances under stable and unstable state conditions.

    Science.gov (United States)

    Villain, Maud; Marrot, Benoît

    2013-01-01

    Food to microorganisms ratio (F/M) and sludge retention time (SRT) are known to affect in different ways biomass growth, bioactivities and foulants characteristics. Thus the aim of this study was to dissociate the effects of SRT from those of F/M ratio on lab-scale membrane bioreactors performances during stable and unstable state. Two acclimations were stabilized at a SRT of either 20 or 50 d with a constant F/M ratio of 0.2 kg(COD)kg(MLVSS)(-1) d(-1). During stable state, a higher N-NH(4)(+) removal rate (78%) was obtained at SRT of 50 d as an easier autotroph development was observed. Soluble microbial products (SMPs) release was double at 50 d with a majority of polysaccharides (49% of total SMP). The unstable conditions consisted in F/M ratio changes and operation without air and nutrient. Autotrophs were highly affected by the tested disturbances and SMP retention on membrane surface exhibited consistent changes during the performed stresses.

  2. Effects of cultivation conditions on the diversity of microbes involved in the conversion of rice straw to fodder

    Institute of Scientific and Technical Information of China (English)

    YANG Hong-yan; GAO Li-juan; WANG Xiao-fen; WANG Wei-dong; CUI Zong-jun

    2007-01-01

    To confirm the optimum cultivation conditions for analyzing lactic acid bacterial communities and to provide the cultivation foundation for lactic acid bacterial communities that were used to convert straw into fodder, fermented rice straw was inoculated into 13 different broths. After 48 h of cultivation, pH values, volatile products, and microbial diversity were analyzed. Except for LAB broth, the pH values of the other broths could decrease to approximately 4.5. GC/MS analysis showed that lactic acid in Tomato MRS broth, MRS broth, LAB broth, and Tomato juice broth was higher than that in the other broths. DNA concentration analysis showed that the counts of microbes in Tomato MRS broth were 2.5 times higher than those in other broths and that tomato juice favored the reproduction of the microbes. Denaturing gradient gel electrophoresis (DGGE) analysis showed that the number of lactic acid bacterial species in HYA broth, Tomato juice broth, and Tomato MRS broth were higher than those in the other broths.

  3. Optimization of Cultivation Conditions for Exopolysaccharide and Mycelial Biomass by Clitocybe sp.Using Box-Behnken Design

    Institute of Scientific and Technical Information of China (English)

    WANG Yun-xiang; L(U) Feng-xia; LU Zhao-xin

    2004-01-01

    Response surface(RSM)methodology based on a three-level three-factor Box-Behnken design of experiment was used to optimize the exopolysaccharide content(EPC)and the mycelium biomass in submerged cultivation by Clitocybe sp. AS 5.112. The critical factors selected for the investigation were cultivation temperature,time and volume of medium,based on the results of previous Plackett-Burman design. By analyzing the response surface plots,the optimum ranges of cultivation temperature,time and medium volume for obtaining over 1 253.00μgmL-1 of EPC lie in 24.3-25.8℃,9.7- 10.2d and 76.0-90.0 mL,respectively. While for obtaining over 8.32 mg mL-1 of dry cell weight(DCW),the above variables would be in the range of 23.8- 24.8℃,9.6- 10.3 d and 71.0 - 98.0mL,respectively. By solving the inverse matrix from the quadratic regression equations,the optimal conditions to gain 1 265.45μgmL-1 of EPC were 25.0℃,9.9d and 83.4mL,to gain8.50mg mL-1 of DCW were 24.4℃,9.9d and 87.1mL. In order to obtain the maximum yield of EPC and DCW at the same time,the above conditions would be 24.5℃,9.9d and 84.7mL,respectively,in this situation,the maximum predicted EPC and DCW were 1 261.60μgmL-1and 8.47 mg mL-1,respectively.. The experimental data under various conditions have validated the theoretical values.

  4. Bioreactor cultivation and mechanical stimulation for regeneration of tissue-engineered cartilage%生物反应器中的力学刺激促进组织工程软骨再生

    Institute of Scientific and Technical Information of China (English)

    余晓明; 孟昊业; 孙振; 尹合勇; 袁雪凌; 郭全义; 彭江; 汪爱媛; 卢世璧

    2016-01-01

    BACKGROUND:Cartilage tissue engineering has been widely used to achieve cartilage regeneration in vitro and repair cartilage defects. Tissue-engineered cartilage mainly consists of chondrocytes, cartilage scaffold and in vitro environment. OBJECTIVE:To mimic the environment of articular cartilage development in vivo, in order to increase the bionic features of tissue-engineered cartilage scaffold and effectiveness of cartilage repair. METHODS: Knee joint chondrocytes were isolated from New Zealand white rabbits, 2 months old, and expanded in vitro. The chondrocytes at passage 2 were seeded onto a scaffold of articular cartilage extracelular matrix in the concentration of 1×106/L to prepare cel-scaffold composites. Cel-scaffold composites were cultivated in an Instron bioreactor with mechanical compression (1 Hz, 3 hours per day, 10% compression) as experimental group for 7, 14, 24, 28 days or cultured staticaly for 1 day as control group. RESULTS AND CONCLUSION:Morphological observations demonstrated that the thickness, elastic modulus and maximum load of the composite in the experimental group were significantly higher than those in the control group, which were positively related to time (P < 0.05). Histological staining showed the proliferation of chondrocytes, formation of cartilage lacuna and synthesis of proteoglycan in the experimental group through hematoxylin-eosin staining and safranin-O staining, which were increased gradualy with mechanical stimulation time. These results were consistent with the findings of proteoglycan kit. Real-time quantitative PCR revealed that mRNA expressions of colagen type I and colagen type II were significantly higher in the experimental group than the control group (P < 0.05). The experimental group showed the highest mRNA expression of colagen type I and colagen type II at 21 and 28 days of mechanical stimulation, respectively (P < 0.05). With the mechanical stimulation of bioreactor, the cel-scaffold composite can produce

  5. Genetic stability of murine pluripotent and somatic hybrid cells may be affected by conditions of their cultivation.

    Science.gov (United States)

    Ivanovna, Shramova Elena; Alekseevich, Larionov Oleg; Mikhailovich, Khodarovich Yurii; Vladimirovna, Zatsepina Olga

    2011-01-01

    Using mouse pluripotent teratocarcinoma PCC4azal cells and proliferating spleen lymphocytes we obtained a new type of hybrids, in which marker lymphocyte genes were suppressed, but expression the Oct-4 gene was not effected; the hybrid cells were able to differentiate to cardiomyocytes. In order to specify the environmental factors which may affect the genetic stability and other hybrid properties, we analyzed the total chromosome number and differentiation potencies of hybrids respectively to conditions of their cultivation. Particular attention was paid to the number and transcription activity of chromosomal nucleolus organizing regions (NORs), which harbor the most actively transcribed - ribosomal - genes. The results showed that the hybrids obtained are characterized by a relatively stable chromosome number which diminished less than in 5% during 27 passages. However, a long-term cultivation of hybrid cells in non-selective conditions resulted in preferential elimination of some NO- chromosomes, whereas the number of active NORs per cell was increased due to activation of latent NORs. On the contrary, in selective conditions, i.e. in the presence of hypoxantine, aminopterin and thymidine, the total number of NOR-bearing chromosomes was not changed, but a partial inactivation of remaining NORs was observed. The higher number of active NORs directly correlated with the capability of hybrid cells for differentiation to cardiomyocytes.

  6. Effect of storage conditions on the quality of cultivated mushrooms (Agaricus bisporus (Lange Sing.

    Directory of Open Access Journals (Sweden)

    Janusz Czapski

    2013-12-01

    Full Text Available A number of quality factors were studied during storage of cultivated mushrooms (Agaricus bisporus at 2°C in controlled atmospheres. A concentration of 15% CO2 and 1.5-2% O2 and an atmosphere with a continuous flow of nitrogen retarded cap expansion and stipe elongation, while 10% CO2 retarded only cap expansion. Controlled atmospheres suppressed the growth of some microorganisms. The toughness of mushrooms stored in a normal atmosphere at 2°C markedly decreased during storage, while 10% CO2 and nitrogen atmosphere did not influence toughness as compared to initial mushrooms. The acceptability value of mushrooms in controlled atmospheres was lower during 13 days of storage as compared to normal atmosphere. Normal atmosphere appeared to keep whiteness of mushrooms longer than did other treatments.

  7. {sup 137}Cs and {sup 9}Sr uptake by sunflower cultivated under hydroponic conditions

    Energy Technology Data Exchange (ETDEWEB)

    Soudek, Petr [Department of Plant Tissue Cultures, Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nam. 2, CZ-166 10 Prague 6 (Czech Republic); Valenova, Sarka [Department of Plant Tissue Cultures, Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nam. 2, CZ-166 10 Prague 6 (Czech Republic); Vavrikova, Zuzana [Department of Plant Tissue Cultures, Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nam. 2, CZ-166 10 Prague 6 (Czech Republic); Vanek, Tomas [Department of Plant Tissue Cultures, Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nam. 2, CZ-166 10 Prague 6 (Czech Republic)]. E-mail: vanek@uochb.cas.cz

    2006-07-01

    The {sup 9}Sr and {sup 137}Cs uptake by the plant Helianthus annuus L. was studied during cultivation in a hydroponic medium. The accumulation of radioactivity in plants was measured after 2, 4, 8, 16 and 32 days of cultivation. About 12% of {sup 137}Cs and 20% of {sup 9}Sr accumulated during the experiments. We did not find any differences between the uptake of radioactive and stable caesium and strontium isotopes. Radioactivity distribution within the plant was determined by autoradiography. {sup 137}Cs was present mainly in nodal segments, leaf veins and young leaves. High activity of {sup 9}Sr was localized in leaf veins, stem, central root and stomata. The influence of stable elements or analogues on the transfer behaviour was investigated. The percentage of non-active caesium and strontium concentration in plants decreased with the increasing initial concentration of Cs or Sr in the medium. The percentage of {sup 9}Sr activity in plants decreased with increasing initial activity of the nuclide in the medium, but the activity of {sup 137}Cs in plants increased. The influence of K{sup +} and NH{sub 4} {sup +} on the uptake of {sup 137}Cs and the influence of Ca{sup 2+} on the uptake of {sup 9}Sr was tested. The highest accumulation of {sup 137}Cs (24-27% of the initial activity of {sup 137}Cs) was found in the presence of 10 mM potassium and 12 mM ammonium ions. Accumulation of about 22% of initial activity of {sup 9}Sr was determined in plants grown on the medium with 8 mM calcium ions.

  8. An innovative membrane bioreactor for methane biohydroxylation.

    Science.gov (United States)

    Pen, N; Soussan, L; Belleville, M-P; Sanchez, J; Charmette, C; Paolucci-Jeanjean, D

    2014-12-01

    In this study, a membrane bioreactor (MBR) was developed for efficient, safe microbial methane hydroxylation with Methylosinus trichosporium OB3b. This innovative MBR, which couples a bioreactor with two gas/liquid macroporous membrane contactors supplying the two gaseous substrates (methane and oxygen) was operated in fed-batch mode. The feasibility and the reproducibility of this new biohydroxylation process were first demonstrated. The mass transfer within this MBR was twice that observed in a batch reactor in similar conditions. The productivity reached with this MBR was 75±25mgmethanol(gdrycell)(-1)h(-1). Compared to the literature, this value is 35times higher than that obtained with the only other fed-batch membrane bioreactor reported, which was run with dense membranes, and is comparable to those obtained with bioreactors fed by bubble-spargers. However, in the latter case, an explosive gas mixture can be formed, a problem that is avoided with the MBR.

  9. Performance of a partially packed charcoal pellet bioreactor for acetic acid fermentation.

    Science.gov (United States)

    Horiuchi, J; Ando, K; Watanabe, S; Tada, K; Kobayashi, M; Kanno, T

    2001-01-01

    The performance of a partially packed charcoal pellet bioreactor was compared to that of a fully packed bioreactor for aerobic acetic acid production. In the fully packed charcoal pellet bioreactor, it was considered that the shortening of an actual retention time of the culture broth limited the bioreactor performance under high dilution rate and high aeration conditions. By reducing the filling ratio of charcoal pellets to 44%, which increased the actual retention time of the culture broth, the maximum productivity increased from 3.9 g/l/h in the fully packed bed bioreactor to 5.7 g/l/h in the partially packed bioreactor without affecting the operational stability.

  10. Effects of cultivation conditions and media composition on cell growth and lipid productivity of indigenous microalga Chlorella vulgaris ESP-31.

    Science.gov (United States)

    Yeh, Kuei-Ling; Chang, Jo-Shu

    2012-02-01

    The growth and lipid productivity of an isolated microalga Chlorella vulgaris ESP-31 were investigated under different media and cultivation conditions, including phototrophic growth (NaHCO(3) or CO(2), with light), heterotrophic growth (glucose, without light), photoheterotrophic growth (glucose, with light) and mixotrophic growth (glucose and CO(2), with light). C. vulgaris ESP-31 preferred to grow under phototrophic (CO(2)), photoheterotrophic and mixotrophic conditions on nitrogen-rich medium (i.e., Basal medium and Modified Bristol's medium), reaching a biomass concentration of 2-5 g/l. The growth on nitrogen-limiting MBL medium resulted in higher lipid accumulation (20-53%) but slower growth rate. Higher lipid content (40-53%) and higher lipid productivity (67-144 mg/l/d) were obtained under mixotrophic cultivation with all the culture media used. The fatty acid composition of the microalgal lipid comprises over 60-68% of saturated fatty acids (i.e., palmitic acid (C16:0), stearic acid (C18:0)) and monounsaturated acids (i.e., oleic acid (C18:1)). This lipid composition is suitable for biodiesel production.

  11. Mechanobiologic Research in a Microgravity Environment Bioreactor

    Science.gov (United States)

    Guidi, A.; Dubini, G.; Tominetti, F.; Raimondi, M.

    A current problem in tissue culturing technology is the unavailability of an effective Bioreactor for the in vitro cultivation of cells and explants. It has, in fact, proved extremely difficult to promote the high-density three-dimensional in vitro growth of human tissues that have been removed from the body and deprived of their normal in vivo vascular sources of nutrients and gas exchange. A variety of tissue explants can be maintained for a short period of time on a supportive collagen matrix surrounded by culture medium. But this system provides only limited mass transfer of nutrients and wastes through the tissue, and gravity-induced sedimentation prevents complete three- dimensional cell-cell and cell-matrix interactions. Several devices presently on the market have been used with only limited success since each has limitations, which restrict usefulness and versatility. Further, no Bioreactor or culture vessel is known that will allow for unimpeded growth of three dimensional cellular aggregates or tissue. Extensive research on the effect of mechanical stimuli on cell metabolism suggests that tissues may respond to mechanical stimulation via loading-induced flow of the interstitial fluids. During the culture, cells are subject to a flow of culture medium. Flow properties such as flow field, flow regime (e.g. turbulent or laminar), flow pattern (e.g. circular), entity and distribution of the shear stress acting on the cells greatly influence fundamental aspects of cell function, such as regulation and gene expression. This has been demonstrated for endothelial cells and significant research efforts are underway to elucidate these mechanisms in various other biological systems. Local fluid dynamics is also responsible of the mass transfer of nutrients and catabolites as well as oxygenation through the tissue. Most of the attempts to culture tissue-engineered constructs in vitro have utilized either stationary cultures or systems generating relatively small

  12. Spiral vane bioreactor

    Science.gov (United States)

    Morrison, Dennis R. (Inventor)

    1991-01-01

    A spiral vane bioreactor of a perfusion type is described in which a vertical chamber, intended for use in a microgravity condition, has a central rotating filter assembly and has flexible membranes disposed to rotate annularly about the filter assembly. The flexible members have end portions disposed angularly with respect to one another. A fluid replenishment medium is input from a closed loop liquid system to a completely liquid filled chamber containing microcarrier beads, cells and a fluid medium. Output of spent medium is to the closed loop. In the closed loop, the output and input parameters are sensed by sensors. A manifold permits recharging of the nutrients and pH adjustment. Oxygen is supplied and carbon dioxide and bubbles are removed and the system is monitored and controlled by a microprocessor.

  13. 膜生物反应器脱氮除碳环境的研究%Study on Conditions for Denitrification-decarbonization Using Membrane Bioreactor

    Institute of Scientific and Technical Information of China (English)

    宁桂兴; 黄周满; 吴迪; 王凯; 刘宇; 魏婧娟

    2011-01-01

    利用膜生物反应器研究垃圾填埋场垃圾渗滤运行环境,在常温环境下,运行结果表明:膜能够截留大量并使世代时间长的硝化菌在最短的时间富集成为优势菌种,对垃圾渗滤液中氨氮具有高效的去除效率;氨氮负荷0.082~0.109gN/g MLSS·d,CODCr负荷0.136~0.192g CODCr/g MLSS·d,DO 2.0~3.5mg/L,脱除氨氮的效果较好,去除率在95%~98%,CODCr去除率60%~70%.%An experiment was conducted to study optimization of operation conditions for membrane bioreactor at ambient temperatureto treat landfill leachate. The result showed that the membrane intercepted and enriched a lot of nitrobacteria with long generation time to make them as dominate bacteria in short time. So it was able to remove NH3-N from leachate with high rate. The process was operated with parameters as NH3-N load rate of 0. 082 ~ 0. 109 g/g MLSS · d; CODcr load rate of 0.136 ~ 0. 192 g/g MLSS · d and DO of 2.0 ~3.5mg/L The removal rates of NH3-N and CODCr were 95% ~98% and 60% ~70%,respectively, being good effects of denitrification.

  14. Glyco-engineering for biopharmaceutical production in moss bioreactors

    Directory of Open Access Journals (Sweden)

    Eva L. Decker

    2014-07-01

    Full Text Available The production of recombinant biopharmaceuticals (pharmaceutical proteins is a strongly growing area in the pharmaceutical industry. While most products to date are produced in mammalian cell cultures, namely CHO cells, plant-based production systems gained increasing acceptance over the last years. Different plant systems have been established which are suitable for standardization and precise control of cultivation conditions, thus meeting the criteria for pharmaceutical production.The majority of biopharmaceuticals comprise glycoproteins. Therefore, differences in protein glycosylation between humans and plants have to be taken into account and plant-specific glycosylation has to be eliminated to avoid adverse effects on quality, safety and efficacy of the products.The basal land plant Physcomitrella patens (moss has been employed for the recombinant production of high-value therapeutic target proteins (e.g., Vascular Endothelial Growth Factor, Complement Factor H, monoclonal antibodies, Erythropoietin. Being genetically excellently characterized and exceptionally amenable for precise gene targeting via homologous recombination, essential steps for the optimization of moss as a bioreactor for the production of recombinant proteins have been undertaken.Here, we discuss the glyco-engineering approaches to avoid non-human N- and O-glycosylation on target proteins produced in moss bioreactors.

  15. Cultivation technique for marine seaweeds allowing controlled and optimized conditions in the laboratory and on a pilotscale

    Energy Technology Data Exchange (ETDEWEB)

    Lignell, A.; Ekman, P.; Pedersen, M.

    1987-09-01

    A method for cultivation of Gracilaria secundata Harv. in plexiglass cylinders and in tanks with submerged light sources is described. Growth rates up to 47% d/sup -1/ were recorded in the system, compared to growth rates of 6% d/sup -1/ in a spray system and less than 3% d/sup -1/ in tank cultivation of G. secundata. The advantage of the system described is the greater availability of light to the seaweeds. Growth of G. secundata was not light saturated up to the highest photon irradiance used (1450 ..mu..Em/sup -2/ s/sup -1/). Self-shading followed a hyperbolic function with increasing biomass and had a great impact on the growth rates of the seaweed. At a distance of 15 cm from the light source 90% of the irradiance was absorbed at a plant density of 3 kg fresh-weight per m/sup -3/. The water in the tanks was agitated by injection of compressed air at the bottom. The constant rubbing of the seaweeds against each other and on the walls, together with recirculation of the water through filters, kept the system free from epiphytes and debris. In this way the cultures were stable, thus allowing growth under optimal conditions without any infection by bacteria or epiphytes. High growth rates could be achieved for long periods of time. An annual yield of 80 kg dry-weight per m/sup -3/ of G. secundata was estimated after pilot-scale studies. Ectocarpus siliculosus (Dillw.) Lyngb. and Ceramium rubrum (Huds.) C. Ag. also showed high potentials for growth in the system. Enteromorpha intestinalis (L.) Link and other species that easily form dense mats are not suitable for the cultivation technique described.

  16. Soil Nutrient Condition of Coffee Cultivation with Industrial Woody-crops

    Directory of Open Access Journals (Sweden)

    Rudy Erwiyo

    2008-05-01

    Full Text Available Black pod rot disease (BPRD which is caused by Phytophthora palmivora is one of the main diseases of cocoa cultivations particularly in plantations with wet climate. Black pod rot can develop rapidly under high humidity environments, particularly during rainy seasons. This disease can cause loss of harvest of up to 46.63% in East Java. The various control efforts attempted so far have not resulted in significant improvements. Urea, in addition to functioning as fertilizer, can also produce the ammonia gas which is believed to be able to suppress black pod rot. This research aims to determine the effectiveness of black pod rot control using the combination of lime and urea. This research was conducted from June to September 2013. The materials used in test included sterile soil, black pod rot infected cocoa, urea, and agricultural lime. Observation results showed that ammonia could form from urea. Lime can increase the speed of the formation. The ammonia gas forming from 0.06% urea and 0.3% lime can control the P. palmivora fungus inside the soil. Key words: Pod rot, P. palmivora, urea, lime, ammonia

  17. Reasons of leaves withering in tropical plants cultivated under greenhouse conditions

    Directory of Open Access Journals (Sweden)

    Z. Czerwiński

    2013-12-01

    Full Text Available In order to determine the reasons of necrosis of exotic plants leaves cultivated in greenhouses plants belonging to ten following species were examined: Ceratozamia mexicana Brongn., Stangeria eriopus (Kunze Nash (Cycadaceae, Eriobotrya japonica Lindl. (Rosaceae, Camellia japonica L. (Theaceae, Phoenix roebeleni O'Brien (Palmae, Sequoia sempervirens Endl. (Taxodiaceae, Calathea bachemiana Morr. (Marantaceae, Cordyline terminalis Kunth (Agavaceae, Spathiphyllum wallisii Reg. and Anthurium magnificum Lind. (Araceae. Chemical analysis were performed in soil samples in which these plants grow, in samples of tap-water applied for watering and in samples of decaying and healthy leaves. In order to examine the process of withdrawal of mineral components from necrotic leaves, both: necrotic and green parts of decaying leaves were subjected to examination. On the basis of the research it was concluded, that - in spite of generally low level of salinity of the water used for watering - some ions content, particularity that of CI-, was unfavourable to plants. Unfavourable ionic composition was discovered in water extracts derived from some of the breeding-ground soils. A comparison of healthy and decaying, necrotic leaves chemism proves that CI- assimilated by the plants from the breeding-grounds and accumulated in leaves, affects them toxically.

  18. Bioreactor Mass Transport Studies

    Science.gov (United States)

    Kleis, Stanley J.; Begley, Cynthia M.

    1997-01-01

    The objectives of the proposed research efforts were to develop both a simulation tool and a series of experiments to provide a quantitative assessment of mass transport in the NASA rotating wall perfused vessel (RWPV) bioreactor to be flown on EDU#2. This effort consisted of a literature review of bioreactor mass transport studies, the extension of an existing scalar transport computer simulation to include production and utilization of the scalar, and the evaluation of experimental techniques for determining mass transport in these vessels. Since mass transport at the cell surface is determined primarily by the relative motion of the cell assemblage and the surrounding fluid, a detailed assessment of the relative motion was conducted. Results of the simulations of the motion of spheres in the RWPV under microgravity conditions are compared with flight data from EDU#1 flown on STS-70. The mass transport across the cell membrane depends upon the environment, the cell type, and the biological state of the cell. Results from a literature review of cell requirements of several scalars are presented. As a first approximation, a model with a uniform spatial distribution of utilization or production was developed and results from these simulations are presented. There were two candidate processes considered for the experimental mass transport evaluations. The first was to measure the dissolution rate of solid or gel beads. The second was to measure the induced fluorescence of beads as a stimulant (for example hydrogen peroxide) is infused into the vessel. Either technique would use video taped images of the process for recording the quantitative results. Results of preliminary tests of these techniques are discussed.

  19. Optimization of Cultivation Conditions for Inoculums of Cordyceps sinensis%冬虫夏草液体种子培养条件的优化

    Institute of Scientific and Technical Information of China (English)

    殷海松; 秦韶燕; 乔长晟; 贾士儒

    2013-01-01

    At the submerged cultivation conditions in the shake flask , to optimize the cultivation conditions for Inoculums of Cordyceps sinensis,single factor and orthogonal array design were employed for investigating the effects of different cultivation conditions on cell growth of Cordyceps sinensis. The result indicated that the optimum cultivation conditions for Cordyceps sinensis were as follows:cultivation temperature 24 ℃,rotation speed 150 r/min, the medium volume in the flask 30%(75 mL/250 mL). At the cultivation conditions, we get 10.45 g DW/L at the submerged cultivation in the shake flask.%  在摇瓶液体培养条件下,优化冬虫夏草液体种子培养条件,采用单因素和正交试验法,研究了不同培养条件对冬虫夏草液体细胞生长的影响。结果表明,在所考察的因素中,冬虫夏草液体种子最优培养条件为:培养温度24℃,转速150 r/min,装液量30%(75 mL/250 mL)。在此条件下,经摇瓶培养,获得最大菌体干重为10.45 g/L。

  20. Oxygen Sensors Monitor Bioreactors and Ensure Health and Safety

    Science.gov (United States)

    2014-01-01

    In order to cultivate healthy bacteria in bioreactors, Kennedy Space Center awarded SBIR funding to Needham Heights, Massachusetts-based Polestar Technologies Inc. to develop sensors that could monitor oxygen levels. The result is a sensor now widely used by pharmaceutical companies and medical research universities. Other sensors have also been developed, and in 2013 alone the company increased its workforce by 50 percent.

  1. BIOFERTILISERS - EFFICIENCY ON THE VEGETATIVE GROWTH AND PRODUCTIVITY OF TOMATOES CULTIVATED UNDER THE CONDITIONS OF ORGANIC FARMING

    Directory of Open Access Journals (Sweden)

    Veselka Vlahova

    2014-12-01

    Full Text Available The objective of this study was to examine the influence of biofertilisers on the vegetative growth, yield and productivity of tomatoes cultivated under the conditions of organic farming. The research was carried out in the polyethylene greenhouse and experimental field of the Agroecological centre at the Agricultural University- Plovdiv (Bulgaria, in the period from 2013 until 2014 with on determinant tomatoes (solanum lycopersicum l., of the variety of “Trapezitsa”. The standard yield had its highest value upon the application of biofertiliser Emosan on the basic fertilization Boneprot- 3140 kg/da (2013 and 3116 kg/da (2014, thus determining the positive impact of the tested combination of biofertilisers. The combined application of biofertilisers in the form of basic fertilisation and additional vegetation feeding had a positive impact on the formation of the standard yield, the number and the mass of fruits.

  2. Selection of optimum conditions of medium acidity and aeration for submerget cultivation of Bacillus thuringiensis and Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    O. A. Dregval

    2010-06-01

    Full Text Available The paper deals with the influence of medium pH and aeration rate on growth and sporulation of Bacillus thuringiensis and Вeauveria bassiana, which are main constituents of the complex microbial insecticide. It was established optimal medium pH for B. thuringiensis – 6.0 and for В. bassiana – 6.0–7.0. The maximum productivity of the studied microorganisms was observed in the same range of aeration – 7– 14 mmol O2/l/h. The selected conditions of cultivation are necessary for the production of complex biological insecticide based on the association of B. thuringiensis and B. bassiana.

  3. Bioreactor rotating wall vessel

    Science.gov (United States)

    2001-01-01

    The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells.

  4. A novel process for production of hepatitis A virus in Vero cells grown on microcarriers in bioreactor

    Institute of Scientific and Technical Information of China (English)

    Ming-Bo Sun; Yan-Jun Jiang; Wei-Dong Li; Ping-Zhong Li; Guo-Liang Li; Shu-De Jiang; Guo-Yang Liao

    2004-01-01

    AIM: To develop a novel process for production of HAV in Vero cells grown on microcarriers in a bioreactor.METHODS: Vero cells infected with HAV strain W were seeded at an initial density of 1×105 cells/mL into a 7-L bioreactor containing Cytodex-I microcarriers. During the stage of cell proliferation, the following conditions were applied: pH 7.2±0.2, temperature 37±0.2 ℃, dissolved oxygen 40% of air saturation and agitation rate 40 r/min.After the stage of virus culture started, the culture conditions were altered to pH 7.2±0.2, temperature 35±0.2 ℃,dissolved oxygen 25% of air saturation, agitation rate 50 r/min and perfusion of fresh medium at a flux of 20 mL/h. During the course of fermentation, cell density, HAV antigen titre,glucose, lactate and ammonia levels were monitored. A control experiment using conventional static culture was conducted in the T150 flask.RESULTS: After a 28-d cultivation, cell density increased to 14.0×105 cells/mL in the bioreactor, 5.6×109viable cells and 4 000 mL virus suspension with a titre of 1:64 were harvested.The viral antigen output per cell unit in the bioreactor was 3-fold higher than that in the T150 flask. Meanwhile the metabolic mode of Vero cells did not change after the infection with HAV strain W.CONCLUSION: The process for production of HAV in Veto cells grown on microcarriers in a bioreactor is a novel,efficient and practical way to obtain virus antigen for vaccine purpose. This approach produces more cells and HAV antigen than the conventional static culture. With futher improvement, it is possible to be used for the production of hepatitis A vaccine.

  5. Enhanced production of elastase by Bacillus licheniformis ZJUEL31410: optimization of cultivation conditions using response surface methodology

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Sequential methodology based on the application of three types of experimental designs was used to optimize the fermentation conditions for elastase production from mutant strain ZJUEL31410 of Bacillus licheniformis in shaking flask cultures. The optimal cultivation conditions stimulating the maximal elastase production consist of 220 r/min shaking speed, 25 h fermentation time, 5% (v/v) inoculums volume, 25 ml medium volume in 250 ml Erlenmeyer flask and 18 h seed age. Under the optimized conditions, the predicted maximal elastase activity was 495 U/ml. The application of response surface methodology resulted in a significant enhancement in elastase production. The effects of other factors such as elastin and the growth factor (corn steep flour) on elastase production and cell growth were also investigated in the current study. The elastin had no significant effect on enzyme-improved production. It is still not clear whether the elastin plays a role as a nitrogen source or not. Corn steep flour was verified to be the best and required factor for elastase production and cell growth by Bacillus licheniformis ZJUEL31410.

  6. Cultivating conditions effects on kefiran production by the mixed culture of lactic acid bacteria imbedded within kefir grains.

    Science.gov (United States)

    Zajšek, Katja; Goršek, Andreja; Kolar, Mitja

    2013-08-15

    The influence of fermentation temperature, agitation rate, and additions of carbon sources, nitrogen sources, vitamins and minerals on production of kefiran by kefir grains lactic acid bacteria was studied in a series of experiments. The main aim of the work was to increase the exopolysaccharide (EPS) production where customised milk was used as fermentation medium. It was proved that the controlling of culturing conditions and the modifying of fermentation medium conditions (i.e., carbon, nitrogen, mineral sources and vitamins) can dramatically enhance the production of the EPS. The temperature and agitation rate were critical for kefiran production during the 24 h cultivation of grains; our optimised conditions being 25°C and 80 rpm, respectively. In addition, when optimising the effects of additional nutrition, it was found that 5% (w/v) lactose, 0.1% (w/v) thiamine, and 0.1% (w/v) FeCl3 led to the maximal production of EPS. The results indicate that nutrients can be utilised to improve the production of EPS and that good kefir grains growth does not appear to be a determining factor for a high production yield of EPS.

  7. Novel micro-bioreactor high throughput technology for cell culture process development: Reproducibility and scalability assessment of fed-batch CHO cultures.

    Science.gov (United States)

    Amanullah, Ashraf; Otero, Jose Manuel; Mikola, Mark; Hsu, Amy; Zhang, Jinyou; Aunins, John; Schreyer, H Brett; Hope, James A; Russo, A Peter

    2010-05-01

    With increasing timeline pressures to get therapeutic and vaccine candidates into the clinic, resource intensive approaches such as the use of shake flasks and bench-top bioreactors may limit the design space for experimentation to yield highly productive processes. The need to conduct large numbers of experiments has resulted in the use of miniaturized high-throughput (HT) technology for process development. One such high-throughput system is the SimCell platform, a robotically driven, cell culture bioreactor system developed by BioProcessors Corp. This study describes the use of the SimCell micro-bioreactor technology for fed-batch cultivation of a GS-CHO transfectant expressing a model IgG4 monoclonal antibody. Cultivations were conducted in gas-permeable chambers based on a micro-fluidic design, with six micro-bioreactors (MBs) per micro-bioreactor array (MBA). Online, non-invasive measurement of total cell density, pH and dissolved oxygen (DO) was performed. One hundred fourteen parallel MBs (19 MBAs) were employed to examine process reproducibility and scalability at shake flask, 3- and 100-L bioreactor scales. The results of the study demonstrate that the SimCell platform operated under fed-batch conditions could support viable cell concentrations up to least 12 x 10(6) cells/mL. In addition, both intra-MB (MB to MB) as well as intra-MBA (MBA to MBA) culture performance was found to be highly reproducible. The intra-MB and -MBA variability was calculated for each measurement as the coefficient of variation defined as CV (%) = (standard deviation/mean) x 100. The % CV values for most intra-MB and intra-MBA measurements were generally under 10% and the intra-MBA values were slightly lower than those for intra-MB. Cell growth, process parameters, metabolic and protein titer profiles were also compared to those from shake flask, bench-top, and pilot scale bioreactor cultivations and found to be within +/-20% of the historical averages.

  8. Tapered bed bioreactor

    Science.gov (United States)

    Scott, Charles D.; Hancher, Charles W.

    1977-01-01

    A vertically oriented conically shaped column is used as a fluidized bed bioreactor wherein biologically catalyzed reactions are conducted in a continuous manner. The column utilizes a packing material a support having attached thereto a biologically active catalytic material.

  9. Cultivation of juvenile fat snook (Centropomus parallelus Poey, 1860 fed probiotic in laboratory conditions

    Directory of Open Access Journals (Sweden)

    Moysés Cavichioli Barbosa

    2011-08-01

    Full Text Available The objective of this study was to investigate the growth of juvenile fat snook (Centropomus parallelus in laboratory conditions when fed a diet supplemented with the probiotic Lactobacillus plantarum. Changes in the intestinal flora, haematological parameters and growth performance were assessed using 180 fishes (54.2 ± 13.4 g each. The fishes were subjected to two treatments divided into six cages: 1 feed supplemented with probiotic, and 2 feed without probiotic (control. The temperature, dissolved oxygen and salinity were maintained at 25 ± 1 ° C, 4.0 mg.L-1 and 33 ‰, respectively. After 10 weeks of culture, the fishes fed probiotic had reduced viable culturable heterotrophic bacteria and Vibrio spp. and increased lactic acid bacteria in the intestinal tract, as well as a higher number of thrombocytes, leukocytes and lymphocytes in the blood. No significant difference was observed in the growth, survival or body composition, but the hepatosomatic index was significantly higher in the fishes fed with probiotic and control.

  10. Cultivation conditions and selenium fertilization alter the phenolic profile, glucosinolate, and sulforaphane content of broccoli.

    Science.gov (United States)

    Robbins, Rebecca J; Keck, Anna-Sigrid; Banuelos, Gary; Finley, John W

    2005-01-01

    Broccoli is a food often consumed for its potential health-promoting properties. The health benefits of broccoli are partly associated with secondary plant compounds that have bioactivity; glucosinolates and phenolic acids are two of the most abundant and important in broccoli. In an effort to determine how variety, stress, and production conditions affect the production of these bioactive components broccoli was grown in the greenhouse with and without selenium (Se) fertilization, and in the field under conventional or organic farming procedures and with or without water stress. High-performance liquid chromatography/mass spectrometry was used to separate and identify 12 primary phenolic compounds. Variety had a major effect: There was a preponderance of flavonoids in the Majestic variety, but hydroxycinnamic esters were relatively more abundant in the Legacy variety. Organic farming and water stress decreased the overall production of phenolics. Se fertilization increased glucosinolates in general, and sulforaphane in particular, up to a point; above that Se fertilization decreased glucosinolate production. Organic farming and water stress also decreased glucosinolate production. These data show environmental and genetic variation in phenolics and glucosinolates in broccoli, and warn that not all broccoli may contain all health-promoting bioactive components. They further show that selection for one bioactive component (Se) may decrease the content of other bioactive components such as phenolics and glucosinolates.

  11. Effect of cultivation conditions on ligninolytic enzyme production by Ganoderma carnosum

    Directory of Open Access Journals (Sweden)

    Simonić Jasmina Lj.

    2009-01-01

    Full Text Available Ganoderma carnosum has been found in Europe only, at coniferous trees and it is difficult to distinguish it morphologically from G. lucidum. Since G. carnosum has not been studied biochemically yet, the aim of this study was to analyse its ability to produce Mn-dependent peroxidase (MnP, versatile peroxidase (VP and laccase (Lac under conditions of solid-state fermentation of selected plant raw materials as carbon sources, in the presence of different nitrogen concentrations in the medium. Wheat straw, corn stem, oak and grapevine sawdust were the analysed plant raw materials. Nitrogen source in synthetic medium was NH4NO3 and its concentrations were: 10mM N and 20 mM N. Enzyme activity was determined spectrophotometrically, using ABTS and phenol red, as the substrates for Lac and Mn-oxidizing peroxidases, respectively. Maximum level of MnP activity (56.82 U/l was obtained in the medium with wheat straw and nitrogen concentration of 10 mM. Best carbon source for VP production was grapevine sawdust at nitrogen concentration of 10 mM (80.80 U/l. The obtained Lac activity was very low in the medium with wheat straw (1.80 U/l, while it was not detected in the presence of other three analyzed plant raw materials. Maximum of total protein content (0.06 mgml-1 was noted in the medium where oak sawdust was carbon source and nitrogen concentration was 20 mM.

  12. Bioprocess development for the cultivation of human T-lymphocytes in a clinical scale.

    Science.gov (United States)

    Bohnenkamp, H; Hilbert, U; Noll, T

    2002-01-01

    Adoptive transfer of large numbers of donor-derived T-lymphocytesmay offer a promising treatment of a variety of viral and malignant diseases. The key step in this approach is the ex vivo generation of sufficient quantities of these cells in a short time.We have investigated the influence of several important cultivation parameters on the proliferation of human T-lymphocytes to develop a large-scale fermentation process usingdifferent types of stirred bioreactors. Such systems offer manypotential advantages over the static culture systems commonlyused today.Peripheral blood mononuclear cells of healthy but CMV positive donors were stimulated with monoclonal antibodies (anti-CD3 and anti-CD28) and Interleukin-2. The influence of osmolality, Interleukin-2 concentration, pH, oxygen tension, feeding strategyand temperature on T-cell proliferation was investigated and theoptimised conditions were transferred to a novel stirred suspension bioreactor with an especially designed magnetic stirrbar to minimize the shear force (working volume 550 ml) and a standard stirred vessel (working volume 1000 ml).Preferable conditions for the cultivation of primary T-lymphocytes were an osmolality of 276-330 mOsmol kg(-1),an Interleukin-2 concentration of 100 U ml(-1), a pH rangeof 7.0 to 7.3, an oxygen tension of 5-50% and a temperature of 38.5 degrees C. After 238 h of cultivation 2.8 x 10(9) cells in the stirred vesseland 1.5 x 10(9) cells in the suspension bioreactor were obtained with a percentage of T-cells >94%. The specificity of the cells wasmaintained during cultivation as proven by IFN-gamma secretionafter exposure to a hCMV protein.

  13. Open source software to control Bioflo bioreactors.

    Science.gov (United States)

    Burdge, David A; Libourel, Igor G L

    2014-01-01

    Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW.

  14. Open source software to control Bioflo bioreactors.

    Directory of Open Access Journals (Sweden)

    David A Burdge

    Full Text Available Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW.

  15. Lactobacillus plantarum BL011 cultivation in industrial isolated soybean protein acid residue

    Directory of Open Access Journals (Sweden)

    Chaline Caren Coghetto

    Full Text Available Abstract In this study, physiological aspects of Lactobacillus plantarum BL011 growing in a new, all-animal free medium in bioreactors were evaluated aiming at the production of this important lactic acid bacterium. Cultivations were performed in submerged batch bioreactors using the Plackett-Burman methodology to evaluate the influence of temperature, aeration rate and stirring speed as well as the concentrations of liquid acid protein residue of soybean, soy peptone, corn steep liquor, and raw yeast extract. The results showed that all variables, except for corn steep liquor, significantly influenced biomass production. The best condition was applied to bioreactor cultures, which produced a maximal biomass of 17.87 g L-1, whereas lactic acid, the most important lactic acid bacteria metabolite, peaked at 37.59 g L-1, corresponding to a productivity of 1.46 g L-1 h-1. This is the first report on the use of liquid acid protein residue of soybean medium for L. plantarum growth. These results support the industrial use of this system as an alternative to produce probiotics without animal-derived ingredients to obtain high biomass concentrations in batch bioreactors.

  16. Lactobacillus plantarum BL011 cultivation in industrial isolated soybean protein acid residue.

    Science.gov (United States)

    Coghetto, Chaline Caren; Vasconcelos, Carolina Bettker; Brinques, Graziela Brusch; Ayub, Marco Antônio Záchia

    In this study, physiological aspects of Lactobacillus plantarum BL011 growing in a new, all-animal free medium in bioreactors were evaluated aiming at the production of this important lactic acid bacterium. Cultivations were performed in submerged batch bioreactors using the Plackett-Burman methodology to evaluate the influence of temperature, aeration rate and stirring speed as well as the concentrations of liquid acid protein residue of soybean, soy peptone, corn steep liquor, and raw yeast extract. The results showed that all variables, except for corn steep liquor, significantly influenced biomass production. The best condition was applied to bioreactor cultures, which produced a maximal biomass of 17.87gL(-1), whereas lactic acid, the most important lactic acid bacteria metabolite, peaked at 37.59gL(-1), corresponding to a productivity of 1.46gL(-1)h(-1). This is the first report on the use of liquid acid protein residue of soybean medium for L. plantarum growth. These results support the industrial use of this system as an alternative to produce probiotics without animal-derived ingredients to obtain high biomass concentrations in batch bioreactors.

  17. Engineering solutions for open microalgae mass cultivation and realistic indoor simulation of outdoor environments.

    Science.gov (United States)

    Apel, Andreas Christoph; Weuster-Botz, Dirk

    2015-06-01

    Microalgae could become an important renewable source for chemicals, food, and energy if process costs can be reduced. In the past 60 years, relevant factors in open outdoor mass cultivation of microalgae were identified and elaborate solutions regarding bioprocesses and bioreactors were developed. An overview of these solutions is presented. Since the cost of most microalgal products from current mass cultivation systems is still prohibitively high, further development is required. The application of complex computational techniques for cost-effective process and reactor development will become more important if experimental validation of simulation results can easily be achieved. Due to difficulties inherent to outdoor experimentation, it can be useful to conduct validation experiments indoors. Considerations and approaches for realistic indoor reproduction of the most important environmental conditions in microalgae cultivation experiments-light, temperature, and substance concentrations, are discussed.

  18. [Proliferative activity parameters and their correlation with genetic damage of blood lymphocytes during cultivation under the conditions of cytokinetic block].

    Science.gov (United States)

    Ingel', F I; Iurchenko, V V; Gus'kov, A S; Krivtsova, E K; Iurtseva, N A

    2006-01-01

    The subjects of the study were 15 volunteers aged 22 to 25 years, who underwent 25 air ionization sessions. The effects of genome instability were evaluated, and correlations between indicators of genome damage (lesions of micronuclei and nucleoplasmatic bridges) and parameters of proliferative and replicative activity (mitotic index, proliferative pool, the fraction of rapidly dividing cells, and replication index) of blood lymphocytes in the culture were studied. In order to establish the associations between the parameters, the parallel cultures were exposed to 0.07 mM of the standard mutagen MNNG during 5 hours. The study showed that the course of air ionization did not induce the micronuclei and nucleoplasmatic bridges in binuclear cells, but increased proliferative cell activity. This effect was accompanied by an increase in the fraction of rapidly dividing cells among all the dividing cells, and an increase in the dispersion of all proliferation parameters. MNNG induced a constant level of micronuclei in binuclear cells during the whole course, but not before the beginning of air ionization. The changes in the parameter "the fraction of dividing cells" (proliferative pool) were the most prominent manifestation of the suppression of proliferation by MNNG. MNNG loading inhibited the formation of binuclear cells most of all. The results demonstrate a non-random character of the correlation between the level of micronuclei in binuclear cells and proliferative activity parameters during cell cultivation under the conditions of cytokinetic block.

  19. Steviol glycosides targeted analysis in leaves of Stevia rebaudiana (Bertoni) from plants cultivated under chilling stress conditions.

    Science.gov (United States)

    Soufi, Sihem; D'Urso, Gilda; Pizza, Cosimo; Rezgui, Salah; Bettaieb, Taoufik; Montoro, Paola

    2016-01-01

    Stevia rebaudiana is an important agricultural crop for the production of a high-potency natural sweetener, sensitive to low temperature during the developmental stage. Stimulation of chilling stress with a pre-treatment with endogenous signalling components and in particular with salicylic acid (SA), hydrogen peroxide (H2O2), 6-benzylaminopurine (BAP) and calcium chloride (CaCl2) could induce tolerance to chilling and could constitute a suitable way to maintain quality and quantity of steviol glycosides under controlled artificial environment. In the present work the effects of different putative signalling molecules on the morpho-physiological parameters were evaluated, and a specific method for the quali-quantitative analysis of steviol glycosides in S. rebaudiana plants cultivated under controlled conditions was developed, by using LC-ESI-FT (Orbitrap) MS, LC-ESI-QqQ-MS/MS and multivariate data analysis. This approach underlined that the pre-treatment has influence on the production of secondary metabolites. In particular Stevia plants characterised by higher contents of rebaudioside A and stevioside, were identified.

  20. The flowering pattern of Arnica montana L. and A. chamissonis Less. under field cultivation conditions with successive flower head collection

    Directory of Open Access Journals (Sweden)

    Danuta Sugier

    2012-12-01

    Full Text Available The research on the flowering of Arnica montana L. and A. chamissonis Less. was carried out in the years 2005-2006 on two- and three-year plantations on grey-brown podsolic soil with the granulometric composition of heavy clay sand. The aim of the work was to determine the pattern and length of flowering of two- and three-year-old plants under field cultivation conditions with successive flower head collection. The flowering of Arnica montana in both years of study lasted 26 days (from the 28th of May to the 22nd of June. In this period, eight collections of flower heads were made. When analyzing the number of inflorescences gathered during particular collections, it was concluded that at the initial stage of flowering it was not great, but it successively increased with the passage of time up to the seventh collection. The pattern of flowering of the two- and three-year-old Arnica montana plants was similar. The inflorescences of Arnica chamissonis were collected six times, and the most abundant flowering was found in the middle period of this stage, which lasted 20 days.

  1. Experimental and CFD-PBM Study of Oxygen Mass Transfer Coefficient in Different Impeller Configurations and Operational Conditions of a Two-Phase Partitioning Bioreactor.

    Science.gov (United States)

    Moradkhani, Hamed; Izadkhah, Mir-Shahabeddin; Anarjan, Navideh

    2017-02-01

    In this work, gas dispersion in a two-phase partitioning bioreactor is analyzed by calculating volumetric oxygen mass transfer coefficient which is modeled using a commercial computational fluid dynamics (CFD), code FLUENT 6.2. Dispersed oxygen bubbles dynamics is based on standard "k-ε" Reynolds-averaged Navier-Stokes (RANS) model. This paper describes a three-dimensional CFD model coupled with population balance equations (PBE) in order to get more confirming results of experimental measurements. Values of k L a are obtained using dynamic gassing-out method. Using the CFD simulation, the volumetric mass transfer coefficient is calculated based on Higbie's penetration theory. Characteristics of mass transfer coefficient are investigated for five configurations of impeller and three different aeration flow rates. The pitched six blade type, due to the creation of downward flow direction, leads to higher dissolved oxygen (DO) concentrations, thereby, higher values of k L a compared with other impeller compositions. The magnitude of dissolved oxygen percentage in the aqueous phase has direct correlation with impeller speed and any increase of the aeration magnitude leads to faster saturation in shorter periods of time. Agitation speeds of 300 to 800 rpm are found to be the most effective rotational speeds for the mass transfer of oxygen in two-phase partitioning bioreactors (TPPB).

  2. BIOPREPARATIONS AS THE NEW ELEMENT OF CULTIVATION OF AN APPLE-TREE GARDEN IN THE CONDITIONS OF NIZHNEE POVOLZHYE

    OpenAIRE

    2013-01-01

    In the article there were presented the results of cultivation of an apple-tree garden with application of growth stimulators. Positive action of Buton and Mival-Agro preparations on grades of all terms of maturing is noted

  3. NASA Classroom Bioreactor

    Science.gov (United States)

    Scully, Robert

    2004-01-01

    Exploration of space provides a compelling need for cell-based research into the basic mechanisms that underlie the profound changes that occur in terrestrial life that is transitioned to low gravity environments. Toward that end, NASA developed a rotating bioreactor in which cells are cultured while continuously suspended in a cylinder in which the culture medium rotates with the cylinder. The randomization of the gravity vector accomplished by the continuous rotation, in a low shear environment, provides an analog of microgravity. Because cultures grown in bioreactors develop structures and functions that are much closer to those exhibited by native tissue than can be achieved with traditional culture methods, bioreactors have contributed substantially to advancing research in the fields of cancer, diabetes, infectious disease modeling for vaccine production, drug efficacy, and tissue engineering. NASA has developed a Classroom Bioreactor (CB) that is built from parts that are easily obtained and assembled, user-friendly and versatile. It can be easily used in simple school settings to examine the effect cultures of seeds or cells. An educational brief provides assembly instructions and lesson plans that describes activities in science, math and technology that explore free fall, microgravity, orbits, bioreactors, structure-function relationships and the scientific method.

  4. NASA Bioreactor Schematic

    Science.gov (United States)

    2001-01-01

    The schematic depicts the major elements and flow patterns inside the NASA Bioreactor system. Waste and fresh medium are contained in plastic bags placed side-by-side so the waste bag fills as the fresh medium bag is depleted. The compliance vessel contains a bladder to accommodate pressure transients that might damage the system. A peristolic pump moves fluid by squeezing the plastic tubing, thus avoiding potential contamination. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  5. Propagation and Dissolution of CO2 bubbles in Algae Photo-bioreactors

    Science.gov (United States)

    Kosaraju, Srinivas

    2015-11-01

    Research grade photo-bioreactors are used to study and cultivate different algal species for biofuel production. In an attempt to study the growth properties of a local algal species in rain water, a custom made bioreactor is designed and being tested. Bio-algae consumes dissolved CO2 in water and during its growth cycle, the consumed CO2 must be replenished. Conventional methods use supply of air or CO2 bubbles in the growth medium. The propagation and dissolution of the bubbles, however, are strongly dependent on the design parameters of the photo-bioreactor. In this paper, we discuss the numerical modeling of the air and CO2 bubble propagation and dissolution in the photo-bioreactor. Using the results the bioreactor design will be modified for maximum productivity.

  6. Production of Calcaride A by Calcarisporium sp. in Shaken Flasks and Stirred Bioreactors.

    Science.gov (United States)

    Tamminen, Anu; Wang, Yanming; Wiebe, Marilyn G

    2015-06-24

    Increased interest in marine resources has led to increased screening of marine fungi for novel bioactive compounds and considerable effort is being invested in discovering these metabolites. For compound discovery, small-scale cultures are adequate, but agitated bioreactors are desirable for larger-scale production. Calcarisporium sp. KF525 has recently been described to produce calcaride A, a cyclic polyester with antibiotic activity, in agitated flasks. Here, we describe improvements in the production of calcaride A in both flasks (13-fold improvement) and stirred bioreactors (200-fold improvement). Production of calcaride A in bioreactors was initially substantially lower than in shaken flasks. The cultivation pH (reduced from 6.8 to bioreactor, which was only slightly less than in shaken flasks (14 mg·g-1, 100 mg·L-1). The results demonstrate that a scalable process for calcaride A production could be developed using an iterative approach with flasks and bioreactors.

  7. Optimizing of Bioreactor Heat Supply and Material Feeding by Numerical Calculation

    Science.gov (United States)

    Zhou, Zhiwei; Song, Boyan; Zhu, Likuan; Li, Zuntao; Wang, Yang

    Cell culture at large scale normally uses stirred structure. And the situation of temperature field distribution is very important to the cell culture at large scale. Some cells are very sensitive to the circumstances. The local temperature is too high or too low all influences the cell survival and low the cell quantity at unit volume. This paper simulates the temperature field under three different heating conditions. Then analysis and contrast the simulation results. The mixed situation in bioreactor is extremely significant for nutrition transmit. Usually, use ways to measure the average mixture time in bioreactor, and improve the mixture circumstance in the bioreactor through stirred impeller and bioreactor structure change. This paper adopts numerical calculation method to investigate the flow field in bioreactor. It gets the mixture time of bioreactor through virtual tracer in simulate flow field and detects the tracer density time variation curve in the bioreactor.

  8. Production of Limonoids with Insect Antifeedant Activity in a Two-Stage Bioreactor Process with Cell Suspension Culture of Azadirachta indica.

    Science.gov (United States)

    Vásquez-Rivera, Andrés; Chicaiza-Finley, Diego; Hoyos, Rodrigo A; Orozco-Sánchez, Fernando

    2015-09-01

    Neem tree (Azadirachta indica) cell suspension culture is an alternative for the production of limonoids for insect control that overcomes limitations related to the supply of neem seeds. To establish conditions for cell growth and azadiracthin-related limonoid production, the effect of different sucrose concentrations, nitrate and phosphate in Murashige and Skoog (MS) medium, and the addition of one precursor and three elicitors was evaluated in shake flasks. The process was scaled up to a 3-l stirred tank bioreactor in one- and two-stage batch cultivation. In shake flasks, more than fivefold increase in the production of limonoids with the modified MS medium was observed (increase from 0.77 to 4.52 mg limonoids/g dry cell weight, DCW), while an increase of more than fourfold was achieved by adding the elicitors chitosan, salicylic acid, and jasmonic acid together (increase from 1.03 to 4.32 mg limonoids/g DCW). In the bioreactor, the volumetric production of limonoids was increased more than threefold with a two-stage culture in day 18 (13.82 mg limonoids/l in control single-stage process and 41.44 mg/l in two-stage process). The cultivation and operating mode of the bioreactor reported in this study may be adapted and used in optimization and process plant development for production of insect antifeedant limonoids with A. indica cell suspension cultures.

  9. Bioreactor technology in marine microbiology: from design to future application.

    Science.gov (United States)

    Zhang, Yu; Arends, Jan B A; Van de Wiele, Tom; Boon, Nico

    2011-01-01

    Marine micro-organisms have been playing highly diverse roles over evolutionary time: they have defined the chemistry of the oceans and atmosphere. During the last decades, the bioreactors with novel designs have become an important tool to study marine microbiology and ecology in terms of: marine microorganism cultivation and deep-sea bioprocess characterization; unique bio-chemical product formation and intensification; marine waste treatment and clean energy generation. In this review we briefly summarize the current status of the bioreactor technology applied in marine microbiology and the critical parameters to take into account during the reactor design. Furthermore, when we look at the growing population, as well as, the pollution in the coastal areas of the world, it is urgent to find sustainable practices that beneficially stimulate both the economy and the natural environment. Here we outlook a few possibilities where innovative bioreactor technology can be applied to enhance energy generation and food production without harming the local marine ecosystem.

  10. Evaluation of the cultivation conditions of marine microalgae Chlorella sp. to be used as feedstock in ultrasound-assisted ethanolysis

    Directory of Open Access Journals (Sweden)

    Mateus S. Amaral

    2015-09-01

    Full Text Available A total of 8 assays was conducted to study the influence of different variables namely, light intensity, CO2 level, NaNO3 concentration and aeration rate, on the cultivation of the marine microalgae Chlorella sp. to enhance the biomass feedstock availability for biodiesel. The experiments were designed using a Taguchi L8 experimental array set at two levels of operation, having light intensity (0.85 and 14.5 klux, CO2 (5 and 10%, NaNO3 (0.025 and 0.075 g L-1 and aeration rate (3:33 and 1.67 vvm as independent variables and considering biomass productivity and lipid content as response variables. All the experiments were performed in six photobioreactor vessels connected in series with a total volume of 8.4 L and working volumes of 2 L and 4 L, depending on the conditions assessed. The highest biomass productivity was 210.9 mg L-1day-1, corresponding to a lipid content of 8.2%. Such results were attained when the culture conditions were set at 0.85 klux light intensity, 5% CO2 and 0.075 g L-1 NaNO3. The aeration rate showed no significant influence on the biomass productivity. On the other hand, the highest lipid content was achieved when the cultures were grown using the lowest concentration of NaNO3 (0.025 g L-1 and an aeration rate of 1.67 vvm, while the other factors had no statistical significance. Under these conditions, the lipid content obtained was 19.8%, at the expense of reducing the biomass productivity to 85.9 mg L-1day-1.The fatty acid profile of the lipid material characterized by gas chromatography identified fourteen fatty acids with carbon chain ranging from C8 to C20 in which most of the fatty acids present were saturated (58.7 % and monounsaturated (36.1% fatty acids. Those obtained at higher proportions were the oleic (22.8%, palmitic (20.7% and lauric (17.7 % acids, indicating a suitable composition for fatty acid ethyl esters (FAEE synthesis. This was confirmed by acid catalysis performed under ultrasound irradiations

  11. Polymer characterization and optimization of conditions for the enhanced bioproduction of benzaldehyde by Pichia pastoris in a two-phase partitioning bioreactor.

    Science.gov (United States)

    Craig, Tom; Daugulis, Andrew J

    2013-04-01

    Benzaldehyde, with its apricot and almond-like aroma, is the second most abundantly used molecule in the flavor industry, and is most commonly produced via chemical routes, such as by the oxidation of toluene. Biologically produced benzaldehyde, whether by extraction of plant material or via microbial biotransformation, commands a substantial price advantage, and greater consumer acceptance. Methylotrophic yeast, such as Pichia pastoris, contain the enzyme alcohol oxidase (AOX), which, in the presence of alcohols other than methanol, are able to yield aldehydes as dead-end products, for example, benzaldehyde from benzyl alcohol. In this work, we have determined that benzaldehyde, and not benzyl alcohol, is inhibitory to the transformation reaction by P. pastoris, prompting the development of a selection strategy for identifying sequestering polymers for use in a partitioning bioreactor that was based on the ratio of partition coefficients (PCs) for the two target molecules. Additionally, we have now confirmed for the first time, that the mechanism of solute uptake by amorphous polymers is via absorption, not adsorption. Finally, we have adopted a common strategy used for the production of heterologous proteins by P. pastoris, namely the use of a mixed methanol/glycerol feed for inducing the required AOX enzyme, while reducing the time required for high density biomass generation. All of these components were combined in a final experiment in which 10% of the polymer Kraton D1102K, whose PC ratio of benzaldehyde to benzyl alcohol was 14.9, was used to detoxify the biotransformation in a 5 L partitioning bioreactor, resulting in a 3.4-fold increase in benzaldehyde produced (14.4 g vs. 4.2 g) relative to single phase operation, at more than double the volumetric productivity (97 mg L(-1) h(-1) vs. 41 mg L(-1) h(-1) ).

  12. [Research on ursolic acid production of Eriobotrya japonica cell suspension culture in WAVE bioreactor].

    Science.gov (United States)

    Li, Hui-hua; Yao, De-heng; Xu, Jian; Wang, Wei; Chang, Qiang; Su, Ming-hua

    2015-05-01

    Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.

  13. Quality of mixing in a stired bioreactor used for animal cells culture: heterogeneities in a lab scale bioreactor and evolution of mixing time with scale up

    Directory of Open Access Journals (Sweden)

    Collignon, ML.

    2010-01-01

    Full Text Available Animal cells are industrially cultivated inside stirred bioreactors to produce proteinic compounds. Due to the use of mild agitation conditions in order to limit mechanical constraints, the homogeneity of the culture medium can be far from perfect. This study has therefore two objectives: the global characterization of the mixing via the mixing time and the local description of concentration fields. The mixing time is measured by conductimetry inside 20 l, 80 l, 600 l tanks. The Grenville correlation is adjusted on these experimental measurements to improve the prediction of the mixing time during the scale-up of the process. The concentration fields are visualized by the Planar Laser Induced Fluorescence (P.L.I.F. technique in the 20 l tank. This part of the study is focused on the time evolution of the maximum value of the tracer concentration inside measurement planes and of the numerical distribution of theses concentration fields.

  14. Conditions for vigorous growth on sulfide and reactor-scale cultivation protocols for the thermophilic green sulfur bacterium Chlorobium tepidum

    Energy Technology Data Exchange (ETDEWEB)

    Mukhopadhyay, B.; Johnson, E.F.; Ascano, M. Jr. [Univ. of Illinois, Urbana, IL (United States). Dept. of Microbiology

    1999-01-01

    The authors describe a reactor-scale cultivation protocol for the fastest-growing and only known thermophilic member of the family Chlorobiaceae, Chlorobium tepidum. They discovered that C. tepidum would grow with sulfide as the sole electron source at rates and with final cell yields comparable to those found with thiosulfate only if the sulfide concentration was maintained below 0.1 mM and the culture redox potential was at {minus}300 {+-} 20 mV. Such was also the requirement for growth in a photobioreactor when thiosulfate (optimum level, 12 mM) was used as the preferred electron source. For cultivation of C. tepidum on a 5- to 500-ml scale, the authors used the system of Balch and Wolfe using stopper-sealed serum tubes and bottles as an alternative to the methods commonly used for the cultivation of phototrophic anaerobes and obtained consistent results.

  15. NMR and GC-MS based metabolic profiling and free-radical scavenging activities of Cordyceps pruinosa mycelia cultivated under different media and light conditions.

    Directory of Open Access Journals (Sweden)

    Taek-Joo Oh

    Full Text Available Variation of metabolic profiles in Cordyceps pruinosa mycelia cultivated under various media and light conditions was investigated using 1H nuclear magnetic resonance (NMR analysis and gas chromatography mass spectrometry (GC-MS with multivariate statistical analysis. A total of 71 metabolites were identified (5 alcohols, 21 amino acids, 15 organic acids, 4 purines, 3 pyrimidines, 7 sugars, 11 fatty acids, and 5 other metabolites by NMR and GC-MS analysis. The mycelia grown in nitrogen media and under dark conditions showed the lowest growth and ergosterol levels, essential to a functional fungal cell membrane; these mycelia, however, had the highest levels of putrescine, which is involved in abiotic stress tolerance. In contrast, mycelia cultivated in sabouraud dextrose agar with yeast extract (SDAY media and under light conditions contained relatively higher levels of fatty acids, including valeric acid, stearic acid, lignoceric acid, myristic acid, oleic acid, palmitoleic acid, hepadecenoic acid, and linoleic acid. These mycelia also had the highest phenolic content and antioxidant activity, and did not exhibit growth retardation due to enhanced asexual development caused by higher levels of linoleic acid. Therefore, we suggested that a light-enriched environment with SDAY media was more optimal than dark condition for cultivation of C. pruinosa mycelia as biopharmaceutical or nutraceutical resources.

  16. NMR and GC-MS Based Metabolic Profiling and Free-Radical Scavenging Activities of Cordyceps pruinosa Mycelia Cultivated under Different Media and Light Conditions

    Science.gov (United States)

    Oh, Taek-Joo; Hyun, Sun-Hee; Lee, Seul-Gi; Chun, Young-Jin; Sung, Gi-Ho; Choi, Hyung-Kyoon

    2014-01-01

    Variation of metabolic profiles in Cordyceps pruinosa mycelia cultivated under various media and light conditions was investigated using 1H nuclear magnetic resonance (NMR) analysis and gas chromatography mass spectrometry (GC-MS) with multivariate statistical analysis. A total of 71 metabolites were identified (5 alcohols, 21 amino acids, 15 organic acids, 4 purines, 3 pyrimidines, 7 sugars, 11 fatty acids, and 5 other metabolites) by NMR and GC-MS analysis. The mycelia grown in nitrogen media and under dark conditions showed the lowest growth and ergosterol levels, essential to a functional fungal cell membrane; these mycelia, however, had the highest levels of putrescine, which is involved in abiotic stress tolerance. In contrast, mycelia cultivated in sabouraud dextrose agar with yeast extract (SDAY) media and under light conditions contained relatively higher levels of fatty acids, including valeric acid, stearic acid, lignoceric acid, myristic acid, oleic acid, palmitoleic acid, hepadecenoic acid, and linoleic acid. These mycelia also had the highest phenolic content and antioxidant activity, and did not exhibit growth retardation due to enhanced asexual development caused by higher levels of linoleic acid. Therefore, we suggested that a light-enriched environment with SDAY media was more optimal than dark condition for cultivation of C. pruinosa mycelia as biopharmaceutical or nutraceutical resources. PMID:24608751

  17. High cell density media for Escherichia coli are generally designed for aerobic cultivations – consequences for large-scale bioprocesses and shake flask cultures

    Directory of Open Access Journals (Sweden)

    Neubauer Peter

    2008-08-01

    Full Text Available Abstract Background For the cultivation of Escherichia coli in bioreactors trace element solutions are generally designed for optimal growth under aerobic conditions. They do normally not contain selenium and nickel. Molybdenum is only contained in few of them. These elements are part of the formate hydrogen lyase (FHL complex which is induced under anaerobic conditions. As it is generally known that oxygen limitation appears in shake flask cultures and locally in large-scale bioreactors, function of the FHL complex may influence the process behaviour. Formate has been described to accumulate in large-scale cultures and may have toxic effects on E. coli. Although the anaerobic metabolism of E. coli is well studied, reference data which estimate the impact of the FHL complex on bioprocesses of E. coli with oxygen limitation have so far not been published, but are important for a better process understanding. Results Two sets of fed-batch cultures with conditions triggering oxygen limitation and formate accumulation were performed. Permanent oxygen limitation which is typical for shake flask cultures was caused in a bioreactor by reduction of the agitation rate. Transient oxygen limitation, which has been described to eventually occur in the feed-zone of large-scale bioreactors, was mimicked in a two-compartment scale-down bioreactor consisting of a stirred tank reactor and a plug flow reactor (PFR with continuous glucose feeding into the PFR. In both models formate accumulated up to about 20 mM in the culture medium without addition of selenium, molybdenum and nickel. By addition of these trace elements the formate accumulation decreased below the level observed in well-mixed laboratory-scale cultures. Interestingly, addition of the extra trace elements caused accumulation of large amounts of lactate and reduced biomass yield in the simulator with permanent oxygen limitation, but not in the scale-down two-compartment bioreactor. Conclusion The

  18. Nutrient Deficiency Affects Root Architecture of Young Seedlings of Malus hupehensis (Pamp) Rehd. Under Conditions of Artificial Medium Cultivation

    Institute of Scientific and Technical Information of China (English)

    FAN Wei-guo; YANG Hong-qiang

    2007-01-01

    What the researchers go in for is to establish models between root architecture (RA) changes and nutrition, mold ideal root architecture of apple trees, improve the nutrient uptake efficiency, and further explore the functional mechanism of nutrient elements during the course of RA construction. The cultivation system of filter paper is utilized to research the effect of nutrient deficiency on the RA of Malus hupehensis (Pamp.) Rehd. There may be eight types of RA. In complete Hogland solution, the main type of RA is "lateral roots clustering in the upper and middle regions of primary root". With the lack of P, K or Ca, the main type of RA is "lateral roots clustering in the upper region primary root", and the "lateral roots clustering in the upper and middle regions of primary root" types of RA decrease. But with shortage of P, the type of lateral roots clustering in the upper and lower regions of primary root increases, and the type of lateral roots clustering in the middle region of primary root decreases, with the types of RA diversified. Under the condition of K deficiency, the type of no lateral root increases and types of lateral roots clustering in the middle region of primary root decrease, and the percentage of such types as "no lateral root", "lateral roots clustering in the upper region of primary root", and "lateral roots clustering in the upper and middle regions of primary root" accounts for 97.9% in all, with the types of RA simplified.With lack of Fe, Mg or Zn, the main type of RA is "lateral roots clustering in the upper and middle regions of primary root",but the type of lateral roots evenly-distributed on primary root increases. The main type of RA is "lateral roots evenlydistributed on primary root", under the condition of N deficiency, and the types of RA turn out to be diversified. There exists a close relation between nutrient deficiency and RA changes. Owing to various forms of nutrient deficiency,correspondingly different types of RA have

  19. Production of ethanol directly from potato starch by mixed culture of Saccharomyces cerevisiae and Aspergillus niger using electrochemical bioreactor.

    Science.gov (United States)

    Jeon, Bo Young; Kim, Dae Hee; Na, Byung Kwan; Ahn, Dae Hee; Park, Doo Hyun

    2008-03-01

    When cultivated aerobically, Aspergillus niger hyphae produced extracellular glucoamylase, which catalyzes the saccharification of unliquified potato starch into glucose, but not when grown under anaerobic conditions. The Km and Vmax of the extracellular glucoamylase were 652.3 mg starch l-1 and 253.3 mg glucose l-1 min-1, respectively. In mixed culture of A. niger and Saccharomyces cerevisiae, oxygen had a negative influence on the alcohol fermentation of yeast, but activated fungal growth. Therefore, oxygen is a critical factor for ethanol production in the mixed culture, and its generation through electrolysis of water in an electrochemical bioreactor needs to be optimized for ethanol production from starch by coculture of fungal hyphae and yeast cells. By applying pulsed electric fields (PEF) into the electrochemical bioreactor, ethanol production from starch improved significantly: Ethanol produced from 50 g potato starch l-1 by a mixed culture of A. niger and S. cerevisiae was about 5 g l-1 in a conventional bioreactor, but was 9 g l-1 in 5 volts of PEF and about 19 g l-1 in 4 volts of PEF for 5 days.

  20. Cascades of bioreactors.

    NARCIS (Netherlands)

    Gooijer, de C.D.

    1995-01-01

    In this thesis a common phenomenon in bioprocess engineering is described : the execution of a certain bioprocess in more than one bioreactor. Chapter 1, a review, classifies bioprocesses by means of a number of characteristics :i) processes with a variable stoichiometry ,ii) processes with a consta

  1. NASA Bioreactor tissue culture

    Science.gov (United States)

    1998-01-01

    Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  2. Basic bioreactor design.

    NARCIS (Netherlands)

    Riet, van 't K.; Tramper, J.

    1991-01-01

    Based on a graduate course in biochemical engineering, provides the basic knowledge needed for the efficient design of bioreactors and the relevant principles and data for practical process engineering, with an emphasis on enzyme reactors and aerated reactors for microorganisms. Includes exercises.

  3. Pyrosequence analysis of bacterial communities in aerobic bioreactors treating polycyclic aromatic hydrocarbon-contaminated soil.

    Science.gov (United States)

    Singleton, David R; Richardson, Stephen D; Aitken, Michael D

    2011-11-01

    Two aerobic, lab-scale, slurry-phase bioreactors were used to examine the biodegradation of polycyclic aromatic hydrocarbons (PAHs) in contaminated soil and the associated bacterial communities. The two bioreactors were operated under semi-continuous (draw-and-fill) conditions at a residence time of 35 days, but one was fed weekly and the other monthly. Most of the quantified PAHs, including high-molecular-weight compounds, were removed to a greater extent in the weekly-fed bioreactor, which achieved total PAH removal of 76%. Molecular analyses, including pyrosequencing of 16S rRNA genes, revealed significant shifts in the soil bacterial communities after introduction to the bioreactors and differences in the abundance and types of bacteria in each of the bioreactors. The weekly-fed bioreactor displayed a more stable bacterial community with gradual changes over time, whereas the monthly-fed bioreactor community was less consistent and may have been more strongly influenced by the influx of untreated soil during feeding. Phylogenetic groups containing known PAH-degrading bacteria previously identified through stable-isotope probing of the untreated soil were differentially affected by bioreactor conditions. Sequences from members of the Acidovorax and Sphingomonas genera, as well as the uncultivated "Pyrene Group 2" were abundant in the bioreactors. However, the relative abundances of sequences from the Pseudomonas, Sphingobium, and Pseudoxanthomonas genera, as well as from a group of unclassified anthracene degraders, were much lower in the bioreactors compared to the untreated soil.

  4. Bioreactor technology in marine microbiology: From design to future application

    OpenAIRE

    Zhang, Yu; Arends, Jan B.A.; van de Wiele, Tom; Boon, Nico

    2011-01-01

    Marine micro-organisms have been playing highly diverse roles over evolutionary time: they have defined the chemistry of the oceans and atmosphere. During the last decades, the bioreactors with novel designs have become an important tool to study marine microbiology and ecology in terms of: marine microorganism cultivation and deep-sea bioprocess characterization; unique bio-chemical product formation and intensification; marine waste treatment and clean energy generation. In this review we b...

  5. Influence of fermentation conditions on glucosinolates, ascorbigen, and ascorbic acid content in white cabbage (Brassica oleracea var. capitata cv. Taler) cultivated in different seasons.

    Science.gov (United States)

    Martinez-Villaluenga, C; Peñas, E; Frias, J; Ciska, E; Honke, J; Piskula, M K; Kozlowska, H; Vidal-Valverde, C

    2009-01-01

    The content of glucosinolates (GLS), ascorbigen, and ascorbic acid in white cabbage (Brassica oleracea var. capitata cv. Taler) cultivated in different seasons (summer and winter) was determined, before and after spontaneous and starter-induced fermentation. Different salt concentrations (0.5% NaCl or 1.5% NaCl) were used for sauerkraut production. Glucoiberin, sinigrin, and glucobrassicin were dominating in raw white cabbage cultivated either in winter or summer seasons. Ascorbigen precursor, glucobrassicin, was found higher in cabbage cultivated in winter (2.54 micromol/g dw) than those grown in summer (1.83 micromol/g dw). Cabbage fermented for 7 d was found to contain only traces of some GLS irrespective of the fermentation conditions used. Ascorbigen synthesis occurred during white cabbage fermentation. Brining cabbage at low salt concentration (0.5% NaCl) improved ascorbigen content in sauerkraut after 7 d of fermentation at 25 degrees C. The highest ascorbigen concentration was observed in low-sodium (0.5% NaCl) sauerkraut produced from cabbage cultivated in winter submitted to either natural (109.0 micromol/100 g dw) or starter-induced fermentation (108.3 and 104.6 micromol/100 g dw in cabbages fermented by L. plantarum and L. mesenteroides, respectively). Ascorbic acid content was found higher in cabbage cultivated in summer and fermentation process led to significant reductions. Therefore, the selection of cabbages with high glucobrassicin content and the production of low-sodium sauerkrauts may provide enhanced health benefits towards prevention of chronic diseases.

  6. Human cell culture in a space bioreactor

    Science.gov (United States)

    Morrison, Dennis R.

    1988-01-01

    Microgravity offers new ways of handling fluids, gases, and growing mammalian cells in efficient suspension cultures. In 1976 bioreactor engineers designed a system using a cylindrical reactor vessel in which the cells and medium are slowly mixed. The reaction chamber is interchangeable and can be used for several types of cell cultures. NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first Space Bioreactor was designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small Bioreactor is being constructed for flight experiments in the Shuttle Middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption and control of low shear stress on cells.

  7. Establishing Liver Bioreactors for In Vitro Research.

    Science.gov (United States)

    Rebelo, Sofia P; Costa, Rita; Sousa, Marcos F Q; Brito, Catarina; Alves, Paula M

    2015-01-01

    In vitro systems that can effectively model liver function for long periods of time are fundamental tools for preclinical research. Nevertheless, the adoption of in vitro research tools at the earliest stages of drug development has been hampered by the lack of culture systems that offer the robustness, scalability, and flexibility necessary to meet industry's demands. Bioreactor-based technologies, such as stirred tank bioreactors, constitute a feasible approach to aggregate hepatic cells and maintain long-term three-dimensional cultures. These three-dimensional cultures sustain the polarity, differentiated phenotype, and metabolic performance of human hepatocytes. Culture in computer-controlled stirred tank bioreactors allows the maintenance of physiological conditions, such as pH, dissolved oxygen, and temperature, with minimal fluctuations. Moreover, by operating in perfusion mode, gradients of soluble factors and metabolic by-products can be established, aiming at resembling the in vivo microenvironment. This chapter provides a protocol for the aggregation and culture of hepatocyte spheroids in stirred tank bioreactors by applying perfusion mode for the long-term culture of human hepatocytes. This in vitro culture system is compatible with feeding high-throughput screening platforms for the assessment of drug elimination pathways, being a useful tool for toxicology research and drug development in the preclinical phase.

  8. Determination of CO₂ sensitivity of micro-organisms in shaken bioreactors. II. Novel online monitoring method.

    Science.gov (United States)

    Amoabediny, Ghassem; Abbas, Mahdi Pesaran Haji; Büchs, Jochen

    2010-12-01

    In the present study, a new online monitoring method for the determination of the CO₂ sensitivity of micro-organisms, based on the values of the respiration factors [OTR (oxygen transfer rate) and CTR (carbon dioxide transfer rate)], obtained by using the RAMOS (respiratory activity monitoring system) device considering a variety of aeration rates in the measuring flask, is investigated. Based on the data of the OTR, obtained by RAMOS under a variety of specific aeration rates, the proposed new method was developed as an online monitoring method for CO₂ sensitivity of micro-organisms in shaken bioreactors. A maximum accumulated CO₂ concentration of 12% was derived in applied methods, provided that the cultivation system is carried out under optimal conditions. Additionally, to predict these conditions, an unsteady-state gas transfer model in shaken bioreactors would be very advantageous. The data of OTR obtained using the RAMOS device were analysed and recalculated by a programme considering the calibration factor (Cf). The major advantage of the new method is the possibility to determine the metabolic activity, regardless of manual sampling.

  9. Scaled-up manufacturing of recombinant antibodies produced by plant cells in a 200-L orbitally-shaken disposable bioreactor

    NARCIS (Netherlands)

    Raven, N.; Rasche, F.; Kuehn, C.; Anderlei, T.; Klöckner, W.; Schuster, F.; Henquet, M.G.L.; Bosch, H.J.; Büchs, J.; Fischer, R.; Schillberg, S.

    2015-01-01

    Tobacco BY-2 cells have emerged as a promising platform for the manufacture of biopharmaceutical proteins, offering efficient protein secretion, favourable growth characteristics and cultivation in containment under a controlled environment. The cultivation of BY-2 cells in disposable bioreactors is

  10. Comparative study of emerging micropollutants removal by aerobic activated sludge of large laboratory-scale membrane bioreactors and sequencing batch reactors under low-temperature conditions.

    Science.gov (United States)

    Kruglova, Antonina; Kråkström, Matilda; Riska, Mats; Mikola, Anna; Rantanen, Pirjo; Vahala, Riku; Kronberg, Leif

    2016-08-01

    Four emerging micropollutants ibuprofen, diclofenac, estrone (E1) and 17α-ethinylestradiol (EE2) were studied in large laboratory-scale wastewater treatment plants (WWTPs) with high nitrifying activity. Activated sludge (AS) with sludge retention times (SRTs) of 12days and 14days in sequencing batch reactors (SBRs) and 30days, 60days and 90days in membrane bioreactors (MBRs) were examined at 8°C and 12°C. Concentrations of pharmaceuticals and their main metabolites were analysed in liquid phase and solid phase of AS by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A remarkable amount of contaminants were detected in solids of AS, meaning the accumulation of micropollutants in bacterial cells. The biodegradation rate constants (Kbiol) were affected by SRT and temperature. MBR with a 90-day SRT showed the best results of removal. Conventional SBR process was inefficient at 8°C showing Kbiol values lower than 0.5lgSS(-1)d(-1) for studied micropollutants.

  11. IMPROVED CELLULASE PRODUCTION BY Aspergillus terreus USING OIL PALM EMPTY FRUIT BUNCH FIBRE AS SUBSTRATE IN A STIRRED TANK BIOREACTOR THROUGH OPTIMIZATION OF THE FERMENTATION CONDITIONS

    Directory of Open Access Journals (Sweden)

    Mahdi Shahriarinour

    2011-05-01

    Full Text Available Response surface methodology (RSM was performed to evaluate the effects of dissolved oxygen tension (DOT and initial pH on the production of carboxymethyl cellulase (CMCase, filter-paper hydrolase (FPase, and β-glucosidase by Aspergillus terreus in a 2 L stirred tank bioreactor. Delignified oil palm empty fruit bunch (OPEFB fibre was used as the main substrate under submerged fermentation. Growth of A. terreus and the production of three main components of cellulase were optimized by central composite design (CCD design. Statistical analysis of results showed that the individual terms of these two variables (DOT and pH had significant effects on growth and the production of all components of cellulase. Maximum growth (13.07 g/L and cellulase activity (CMCase = 50.33 U/mL, FPase = 2.29 U/mL and β-glucosidase = 15.98 U/ml were obtained when the DOT and initial culture pH were set at 55% and 5.5, respectively. A high proportion of β-glucosidase to FPase (8:1 in cellulase of A. terreus could be beneficial for efficient hydrolysis of cellulosic materials. The use of OPEFB as a main substrate would reduce the cost of fermentation for the production of cellulase.

  12. Trophic relations of Opatrum sabulosum (Coleoptera, Tenebrionidae with leaves of cultivated and uncultivated species of herbaceous plants under laboratory conditions

    Directory of Open Access Journals (Sweden)

    Viktor Brygadyrenko

    2015-02-01

    Full Text Available We carried out a quantitative assessment of the consumption of herbaceous plants by Opatrum sabulosum (Linnaeus, 1761 – a highly significant agricultural pest species. We researched the feeding preferences of this pest species with respect to 33 uncultivated and 22 cultivated plant species. This species of darkling beetle feeds on many uncultivated plant species, including those with hairy leaves and bitter milky sap, such as Scabiosa ucrainca (5.21 mg/specimen/24 hours, Euphorbia virgata (3.45, Solanum nigrum (3.32, Centauria scabiosa (2.47, Lamium album (2.41, Aristolochia clematitis (1.76, Chenopodium album (1.73, Arctium lappa (1.51, Asperula odorata (1.20. A high rate of leaf consumption is also characteristic for cultivated species, for example, Perilla nankinensis (5.05 mg/specimen/24 hours, Lycopersicon esculentum (3.75, Tropaeolum majus (3.29, Nicotiana tabacum (2.66, Rumex acetosa (1.96, Beta vulgaris (1.27. O. sabulosum is capable of feeding on plants which are poisonous to cattle. This species of darkling beetle consumes 95.5% of the cultivated and 48.5% of the uncultivated herbaceous plants researched.

  13. Microliter-bioreactor array with buoyancy-driven stirring for human hematopoietic stem cell culture

    OpenAIRE

    Luni, Camilla; Feldman, Hope C.; Pozzobon, Michela; De Coppi, Paolo; Meinhart, Carl D.; Elvassore, Nicola

    2010-01-01

    This work presents the development of an array of bioreactors where finely controlled stirring is provided at the microliter scale (100–300 μl). The microliter-bioreactor array is useful for performing protocol optimization in up to 96 parallel experiments of hematopoietic stem cell (HSC) cultures. Exploring a wide range of experimental conditions at the microliter scale minimizes cost and labor. Once the cell culture protocol is optimized, it can be applied to large-scale bioreactors for ste...

  14. Influence of temperature in phosphate removal by microalgae in heterotrophic bioreactors

    Directory of Open Access Journals (Sweden)

    Maria Isabel Queiroz

    2012-08-01

    Full Text Available The removal of total dissolved phosphate by the cyanobacteria Aphanothece microscopica Nägeli cultivated heterotrophically in dairy processing wastewater was investigated in this work. The experiments were carried out in bioreactors operating in a batch mode, fed with 4.5 L of wastewater. The experimental conditions were as follows: initial cell concentration of 0.2 g/L, pH adjusted to 7.6, isothermal reactor operating at temperatures of 10, 20 and 30ºC with absence of light and continuous aeration of 1 VVM. The results showed that phosphate removal is strongly dependent on process temperature. A. microscopica Nägeli was effective in the removal phosphate achieving removal rates of 3.77 mg/L.h, which reflected in the conversion of 98.4% in hydraulic detention times of 24 h.

  15. Pilot scale harvesting, separation and drying of microalgae biomass from compact photo-bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Cardoso, Alberto Tadeu Martins; Luz Junior, Luiz Fernando de Lima [Dept. de Engenharia Quimica. Universidade Federal do Parana, Curitiba, PR (Brazil)], e-mail: luzjr@ufpr.br; Mariano, Andre Bellin; Ghidini, Luiz Francisco Correa; Gnoatto, Victor Eduardo; Locatelli Junior, Vilson; Mello, Thiago Carvalho de; Vargas, Jose Viriato Coelho [Nucleo de Pesquisa e Desenvolvimento em Energia Autossustentavel (NPDEAS). Dept. de Engenharia Mecanica. Universidade Federal do Parana, Curitiba (Brazil)], E-mail: jvargas@demec.ufpr.br

    2010-07-01

    Bio diesel produced from microalgae lipids is gaining a substantial ground in the search for renewable energy sources. In order to optimize the operating conditions of a continuous process, several experiments were realized, both in laboratory and pilot scale. The microalgae cultivation can be conducted in a photo-bioreactor, a closed system which allows parameters control and necessarily involves the aquatic environment. Because of that, the use of separation unit operations is required. The process starts in a proposed compact photo-bioreactor, which consist of a chain of transparent tubes with 6 cm of diameter arranged in parallel where the cultivation media circulate with the help of a pump. This arrangement offers a closed culture with less risk of contamination and maintains a minimum contact with the environment. The microalgae grow inside the pipes under incidence of ambient light. In this paper, harvesting, separation and drying were studied, as part of the processes of a sustainable energy plant under construction at UFPR, as shown in Fig. 1. To control the production in a photo-bioreactor in continuous system, it is necessary to monitor the concentration of microalgae growth in suspension. To measure the cell concentration in this equipment, an optic sensor has been developed. The microalgae biomass separation from the culture media is achieved by microalgae flocculation. Several cultivation situations have been tested with different NaOH concentrations, increasing the pH to 10. The system was kept under agitation during the addition by an air pump into the tank. Thereafter the system was maintained static. After a short time, it was observed that the microalgae coagulated and settled. The clarified part water was removed, remaining a concentrated microalgae suspension. Our results suggest that pH increase is a suitable methodology for microalgae separation from the growth suspension. The microalgae sedimentation time was recorded, which allowed the

  16. Energy efficiency in membrane bioreactors.

    Science.gov (United States)

    Barillon, B; Martin Ruel, S; Langlais, C; Lazarova, V

    2013-01-01

    Energy consumption remains the key factor for the optimisation of the performance of membrane bioreactors (MBRs). This paper presents the results of the detailed energy audits of six full-scale MBRs operated by Suez Environnement in France, Spain and the USA based on on-site energy measurement and analysis of plant operation parameters and treatment performance. Specific energy consumption is compared for two different MBR configurations (flat sheet and hollow fibre membranes) and for plants with different design, loads and operation parameters. The aim of this project was to understand how the energy is consumed in MBR facilities and under which operating conditions, in order to finally provide guidelines and recommended practices for optimisation of MBR operation and design to reduce energy consumption and environmental impacts.

  17. Thin film bioreactors in space

    Science.gov (United States)

    Hughes-Fulford, M.; Scheld, H. W.

    Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers us an opportunity to learn more about basic biological systems with one important variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would enable us to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.

  18. Thin film bioreactors in space

    Science.gov (United States)

    Hughes-Fulford, M.; Scheld, H. W.

    1989-01-01

    Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization, and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers an opportunity to learn more about basic biological systems with one inmportant variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would make it possible to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.

  19. 竹粉灵芝仿野生栽培技术研究%Studies of Cultivation Technique on Bamboo Powder Ganoderma Lucidum Planted by Imitating Wild Condition

    Institute of Scientific and Technical Information of China (English)

    吴德峰; 涂建生; 叶财旺; 邱思锋; 蒙绍权; 韩娜

    2016-01-01

    采用竹制品下脚料诸如竹粉等作为培养基主要原料,替代木材进行仿野生栽培灵芝,结果表明,仿野生栽培的竹粉灵芝所含的有效成分与纯天然野生灵芝相似,竹粉灵芝仿野生栽培技术研究为灵芝人工栽培节约了大量的林木资源,对于保护自然资源,开发野生灵芝产业有着广阔的前景和意义。%Abstrct:The cultivation technique of bamboo powder Ganoderma lucidum was an fungus planting technology imitating wild condition, which are drawn from bamboo products waste as culture medium to replace wood for producing Ganoderma lucidum. The result showed that effective components of bamboo powder Ganoderma lucidum was very similar to that of pure natural wild Ganoderma lucidum, which also save a lot of forest resources in Ganoderma lucidum cultivation. There was the wide prospects and important significance in cultivation technique of bamboo powder Ganoderma lucidum, which also opened up a new way in development of wild Ganoderma lucidum industry.

  20. Bioreactor Yields Extracts for Skin Cream

    Science.gov (United States)

    2015-01-01

    Johnson Space Flight Center researchers created a unique rotating-wall bioreactor that simulates microgravity conditions, spurring innovations in drug development and medical research. Renuèll Int'l Inc., based in Aventure, Florida, licensed the technology and used it to produce a healing skin care product, RE`JUVEL. In a Food and Drug Administration test, RE`JUVEL substantially increased skin moisture and elasticity while reducing dark blotches and wrinkles.

  1. Human Colon Cancer Cells Cultivated in Space

    Science.gov (United States)

    1995-01-01

    Within five days, bioreactor cultivated human colon cancer cells (shown) grown in Microgravity on the STS-70 mission in 1995, had grown 30 times the volume of the control specimens on Earth. The samples grown in space had a higher level of cellular organization and specialization. Because they more closely resemble tumors found in the body, microgravity grown cell cultures are ideal for research purposes.

  2. A novel bioreactor to simulate urinary bladder mechanical properties and compliance for bladder functional tissue engineering

    Institute of Scientific and Technical Information of China (English)

    WEI Xin; LI Dao-bing; XU Feng; WANG Yan; ZHU Yu-chun; LI Hong; WANG Kun-jie

    2011-01-01

    Background Bioreactors are pivotal tools for generating mechanical stimulation in functional tissue engineering study.This study aimed to create a bioreactor that can simulate urinary bladder mechanical properties, and to investigate the effects of a mechanically stimulated culture on urothelial cells and bladder smooth muscle cells.Methods We designed a bioreactor to simulate the mechanical properties of bladder. A pressure-record system was used to evaluate the mechanical properties of the bioreactor by measuring the pressure in culture chambers. To test the biocompatibility of the bioreactor, viabilities of urothelial cells and smooth muscle cells cultured in the bioreactor under static and mechanically changed conditions were measured after 7-day culture. To evaluate the effect of mechanical stimulations on the vital cells, urethral cells and smooth muscle cells were cultured in the simulated mechanical conditions. After that, the viability and the distribution pattern of the cells were observed and compared with cells cultured in non-mechanical stimulated condition.Results The bioreactor system successfully generated waveforms similar to the intended programmed model while maintaining a cell-seeded elastic membrane between the chambers. There were no differences between viabilities of urothelial cells ((91.90±1.22)% vs. (93.14±1.78)%, P >0.05) and bladder smooth muscle cells ((93.41±1.49)% vs.(92.61±1.34)%, P >0.05). The viability of cells and tissue structure observation after cultured in simulated condition showed that mechanical stimulation was the only factor affected cells in the bioreactor and improved the arrangement of cells on silastic membrane.Conclusions This bioreactor can effectively simulate the physiological and mechanical properties of the bladder.Mechanical stimulation is the only factor that affected the viability of cells cultured in the bioreactor. The bioreactor can change the growth behavior of urothelial cells and bladder smooth

  3. Biodiversity of Dominant Cultivable Endophytic Bacteria Inhabiting Tissues of Six Different Cultivars of Maize (Zea mays L. ssp. mays) Cropped under Field Conditions.

    Science.gov (United States)

    Pisarska, Katarzyna; Pietr, Stanisław Jerzy

    2015-01-01

    Endophytic bacteria (EnB) play a crucial role in plant development. This study was an attempt to isolate and identify dominant cultivable EnB inhabiting young seedlings germinated in vitro and leaves of six maize cultivars grown under field conditions at temperate climate zone with culture-dependent approach. We isolated bacteria from field cropped maize only. Strains were identified based on 16S rRNA gene sequencing. In particular, members of Actinobacteria, Bacteroidetes, Firmicutes and α- and γ-Proteobacteria were found. Species of two genus Pseudomonas and Bacillus were dominant among them. Higher diversity of EnB was found in plants collected from Kobierzyce, where we identified 35 species from 16 genera with 22 species uniquely found at this field. On the contrary, from maize leaves collected at Smolice we identified 24 species representing 10 genera with 10 species uniquely isolated from this field. However, none of species was common for all cultivars at both locations. Among isolated EnB six species only, Pseudomonas clemancea, Pseudomonasfluorescens, Bacillus megaterium, Bacillus simplex, Arthrobacter nicotinovorans and Arthrobacter nitroguajacolicus, were found in aboveground parts of the same cultivar grown on both tested fields. The fact that the same cultivars, sown from the same lots of seeds, under field conditions on two different locations were colonized with noticeably different associations of cultivable EnB suggest that cultivar genotype is an important factor selecting endophytic bacteria from local agro-environment. To our knowledge this is first report about the significant variation of diversity of cultivable endophytic bacteria inhabiting aboveground parts of the same maize cultivars grown at different locations.

  4. High-throughput miniaturized bioreactors for cell culture process development: reproducibility, scalability, and control.

    Science.gov (United States)

    Rameez, Shahid; Mostafa, Sigma S; Miller, Christopher; Shukla, Abhinav A

    2014-01-01

    Decreasing the timeframe for cell culture process development has been a key goal toward accelerating biopharmaceutical development. Advanced Microscale Bioreactors (ambr™) is an automated micro-bioreactor system with miniature single-use bioreactors with a 10-15 mL working volume controlled by an automated workstation. This system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody in a recombinant Chinese Hamster Ovary cell line. The miniaturized bioreactor system was found to produce cell culture profiles that matched across scales to 3 L, 15 L, and 200 L stirred tank bioreactors. The processes used in this article involve complex feed formulations, perturbations, and strict process control within the design space, which are in-line with processes used for commercial scale manufacturing of biopharmaceuticals. Changes to important process parameters in ambr™ resulted in predictable cell growth, viability and titer changes, which were in good agreement to data from the conventional larger scale bioreactors. ambr™ was found to successfully reproduce variations in temperature, dissolved oxygen (DO), and pH conditions similar to the larger bioreactor systems. Additionally, the miniature bioreactors were found to react well to perturbations in pH and DO through adjustments to the Proportional and Integral control loop. The data presented here demonstrates the utility of the ambr™ system as a high throughput system for cell culture process development.

  5. EFFICIENCY OF CULTIVATION OF DIFFERENT WINTER WHEAT CULTIVARS IN THE CONDITIONS OF INSUFFICIENT HUMIDIFYING IN THE KRASNODAR REGION

    Directory of Open Access Journals (Sweden)

    Bershatskaya S. I.

    2016-06-01

    Full Text Available The influence on the predecessor, on the time of sowing and various mineral nutrition levels, on the productivity of seven cultivars of winter soft wheat of Krasnodar Agricultural Research Institute selection was investigated. On the basis of the obtained experimental data and production audit it was found that all studied cultivars under the total complex of cultivation technological methods are able to form the yield of 6-7 t / ha with the payment of one kg of active ingredient of fertilizer by the grain within 6.50-9.86, and 8.56- 10.71 kg. Soil was ordinary chernozem with a low humus content powerful humus content, depending on the power of backgrounds in the arable (0-30sm soil layer is 3,95-4,00%, of mineral nitrogen is 5,9-8,3 mg / kg of soil, mobile phosphorus is 22,4 - 26,6 mg / kg of soil, of exchangeable potassium is 330-360 mg / kg of soil. Predecessors are winter wheat, sunflower, white mustard cultivated on seeds. The main tillage is resource-saving with the mulching of 0-10 cm layer. Studied sowing terms are the 20th of September, the 1st and 10th of October. The backgrounds of mineral nutrition: 1-without fertilizer N0P0К0; 2-minimum dose of complete mineral fertilizer N60P30К30; 3-average dose N90P60К60; 4-high dose N120- 160P90К60. Seeding rate is 5 million germinating seeds per hectare. The objects of the research were seven winter soft wheat cultivars of Krasnodar Agricultural Research Institute selection. We studied winter wheat cultivars with different combination of biological characteristics and predecessors. This combination is contributed to the yield growing from 4,26-5,82 t / ha up to 6,97-7,2 4,26-5,82 t / ha with the payment of one kilogram of active fertilizer substance with the output of grain within 8,52-12,0kg

  6. Degradation of Refuse in Hybrid Bioreactor Landfill

    Institute of Scientific and Technical Information of China (English)

    YAN LONG; Yu-YANG LONG; HAI-CHUN LIU; DONG-SHENG SHEN

    2009-01-01

    Objectivess To explore the process of refuse decomposition in hybrid bioreactor landfill. Methods The bioreactor landfill was operated in sequencing of facultative-anaerobic and aerobic conditions with leachate recireulation, pH, COD, and ammonia in the leachate and pH, biodegradable organic matter (BDM), and cation exchange capacity (CEC) in refuse were detected. Results CEC increased gradually with the degradation of refuse, which was negatively correlad, With BDM. COD and ammonia in the leachate was declined to 399.2 mg L-1 and 20.6 mg N L-1, respectively, during the 357-day operation. The respective concentrations of ammonia and COD were below the second and the third levels of current discharge standards in China. Conclusion The refuse is relatively stable at the end of hybrid bioreactor landfill operation. Most of the readily biodegradable organic matter is mineralized in the initial phase of refuse degradation, whereas the hard-biodegradable organic matter is mainly humidified in the maturity phase of refuse degradation.

  7. Mycelium growth kinetics and optimal temperature conditions for the cultivation of edible mushroom species on lignocellulosic substrates.

    Science.gov (United States)

    Zervakis, G; Philippoussis, A; Ioannidou, S; Diamantopoulou, P

    2001-01-01

    The influence of environmental parameters on mycelial linear growth of Pleurotus ostreatus, P. eryngii, P. pulmonarius, Agrocybe aegerita, Lentinula edodes, Volvariella volvacea and Auricularia auricula-judae was determined in two different nutrient media in a wide range of temperature, forming the basis for the assessment of their temperature optima. V. volvacea grew faster at 35 degrees C, P. eryngii at 25 degrees C, P. ostreatus and P. pulmonarius at 30 degrees C, A. aegerita at 25 or 30 degrees C and A. auricula-judae at 20 or 25 degrees C depending on the nutrient medium used and L. edodes at 20 or 30 degrees C depending on the strain examined. The mycelium extension rates were evaluated on seven mushroom cultivation substrates: wheat straw, cotton gin-trash, peanut shells, poplar sawdust, oak sawdust, corn cobs and olive press-cake. The mycelium extension rates (linear growth and colonization rates) were determined by the 'race-tube' technique, and were found to be the highest on cotton gin-trash, peanut shells and poplar sawdust for Pleurotus spp. and A. aegerita. Wheat straw, peanut shells and particularly cotton gin-trash supported fast growth of V. volvacea, whereas wheat straw was the most suitable substrate for L. edodes and A. auricula-judae. Supplemented oak sawdust and olive press-cake were poor substrates for most species examined, while almost all strains performed adequately on corn cobs.

  8. De novo assembly and transcriptome analysis of osmoregulation in Litopenaeus vannamei under three cultivated conditions with different salinities.

    Science.gov (United States)

    Zhang, Dan; Wang, Fang; Dong, Shuanglin; Lu, Yunliang

    2016-03-10

    Litopenaeus vannamei, one of the most important euryhaline crustaceans, is cultured in seawater, brackish water, and freshwater worldwide. We performed Illumina RNA sequencing of L. vannamei gills, generating 124,914,870; 119,250,450; and 105,487,350 raw reads from the shrimps cultured in seawater, brackish water, and freshwater, respectively. From these reads, 466,293 transcripts were de novo assembled and annotated. Comparative genomic analysis showed that 1752 genes were significantly differentially expressed in the freshwater group compared with the seawater group, including 1242 upregulated and 510 downregulated genes. In addition, 1246 genes were differentially expressed in the brackish group vs. the seawater water group, including 659 upregulated and 587 downregulated genes. These differentially expressed genes were mainly involved in energy metabolism, substance metabolism, ion transport and signal transduction, and genetic process. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to analyze the functional significance of the differentially expressed genes, included those responding to salinity through diverse biological functions and processes and numerous potential genes associated with the osmotic response. L. vannamei responses to the three cultivated salinities were analyzed using next-generation sequencing. The transcriptional database established from the current research adds to the information available on L. vannamei and the findings expand our knowledge of the molecular basis of osmoregulation mechanisms in this species.

  9. Optimal conditions of different flocculation methods for harvesting Scenedesmus sp. cultivated in an open-pond system.

    Science.gov (United States)

    Chen, Lu; Wang, Cunwen; Wang, Weiguo; Wei, Jiang

    2013-04-01

    The effects of culture medium pH, flocculant type (FeCl3, Al2(SO4)3, Alum, Ca(OH)2, chitosan, polyacrylamide), dosage and sedimental time on flocculation efficiency of harvesting Scenedesmus sp. cultivated in an open-pond system were investigated. Meanwhile, the relation between initial biomass concentration and the flocculant dosage needed was also investigated. The results from this work indicated that the flocculation efficiency achieved 97.4% after 10 min of sedimentation when the pH was adjusted to be 11.5, without adding flocculants. FeCl3 and chitosan showed a good flocculation efficiency at dosage of 0.15 and 0.08 g/L, respectively without pH adjustment. The flocculation efficiency increased from 49.74% to 90.63% when the final medium pH was adjusted to 6 after adding 0.1 g/L Alum. An increment from 68.18% to 92.84% was observed after adding 0.1 g/L Al2(SO4)3 followed by pH adjustment. Finally, the most suitable flocculation method was discussed in this paper.

  10. D-Root: a system for cultivating plants with the roots in darkness or under different light conditions.

    Science.gov (United States)

    Silva-Navas, Javier; Moreno-Risueno, Miguel A; Manzano, Concepción; Pallero-Baena, Mercedes; Navarro-Neila, Sara; Téllez-Robledo, Bárbara; Garcia-Mina, Jose M; Baigorri, Roberto; Gallego, Francisco Javier; del Pozo, Juan C

    2015-10-01

    In nature roots grow in the dark and away from light (negative phototropism). However, most current research in root biology has been carried out with the root system grown in the presence of light. Here, we have engineered a device, called Dark-Root (D-Root), to grow plants in vitro with the aerial part exposed to the normal light/dark photoperiod while the roots are in the dark or exposed to specific wavelengths or light intensities. D-Root provides an efficient system for cultivating a large number of seedlings and easily characterizing root architecture in the dark. At the morphological level, root illumination shortens root length and promotes early emergence of lateral roots, therefore inducing expansion of the root system. Surprisingly, root illumination also affects shoot development, including flowering time. Our analyses also show that root illumination alters the proper response to hormones or abiotic stress (e.g. salt or osmotic stress) and nutrient starvation, enhancing inhibition of root growth. In conclusion, D-Root provides a growing system closer to the natural one for assaying Arabidopsis plants, and therefore its use will contribute to a better understanding of the mechanisms involved in root development, hormonal signaling and stress responses.

  11. 荔枝果汁红茶菌发酵工艺条件的探讨%Investigation on the cultivation conditions of kombucha fermentation of Litchi Juice

    Institute of Scientific and Technical Information of China (English)

    王柳玲; 胡卓炎; 余小林; 赵雷; 方祥; 梁芹

    2013-01-01

    为了探索荔枝果汁红茶菌发酵果醋的适宜条件,探讨了红茶菌接种量、荔枝果汁初始可溶性固形物含量、发酵温度和初始pH值等因素对荔枝果汁红茶菌发酵液的总酸含量、pH、可溶性固形物含量、还原糖转化率等指标的影响,并通过响应曲面分析和期望函数优化途径,获得荔枝果汁红茶菌发酵优化的工艺条件.结果表明:发酵温度32℃,荔枝果汁初始可溶性固形物浓度为14°Brix,红茶菌接种量10%,发酵6d后,发酵液总酸含量可达2.48 g/100 mL,pH值为3.04.%In order to obtain the cultivation conditions of kombucha fermentation for litchi juice,effects of fermentation conditions including kombucha inoculation concentration,initial total suluble solid content,fermentation temperature,and initial pH value of litchi juice on acid content,pH,total suluble solids content,reducing sugar consumption rate of the fermented litchi juice were investigated.Response surface methodology and desirability function approach was applied to obtain the optimal cultivation conditions.Results showed that the kombucha cultivation conditions for litchi juice were selected as 10% of inoculums concentration,14°Brix of initial soluble solid content,30℃ of fermentation temperature,after 6 days of kombucha fermentation of litchi juice,the content of acid was increased to 2.48g/100 mL,pH value was 3.04.

  12. Imaging glucose metabolism in perfluorocarbon-perfused hepatocyte bioreactors using positron emission tomography.

    Science.gov (United States)

    Nieuwoudt, Martin; Wiggett, Scholtz; Malfeld, Susan; van der Merwe, Schalk W

    2009-01-01

    In vitro hepatocyte bioreactor functionality depends particularly on maintaining appropriate oxygen levels and exposure to nonparenchymal cells. An attractive solution without immunological consequences to the patient is incorporating a perfluorocarbon oxygen carrier in the circulating medium and co-culturing hepatocytes with stellate cells. Since bioreactors are normally sealed sterile units, demonstrating metabolic functionality is hindered by limited access to the cells after their aggregation in the matrix. A novel possibility is to use positron emission tomography (PET) to image cellular radioactive glucose uptake under O(2)-limited conditions. In this study, primary cell isolation procedures were carried out on eight pigs. Pairs of cell-seeded and cell-free (control) bioreactors with and without perfluorocarbon were cultured under identical conditions and were oxygenated using hypoxic (5% O(2)) and ambient (20% O(2)) gas mixes. Sixteen PET scans were conducted 24 h after cell isolation, the same timescale as that involved in treating a liver failure patient with a primary-cell bioreactor. In all cases, cell-seeded bioreactors without perfluorocarbon were more radioactive, i.e., were more glycolytic, than those with perfluorocarbon. This difference was significant in the hypoxic pair of bioreactors but not in the ambient pair of bioreactors. Additionally, in the same hypoxic bioreactors, circulating extracellular steady-state glucose levels were significantly lower and lactate levels were higher than those in the ambient bioreactors. Similar findings have been made in other in vitro hepatocyte studies investigating the effects of perfluorocarbons. PET is attractive for studying in situ O(2)-dependent bioreactor metabolism because of its visual and numerically quantifiable outputs. Longer-term metabolic studies (e.g., 5-10 days) investigating the effect of perfluorocarbon on bioreactor longevity will complement these findings in the future.

  13. ANAEROBIC MEMBRANE BIOREACTORS FOR DOMESTIC WASTEWATER TREATMENT. PRELIMINARY STUDY

    Directory of Open Access Journals (Sweden)

    Luisa Vera

    2014-12-01

    Full Text Available The operation of submerged anaerobic membrane bioreactors (SAnMBRs for domestic wastewaters treatment was studied in laboratory scale, with the objective to define sustainable filtration conditions of the suspensions along the process. During continuous experiments, the organic matter degradation by anaerobic way showed an average DQOT removal of 85% and 93%. Indeed, the degradation generated biogas after 12 days of operation and its relative methane composition was of 60% after 25 days of operation. Additionally, the comparison between membrane bioreactors (MBRs performance in aerobic and anaerobic conditions in filterability terms, reported that both systems behave similarly once reached the stationary state.

  14. Dynamic Single-Use Bioreactors Used in Modern Liter- and m(3)- Scale Biotechnological Processes: Engineering Characteristics and Scaling Up.

    Science.gov (United States)

    Löffelholz, Christian; Kaiser, Stephan C; Kraume, Matthias; Eibl, Regine; Eibl, Dieter

    2014-01-01

    During the past 10 years, single-use bioreactors have been well accepted in modern biopharmaceutical production processes targeting high-value products. Up to now, such processes have mainly been small- or medium-scale mammalian cell culture-based seed inoculum, vaccine or antibody productions. However, recently first attempts have been made to modify existing single-use bioreactors for the cultivation of plant cells and tissue cultures, and microorganisms. This has even led to the development of new single-use bioreactor types. Moreover, due to safety issues it has become clear that single-use bioreactors are the "must have" for expanding human stem cells delivering cell therapeutics, the biopharmaceuticals of the next generation. So it comes as no surprise that numerous different dynamic single-use bioreactor types, which are suitable for a wide range of applications, already dominate the market today. Bioreactor working principles, main applications, and bioengineering data are presented in this review, based on a current overview of greater than milliliter-scale, commercially available, dynamic single-use bioreactors. The focus is on stirred versions, which are omnipresent in R&D and manufacturing, and in particular Sartorius Stedim's BIOSTAT family. Finally, we examine development trends for single-use bioreactors, after discussing proven approaches for fast scaling-up processes.

  15. A novel membrane distillation-thermophilic bioreactor system: biological stability and trace organic compound removal.

    Science.gov (United States)

    Wijekoon, Kaushalya C; Hai, Faisal I; Kang, Jinguo; Price, William E; Guo, Wenshan; Ngo, Hao H; Cath, Tzahi Y; Nghiem, Long D

    2014-05-01

    The removal of trace organic compounds (TrOCs) by a novel membrane distillation-thermophilic bioreactor (MDBR) system was examined. Salinity build-up and the thermophilic conditions to some extent adversely impacted the performance of the bioreactor, particularly the removal of total nitrogen and recalcitrant TrOCs. While most TrOCs were well removed by the thermophilic bioreactor, compounds containing electron withdrawing functional groups in their molecular structure were recalcitrant to biological treatment and their removal efficiency by the thermophilic bioreactor was low (0-53%). However, the overall performance of the novel MDBR system with respect to the removal of total organic carbon, total nitrogen, and TrOCs was high and was not significantly affected by the conditions of the bioreactor. All TrOCs investigated here were highly removed (>95%) by the MDBR system. Biodegradation, sludge adsorption, and rejection by MD contribute to the removal of TrOCs by MDBR treatment.

  16. Construction and evaluation of urinary bladder bioreactor for urologic tissue-engineering purposes.

    LENUS (Irish Health Repository)

    Davis, Niall F

    2012-01-31

    OBJECTIVE: To design and construct a urinary bladder bioreactor for urologic tissue-engineering purposes and to compare the viability and proliferative activity of cell-seeded extracellular matrix scaffolds cultured in the bioreactor with conventional static growth conditions. MATERIALS AND METHODS: A urinary bladder bioreactor was designed and constructed to replicate physiologic bladder dynamics. The bioreactor mimicked the filling pressures of the human bladder by way of a cyclical low-delivery pressure regulator. In addition, cell growth was evaluated by culturing human urothelial cells (UCs) on porcine extracellular matrix scaffolds in the bioreactor and in static growth conditions for 5 consecutive days. The attachment, viability, and proliferative potential were assessed and compared with quantitative viability indicators and by fluorescent markers for intracellular esterase activity and plasma membrane integrity. Scaffold integrity was characterized with scanning electron microscopy and 4\\

  17. Optimization of substrate preparation for oyster mushroom (Pleurotus ostreatus) cultivation by studying different raw materials and substrate preparation conditions (composting: phases I and II).

    Science.gov (United States)

    Vieira, Fabrício Rocha; de Andrade, Meire Cristina Nogueira

    2016-11-01

    In recent years, oyster mushroom (Pleurotus ostreatus) has become one of the most cultivated mushrooms in the world, mainly in Brazil. Among many factors involved in a mushroom production, substrate preparation is the most critical step, which can be influenced by composting management techniques. Looking forward to optimizing the substrate preparation process, were tested different composting conditions (7 and 14 days of composting with or without conditioning), potential raw materials (decumbens grass, brizantha grass and sugarcane straw) and nitrogen supplementation (with or without wheat bran) on oyster mushroom yield and biological efficiency (BE). The substrate composted for 7 days with conditioning showed higher yield and biological efficiency of mushroom (24.04 and 100.54 %, respectively). Substrates without conditioning (7 and 14 days of composting) showed smaller mushroom yield and biological efficiency. Among the raw materials tested, brizantha grass showed higher mushroom yield followed by decumbens grass, sugarcane straw and wheat straw (28.5, 24.32, 23.5 and 19.27 %, respectively). Brizantha grass also showed higher biological efficiency followed by sugarcane straw, decumbens grass and wheat straw (123.95, 103.70, 96.90 and 86.44 %, respectively). Supplementation with wheat bran improved yield and biological efficiency in all substrate formulations tested; thus, oyster mushroom yield and biological efficiency were influenced by substrate formulation (raw materials), supplementation and composting conditions.

  18. A simple eccentric stirred tank mini-bioreactor: mixing characterization and mammalian cell culture experiments.

    Science.gov (United States)

    Bulnes-Abundis, David; Carrillo-Cocom, Leydi M; Aráiz-Hernández, Diana; García-Ulloa, Alfonso; Granados-Pastor, Marisa; Sánchez-Arreola, Pamela B; Murugappan, Gayathree; Alvarez, Mario M

    2013-04-01

    In industrial practice, stirred tank bioreactors are the most common mammalian cell culture platform. However, research and screening protocols at the laboratory scale (i.e., 5-100 mL) rely primarily on Petri dishes, culture bottles, or Erlenmeyer flasks. There is a clear need for simple-easy to assemble, easy to use, easy to clean-cell culture mini-bioreactors for lab-scale and/or screening applications. Here, we study the mixing performance and culture adequacy of a 30 mL eccentric stirred tank mini-bioreactor. A detailed mixing characterization of the proposed bioreactor is presented. Laser induced fluorescence (LIF) experiments and computational fluid dynamics (CFD) computations are used to identify the operational conditions required for adequate mixing. Mammalian cell culture experiments were conducted with two different cell models. The specific growth rate and the maximum cell density of Chinese hamster ovary (CHO) cell cultures grown in the mini-bioreactor were comparable to those observed for 6-well culture plates, Erlenmeyer flasks, and 1 L fully instrumented bioreactors. Human hematopoietic stem cells were successfully expanded tenfold in suspension conditions using the eccentric mini-bioreactor system. Our results demonstrate good mixing performance and suggest the practicality and adequacy of the proposed mini-bioreactor.

  19. Controlled-Turbulence Bioreactors

    Science.gov (United States)

    Wolf, David A.; Schwartz, Ray; Trinh, Tinh

    1989-01-01

    Two versions of bioreactor vessel provide steady supplies of oxygen and nutrients with little turbulence. Suspends cells in environment needed for sustenance and growth, while inflicting less damage from agitation and bubbling than do propeller-stirred reactors. Gentle environments in new reactors well suited to delicate mammalian cells. One reactor kept human kidney cells alive for as long as 11 days. Cells grow on carrier beads suspended in liquid culture medium that fills cylindrical housing. Rotating vanes - inside vessel but outside filter - gently circulates nutrient medium. Vessel stationary; magnetic clutch drives filter cylinder and vanes. Another reactor creates even less turbulence. Oxygen-permeable tubing wrapped around rod extending along central axis. Small external pump feeds oxygen to tubing through rotary coupling, and oxygen diffuses into liquid medium.

  20. Potential use of the facultative halophyte Chenopodium quinoa Willd. as substrate for biogas production cultivated with different concentrations of sodium chloride under hydroponic conditions.

    Science.gov (United States)

    Turcios, Ariel E; Weichgrebe, Dirk; Papenbrock, Jutta

    2016-03-01

    This project analyses the biogas potential of the halophyte Chenopodium quinoa Willd. In a first approach C. quinoa was grown with different concentrations of NaCl (0, 10 and 20 ppt NaCl) and the crop residues were used as substrate for biogas production. In a second approach, C. quinoa was grown with 0, 10, 20 and 30 ppt NaCl under hydroponic conditions and the fresh biomass was used as substrate. The more NaCl is in the culture medium, the higher the sodium, potassium, crude ash and hemicellulose content in the plant tissue whereas the calcium, sulfur, nitrogen and carbon content in the biomass decrease. According to this study, it is possible to produce high yields of methane using biomass of C. quinoa. The highest specific methane yields were obtained using the substrate from the plants cultivated at 10 and 20 ppt NaCl in both experiments.

  1. Bioreactors for lignocellulose conversion into fermentable sugars for production of high added value products.

    Science.gov (United States)

    Liguori, Rossana; Ventorino, Valeria; Pepe, Olimpia; Faraco, Vincenza

    2016-01-01

    Lignocellulosic biomasses derived from dedicated crops and agro-industrial residual materials are promising renewable resources for the production of fuels and other added value bioproducts. Due to the tolerance to a wide range of environments, the dedicated crops can be cultivated on marginal lands, avoiding conflict with food production and having beneficial effects on the environment. Besides, the agro-industrial residual materials represent an abundant, available, and cheap source of bioproducts that completely cut out the economical and environmental issues related to the cultivation of energy crops. Different processing steps like pretreatment, hydrolysis and microbial fermentation are needed to convert biomass into added value bioproducts. The reactor configuration, the operative conditions, and the operation mode of the conversion processes are crucial parameters for a high yield and productivity of the biomass bioconversion process. This review summarizes the last progresses in the bioreactor field, with main attention on the new configurations and the agitation systems, for conversion of dedicated energy crops (Arundo donax) and residual materials (corn stover, wheat straw, mesquite wood, agave bagasse, fruit and citrus peel wastes, sunflower seed hull, switchgrass, poplar sawdust, cogon grass, sugarcane bagasse, sunflower seed hull, and poplar wood) into sugars and ethanol. The main novelty of this review is its focus on reactor components and properties.

  2. Glycoprofiling effects of media additives on IgG produced by CHO cells in fed-batch bioreactors

    DEFF Research Database (Denmark)

    Kildegaard, Helene Faustrup; Fan, Yuzhou; Wagtberg Sen, Jette

    2016-01-01

    . In this study, the effect on IgG N-glycosylation from feeding CHO cells with eight glycosylation precursors during cultivation was investigated. The study was conducted in fed-batch mode in bioreactors with biological replicates to obtain highly controlled and comparable conditions. We assessed charge......Therapeutic monoclonal antibodies (mAbs) are mainly produced by heterogonous expression in Chinese hamster ovary (CHO) cells. The glycosylation profile of the mAbs has major impact on the efficacy and safety of the drug and is therefore an important parameter to control during production...... heterogeneity and glycosylation patterns of IgG. None of the eight feed additives caused statistically significant changes to cell growth or IgG productivity, compared to controls. However, the addition of 20 mM galactose did result in a reproducible increase of galactosylated IgG from 14% to 25%. On the other...

  3. Treatment of phenolics, aromatic hydrocarbons, and cyanide-bearing wastewater in individual and combined anaerobic, aerobic, and anoxic bioreactors.

    Science.gov (United States)

    Sharma, Naresh K; Philip, Ligy

    2015-01-01

    Studies were conducted on a mixture of pollutants commonly found in coke oven wastewater (CWW) to evaluate the biodegradation of various pollutants under anaerobic, aerobic, and anoxic conditions. The removal of the pollutants was monitored during individual bioreactor operation and using a combination of bioreactors operating in anaerobic-aerobic-anoxic sequence. While studying the performance of individual reactors, it was observed that cyanide removal (83.3 %) was predominant in the aerobic bioreactor, while much of the chemical oxygen demand (COD) (69 %) was consumed in the anoxic bioreactor. With the addition of cyanide, the COD removal efficiency was affected in all the bioreactors, and several intermediates were detected. While treating synthetic CWW using the combined bioreactor system, the overall COD removal efficiency was 86.79 % at an OLR of 2.4 g COD/L/day and an HRT of 96 h. The removal efficiency of 3,5-xylenol and cyanide, with inlet concentration of 150 and 10 mg/L, was found to be 91.8 and 93.6 % respectively. It was found that the impact of xylenol on the performance of the bioreactors was less than cyanide toxicity. Molecular analysis using T-RFLP revealed the dominance of strictly aerobic, mesophilic proteobacterium, Bosea minatitlanensis, in the aerobic bioreactor. The anoxic bioreactor was dominant with Rhodococcus pyridinivorans, known for its remarkable aromatic decomposing activity, while an unclassified Myxococcales bacterium was identified as the predominant bacterial species in the anaerobic bioreactor.

  4. Polysaccharide production in batch process of Neisseria meningitidis serogroup C comparing Frantz, modified Frantz and Cartlin 6 cultivation media

    Directory of Open Access Journals (Sweden)

    Paz Marcelo Fossa da

    2003-01-01

    Full Text Available Polysaccharide of N. meningitidis serogroup C constitutes the antigen for the vaccine against meningitis. The goal of this work was to compare three cultivation media for production of this polysaccharide: Frantz, modified Frantz medium (with replacement of glucose by glycerol, and Catlin 6 (a synthetic medium with glucose. The comparative criteria were based on the final polysaccharide concentrations and the yield coefficient cell/polysaccharide (Y P/X. The kinetic parameters: pH, substrate consumption and cell growth were also determined. For this purpose, 9 cultivation runs were carried out in a 80 L New Brunswick bioreactor, under the following conditions: 42 L of culture medium, temperature 35ºC, air flow 5 L/min, agitation frequency 120 rpm and vessel pressure 6 psi, without dissolved oxygen or pH controls. The cultivation runs were divided in three groups, with 3 repetitions each. The cultivation using the Frantz medium presented the best results: average of final polysaccharide concentration = 0.134 g/L and Y P/X=0.121, followed by Catlin 6 medium, with results of 0.095 g/L and 0.067 respectively. Considering the principal advantages in the use of the synthetic medium, i.e. facilitation of a cultivation and purification steps of the polysaccharide production process, there is a possibility that in the near future, Catlin 6 will replace the traditional Frantz medium.

  5. Bioconversion of high concentrations of hydrogen sulfide to elemental sulfur in airlift bioreactor.

    Science.gov (United States)

    Zytoon, Mohamed Abdel-Monaem; AlZahrani, Abdulraheem Ahmad; Noweir, Madbuli Hamed; El-Marakby, Fadia Ahmed

    2014-01-01

    Several bioreactor systems are used for biological treatment of hydrogen sulfide. Among these, airlift bioreactors are promising for the bioconversion of hydrogen sulfide into elemental sulfur. The performance of airlift bioreactors is not adequately understood, particularly when directly fed with hydrogen sulfide gas. The objective of this paper is to investigate the performance of an airlift bioreactor fed with high concentrations of H2S with special emphasis on the effect of pH in combination with other factors such as H2S loading rate, oxygen availability, and sulfide accumulation. H2S inlet concentrations between 1,008 ppm and 31,215 ppm were applied and elimination capacities up to 113 g H2S m(-3) h(-1) were achieved in the airlift bioreactor under investigation at a pH range 6.5-8.5. Acidic pH values reduced the elimination capacity. Elemental sulfur recovery up to 95% was achieved under oxygen limited conditions (DO bioreactor tolerated accumulated dissolved sulfide concentrations >500 mg/L at pH values 8.0-8.5, and near 100% removal efficiency was achieved. Overall, the resident microorganisms in the studied airlift bioreactor favored pH values in the alkaline range. The bioreactor performance in terms of elimination capacity and sulfur recovery was better at pH range 8-8.5.

  6. Biodegradation of toluene using Candida tropicalis immobilized on polymer matrices in fluidized bed bioreactors.

    Science.gov (United States)

    Song, JiHyeon; Namgung, HyeongKyu; Ahmed, Zubair

    2012-11-30

    A yeast strain, Candida tropicalis, was whole-cell-immobilized on polymer matrices of polyethylene glycol (PEG) and polyethylene glycol/activated carbon/alginate (PACA). The polymer matrices were used as fluidized materials in bubble-column bioreactors for the biodegradation of toluene. Simultaneously, another bubble-column bioreactor using granular activated carbon (GAC) and a conventional compost biofilter were operated for comparison. In the compost biofilter, the toluene removal efficiency gradually deteriorated due to the limitation of microbial activity. The toluene removal in the GAC bioreactor was relatively high because of an increase of toluene mass transfer. However, low toluene removal efficiencies were observed in the PEG bioreactor, presumably because the synthetic polymer alone was not suitable for yeast cell immobilization. In the PACA bioreactor, toluene removal was found to be greater than 95% overall. The CO(2) yield coefficient calculated at the highest toluene loading condition for the PACA bioreactor was found to be higher than those observed in the other bioreactors. Furthermore, almost complete elimination capacities were observed in the PACA bioreactor at short-term toluene loading up to 180 g/m(3)/h. In conclusion, the immobilization of C. tropicalis in the PACA matrix resulted in enhanced toluene biodegradation because of the increases of both mass transfer and microbial activity.

  7. Comparing the Bio-Hydrogen Production Potential of Pretreated Rice Straw Co-Digested with Seeded Sludge Using an Anaerobic Bioreactor under Mesophilic Thermophilic Conditions

    Directory of Open Access Journals (Sweden)

    Asma Sattar

    2016-03-01

    Full Text Available Three common pretreatments (mechanical, steam explosion and chemical used to enhance the biodegradability of rice straw were compared on the basis of bio-hydrogen production potential while co-digesting rice straw with sludge under mesophilic (37 °C and thermophilic (55 °C temperatures. The results showed that the solid state NaOH pretreatment returned the highest experimental reduction of LCH (lignin, cellulose and hemi-cellulose content and bio-hydrogen production from rice straw. The increase in incubation temperature from 37 °C to 55 °C increased the bio-hydrogen yield, and the highest experimental yield of 60.6 mL/g VSremoved was obtained under chemical pretreatment at 55 °C. The time required for maximum bio-hydrogen production was found on the basis of kinetic parameters as 36 h–47 h of incubation, which can be used as a hydraulic retention time for continuous bio-hydrogen production from rice straw. The optimum pH range of bio-hydrogen production was observed to be 6.7 ± 0.1–5.8 ± 0.1 and 7.1 ± 0.1–5.8 ± 0.1 under mesophilic and thermophilic conditions, respectively. The increase in temperature was found useful for controlling the volatile fatty acids (VFA under mechanical and steam explosion pretreatments. The comparison of pretreatment methods under the same set of experimental conditions in the present study provided a baseline for future research in order to select an appropriate pretreatment method.

  8. Effects of cultivation conditions on the uptake of arsenite and arsenic chemical species accumulated by Pteris vittata in hydroponics.

    Science.gov (United States)

    Hatayama, Masayoshi; Sato, Takahiko; Shinoda, Kozo; Inoue, Chihiro

    2011-03-01

    The physiological responses of the arsenic-hyperaccumulator, Pteris vittata, such as arsenic uptake and chemical transformation in the fern, have been investigated. However, a few questions remain regarding arsenic treatment in hydroponics. Incubation conditions such as aeration, arsenic concentration, and incubation period might affect those responses of P. vittata in hydroponics. Arsenite uptake was low under anaerobic conditions, as previously reported. However, in an arsenite uptake experiment, phosphorous (P) starvation-dependent uptake of arsenate was observed under aerobic conditions. Time course-dependent analysis of arsenite oxidation showed that arsenite was gradually oxidized to arsenate during incubation. Arsenite oxidation was not observed in any of the control conditions, such as exposure to a nutrient solution or to culture medium only, or with the use of dried root; arsenite oxidation was only observed when live root was used. This result suggests that sufficient aeration allows the rhizosphere system to oxidize arsenite and enables the fern to efficiently take up arsenite as arsenate. X-ray absorption near edge structure (XANES) analyses showed that long-duration exposure to arsenic using a hydroponic system led to the accumulation of arsenate as the dominant species in the root tips, but not in the whole roots, partly because up-regulation of arsenate uptake by P starvation of the fern was caused and retained by long-time incubation. Analysis of concentration-dependent arsenate uptake by P. vittata showed that the uptake switched from a high-affinity transport system to a low-affinity system at high arsenate concentrations, which partially explains the increased arsenate abundance in the whole root.

  9. A multiphysics/multiscale 2D numerical simulation of scaffold-based cartilage regeneration under interstitial perfusion in a bioreactor.

    Science.gov (United States)

    Sacco, Riccardo; Causin, Paola; Zunino, Paolo; Raimondi, Manuela T

    2011-07-01

    In vitro tissue engineering is investigated as a potential source of functional tissue constructs for cartilage repair, as well as a model system for controlled studies of cartilage development and function. Among the different kinds of devices for the cultivation of 3D cartilage cell colonies, we consider here polymeric scaffold-based perfusion bioreactors, where an interstitial fluid supplies nutrients and oxygen to the growing biomass. At the same time, the fluid-induced shear acts as a physiologically relevant stimulus for the metabolic activity of cells, provided that the shear stress level is appropriately tuned. In this complex environment, mathematical and computational modeling can help in the optimal design of the bioreactor configuration. In this perspective, we propose a computational model for the simulation of the biomass growth, under given inlet and geometrical conditions, where nutrient concentration, fluid dynamic field and cell growth are consistently coupled. The biomass growth model is calibrated with respect to the shear stress dependence on experimental data using a simplified short-time analysis in which the nutrient concentration and the fluid-induced shear stress are assumed constant in time and uniform in space. Volume averaging techniques are used to derive effective parameters that allow to upscale the microscopic structural properties to the macroscopic level. The biomass growth predictions obtained in this way are significant for long times of culture.

  10. Numerical Simulation of Mass Transfer and Three-Dimensional Fabrication of Tissue-Engineered Cartilages Based on Chitosan/Gelatin Hybrid Hydrogel Scaffold in a Rotating Bioreactor.

    Science.gov (United States)

    Zhu, Yanxia; Song, Kedong; Jiang, Siyu; Chen, Jinglian; Tang, Lingzhi; Li, Siyuan; Fan, Jiangli; Wang, Yiwei; Zhao, Jiaquan; Liu, Tianqing

    2017-01-01

    Cartilage tissue engineering is believed to provide effective cartilage repair post-injuries or diseases. Biomedical materials play a key role in achieving successful culture and fabrication of cartilage. The physical properties of a chitosan/gelatin hybrid hydrogel scaffold make it an ideal cartilage biomimetic material. In this study, a chitosan/gelatin hybrid hydrogel was chosen to fabricate a tissue-engineered cartilage in vitro by inoculating human adipose-derived stem cells (ADSCs) at both dynamic and traditional static culture conditions. A bioreactor that provides a dynamic culture condition has received greater applications in tissue engineering due to its optimal mass transfer efficiency and its ability to simulate an equivalent physical environment compared to human body. In this study, prior to cell-scaffold fabrication experiment, mathematical simulations were confirmed with a mass transfer of glucose and TGF-β2 both in rotating wall vessel bioreactor (RWVB) and static culture conditions in early stage of culture via computational fluid dynamic (CFD) method. To further investigate the feasibility of the mass transfer efficiency of the bioreactor, this RWVB was adopted to fabricate three-dimensional cell-hydrogel cartilage constructs in a dynamic environment. The results showed that the mass transfer efficiency of RWVB was faster in achieving a final equilibrium compared to culture in static culture conditions. ADSCs culturing in RWVB expanded three times more compared to that in static condition over 10 days. Induced cell cultivation in a dynamic RWVB showed extensive expression of extracellular matrix, while the cell distribution was found much more uniformly distributing with full infiltration of extracellular matrix inside the porous scaffold. The increased mass transfer efficiency of glucose and TGF-β2 from RWVB promoted cellular proliferation and chondrogenic differentiation of ADSCs inside chitosan/gelatin hybrid hydrogel scaffolds. The

  11. Progress in bioreactors of bioartiifcial livers

    Institute of Scientific and Technical Information of China (English)

    Cheng-Bo Yu; Xiao-Ping Pan; Lan-Juan Li

    2009-01-01

    BACKGROUND: Bioartiifcial liver support systems are becoming an effective therapy for hepatic failure. Bioreactors, as key devices in these systems, can provide a favorable growth and metabolic environment, mass exchange, and immunological isolation as a platform. Currently, stagnancy in bioreactor research is the main factor restricting the development of bioartiifcial liver support systems. DATA SOURCES: A PubMed database search of English-language literature was performed to identify relevant articles using the keywords "bioreactor", "bioartiifcial liver", "hepatocyte", and "liver failure". More than 40 articles related to the bioreactors of bioartiifcial livers were reviewed. RESULTS: Some progress has been made in the improvement of structures, functions, and modiifed macromolecular materials related to bioreactors in recent years. The current data on the improvement of bioreactor conifgurations for bioartiifcial livers or on the potential of the use of certain scaffold materials in bioreactors, combined with the clinical efifcacy and safety evaluation of cultured hepatocytesin vitro, indicate that the AMC (Academic Medical Center) BAL bioreactor and MELS (modular extracorporeal liver support) BAL bioreactor system can partly replace the synthetic and metabolic functions of the liver in phaseⅠ clinical studies. In addition, it has been indicated that the microlfuidic PDMS (polydimethylsiloxane) bioreactor, or SlideBioreactor, and the microfabricated grooved bioreactor are appropriate for hepatocyte culture, which is also promising for bioartiifcial livers. Similarly, modiifed scaffolds can promote the adhesion, growth, and function of hepatocytes, and provide reliable materials for bioreactors.CONCLUSIONS: Bioreactors, as key devices in bioartiifcial livers, play an important role in the therapy for liver failure both now and in the future. Bioreactor conifgurations are indispensable for the development of bioartiifcial livers used for liver

  12. Reduced Order Dead-Beat Observers for a Bioreactor

    CERN Document Server

    Karafyllis, Iasson

    2010-01-01

    This paper studies the strong observability property and the reduced-order dead-beat observer design problem for a continuous bioreactor. New relationships between coexistence and strong observability, and checkable sufficient conditions for strong observability, are established for a chemostat with two competing microbial species. Furthermore, the dynamic output feedback stabilization problem is solved for the case of one species.

  13. Glass bead cultivation of fungi

    DEFF Research Database (Denmark)

    Droce, Aida; Sørensen, Jens Laurids; Giese, H.

    2013-01-01

    Production of bioactive compounds and enzymes from filamentous fungi is highly dependent on cultivation conditions. Here we present an easy way to cultivate filamentous fungi on glass beads that allow complete control of nutrient supply. Secondary metabolite production in Fusarium graminearum and...

  14. Preliminary Study on Airlift Membran—Bioreactor

    Institute of Scientific and Technical Information of China (English)

    XUNong; XINGWeihong; 等

    2002-01-01

    A new type of membrane bioreactor named “airlift membrane-bioreactor”is discussed.For municipal wastewater reclamation,the preliminary study on airlift membrane-bioreactor shows its good performance such as high flux and lower energy consumption.The airlift membrane-bioreactor is potentially applicable in bioengineering and environmental protection fields.

  15. Cultivation and Differentiation of Encapsulated hMSC-TERT in a Disposable Small-Scale Syringe-Like Fixed Bed Reactor

    DEFF Research Database (Denmark)

    Weber, Christian; Pohl, Sebastian; Pörtner, Ralf

    2007-01-01

    The use of commercially available plastic syringes is introduced as disposable small-scale fixed bed bioreactors for the cultivation of implantable therapeutic cell systems on the basis of an alginate-encapsulated human mesenchymal stem cell line. The system introduced is fitted with a noninvasiv...... the fixed bed reactor an interesting option for GMP processes. The cultivation of the encapsulated cells in the fixed bed bioreactor system offered vitalities and adipogenic differentiation similar to well-mixed suspension cultures....

  16. Continuous gluconic acid production by the yeast-like Aureobasidium pullulans in a cascading operation of two bioreactors.

    Science.gov (United States)

    Anastassiadis, Savas; Rehm, Hans-Jürgen

    2006-12-01

    The application of a new developed process for the continuous production of gluconic acid using a cascade of two bioreactors in a continuous process is shown reaching the highest concentration of gluconic acid described in the literature for continuous culture fermentation. Very high gluconic acid concentrations of 272-308 g/l have been achieved under continuous cultivation of free-growing cells of Aureobasidium pullulans in the first bioreactor at residence times (RT) between 19.5 and 24 h with formation rates for the generic product between 12.7 and 13.9 g/(l h). Gluconic acid, 350-370 g/l, was continuously reached in the second bioreactor at a total RT of 30.8-37 h with R (j) of 9.2-12 g/(l h). The highest specific gluconic acid production (m (p)) of 3.6 g/(g h) was found in the first bioreactor at the lowest RT of 19.5 h. The highest selectivity of 93.6% was determined in the first bioreactor as well. Complete glucose consumption was obtained at 37 h total residence time in the second bioreactor. Gluconic acid, 433 g/l, was continuously produced in the second bioreactor at a total RT of 37 h.

  17. Bioreactor engineering using disposable technology for enhanced production of hCTLA4Ig in transgenic rice cell cultures.

    Science.gov (United States)

    Kwon, Jun-Young; Yang, Yong-Suk; Cheon, Su-Hwan; Nam, Hyung-Jin; Jin, Gi-Hong; Kim, Dong-Il

    2013-09-01

    Two kinds of disposable bioreactors, air-lift disposable bioreactors (ADB) and wave disposable bioreactors (WDB) were compared with stirred-tank reactors (5-L STR). These bioreactors were successfully applied to transgenic rice cell cultures for the production of recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig). In both systems, a fed-batch culture method was used to produce hCTLA4Ig efficiently by feeding concentrated amino acids and production levels were enhanced when dissolved oxygen (DO) level was regulated at 30% using pure oxygen sparging. Agitation and aeration rate during cultivation in ADB and WDB were determined by the same mixing time. The results in both disposable bioreactors showed similar values in maximum cell density (11.9 gDCW/L and 12.6 gDCW/L), doubling time (4.8- and 5.0-day), and maximum hCTLA4Ig concentration (43.7 and 43.3 mg/L). Relatively higher cell viability was sustained in the ADB whereas hCTLA4Ig productivity was 1.2-fold higher than that in WDB. The productivity was improved by increasing aeration rate (0.2 vvm). Overall, our experiments demonstrate pneumatically driven disposable bioreactors are applicable for the production of recombinant proteins in plant cell cultures. These results will be useful for development and scale-up studies of disposable bioreactor systems for transgenic plant cell cultures.

  18. Tissue grown in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Cells from kidneys lose some of their special features in conventional culture but form spheres replete with specialized cell microvilli (hair) and synthesize hormones that may be clinically useful. Ground-based research studies have demonstrated that both normal and neoplastic cells and tissues recreate many of the characteristics in the NASA bioreactor that they display in vivo. Proximal kidney tubule cells that normally have rich apically oriented microvilli with intercellular clefts in the kidney do not form any of these structures in conventional two-dimensional monolayer culture. However, when normal proximal renal tubule cells are cultured in three-dimensions in the bioreactor, both the microvilli and the intercellular clefts form. This is important because, when the morphology is recreated, the function is more likely also to be rejuvenated. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC).

  19. Growth of Steptomyces hygroscopicus in rotating-wall bioreactor under simulated microgravity inhibits rapamycin production

    Science.gov (United States)

    Fang, A.; Pierson, D. L.; Mishra, S. K.; Demain, A. L.

    2000-01-01

    Growth of Streptomyces hygroscopicus under conditions of simulated microgravity in a rotating-wall bioreactor resulted in a pellet form of growth, lowered dry cell weight, and inhibition of rapamycin production. With the addition of Teflon beads to the bioreactor, growth became much less pelleted, dry cell weight increased but rapamycin production was still markedly inhibited. Growth under simulated microgravity favored extracellular production of rapamycin, in contrast to a greater percentage of cell-bound rapamycin observed under normal gravity conditions.

  20. Growth of Streptomyces Hygroscopicus in Rotating-Wall Bioreactor Under Simulated Microgravity Inhibits Rapamycin Production

    Science.gov (United States)

    Fang, A.; Pierson, D. L.; Mishra, S. K.; Demain, A. L.

    2000-01-01

    Growth of Streptomyces hygroscopicus under conditions of simulated microgravity in a rotating-wall bioreactor resulted in a pellet form of growth, lowered dry cell weight, and inhibition of rapamycin production. With the addition of Teflon beads to the bioreactor, growth became much less pelleted, dry cell weight increased but rapamycin production was still markedly inhibited. Growth under simulated microgravity favored extracellular production of rapamycin in contrast to a greater percentage of cell-bound rapamycin observed under normal gravity conditions.

  1. Streamlined bioreactor-based production of human cartilage tissues.

    Science.gov (United States)

    Tonnarelli, B; Santoro, R; Adelaide Asnaghi, M; Wendt, D

    2016-05-27

    Engineered tissue grafts have been manufactured using methods based predominantly on traditional labour-intensive manual benchtop techniques. These methods impart significant regulatory and economic challenges, hindering the successful translation of engineered tissue products to the clinic. Alternatively, bioreactor-based production systems have the potential to overcome such limitations. In this work, we present an innovative manufacturing approach to engineer cartilage tissue within a single bioreactor system, starting from freshly isolated human primary chondrocytes, through the generation of cartilaginous tissue grafts. The limited number of primary chondrocytes that can be isolated from a small clinically-sized cartilage biopsy could be seeded and extensively expanded directly within a 3D scaffold in our perfusion bioreactor (5.4 ± 0.9 doublings in 2 weeks), bypassing conventional 2D expansion in flasks. Chondrocytes expanded in 3D scaffolds better maintained a chondrogenic phenotype than chondrocytes expanded on plastic flasks (collagen type II mRNA, 18-fold; Sox-9, 11-fold). After this "3D expansion" phase, bioreactor culture conditions were changed to subsequently support chondrogenic differentiation for two weeks. Engineered tissues based on 3D-expanded chondrocytes were more cartilaginous than tissues generated from chondrocytes previously expanded in flasks. We then demonstrated that this streamlined bioreactor-based process could be adapted to effectively generate up-scaled cartilage grafts in a size with clinical relevance (50 mm diameter). Streamlined and robust tissue engineering processes, as the one described here, may be key for the future manufacturing of grafts for clinical applications, as they facilitate the establishment of compact and closed bioreactor-based production systems, with minimal automation requirements, lower operating costs, and increased compliance to regulatory guidelines.

  2. Novel bacterial sulfur oxygenase reductases from bioreactors treating gold-bearing concentrates

    DEFF Research Database (Denmark)

    Chen, Z-W; Liu, Y-Y; Wu, J-F;

    2007-01-01

    The microbial community and sulfur oxygenase reductases of metagenomic DNA from bioreactors treating gold-bearing concentrates were studied by 16S rRNA library, real-time polymerase chain reaction (RT-PCR), conventional cultivation, and molecular cloning. Results indicated that major bacterial......) of bacteria and archaea were 4.59 x 10(9) and 6.68 x 10(5), respectively. Bacterial strains representing Acidithiobacillus, Leptospirillum, and Sulfobacillus were isolated from the bioreactors. To study sulfur oxidation in the reactors, pairs of new PCR primers were designed for the detection of sulfur...... oxygenase reductase (SOR) genes. Three sor-like genes, namely, sor (Fx), sor (SA), and sor (SB) were identified from metagenomic DNAs of the bioreactors. The sor (Fx) is an inactivated SOR gene and is identical to the pseudo-SOR gene of Ferroplasma acidarmanus. The sor (SA) and sor (SB) showed...

  3. Evaluation of a novel choanoid fluidized bed bioreactor for future bioartificial livers

    Science.gov (United States)

    Yu, Cheng-Bo; Pan, Xiao-Ping; Yu, Liang; Yu, Xiao-Peng; Du, Wei-Bo; Cao, Hong-Cui; Li, Jun; Chen, Ping; Li, Lan-Juan

    2014-01-01

    AIM: To construct and evaluate the functionality of a choanoid-fluidized bed bioreactor (CFBB) based on microencapsulated immortalized human hepatocytes. METHODS: Encapsulated hepatocytes were placed in the constructed CFBB and circulated through Dulbecco’s Modified Eagle’s Medium (DMEM) for 12 h, and then through exchanged plasma for 6 h, and compared with encapsulated cells cultivated under static conditions in a spinner flask. Levels of alanine aminotransferase (ALT) and albumin were used to evaluate the CFBB during media circulation, whereas levels of ALT, total bilirubin (TBil), and albumin were used to evaluate it during plasma circulation. Mass transfer and hepatocyte injury were evaluated by comparing the results from the two experimental conditions. In addition, the viability and microstructure of encapsulated cells were observed in the different environments. RESULTS: The bioartificial liver model based on a CFBB was verified by in vitro experiments. The viability of encapsulated cells accounting for 84.6% ± 3.7% in CFBB plasma perfusion was higher than the 74.8% ± 3.1% in the static culture group (P circulation and static medium culture groups, respectively. Albumin secretion from cells was 234.2 ± 27.8 μg/1 × 107 cells vs 167.8 ± 29.3 μg/1 × 107 cells at 6 h (P circulation/culture groups, respectively. Furthermore, ALT and TBil levels were 172.3 ± 24.1 U/L vs 236.3 ± 21.5 U/L (P 0.05) at 6 h in the CFBB plasma perfusion and static plasma culture groups, respectively. There was no significant difference in albumin concentration between the two experimental plasma groups at any time point. The microstructure of the encapsulated hepatocytes remained healthier in the CFBB group compared with the static culture group after 6 h of plasma perfusion. CONCLUSION: The CFBB can function as a bioartificial liver based on a bioreactor. The efficacy of this novel bioreactor is promising for the study of liver failure. PMID:24944477

  4. Reducing conditions are the key for efficient production of active ribonuclease inhibitor in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Neubauer Peter

    2011-05-01

    Full Text Available Abstract Background The eukaryotic RNase ribonuclease/angiogenin inhibitors (RI are a protein group distinguished by a unique structure - they are composed of hydrophobic leucine-rich repeat motifs (LRR and contain a high amount of reduced cysteine residues. The members of this group are difficult to produce in E. coli and other recombinant hosts due to their high aggregation tendency. Results In this work dithiothreitol (DTT was successfully applied for improving the yield of correctly folded ribonuclease/angiogenin inhibitor in E. coli K12 periplasmic and cytoplasmic compartments. The feasibility of the in vivo folding concepts for cytoplasmic and periplasmic production were demonstrated at batch and fed-batch cultivation modes in shake flasks and at the bioreactor scale. Firstly, the best secretion conditions of RI in the periplasmic space were evaluated by using a high throughput multifactorial screening approach of a vector library, directly with the Enbase fed-batch production mode in 96-well plates. Secondly, the effect of the redox environment was evaluated in isogenic dsbA+ and dsbA- strains at the various cultivation conditions with reducing agents in the cultivation medium. Despite the fusion to the signal peptide, highest activities were found in the cytoplasmic fraction. Thus by removing the signal peptide the positive effect of the reducing agent DTT was clearly proven also for the cytoplasmic compartment. Finally, optimal periplasmic and cytoplasmic RI fed-batch production processes involving externally added DTT were developed in shake flasks and scaled up to the bioreactor scale. Conclusions DTT highly improved both, periplasmic and cytoplasmic accumulation and activity of RI at low synthesis rate, i.e. in constructs harbouring weak recombinant synthesis rate stipulating genetic elements together with cultivation at low temperature. In a stirred bioreactor environment RI folding was strongly improved by repeated pulse addition

  5. Use Alkalinity Monitoring to Optimize Bioreactor Performance.

    Science.gov (United States)

    Jones, Christopher S; Kult, Keegan J

    2016-05-01

    In recent years, the agricultural community has reduced flow of nitrogen from farmed landscapes to stream networks through the use of woodchip denitrification bioreactors. Although deployment of this practice is becoming more common to treat high-nitrate water from agricultural drainage pipes, information about bioreactor management strategies is sparse. This study focuses on the use of water monitoring, and especially the use of alkalinity monitoring, in five Iowa woodchip bioreactors to provide insights into and to help manage bioreactor chemistry in ways that will produce desirable outcomes. Results reported here for the five bioreactors show average annual nitrate load reductions between 50 and 80%, which is acceptable according to established practice standards. Alkalinity data, however, imply that nitrous oxide formation may have regularly occurred in at least three of the bioreactors that are considered to be closed systems. Nitrous oxide measurements of influent and effluent water provide evidence that alkalinity may be an important indicator of bioreactor performance. Bioreactor chemistry can be managed by manipulation of water throughput in ways that produce adequate nitrate removal while preventing undesirable side effects. We conclude that (i) water should be retained for longer periods of time in bioreactors where nitrous oxide formation is indicated, (ii) measuring only nitrate and sulfate concentrations is insufficient for proper bioreactor operation, and (iii) alkalinity monitoring should be implemented into protocols for bioreactor management.

  6. A photosynthetic rotating annular bioreactor (Taylor-Couette type flow) for phototrophic biofilm cultures.

    Science.gov (United States)

    Paule, A; Lauga, B; Ten-Hage, L; Morchain, J; Duran, R; Paul, E; Rols, J L

    2011-11-15

    In their natural environment, the structure and functioning of microbial communities from river phototrophic biofilms are driven by biotic and abiotic factors. An understanding of the mechanisms that mediate the community structure, its dynamics and the biological succession processes during phototrophic biofilm development can be gained using laboratory-scale systems operating with controlled parameters. For this purpose, we present the design and description of a new prototype of a rotating annular bioreactor (RAB) (Taylor-Couette type flow, liquid working volume of 5.04 L) specifically adapted for the cultivation and investigation of phototrophic biofilms. The innovation lies in the presence of a modular source of light inside of the system, with the biofilm colonization and development taking place on the stationary outer cylinder (onto 32 removable polyethylene plates). The biofilm cultures were investigated under controlled turbulent flowing conditions and nutrients were provided using a synthetic medium (tap water supplemented with nitrate, phosphate and silica) to favour the biofilm growth. The hydrodynamic features of the water flow were characterized using a tracer method, showing behaviour corresponding to a completely mixed reactor. Shear stress forces on the surface of plates were also quantified by computer simulations and correlated with the rotational speed of the inner cylinder. Two phototrophic biofilm development experiments were performed for periods of 6.7 and 7 weeks with different inoculation procedures and illumination intensities. For both experiments, biofilm biomasses exhibited linear growth kinetics and produced 4.2 and 2.4 mg cm(-)² of ash-free dry matter. Algal and bacterial community structures were assessed by microscopy and T-RFLP, respectively, and the two experiments were different but revealed similar temporal dynamics. Our study confirmed the performance and multipurpose nature of such an innovative photosynthetic bioreactor

  7. Engineering considerations for process development in mammalian cell cultivation.

    Science.gov (United States)

    Zhang, Hu; Wang, Weixiang; Quan, Chunshan; Fan, Shengdi

    2010-01-01

    Mammalian cell cultivation plays a great role in producing protein therapeutics in the last decades. Many engineering parameters are considered for optimization during process development in mammalian cell cultivation, only shear and mixing are especially highlighted in this paper. It is believed that shear stress due to agitation has been over-estimated to damage cells, but shear may result in nonlethal physiological responses. There is no cell damage in the regions where bubbles form, break up and coalescence, but shear stress becomes significant in the wake of rising bubbles and causes great damage to cells in bubble burst regions. Mixing is not sufficient to provide homogeneous dissolved oxygen tension, pH, CO2 and nutrients in large-scale bioreactors, which can bring severe problems for cell growth, product formation and process control. Scale-down reactors have been developed to address mixing and shear problems for parallel operations. Engineering characterization in conventional and recently developed scale-down bioreactors has been briefly introduced. Process challenges for cultivation of industrial cell lines in high cell densities as well as cultivation of stem cells and other human cells for regenerative medicine, tissue engineering and gene therapy are prospected. Important techniques, such as micromanipulation and nanomanipulation (optical tweezers) for single cell analysis, computational fluid dynamics (CFD) for shear and mixing characterization, and miniaturized bioreactors, are being developed to address those challenges.

  8. Cultivation of two thermotolerant microalgae under tropical conditions: Influences of carbon sources and light duration on biomass and lutein productivity in four seasons.

    Science.gov (United States)

    Chiu, Pai-Ho; Soong, Keryea; Chen, Ching-Nen Nathan

    2016-07-01

    Biomass and lutein productivities of two thermotolerant microalgae were assessed in tropical outdoor conditions in all four seasons. Generally, addition of bicarbonate at 0.2g/L every two days or 2% CO2 did not enhance the productivities compared to the controls, and the productivities in the spring were higher than in the fall. Durations of effective irradiance positively correlated to the productivity of Coelastrella sp. F50 well, but not for Desmodesmus sp. F2. The ineffective light intensity was below 5000 lux (about 106μmol/m(2)s). The productivities produced in the 17cm diameter bottles were 1.5- to 1.9-fold higher than that in the 27cm ones. Lutein content, about 0.5% in biomass on average, did not change significantly grown in different carbon sources or seasons. The annual productivities of the microalgal biomass and lutein in one hectare were estimated to be 33tons and 180kg, respectively, using the non-optimized photobioreactor cultivation.

  9. 聊城灵芝种植的气象条件适宜性分析%Analysis of Weather Conditions for Ganoderma lucidum Cultivation in Liaocheng

    Institute of Scientific and Technical Information of China (English)

    孙青然; 赵芬; 郭晓霞; 王丽; 孙瑞英

    2011-01-01

    [目的分析灵芝(Ganoderma Lucidum)对气象条件的要求,并探讨聊城市适宜灵芝种植的气象因子灾害性天气.[方法]根据灵芝对气象条件的要求,通过分析聊城市4~10月的月平均气温和月平均相对湿度;最后针对聊城市每年4~10月发生的灾害性天气提出了防御措施.[结果]在聊城4月下旬比较适宜灵芝种植接种.[结论]该研究可为地方政府部门决策及灵芝的种植者提供参考.%[ Objective ] To investigate the suitable weather conditions for the cultivation of Ganoderma lucidum in Liaocheng, and propose the defensive measures against disastrous weather. [ Method ] Monthly average temperature and relative humidity from April to October in Liaocheng have been analyzed in this paper. [ Result] The rational planting period of Ganoderma lucidum in Liaocheng was the last ten-day of April.

  10. Induction of secondary metabolism of Aspergillus terreus ATCC 20542 in the batch bioreactor cultures.

    Science.gov (United States)

    Boruta, Tomasz; Bizukojc, Marcin

    2016-04-01

    Cultivation of Aspergillus terreus ATCC 20542 in a stirred tank bioreactor was performed to induce the biosynthesis of secondary metabolites and provide the bioprocess-related insights into the metabolic capabilities of the investigated strain. The activation of biosynthetic routes was attempted by the diversification of process conditions and growth media. Several strategies were tested, including the addition of rapeseed oil or inulin, changing the concentration of nitrogen source, reduction of chlorine supply, cultivation under saline conditions, and using various aeration schemes. Fifteen secondary metabolites were identified in the course of the study by using ultra-high performance liquid chromatography coupled with mass spectrometry, namely mevinolinic acid, 4a,5-dihydromevinolinic acid, 3α-hydroxy-3,5-dihydromonacolin L acid, terrein, aspulvinone E, dihydroisoflavipucine, (+)-geodin, (+)-bisdechlorogeodin, (+)-erdin, asterric acid, butyrolactone I, desmethylsulochrin, questin, sulochrin, and demethylasterric acid. The study also presents the collection of mass spectra that can serve as a resource for future experiments. The growth in a salt-rich environment turned out to be strongly inhibitory for secondary metabolism and the formation of dense and compact pellets was observed. Generally, the addition of inulin, reducing the oxygen supply, and increasing the content of nitrogen source did not enhance the production of examined molecules. The most successful strategy involved the addition of rapeseed oil to the chlorine-deficient medium. Under these conditions, the highest levels of butyrolactone I, asterric acid, and mevinolinic acid were achieved and the presence of desmethylsulochrin and (+)-bisdechlorogeodin was detected in the broth. The constant and relatively high aeration rate in the idiophase was shown to be beneficial for terrein and (+)-geodin biosynthesis.

  11. Analysis of drug metabolism activities in a miniaturized liver cell bioreactor for use in pharmacological studies.

    Science.gov (United States)

    Hoffmann, Stefan A; Müller-Vieira, Ursula; Biemel, Klaus; Knobeloch, Daniel; Heydel, Sandra; Lübberstedt, Marc; Nüssler, Andreas K; Andersson, Tommy B; Gerlach, Jörg C; Zeilinger, Katrin

    2012-12-01

    Based on a hollow fiber perfusion technology with internal oxygenation, a miniaturized bioreactor with a volume of 0.5 mL for in vitro studies was recently developed. Here, the suitability of this novel culture system for pharmacological studies was investigated, focusing on the model drug diclofenac. Primary human liver cells were cultivated in bioreactors and in conventional monolayer cultures in parallel over 10 days. From day 3 on, diclofenac was continuously applied at a therapeutic concentration (6.4 µM) for analysis of its metabolism. In addition, the activity and gene expression of the cytochrome P450 (CYP) isoforms CYP1A2, CYP2B6, CYP2C9, CYP2D6, and CYP3A4 were assessed. Diclofenac was metabolized in bioreactor cultures with an initial conversion rate of 230 ± 57 pmol/h/10(6) cells followed by a period of stable conversion of about 100 pmol/h/10(6) cells. All CYP activities tested were maintained until day 10 of bioreactor culture. The expression of corresponding mRNAs correlated well with the degree of preservation. Immunohistochemical characterization showed the formation of neo-tissue with expression of CYP2C9 and CYP3A4 and the drug transporters breast cancer resistance protein (BCRP) and multidrug resistance protein 2 (MRP2) in the bioreactor. In contrast, monolayer cultures showed a rapid decline of diclofenac conversion and cells had largely lost activity and mRNA expression of the assessed CYP isoforms at the end of the culture period. In conclusion, diclofenac metabolism, CYP activities and gene expression levels were considerably more stable in bioreactor cultures, making the novel bioreactor a useful tool for pharmacological or toxicological investigations requiring a highly physiological in vitro representation of the liver.

  12. Production of Calcaride A by Calcarisporium sp. in Shaken Flasks and Stirred Bioreactors

    Directory of Open Access Journals (Sweden)

    Anu Tamminen

    2015-06-01

    Full Text Available Increased interest in marine resources has led to increased screening of marine fungi for novel bioactive compounds and considerable effort is being invested in discovering these metabolites. For compound discovery, small-scale cultures are adequate, but agitated bioreactors are desirable for larger-scale production. Calcarisporium sp. KF525 has recently been described to produce calcaride A, a cyclic polyester with antibiotic activity, in agitated flasks. Here, we describe improvements in the production of calcaride A in both flasks (13-fold improvement and stirred bioreactors (200-fold improvement. Production of calcaride A in bioreactors was initially substantially lower than in shaken flasks. The cultivation pH (reduced from 6.8 to <5.4, carbon source (sucrose replacing glucose, C/N ratio and nature of mycelial growth (pellets or filaments were important in improving calcaride A production. Up to 4.5 mg·g−1 biomass (85 mg·L−1 calcaride A were produced in the bioreactor, which was only slightly less than in shaken flasks (14 mg·g−1, 100 mg·L−1. The results demonstrate that a scalable process for calcaride A production could be developed using an iterative approach with flasks and bioreactors.

  13. Continuous plant cell perfusion culture: bioreactor characterization and secreted enzyme production.

    Science.gov (United States)

    Su, Wei Wen; Arias, Renee

    2003-01-01

    Culture perfusion is widely practiced in mammalian cell processes to enhance secreted antibody production. Here, we report the development of an efficient continuous perfusion process for the cultivation of plant cell suspensions. The key to this process is a perfusion bioreactor that incorporates an annular settling zone into a stirred-tank bioreactor to achieve continuous cell/medium separation via gravitational sedimentation. From washout experiments, we found that under typical operating conditions (e.g., 200 rpm and 0.3 vvm) the liquid phase in the entire perfusion bioreactor was homogeneous despite the presence of the cylindrical baffle. Using secreted acid phosphatase (APase) produced in Anchusa officinalis cell culture as a model we have studied the perfusion cultures under complete or partial cell retention. The perfusion culture was operated under phosphate limitation to stimulate APase production. Successful operation of the perfusion process over four weeks has been achieved in this work. When A. officinalis cells were grown in the perfusion reactor and perfused at up to 0.4 vvd with complete cell retention, a cell dry weight exceeding 20 g/l could be achieved while secreted APase productivity leveled off at approximately 300 units/l/d. The culture became extremely dense with the maximum packed cell volume (PCV) surpassing 70%. In comparison, the maximum cell dry weight and overall secreted APase productivity in a typical batch culture were 10-12 g/l and 100-150 units/l/d, respectively. Operation of the perfusion culture under extremely high PCV for a prolonged period, however, led to declined oxygen uptake and reduced viability. Subsequently, cell removal via a bleed stream at up to 0.11 vvd was tested and shown to stabilize the culture at a PCV below 60%. With culture bleeding, both specific oxygen uptake rate and viability were shown to increase. This also led to a higher cell dry weight exceeding 25 g/l, and further improvement of secreted APase

  14. Thinking beyond the Bioreactor Box: Incorporating Stream Ecology into Edge-of-Field Nitrate Management.

    Science.gov (United States)

    Goeller, Brandon C; Febria, Catherine M; Harding, Jon S; McIntosh, Angus R

    2016-05-01

    Around the world, artificially drained agricultural lands are significant sources of reactive nitrogen to stream ecosystems, creating substantial stream health problems. One management strategy is the deployment of denitrification enhancement tools. Here, we evaluate the factors affecting the potential of denitrifying bioreactors to improve stream health and ecosystem services. The performance of bioreactors and the structure and functioning of stream biotic communities are linked by environmental parameters like dissolved oxygen and nitrate-nitrogen concentrations, dissolved organic carbon availability, flow and temperature regimes, and fine sediment accumulations. However, evidence of bioreactors' ability to improve waterway health and ecosystem services is lacking. To improve the potential of bioreactors to enhance desirable stream ecosystem functioning, future assessments of field-scale bioreactors should evaluate the influences of bioreactor performance on ecological indicators such as primary production, organic matter processing, stream metabolism, and invertebrate and fish assemblage structure and function. These stream health impact assessments should be conducted at ecologically relevant spatial and temporal scales. Bioreactors have great potential to make significant contributions to improving water quality, stream health, and ecosystem services if they are tailored to site-specific conditions and implemented strategically with land-based and stream-based mitigation tools within watersheds. This will involve combining economic, logistical, and ecological information in their implementation.

  15. A xenogeneic-free bioreactor system for the clinical-scale expansion of human mesenchymal stem/stromal cells.

    Science.gov (United States)

    Dos Santos, Francisco; Campbell, Andrew; Fernandes-Platzgummer, Ana; Andrade, Pedro Z; Gimble, Jeffrey M; Wen, Yuan; Boucher, Shayne; Vemuri, Mohan C; da Silva, Cláudia L; Cabral, Joaquim M S

    2014-06-01

    The large cell doses (>1 × 10(6)  cells/kg) used in clinical trials with mesenchymal stem/stromal cells (MSC) will require an efficient production process. Moreover, monitoring and control of MSC ex-vivo expansion is critical to provide a safe and reliable cell product. Bioprocess engineering approaches, such as bioreactor technology, offer the adequate tools to develop and optimize a cost-effective culture system for the rapid expansion of human MSC for cellular therapy. Herein, a xenogeneic (xeno)-free microcarrier-based culture system was successfully established for bone marrow (BM) MSC and adipose tissue-derived stem/stromal cell (ASC) cultivation using a 1L-scale controlled stirred-tank bioreactor, allowing the production of (1.1 ± 0.1) × 10(8) and (4.5 ± 0.2) × 10(7) cells for BM MSC and ASC, respectively, after 7 days. Additionally, the effect of different percent air saturation values (%Airsat ) and feeding regime on the proliferation and metabolism of BM MSC was evaluated. No significant differences in cell growth and metabolic patterns were observed under 20% and 9%Airsat . Also, the three different feeding regimes studied-(i) 25% daily medium renewal, (ii) 25% medium renewal every 2 days, and (iii) fed-batch addition of concentrated nutrients and growth factors every 2 days-yielded similar cell numbers, and only slight metabolic differences were observed. Moreover, the immunophenotype (positive for CD73, CD90 and CD105 and negative for CD31, CD80 and HLA-DR) and multilineage differentiative potential of expanded cells were not affected upon bioreactor culture. These results demonstrated the feasibility of expanding human MSC from different sources in a clinically relevant expansion configuration in a controlled microcarrier-based stirred culture system under xeno-free conditions. The further optimization of this bioreactor culture system will represent a crucial step towards an efficient GMP-compliant clinical-scale MSC

  16. Nutrient removal from an anaerobic membrane bioreactor effluent using microalgae. Study and modeling of the process.

    OpenAIRE

    Ruiz Martínez, Ana

    2016-01-01

    [EN] Anaerobic membrane bioreactors for urban wastewater treatment present interesting advantages when compared with aerobic treatments, such as less sludge production, lower energy demand and biogas generation. However, the generated effluent cannot generally be discharged without further ammonium and phosphate elimination. This thesis studies the removal of these inorganic nutrients by means of microalgae cultivation. The main objective of this work is therefore to obtain an autochthono...

  17. Introducing Textiles as Material of Construction of Ethanol Bioreactors

    Directory of Open Access Journals (Sweden)

    Osagie A. Osadolor

    2014-11-01

    Full Text Available The conventional materials for constructing bioreactors for ethanol production are stainless and cladded carbon steel because of the corrosive behaviour of the fermenting media. As an alternative and cheaper material of construction, a novel textile bioreactor was developed and examined. The textile, coated with several layers to withstand the pressure, resist the chemicals inside the reactor and to be gas-proof was welded to form a 30 L lab reactor. The reactor had excellent performance for fermentative production of bioethanol from sugar using baker’s yeast. Experiments with temperature and mixing as process parameters were performed. No bacterial contamination was observed. Bioethanol was produced for all conditions considered with the optimum fermentation time of 15 h and ethanol yield of 0.48 g/g sucrose. The need for mixing and temperature control can be eliminated. Using a textile bioreactor at room temperature of 22 °C without mixing required 2.5 times longer retention time to produce bioethanol than at 30 °C with mixing. This will reduce the fermentation investment cost by 26% for an ethanol plant with capacity of 100,000 m3 ethanol/y. Also, replacing one 1300 m3 stainless steel reactor with 1300 m3 of the textile bioreactor in this plant will reduce the fermentation investment cost by 19%.

  18. PRACTICE REVIEW OF FIVE BIOREACTOR/RECIRCULATION LANDFILLS

    Science.gov (United States)

    Six bioreactor landfills were analyzed to provide a perspective of current practice and technical issues that differentiate bioreactor landfills from conventional landfills. Five of the bioreactor landfills were anaerobic and one was aerated. In one case, nearly identical cells e...

  19. Bioreactor design and optimization – a future perspective

    DEFF Research Database (Denmark)

    Gernaey, Krist

    2011-01-01

    Bioreactor design and optimisation are essential in translating the experience gained from lab or pilot scale experiments to efficient production processes in industrial scale bioreactors. This article gives a future perspective on bioreactor design and optimisation, where it is foreseen...

  20. Scale-up impacts on mass transfer and bioremediation of suspended naphthalene particles in bead mill bioreactors.

    Science.gov (United States)

    Wang, Yuching; Riess, Ryan; Nemati, Mehdi; Hill, Gordon; Headley, John

    2008-11-01

    Scale-up effects on mass transfer and bioremediation of suspended naphthalene particles have been studied in 20 and 58L bead mill bioreactors and compared to data generated earlier with a laboratory scaled bioreactor. The bead mill bioreactor performance with respect to naphthalene mass transfer rate was dependent on the size and loading of the inert particles, as well as the rotational speed of the roller apparatus. The optimum operating conditions were found to be 15mm glass beads at a loading of 50% (total volume of particles/working volume of bioreactor: v/v%) and a bioreactor rotational speed of 50rpm. The highest naphthalene mass transfer coefficients obtained in the large scale system under these optimum conditions (19.6 and 22.4h(-1) for 20 and 58L vessels, respectively) were higher than those determined previously in a 2.5L bead mill bioreactor (0.7h(-1)). The acute toxicity tests indicated that the bioreactor effluent was less toxic than the untreated naphthalene suspension. Biodegradation rates obtained in these large scale bead mill bioreactors under optimum conditions (36-37.4mgL(-1)h(-1)) were higher than those achieved in the control bioreactors of similar sizes (11.4 and 11.6mgL(-1)h(-1)) but were slower than those previously determined in a 2.5L bead mill bioreactor (59-61.5mgL(-1)h(-1)). The limitation of oxygen in the large scale systems and damage of the bacterial cells due to the crushing effects of the large beads are likely contributing factors in the lower observed biodegradation rates. The optimum conditions with respect to naphthalene mass transfer might not necessarily translate to optimum performance with regard to bioremediation.

  1. Long-term outdoor cultivation by perfusing spent medium for biodiesel production from Chlorella minutissima.

    Science.gov (United States)

    Oh, Sung Ho; Kwon, Min Chul; Choi, Woon Yong; Seo, Yong Chang; Kim, Ga Bin; Kang, Do Hyung; Lee, Shin Young; Lee, Hyeon Yong

    2010-08-01

    A unique perfusion process was developed to maintain high concentrations of marine alga, Chlorella minutissima. This method is based on recycling cells by continuous feeding with warm spent sea water from nuclear power plants, which has very similar properties as sea water. A temperature of at least 30 degrees C in a 200 L photo-bioreactor was maintained in this system by perfusion of the thermal plume for 80 days in the coldest season. The maximum cell concentration and total lipid content was 8.3 g-dry wt./L and 23.2 %, w/w, respectively, under mixotrophic conditions. Lipid production was found to be due to a partially or non-growth related process, which implies that large amounts of biomass are needed for a high accumulation of lipids within the cells. At perfusion rates greater than 1.5 L/h, the temperature of the medium inside the reactor was around 30 degrees C, which was optimal for cell growth. For this system, a perfusion rate of 2.8 L/h was determined to be optimal for maintaining rapid cell growth and lipid production during outdoor cultivation. It was absolutely necessary to maintain the appropriate perfusion rate so that the medium temperature was optimal for cell growth. In addition, the lipids produced using this process were shown to be feasible for biodiesel production since the lipid composition of C. minutissima grown under these conditions consisted of 17 % (w/w) of C(16) and 47% (w/w) of C(18). The combined results of this study clearly demonstrated that the discharged energy of the thermal plume could be reused to cultivate marine alga by maintaining a relatively constant temperature in an outdoor photo-bioreactor without the need for supplying any extra energy, which could allow for cheap production of biodiesel from waste energy.

  2. Lactic acid Production with in situ Extraction in Membrane Bioreactor

    Directory of Open Access Journals (Sweden)

    Hamidreza Ghafouri Taleghani

    2017-01-01

    Full Text Available Background and Objective: Lactic acid is widely used in the food, chemical and pharmaceutical industries. The major problems associated with lactic acid production are substrate and end-product inhibition, and by-product formation. Membrane technologyrepresents one of the most effective processes for lactic acid production. The aim of this work is to increase cell density and lactic acid productivity due to reduced inhibition effect of substrate and product in membrane bioreactor.Material and Methods: In this work, lactic acid was produced from lactose in membrane bioreactor. A laboratory scale membrane bioreactor was designed and fabricated. Five types of commercial membranes were tested at the same operating conditions (transmembrane pressure: 500 KPa and temperature: 25°C. The effects of initial lactose concentration and dilution rate on biomass growth, lactic acid production and substrate utilization were evaluated.Results and Conclusion: The high lactose retention of 79% v v-1 and low lactic acid retention of 22% v v-1 were obtained with NF1 membrane; therefore, this membrane was selected for membrane bioreactor. The maximal productivity of 17.1 g l-1 h-1 was obtainedwith the lactic acid concentration of 71.5 g l-1 at the dilution rate of 0.24 h−1. The maximum concentration of lactic acid was obtained at the dilution rate of 0.04 h−1. The inhibiting effect of lactic acid was not observed at high initial lactose concentration. The critical lactose concentration at which the cell growth severely hampered was 150 g l-1. This study proved that membrane bioreactor had great advantages such as elimination of substrate and product inhibition, high concentration of process substrate, high cell density,and high lactic acid productivity.Conflict of interest: There is no conflict of interest.

  3. Methane production in simulated hybrid bioreactor landfill.

    Science.gov (United States)

    Xu, Qiyong; Jin, Xiao; Ma, Zeyu; Tao, Huchun; Ko, Jae Hac

    2014-09-01

    The aim of this work was to study a hybrid bioreactor landfill technology for landfill methane production from municipal solid waste. Two laboratory-scale columns were operated for about ten months to simulate an anaerobic and a hybrid landfill bioreactor, respectively. Leachate was recirculated into each column but aeration was conducted in the hybrid bioreactor during the first stage. Results showed that leachate pH in the anaerobic bioreactor maintained below 6.5, while in the hybrid bioreactor quickly increased from 5.6 to 7.0 due to the aeration. The temporary aeration resulted in lowering COD and BOD5 in the leachate. The volume of methane collected from the hybrid bioreactor was 400 times greater than that of the anaerobic bioreactor. Also, the methane production rate of the hybrid bioreactor was improved within a short period of time. After about 10 months' operation, the total methane production in the hybrid bioreactor was 212 L (16 L/kgwaste).

  4. Optimizing of Culture Conditionin Horizontal Rotating Bioreactor

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    1 IntroductionBioreactor is the most important equipment in tissue engineering. It can mimic the micro-environment of cell growth in vitro. At present, horizontal rotating bioreactor is the most advanced equipment for cell culture in the world. 2 Rotating bioreactors2.1 Working principleThere are two kinds of horizontal rotating bioreactor: HARV(high aspect ratio vessel) and RCCS (rotary cell culture system). It is drived by step motor with horizontal rotation, the culture medium and cell is filled between ...

  5. Review of nonconventional bioreactor technology

    Energy Technology Data Exchange (ETDEWEB)

    Turick, C.E.; Mcllwain, M.E.

    1993-09-01

    Biotechnology will significantly affect many industrial sectors in the future. Industrial sectors that will be affected include pharmaceutical, chemical, fuel, agricultural, and environmental remediation. Future research is needed to improve bioprocessing efficiency and cost-effectiveness in order to compete with traditional technologies. This report describes recent advances in bioprocess technologies and bioreactor designs and relates them to problems encountered in many industrial bioprocessing operations. The primary focus is directed towards increasing gas and vapor transfer for enhanced bioprocess kinetics as well as unproved by-product separation and removal. The advantages and disadvantages of various conceptual designs such as hollow-fiber, gas-phase, hyperbaric/hypobaric, and electrochemical bioreactors are also discussed. Specific applications that are intended for improved bioprocesses include coal desulfurization, coal liquefaction, soil bioremediation, biomass conversion to marketable chemicals, biomining, and biohydrometallurgy as well as bioprocessing of gases and vapors.

  6. Monolithic Continuous-Flow Bioreactors

    Science.gov (United States)

    Stephanopoulos, Gregory; Kornfield, Julia A.; Voecks, Gerald A.

    1993-01-01

    Monolithic ceramic matrices containing many small flow passages useful as continuous-flow bioreactors. Ceramic matrix containing passages made by extruding and firing suitable ceramic. Pores in matrix provide attachment medium for film of cells and allow free movement of solution. Material one not toxic to micro-organisms grown in reactor. In reactor, liquid nutrients flow over, and liquid reaction products flow from, cell culture immobilized in one set of channels while oxygen flows to, and gaseous reaction products flow from, culture in adjacent set of passages. Cells live on inner surfaces containing flowing nutrient and in pores of walls of passages. Ready access to nutrients and oxygen in channels. They generate continuous high yield characteristic of immobilized cells, without large expenditure of energy otherwise incurred if necessary to pump nutrient solution through dense biomass as in bioreactors of other types.

  7. Optimal Homogenization of Perfusion Flows in Microfluidic Bio-Reactors: A Numerical Study

    DEFF Research Database (Denmark)

    Okkels, Fridolin; Dufva, Martin; Bruus, Henrik

    2011-01-01

    In recent years, the interest in small-scale bio-reactors has increased dramatically. To ensure homogeneous conditions within the complete area of perfused microfluidic bio-reactors, we develop a general design of a continually feed bio-reactor with uniform perfusion flow. This is achieved...... by introducing a specific type of perfusion inlet to the reaction area. The geometry of these inlets are found using the methods of topology optimization and shape optimization. The results are compared with two different analytic models, from which a general parametric description of the design is obtained...... and tested numerically. Such a parametric description will generally be beneficial for the design of a broad range of microfluidic bioreactors used for, e. g., cell culturing and analysis and in feeding bio-arrays....

  8. Biodegradation of the water-soluble gasoline components in a novel hybrid bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Gomez-De-Jesus, A.; Lara-Rodriguez, A.; Santoyo-Tepole, F.; Juarez-Ramirez, C.; Cristiani-Urbina, E.; Ruiz-Ordaz, N.; Galindez Mayer, J. [Escuela Nacional de Ciencias Biologicas, del Instituto Politecnico Nacional, Departamento de Ingenieria Bioquimica, Carpio y Plan de Ayala, ' ' Centro Operativo Naranjo' ' , Mexico, D.F. (Mexico)

    2003-07-01

    A novel hybrid bioreactor was designed to remove volatile organic compounds from water contaminated with water-soluble gasoline components, and the performance of this new bioreactor was investigated. It was composed of two biotrickling filter sections and one biofilter section. The liquid phase pollutants were removed by a mixed culture in the biotrickling filter sections and the gas phase pollutants stripped by air injection in the biofilter section. The specific rates of chemical oxygen demand (COD) removal obtained in the reactor were directly proportional to the pollutant-loading rate. A stable operation of the hybrid bioreactor was attained for long periods of time. The bioreactor had the potential to simultaneously treat a complex mixture of volatile organic compounds, e.g., those present in the water-soluble fraction of gasoline, as well as the capacity to readily adapt to changing operational conditions, such as an increased contaminant loading, and variations in the airflow rate. (Abstract Copyright [2003], Wiley Periodicals, Inc.)

  9. Three-Dimensional Modelling inside a Differential Pressure Laminar Flow Bioreactor Filled with Porous Media

    Directory of Open Access Journals (Sweden)

    Birgit Weyand

    2015-01-01

    Full Text Available A three-dimensional computational fluid dynamics- (CFD- model based on a differential pressure laminar flow bioreactor prototype was developed to further examine performance under changing culture conditions. Cell growth inside scaffolds was simulated by decreasing intrinsic permeability values and led to pressure build-up in the upper culture chamber. Pressure release by an integrated bypass system allowed continuation of culture. The specific shape of the bioreactor culture vessel supported a homogenous flow profile and mass flux at the scaffold level at various scaffold permeabilities. Experimental data showed an increase in oxygen concentration measured inside a collagen scaffold seeded with human mesenchymal stem cells when cultured in the perfusion bioreactor after 24 h compared to static culture in a Petri dish (dynamic: 11% O2 versus static: 3% O2. Computational fluid simulation can support design of bioreactor systems for tissue engineering application.

  10. Bioreactors Drive Advances in Tissue Engineering

    Science.gov (United States)

    2012-01-01

    It was an unlikely moment for inspiration. Engineers David Wolf and Ray Schwarz stopped by their lab around midday. Wolf, of Johnson Space Center, and Schwarz, with NASA contractor Krug Life Sciences (now Wyle Laboratories Inc.), were part of a team tasked with developing a unique technology with the potential to enhance medical research. But that wasn t the focus at the moment: The pair was rounding up colleagues interested in grabbing some lunch. One of the lab s other Krug engineers, Tinh Trinh, was doing something that made Wolf forget about food. Trinh was toying with an electric drill. He had stuck the barrel of a syringe on the bit; it spun with a high-pitched whirr when he squeezed the drill s trigger. At the time, a multidisciplinary team of engineers and biologists including Wolf, Schwarz, Trinh, and project manager Charles D. Anderson, who formerly led the recovery of the Apollo capsules after splashdown and now worked for Krug was pursuing the development of a technology called a bioreactor, a cylindrical device used to culture human cells. The team s immediate goal was to grow human kidney cells to produce erythropoietin, a hormone that regulates red blood cell production and can be used to treat anemia. But there was a major barrier to the technology s success: Moving the liquid growth media to keep it from stagnating resulted in turbulent conditions that damaged the delicate cells, causing them to quickly die. The team was looking forward to testing the bioreactor in space, hoping the device would perform more effectively in microgravity. But on January 28, 1986, the Space Shuttle Challenger broke apart shortly after launch, killing its seven crewmembers. The subsequent grounding of the shuttle fleet had left researchers with no access to space, and thus no way to study the effects of microgravity on human cells. As Wolf looked from Trinh s syringe-capped drill to where the bioreactor sat on a workbench, he suddenly saw a possible solution to both

  11. Modeling bioaugmentation with nitrifiers in membrane bioreactors.

    Science.gov (United States)

    Mannucci, Alberto; Munz, Giulio; Mori, Gualtiero; Makinia, Jacek; Lubello, Claudio; Oleszkiewicz, Jan A

    2015-01-01

    Bioaugmentation with nitrifiers was studied using two pilot-scale membrane bioreactors, with the purpose of assessing the suitability of state-of-the-art activated sludge models (ASMs) in predicting the efficiency of bioaugmentation as a function of operating conditions. It was demonstrated that the temperature difference between seeding and seeded reactors (ΔT) affects bioaugmentation efficiency. Experimental data were accurately predicted when ΔT was within a range of up to 10 °C at the higher range, and when the temperature was significantly lower in the seeded reactor compared to the seeding one, standard ASMs overestimated the efficiency of bioaugmentation. A modified ASM, capable of accurately representing the behavior of seeded nitrifying biomass in the presence of high ΔT, would require the inclusion of the effect of temperature time gradients on nitrifiers. A simple linear correlation between ΔT and the Arrhenius coefficient was proposed as a preliminary step.

  12. Start-up Strategy for Continuous Bioreactors

    Directory of Open Access Journals (Sweden)

    A.C. da Costa

    1997-06-01

    Full Text Available Abstract - The start-up of continuous bioreactors is solved as an optimal control problem. The choice of the dilution rate as the control variable reduces the dimension of the system by making the use of the global balance equation unnecessary for the solution of the optimization problem. Therefore, for systems described by four or less mass balance equations, it is always possible to obtain an analytical expression for the singular arc as a function of only the state variables. The steady state conditions are shown to satisfy the singular arc expression and, based on this knowledge, a feeding strategy is proposed which leads the reactor from an initial state to the steady state of maximum productivity

  13. Moving Denitrifying Bioreactors beyond Proof of Concept: Introduction to the Special Section.

    Science.gov (United States)

    Christianson, Laura E; Schipper, Louis A

    2016-05-01

    Denitrifying bioreactors are organic carbon-filled excavations designed to enhance the natural process of denitrification for the simple, passive treatment of nitrate-nitrogen. Research on and installation of these bioreactors has accelerated within the past 10 years, particularly in watersheds concerned about high nonpoint-source nitrate loads and also for tertiary wastewater treatment. This special section, inspired by the meeting of the Managing Denitrification in Agronomic Systems Community at the 2014 Annual Meeting of the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, aims to firmly establish that denitrifying bioreactors for treatment of nitrate in drainage waters, groundwater, and some wastewaters have moved beyond the proof of concept. This collection of 14 papers expands the peer-reviewed literature of denitrifying bioreactors into new locations, applications, and environmental conditions. There is momentum behind the pairing of wood-based bioreactors with other media (biochar, corn cobs) and in novel designs (e.g., use within treatment trains or use of baffles) to broaden applicability into new kinds of waters and pollutants and to improve performance under challenging field conditions such as cool early season agricultural drainage. Concerns about negative bioreactor by-products (nitrous oxide and hydrogen sulfide emissions, start-up nutrient flushing) are ongoing, but this translates into a significant research opportunity to develop more advanced designs and to fine tune management strategies. Future research must think more broadly to address bioreactor impacts on holistic watershed health and greenhouse gas balances and to facilitate collaborations that allow investigation of mechanisms within the bioreactor "black box."

  14. 栽培措施对半夏倒苗率和产量的影响%Effect of Different Cultivation Conditions on Yield and Lodge Rate of Pinellia ternata ( Thunb. ) Breit.

    Institute of Scientific and Technical Information of China (English)

    章艳玲

    2011-01-01

    [ Objective ] To find out a cultivation method that can lower the rate of lodge Pinellia ternata ( Thunb. ) Breit. and increase the yield of it. [ Method ] Cultured in perlite and soil respectively, nine treatments for each culture matrix were designed to cultivate Pinellia ternata (Thunb.) Breit. to analyze effect of cultivation method on vield and rate of lodge Pinellia ternata (Thunb.) Breit.. [ Result] The yield of Pinellia temata ( Thunb. ) Breit. cultured in perlite was significantly higher than that cultivated in soil; effects of fertilizing amount on yield and lodge rate of Pinellia ternata ( Thunb. ) Breit. were significant. [ Conclusion] The optimum artificial cultivation conditions for Pinellia ternata (Thunb.) Breit. are 1/5MS + 100 mg/L KH2PO4 in perlite.%[目的]探索能够降低半夏倒苗率、提高半夏产量的栽培措施.[方法]用珍珠岩和土壤为基质,分别设9个处理栽培半夏,分析施肥措施对其倒苗率和产量的影响.[结果]以珍珠岩为基质的半夏产量极显著高于以土壤为基质的半夏;不同的施肥量对半夏的产量和倒苗率有极显著的影响.[结论]种植半夏以珍珠岩为基质, 基本营养液浓度为1/5MS, 施以浓度为100 mg/L的KH2PO4效果最佳.

  15. BIOREACTOR DESIGN - OUTER LOOP LANDFILL, LOUISVILLE, KY

    Science.gov (United States)

    Bioreactor field demonstration projects are underway at the Outer Loop Landfill in Louisville, KY, USA. The research effort is a cooperative research effort between US EPA and Waste Management Inc. Two primary kinds of municipal waste bioreactors are under study at this site. ...

  16. Production of Validamycins from Crude Substrates by Streptomyces hygroscopicus in an External-loop Airlift Bioreactor with a Low Height-to-Diameter Ratio

    Institute of Scientific and Technical Information of China (English)

    郑裕国; 陈小龙; 汪钊; 沈寅初

    2004-01-01

    Fermentation experiments to produce validamycins from crude substrates by Streptomyces hygroscopicus were carried out in an external-loop airlift bioreactor (0.0115 m3 ) with a low ratio of height to diameter of the riser of 2.9 and a ratio of riser to downcomer diameter of 6.6. The influences of gas flow rate and liquid volume on fermentation of validamycins were investigated. Comparisons of validamycin fermentation were made among the external-loop airlift bioreactor, a mechanically stirred tank bioreactor (0.010m3 ) and shaking flasks. Under the same operation conditions including fermentation medium composition, inoculum ratio and culture temperature, the fermentation time in the external-loop airlift bioreactor (45 h) was shorter than that in the shaking flasks (100 h) and the same as that in the mechanically stirred tank bioreactor. After a total fermentation time of 45 h under optimized operation conditions, average validamycin concentration obtained in the external-loop airlift bioreactor was close to 19630 μg·ml-1 validamycin concentration in the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the mechanically stirred tank bioreactor in production of validamycins from crude substrates with dregs by Streptomyces hygroscopicus.

  17. A comparative study of leachate quality and biogas generation in simulated anaerobic and hybrid bioreactors

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Qiyong; Tian, Ying; Wang, Shen; Ko, Jae Hac, E-mail: jaehacko@pkusz.edu.cn

    2015-07-15

    Highlights: • Temporary aeration shortened the initial acid inhibition phase for methanogens. • COD decreased faster in the hybrid bioreactor than that in the anaerobic control. • Methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. • MSW settlement increased with increasing the frequency of intermittent aeration. - Abstract: Research has been conducted to compare leachate characterization and biogas generation in simulated anaerobic and hybrid bioreactor landfills with typical Chinese municipal solid waste (MSW). Three laboratory-scale reactors, an anaerobic (A1) and two hybrid bioreactors (C1 and C2), were constructed and operated for about 10 months. The hybrid bioreactors were operated in an aerobic–anaerobic mode with different aeration frequencies by providing air into the upper layer of waste. Results showed that the temporary aeration into the upper layer aided methane generation by shortening the initial acidogenic phase because of volatile fatty acids (VFAs) reduction and pH increase. Chemical oxygen demand (COD) decreased faster in the hybrid bioreactors, but the concentrations of ammonia–nitrogen in the hybrid bioreactors were greater than those in the anaerobic control. Methanogenic conditions were established within 75 d and 60 d in C1 and C2, respectively. However, high aeration frequency led to the consumption of organic matters by aerobic degradation and resulted in reducing accumulative methane volume. The temporary aeration enhanced waste settlement and the settlement increased with increasing the frequency of aeration. Methane production was inhibited in the anaerobic control; however, the total methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. As for MSW with high content of food waste, leachate recirculation right after aeration stopped was not recommended due to VFA inhibition for methanogens.

  18. Biodegradation of High Concentrations of Benzene Vapors in a Two Phase Partition Stirred Tank Bioreactor

    Directory of Open Access Journals (Sweden)

    Ali Karimi

    2013-01-01

    Full Text Available The present study examined the biodegradation rate of benzene vapors in a two phase stirred tank bioreactor by a bacterial consortium obtained from wastewater of an oil industry refinery house. Initially, the ability of the microbial consortium for degrading benzene was evaluated before running the bioreactor. The gaseous samples from inlet and outlet of bioreactor were directly injected into a gas chromatograph to determine benzene concentrations. Carbone oxide concentration at the inlet and outlet of bioreactor were also measured with a CO2 meter to determine the mineralization rate of benzene. Influence of the second non-aqueous phase (silicon oil has been emphasized, so at the first stage the removal efficiency (RE and elimination capacity (EC of benzene vapors were evaluated without any organic phase and in the second stage, 10% of silicon oil was added to bioreactor media as an organic phase. Addition of silicon oil increased the biodegradation performance up to an inlet loading of 5580?mg/m3, a condition at which, the elimination capacity and removal efficiency were 181?g/m3/h and 95% respectively. The elimination rate of benzene increased by 38% in the presence of 10% of silicone oil. The finding of this study demonstrated that two phase partition bioreactors (TPPBs are potentially effective tools for the treatment of gas streams contaminated with high concentrations of poorly water soluble organic contaminant, such as benzene.

  19. Biodegradation of high concentrations of benzene vapors in a two phase partition stirred tank bioreactor

    Directory of Open Access Journals (Sweden)

    Karimi Ali

    2013-01-01

    Full Text Available Abstract The present study examined the biodegradation rate of benzene vapors in a two phase stirred tank bioreactor by a bacterial consortium obtained from wastewater of an oil industry refinery house. Initially, the ability of the microbial consortium for degrading benzene was evaluated before running the bioreactor. The gaseous samples from inlet and outlet of bioreactor were directly injected into a gas chromatograph to determine benzene concentrations. Carbone oxide concentration at the inlet and outlet of bioreactor were also measured with a CO2 meter to determine the mineralization rate of benzene. Influence of the second non-aqueous phase (silicon oil has been emphasized, so at the first stage the removal efficiency (RE and elimination capacity (EC of benzene vapors were evaluated without any organic phase and in the second stage, 10% of silicon oil was added to bioreactor media as an organic phase. Addition of silicon oil increased the biodegradation performance up to an inlet loading of 5580 mg/m3, a condition at which, the elimination capacity and removal efficiency were 181 g/m3/h and 95% respectively. The elimination rate of benzene increased by 38% in the presence of 10% of silicone oil. The finding of this study demonstrated that two phase partition bioreactors (TPPBs are potentially effective tools for the treatment of gas streams contaminated with high concentrations of poorly water soluble organic contaminant, such as benzene.

  20. A comparative study of leachate quality and biogas generation in simulated anaerobic and hybrid bioreactors.

    Science.gov (United States)

    Xu, Qiyong; Tian, Ying; Wang, Shen; Ko, Jae Hac

    2015-07-01

    Research has been conducted to compare leachate characterization and biogas generation in simulated anaerobic and hybrid bioreactor landfills with typical Chinese municipal solid waste (MSW). Three laboratory-scale reactors, an anaerobic (A1) and two hybrid bioreactors (C1 and C2), were constructed and operated for about 10months. The hybrid bioreactors were operated in an aerobic-anaerobic mode with different aeration frequencies by providing air into the upper layer of waste. Results showed that the temporary aeration into the upper layer aided methane generation by shortening the initial acidogenic phase because of volatile fatty acids (VFAs) reduction and pH increase. Chemical oxygen demand (COD) decreased faster in the hybrid bioreactors, but the concentrations of ammonia-nitrogen in the hybrid bioreactors were greater than those in the anaerobic control. Methanogenic conditions were established within 75d and 60d in C1 and C2, respectively. However, high aeration frequency led to the consumption of organic matters by aerobic degradation and resulted in reducing accumulative methane volume. The temporary aeration enhanced waste settlement and the settlement increased with increasing the frequency of aeration. Methane production was inhibited in the anaerobic control; however, the total methane generations from hybrid bioreactors were 133.4L/kgvs and 113.2L/kgvs. As for MSW with high content of food waste, leachate recirculation right after aeration stopped was not recommended due to VFA inhibition for methanogens.

  1. Cell retention by encapsulation for the cultivation of Jurkat cells in fixed and fluidized bed reactors.

    Science.gov (United States)

    Kaiser, P; Werner, M; Jérôme, V; Hübner, H; Buchholz, R; Freitag, R

    2014-12-01

    Jurkat cells are accepted model cells for primary human T lymphocytes, for example, in medical research. Their growth to tissue-like cell densities (up to 100 × 10(6)  cells/mLcapsule ) in semi-permeable (molecular weight cut off cultivations, that is, under conditions where both encapsulated and non-encapsulated cells can be cultivated under otherwise identical conditions, showed that maximum specific growth rates were higher for the encapsulated than for the non-encapsulated cells. In the subsequent batch and repeated batch bioreactor experiments (only encapsulated cells), growth rates were similar, with the exception of the fixed bed batch reactor, where growth kinetics were significantly slower. Concomitantly, a significant fraction of the cells towards the bottom of the bed were no longer metabolically active, though apparently not dead. In the repeated batch fluidized bed reactor cellular division could be maintained for more than two weeks, albeit with a specific growth rate below the maximum one, leading to final cell densities of approximately 180 × 10(6)  cell/gcapsule . At the same time, the cell cycle distribution of the cells was shifted to the S and G2/M phases.

  2. Characteristics, Process Parameters, and Inner Components of Anaerobic Bioreactors

    Science.gov (United States)

    Abdelgadir, Awad; Chen, Xiaoguang; Liu, Jianshe; Xie, Xuehui; Zhang, Jian; Zhang, Kai; Wang, Heng; Liu, Na

    2014-01-01

    The anaerobic bioreactor applies the principles of biotechnology and microbiology, and nowadays it has been used widely in the wastewater treatment plants due to their high efficiency, low energy use, and green energy generation. Advantages and disadvantages of anaerobic process were shown, and three main characteristics of anaerobic bioreactor (AB), namely, inhomogeneous system, time instability, and space instability were also discussed in this work. For high efficiency of wastewater treatment, the process parameters of anaerobic digestion, such as temperature, pH, Hydraulic retention time (HRT), Organic Loading Rate (OLR), and sludge retention time (SRT) were introduced to take into account the optimum conditions for living, growth, and multiplication of bacteria. The inner components, which can improve SRT, and even enhance mass transfer, were also explained and have been divided into transverse inner components, longitudinal inner components, and biofilm-packing material. At last, the newly developed special inner components were discussed and found more efficient and productive. PMID:24672798

  3. Hydrodynamic Characterization of a Column-type Prototype Bioreactor

    Science.gov (United States)

    Espinosa-Solares, Teodoro; Morales-Contreras, Marcos; Robles-Martínez, Fabián; García-Nazariega, Melvin; Lobato-Calleros, Consuelo

    Agro-food industrial processes produce a large amount of residues, most of which are organic. One of the possible solutions for the treatment of these residues is anaerobic digestion in bioreactors. A novel 18-L bioreactor for treating waste water was designed based on pneumatic agitation and semispherical baffles. Flow patterns were visualized using the particle tracer technique. Circulation times were measured with the particle tracer and the thermal technique, while mixing times were measured using the thermal technique. Newtonian fluid and two non-Newtonian fluids were used to simulate the operational conditions. The results showed that the change from Newtonian to non-Newtonian properties reduces mixed zones and increases circulation and mixing times. Circulation time was similar when evaluated with the thermal and the tracer particle methods. It was possible to predict dimensionless mixing time (θm) using an equivalent Froude number (Fr eq).

  4. Optimization of biological sulfide removal in a CSTR bioreactor.

    Science.gov (United States)

    Roosta, Aliakbar; Jahanmiri, Abdolhossein; Mowla, Dariush; Niazi, Ali; Sotoodeh, Hamidreza

    2012-08-01

    In this study, biological sulfide removal from natural gas in a continuous bioreactor is investigated for estimation of the optimal operational parameters. According to the carried out reactions, sulfide can be converted to elemental sulfur, sulfate, thiosulfate, and polysulfide, of which elemental sulfur is the desired product. A mathematical model is developed and was used for investigation of the effect of various parameters on elemental sulfur selectivity. The results of the simulation show that elemental sulfur selectivity is a function of dissolved oxygen, sulfide load, pH, and concentration of bacteria. Optimal parameter values are calculated for maximum elemental sulfur selectivity by using genetic algorithm as an adaptive heuristic search. In the optimal conditions, 87.76% of sulfide loaded to the bioreactor is converted to elemental sulfur.

  5. Wastewater treatment in a submerged anaerobic membrane bioreactor.

    Science.gov (United States)

    Casu, Stefania; Crispino, Nedda A; Farina, Roberto; Mattioli, Davide; Ferraris, Marco; Spagni, Alessandro

    2012-01-01

    Although most membrane bioreactors are used under aerobic conditions, over the last few years there has been increased interest in their application for anaerobic processes. This paper presents the results obtained when a bench-scale submerged anaerobic membrane bioreactor was used for the treatment of wastewaters generated in the agro-food industry. The reactor was fed with synthetic wastewater consisting of cheese whey and sucrose, and volumetric organic loading rates (OLRs) ranging from 1.5 to 13 kgCOD/(m(3)*d) were applied. Under the operating conditions studied, the maximum applicable OLR was between 6 and 10 gCOD/(g*L), which fell within the ranges of the high-rate anaerobic wastewater treatment systems, while high concentrations of volatile fatty acids were produced at higher OLR rates. With an OLR of 1.5-10 gCOD/(g*L), the reactor showed 94% COD removal, whereas this value dropped to 33% with the highest applied OLR of 13 gCOD/(g*L). The study therefore confirms that membrane bioreactors can be used for anaerobic wastewater treatment.

  6. A bioreactor approach to investigate the linkage between methane oxidation and nitrate/nitrite reduction in the pelagic oxic-anoxic transition zone of the central Baltic Sea

    Directory of Open Access Journals (Sweden)

    Gunnar Jakobs

    2016-08-01

    Full Text Available Evidence of aerobic methane oxidation coupled to denitrification has been provided for different freshwater environments, whereas the significance of this process for the marine realm has not been adequately investigated. The goal of this study was to investigate the methane-related reduction of nitrate/nitrite in a marine environment (salinity 8.5. A water sample was collected from the oxic-anoxic transition zone of the Gotland Deep (central Baltic Sea and the microorganisms contained therein were cultivated in a bioreactor under hypoxic conditions (0.5 µM O2. To enrich the microorganisms involved in the coupled process the bioreactor was continuously sparged with methane as the sole energy and carbon source and simultaneously supplied with a nutrient solution rich in nitrate and nitrite. The bioreactor experiment showed a relationship between the turnover of methane and the concomitant concentration decrease of nitrite and nitrate at the early stage of the experiment. This relationship indicates the role of methanotrophs, which may support heterotrophic denitrifiers by the release of organic compounds as an energy source. Besides, a mixture of uncultured microorganisms, aerobic methanotrophic and heterotrophic denitrifying bacteria were identified in the enrichment culture. Microbial incorporation of nitrite and methane was proven on the cellular and gene levels via 15NO2- / 13CH4 incubation experiments and subsequent analyses with nano secondary ion mass spectrometry (NanoSIMS and stable isotope probing (SIP. The NanoSIMS showed the incorporation of 15N in almost all the bacteria and in 9% of those there was a concomitant enrichment in 13C. The relatively low abundance of methane-consuming bacteria in the bioreactor was further reflected in specific fatty acids indicative for type I methanotrophic bacteria. Based on pmoA gene analyses, this bacterium is different from the one that was identified as the only key player of methane oxidation in

  7. Online monitoring of cartilage tissue in a novel bioreactor

    Science.gov (United States)

    von der Burg, E.; von Buttlar, M.; Grill, W.

    2011-04-01

    Standard techniques for the analysis of biological tissues like immunohistochemical staining are typically invasive and lead to mortification of cells. Non-invasive monitoring is an important element of regenerative medicine because implants and components of implants should be 100% quality-checked with non-invasive and therefore also marker-free methods. We report on a new bioreactor for the production of collagen scaffolds seeded with Mesenchymal Stem Cells (MSCs). It contains a computer controlled mechanical activation and ultrasonic online monitoring and has been constructed for the in situ determination of ultrasonic and rheological parameters. During the cultivation period of about two weeks the scaffold is periodically compressed by two movable pistons for improved differentiation of the MSCs. This periodic compression beneficially ensures the supply with nutrition even inside the sample. During the physiological stimuli, rheological properties are measured by means of highly sensitive load cells. In addition measurements of the speed of sound in the sample and in the culture medium, with frequencies up to 16 MHz, are performed continuously. Therefore piezoceramic transducers are attached to the pistons and emit and detect ultrasonic waves, travelling through the pistons, the sample and the culture medium. The time-of-flight (TOF) of the ultrasonic signals is determined in real time with the aid of chirped excitation and correlation procedures with a resolution of at least 10 ps. The implemented ultrasonic measurement scheme allows beside the speed of sound measurements the detection of the distance between the pistons with a resolution better than 100 nm. The developed monitoring delivers information on rigidity, fluid dynamics and velocity of sound in the sample and in the culture medium. The hermetically sealed bioreactor with its life support system provides a biocompatible environment for MSCs for long time cultivation.

  8. Cultivation of Marine Sponges.

    Science.gov (United States)

    Osinga; Tramper; Wijffels

    1999-11-01

    There is increasing interest in biotechnological production of marine sponge biomass owing to the discovery of many commercially important secondary metabolites in this group of animals. In this article, different approaches to producing sponge biomass are reviewed, and several factors that possibly influence culture success are evaluated. In situ sponge aquacultures, based on old methods for producing commercial bath sponges, are still the easiest and least expensive way to obtain sponge biomass in bulk. However, success of cultivation with this method strongly depends on the unpredictable and often suboptimal natural environment. Hence, a better-defined production system would be desirable. Some progress has been made with culturing sponges in semicontrolled systems, but these still use unfiltered natural seawater. Cultivation of sponges under completely controlled conditions has remained a problem. When designing an in vitro cultivation method, it is important to determine both qualitatively and quantitatively the nutritional demands of the species that is to be cultured. An adequate supply of food seems to be the key to successful sponge culture. Recently, some progress has been made with sponge cell cultures. The advantage of cell cultures is that they are completely controlled and can easily be manipulated for optimal production of the target metabolites. However, this technique is still in its infancy: a continuous cell line has yet to be established. Axenic cultures of sponge aggregates (primmorphs) may provide an alternative to cell culture. Some sponge metabolites are, in fact, produced by endosymbiotic bacteria or algae that live in the sponge tissue. Only a few of these endosymbionts have been cultivated so far. The biotechnology for the production of sponge metabolites needs further development. Research efforts should be continued to enable commercial exploitation of this valuable natural resource in the near future.

  9. Hydrodynamic characteristics of UASB bioreactors.

    Science.gov (United States)

    John, Siby; Tare, Vinod

    2011-10-01

    The hydrodynamic characteristics of UASB bioreactors operated under different organic loading and hydraulic loading rates were studied, using three laboratory scale models treating concocted sucrose wastewater. Residence time distribution (RTD) analysis using dispersion model and tanks-in-series model was directed towards the characterization of the fluid flow pattern in the reactors and correlation of the hydraulic regime with the biomass content and biogas production. Empty bed reactors followed a plug flow pattern and the flow pattern changed to a large dispersion mixing with biomass and gas production. Effect of increase in gas production on the overall hydraulics was insignificant.

  10. Impact of Furfural on Rapid Ethanol Production Using a Membrane Bioreactor

    Directory of Open Access Journals (Sweden)

    Mohammad J. Taherzadeh

    2013-03-01

    Full Text Available A membrane bioreactor was developed to counteract the inhibition effect of furfural in ethanol production. Furfural, a major inhibitor in lignocellulosic hydrolyzates, is a highly toxic substance which is formed from pentose sugars released during the acidic degradation of lignocellulosic materials. Continuous cultivations with complete cell retention were performed at a high dilution rate of 0.5 h−1. Furfural was added directly into the bioreactor by pulse injection or by addition into the feed medium to obtain furfural concentrations ranging from 0.1 to 21.8 g L−1. At all pulse injections of furfural, the yeast was able to convert the furfural very rapidly by in situ detoxification. When injecting 21.8 g L−1 furfural to the cultivation, the yeast converted it by a specific conversion rate of 0.35 g g−1 h−1. At high cell density, Saccharomyces cerevisiae could tolerate very high furfural levels without major changes in the ethanol production. During the continuous cultures when up to 17.0 g L−1 furfural was added to the inlet medium, the yeast successfully produced ethanol, whereas an increase of furfural to 18.6 and 20.6 g L−1 resulted in a rapidly decreasing ethanol production and accumulation of sugars in the permeate. This study show that continuous ethanol fermentations by total cell retention in a membrane bioreactor has a high furfural tolerance and can conduct rapid in situ detoxification of medium containing high furfural concentrations.

  11. Climatic conditions for grapevine cultivation in Boa Vista, Roraima, Brazil = Aptidão climática para o cultivo da videira em Boa Vista, Roraima

    Directory of Open Access Journals (Sweden)

    Marco Antônio Fonseca Conceição

    2014-12-01

    Full Text Available Grapevine plants have been grown in different tropical regions of Brazil. In the state of Roraima, this culture was introduced commercially in 2005 in Boa Vista. Unlike temperate regions, tropical regions present thermal conditions for grape production throughout the year. To evaluate the climatic characteristics on different periods of the year, it can be used different indices. The objective of this study was to characterize climate suitability of Boa Vista, Roraima, Brazil, for grapevine cultivation at different months of the year. For climate classification it was used the Geoviticulture Multicriteria Climatic Classification System (CCM, which is composed by three indices: Heliothermal (IH, Cool Night (IF, and Drought (IS. It was also used the Zuluaga Index (IZ, to assess the risk of incidence of fungal diseases on the vine, especially in relation to the incidence of downy mildew (Plasmopara viticola, one of the main grapevine diseases in humid regions. Based on the evaluated indices, it was found that the most favorable period for grape production goes from October to March because of the lower values of rainfall. During the other months, it can be employed branch pruning without fruit production, as occurs in other tropical locations of Brazil. =A videira tem sido cultivada em diferentes regiões tropicais do Brasil. No estado de Roraima, ela foi implantada comercialmente em 2005 em Boa Vista. Ao contrário das regiões de clima temperado, as regiões tropicais apresentam condições térmicas para a produção de uvas durante o ano inteiro. Para se avaliar as características climáticas dos diferentes períodos do ano pode-se recorrer a diversos índices. Objetivou-se, com o presente trabalho, avaliar a aptidão climática de Boa Vista, Roraima, para o cultivo da videira, considerando-se diferentes períodos de produção ao longo do ano. Na caracterização climática, foi utilizado o Sistema de Classificação Climática Multicrit

  12. Full-scale demonstration of treatment of mechanically separated organic residue in a bioreactor at VAM in Wijster[Netherlands

    Energy Technology Data Exchange (ETDEWEB)

    Oonk, H. [TNO-MEP, Apledoorn (Netherlands); Woelders, H. [VAM, Wijster (Netherlands)

    1999-07-01

    At the VAM waste treatment company in Wijster a demonstration is in progress of bioreactor technology for the treatment of mechanically separated organic residue (MSOR) of a waste separation plant. This bioreactor is an in situ fermentation cell in which physical, chemical and biological processes are controlled by leachate circulation in such a way to ensure obtaining a product with acceptable rest-emissions. A 49 000 tonne demonstration cell has been built and methanogenic conditions are being realized. During operation measures are taken to limit emissions from the bioreactor and to optimize the conversion of waste to energy. (au)

  13. Toward the Standardization of Bioreactors for Space Research

    Science.gov (United States)

    Garcia, Michel; Nebuloni, Stefano; Dainesi, Paolo; Gass, Samuel

    be implemented which do not require major new and lengthy developments. RUAG believes this is pivotal in facilitating access to life-science research in space and thereby increasing scientific output in this research area. The overview includes a description of technical features such as feasible cultivation volumes in highly biocompatible and transparent culture chambers, cultivation method options (batch vs. continuous), associated feed rates, and chemical fixation methods. Feasible regulation and sensing possibilities (i.e. O2, CO2, temperature, pH, cell concentration), based on technologies already used in laboratory application, are also discussed. Important aspects related to flow homogeneity, mass/gas transfer and mixing methods in micro-gravity are also presented. Lastly, the environmental performance of existing bioreactors is also shown and a particular emphasis is placed on safety design aspects for space hardware.

  14. Microliter-bioreactor array with buoyancy-driven stirring for human hematopoietic stem cell culture.

    Science.gov (United States)

    Luni, Camilla; Feldman, Hope C; Pozzobon, Michela; De Coppi, Paolo; Meinhart, Carl D; Elvassore, Nicola

    2010-08-11

    This work presents the development of an array of bioreactors where finely controlled stirring is provided at the microliter scale (100-300 mul). The microliter-bioreactor array is useful for performing protocol optimization in up to 96 parallel experiments of hematopoietic stem cell (HSC) cultures. Exploring a wide range of experimental conditions at the microliter scale minimizes cost and labor. Once the cell culture protocol is optimized, it can be applied to large-scale bioreactors for stem cell production at the clinical level. The controlled stirring inside the wells of a standard 96-well plate is provided by buoyancy-driven thermoconvection. The temperature and velocity fields within the culture volume are determined with numerical simulations. The numerical results are verified with experimental velocity measurements using microparticle image velocimetry (muPIV) and are used to define feasible experimental conditions for stem cell cultures. To test the bioreactor array's functionality, human umbilical cord blood-derived CD34(+) cells were cultured for 7 days at five different stirring conditions (0.24-0.58 mums) in six repeated experiments. Cells were characterized in terms of proliferation, and flow cytometry measurements of viability and CD34 expression. The microliter-bioreactor array demonstrates its ability to support HSC cultures under stirred conditions without adversely affecting the cell behavior. Because of the highly controlled operative conditions, it can be used to explore culture conditions where the mass transport of endogenous and exogenous growth factors is selectively enhanced, and cell suspension provided. While the bioreactor array was developed for culturing HSCs, its application can be extended to other cell types.

  15. Biogas Production from Citrus Waste by Membrane Bioreactor

    Science.gov (United States)

    Wikandari, Rachma; Millati, Ria; Cahyanto, Muhammad Nur; Taherzadeh, Mohammad J.

    2014-01-01

    Rapid acidification and inhibition by d-limonene are major challenges of biogas production from citrus waste. As limonene is a hydrophobic chemical, this challenge was encountered using hydrophilic polyvinylidine difluoride (PVDF) membranes in a biogas reactor. The more sensitive methane-producing archaea were encapsulated in the membranes, while freely suspended digesting bacteria were present in the culture as well. In this membrane bioreactor (MBR), the free digesting bacteria digested the citrus wastes and produced soluble compounds, which could pass through the membrane and converted to biogas by the encapsulated cell. As a control experiment, similar digestions were carried out in bioreactors containing the identical amount of just free cells. The experiments were carried out in thermophilic conditions at 55 °C, and hydraulic retention time of 30 days. The organic loading rate (OLR) was started with 0.3 kg VS/m3/day and gradually increased to 3 kg VS/m3/day. The results show that at the highest OLR, MBR was successful to produce methane at 0.33 Nm3/kg VS, while the traditional free cell reactor reduced its methane production to 0.05 Nm3/kg VS. Approximately 73% of the theoretical methane yield was achieved using the membrane bioreactor. PMID:25167328

  16. CFD Simulation of Fouling by Biological materials in Membrane Bioreactor

    Directory of Open Access Journals (Sweden)

    Raziye, Ahmadi

    2016-06-01

    Full Text Available In recent years membrane bioreactors filtration is increasingly used in wastewater treatment to enhance the quality of wastewater. The main problem in preventing the widespread use of membrane bioreactor is its congestion which has a severe impact on output flux to time ratio. If solid suspensions with high concentrations exist in the wastewater, this influence will be even more severe. In addition to the suspended solids in the liquid mixture, Extracellular polymeric materials (EPS and soluble microbial products (SMP are also known as basic microbial products that cause membrane fouling. EPS can be calculated within and on the membrane which increases the viscosity of suspended solids in the liquid mixture and increases filtration resistance. SMPs cannot penetrate the pores of the ultra filtration membrane due to the limited size of the pores which would cause fouling in membrane processes. According to the above issues, providing a model that indicates the properties and conditions of formation and destruction of SMP and EPS at the same time seems necessary. In this paper, CFD simulation of biological fouling in membrane bioreactor is provided using Fluent software.

  17. Bioreactors for tissue engineering--a new role for perfusionists?

    Science.gov (United States)

    Sistino, Joseph J

    2003-09-01

    Tissue engineering is an exciting new area of medicine with rapid growth and expansion over the last decade. It has the potential to have a profound impact on the practice of medicine and influence the economic development in the industry of biotechnology. In almost every specialty of medicine, the ability to generate replacement cells and develop tissues will change the focus from artificial organs and transplantation to growing replacement organs from the patient's own stem cells. Once these organs are at a size that requires perfusion to maintain oxygen and nutrient delivery, then automated perfusion systems termed "bioreactors" will be necessary to sustain the organ until harvesting. The design of these "bioreactors" will have a crucial role in the maintenance of cellular function throughout the growth period. The perfusion schemes necessary to determine the optimal conditions have not been well elucidated and will undergo extensive research over the next decade. The key to progress in this endeavor will development of long-term perfusion techniques and identifying the ideal pressures, flow rates, type of flow (pulsatile/nonpulsatile), and perfusate solution. Perfusionists are considered experts in the field of whole body perfusion, and it is possible that they can participate in the development and operation of these "bioreactors." Additional education of perfusionists in the area of tissue engineering is necessary in order for them to become integral parts of this exciting new area of medicine.

  18. Biogas Production from Citrus Waste by Membrane Bioreactor

    Directory of Open Access Journals (Sweden)

    Rachma Wikandari

    2014-08-01

    Full Text Available Rapid acidification and inhibition by d-limonene are major challenges of biogas production from citrus waste. As limonene is a hydrophobic chemical, this challenge was encountered using hydrophilic polyvinylidine difluoride (PVDF membranes in a biogas reactor. The more sensitive methane-producing archaea were encapsulated in the membranes, while freely suspended digesting bacteria were present in the culture as well. In this membrane bioreactor (MBR, the free digesting bacteria digested the citrus wastes and produced soluble compounds, which could pass through the membrane and converted to biogas by the encapsulated cell. As a control experiment, similar digestions were carried out in bioreactors containing the identical amount of just free cells. The experiments were carried out in thermophilic conditions at 55 °C, and hydraulic retention time of 30 days. The organic loading rate (OLR was started with 0.3 kg VS/m3/day and gradually increased to 3 kg VS/m3/day. The results show that at the highest OLR, MBR was successful to produce methane at 0.33 Nm3/kg VS, while the traditional free cell reactor reduced its methane production to 0.05 Nm3/kg VS. Approximately 73% of the theoretical methane yield was achieved using the membrane bioreactor.

  19. Development of Fundamental Technologies for Micro Bioreactors

    Science.gov (United States)

    Sato, Kiichi; Kitamori, Takehiko

    This chapter reviews the development of fundamental technologies required for microchip-based bioreactors utilizing living mammalian cells and pressure driven flow. The most important factor in the bioreactor is the cell culture. For proper cell culturing, continuous medium supply from a microfluidic channel and appropriate modification of the channel surface to accommodate cell attachment is required. Moreover, the medium flow rate should be chosen carefully, because shear stress affects cell activity. The techniques presented here could be applied to the development of micro bioreactors such as microlivers, pigment production by plant cells, and artificial insemination.

  20. Spatial Experiment Technologies Suitable for Unreturnable Bioreactor

    Science.gov (United States)

    Zhang, Tao; Zheng, Weibo; Tong, Guanghui

    2016-07-01

    The system composition and main function of the bioreactor piggybacked on TZ cargo transport spacecraft are introduced briefly in the paper.The spatial experiment technologies which are suitable for unreturnable bioreactor are described in detail,including multi-channel liquid transportion and management,multi-type animal cells circuit testing,dynamic targets microscopic observation in situ etc..The feasibility and effectiveness of these technologies which will be used in space experiment in bioreactor are verified in tests and experiments on the ground.

  1. Denitrification 'Woodchip' Bioreactors for Productive and Sustainable Agricultural Systems

    Science.gov (United States)

    Christianson, L. E.; Summerfelt, S.; Sharrer, K.; Lepine, C.; Helmers, M. J.

    2014-12-01

    Growing alarm about negative cascading effects of reactive nitrogen in the environment has led to multifaceted efforts to address elevated nitrate-nitrogen levels in water bodies worldwide. The best way to mitigate N-related impacts, such as hypoxic zones and human health concerns, is to convert nitrate to stable, non-reactive dinitrogen gas through the natural process of denitrification. This means denitrification technologies need to be one of our major strategies for tackling the grand challenge of managing human-induced changes to our global nitrogen cycle. While denitrification technologies have historically been focused on wastewater treatment, there is great interest in new lower-tech options for treating effluent and drainage water from one of our largest reactive nitrogen emitters -- agriculture. Denitrification 'woodchip' bioreactors are able to enhance this natural N-conversion via addition of a solid carbon source (e.g., woodchips) and through designs that facilitate development of anoxic conditions required for denitrification. Wood-based denitrification technologies such as woodchip bioreactors and 'sawdust' walls for groundwater have been shown to be effective at reducing nitrate loads in agricultural settings around the world. Designing these systems to be low-maintenance and to avoid removing land from agricultural production has been a primary focus of this "farmer-friendly" technology. This presentation provides a background on woodchip bioreactors including design considerations, N-removal performance, and current research worldwide. Woodchip bioreactors for the agricultural sector are an accessible new option to address society's interest in improving water quality while simultaneously allowing highly productive agricultural systems to continue to provide food in the face of increasing demand, changing global diets, and fluctuating weather.

  2. Effect of Tetracycline Antibiotics on Performance and Microbial Community of Algal Photo-Bioreactor.

    Science.gov (United States)

    Taşkan, Ergin

    2016-07-01

    Tetracycline antibiotics have been increasingly used in medical applications and have been found in wastewater treatment plants as a result of human and industrial activities. This study investigates the combined effects of tetracycline antibiotics on the performance of an algal photo-bioreactor operated under different antibiotic concentrations in the ranges of 0.25 to 30 mg/L and considers the inhibition of algal growth, carbon and nutrient removal rates, and eukaryotic and cyanobacterial algal community changes. The results indicated that increases in the concentration of tetracycline mixtures have adverse effects on the algal community and the performance of a photo-bioreactor, and the eukaryotic algae species were more sensitive to tetracycline antibiotics than were the cyanobacterial species. Cultivation tests showed that approximately 94 % growth inhibition of mixed algae occurred at 30 mg/L.

  3. Influence of temperature on carbon and nitrogen dynamics during in situ aeration of aged waste in simulated landfill bioreactors.

    Science.gov (United States)

    Tong, Huanhuan; Yin, Ke; Giannis, Apostolos; Ge, Liya; Wang, Jing-Yuan

    2015-09-01

    The effect of temperature on carbon and nitrogen compounds during in situ aeration of aged waste was investigated in lab-scale simulated landfill bioreactors at 35, 45 and 55 °C, respectively. The bioreactor operated at 55 °C presented the highest carbon mineralization rate in the initial stage, suggesting accelerated biodegradation rates under thermophilic conditions. The nitrogen speciation study indicated that organic nitrogen was the dominant species of total N in aerobic bioreactors due to ammonia removal. Leachate organic nitrogen was further fractionated to elucidate the fate of individual constituent. Detailed investigation revealed the higher bioconversion rates of N-humic and N-fulvic compounds compared to hydrophilic compounds in thermophilic conditions. At the end, waste material in 55 °C bioreactor was richer in highly matured humic substances (HS) verifying the high bioconversion rates.

  4. Cardiac tissue engineering: cell seeding, cultivation parameters, and tissue construct characterization.

    Science.gov (United States)

    Carrier, R L; Papadaki, M; Rupnick, M; Schoen, F J; Bursac, N; Langer, R; Freed, L E; Vunjak-Novakovic, G

    1999-09-01

    Cardiac tissue engineering has been motivated by the need to create functional tissue equivalents for scientific studies and cardiac tissue repair. We previously demonstrated that contractile cardiac cell-polymer constructs can be cultivated using isolated cells, 3-dimensional scaffolds, and bioreactors. In the present work, we examined the effects of (1) cell source (neonatal rat or embryonic chick), (2) initial cell seeding density, (3) cell seeding vessel, and (4) tissue culture vessel on the structure and composition of engineered cardiac muscle. Constructs seeded under well-mixed conditions with rat heart cells at a high initial density ((6-8) x 10(6) cells/polymer scaffold) maintained structural integrity and contained macroscopic contractile areas (approximately 20 mm(2)). Seeding in rotating vessels (laminar flow) rather than mixed flasks (turbulent flow) resulted in 23% higher seeding efficiency and 20% less cell damage as assessed by medium lactate dehydrogenase levels (p laminar and dynamic, yielded constructs with a more active, aerobic metabolism as compared to constructs cultured in mixed or static flasks. After 1-2 weeks of cultivation, tissue constructs expressed cardiac specific proteins and ultrastructural features and had approximately 2-6 times lower cellularity (p < 0.05) but similar metabolic activity per unit cell when compared to native cardiac tissue.

  5. Production of diosgenin from Dioscorea zingiberensis with mixed culture in a new tray bioreactor

    Directory of Open Access Journals (Sweden)

    Yutong Cheng

    2016-01-01

    Full Text Available A new tray bioreactor was developed for the production of diosgenin from Dioscorea zingiberensis with Trichoderma reesei and Aspergillus fumigatus. The influence of initial moisture content, temperature, tray bed depth and mixing times was investigated. The best fermentation condition is initial moisture content of 75%, bioreactor temperature of 35°C, solid bed depth of 1.5 cm and three mixings carrying out on the first, third and fifth day. Under the optimized fermentation conditions, after 144 h incubation, maximum diogenin concentration of 68.2 μmol/g was detected.

  6. Alternative cultivation systems for energy crops. Exploitation of phosphor and nitrogen in the cultivation of mixed fruits with leguminous plants under the conditions of drought stress; Alternative Anbausysteme fuer Energiepflanzen. Phosphor- und Stickstoffausnutzung im Mischfruchtanbau mit Leguminosen unter Trockenstressbedingungen

    Energy Technology Data Exchange (ETDEWEB)

    Busch, Stefanie; Eichler-Loebermann, Bettina [Rostock Univ. (Germany). Professur Pflanzenbau

    2013-10-01

    Alternative cropping systems with an efficient utilization of resources are particularly interesting for energy cropping. The P- and N- uptake of phosphorus (P) and nitrogen (N) of maize and sorghum (here called ''energy crops'') intercropped with legumes (substitutive, 50:50) under drought conditions were investigated in a eight week pot experiment. Yield, P- and N- uptake of all species and mixtures were significantly lower under drought conditions than when well watered. The yield and the P-uptake of the mixtures was lower than of the sole cropped energy crops when well watered, but comparable under water deficit with exception of the sorghum mixtures, which reached a lower yield than sole sorghum. Despite the lower N-fertilization the N-uptake of the mixtures was comparable to the sole cropped maize or sorghum when well watered, but under drought N uptake of the mixtures was decreased in comparison to sole cropping. Under drought conditions the N-uptake of maize and sorghum plants in mixtures was not decreased, while the N-uptake of the legumes decreased in comparison to the well watered treatment. This may be an evidence for the benefit of the non-legumes in the investigated intercropping system under drought conditions. (orig.)

  7. A new microfluidic concept for parallel operated milliliter-scale stirred tank bioreactors.

    Science.gov (United States)

    Gebhardt, Gabi; Hortsch, Ralf; Kaufmann, Klaus; Arnold, Matthias; Weuster-Botz, Dirk

    2011-01-01

    Parallel miniaturized stirred tank bioreactors are an efficient tool for "high-throughput bioprocess design." As most industrial bioprocesses are pH-controlled and/or are operated in a fed-batch mode, an exact scale-down of these reactions with continuous dosing of fluids into the miniaturized bioreactors is highly desirable. Here, we present the development, characterization, and application of a novel concept for a highly integrated microfluidic device for a bioreaction block with 48 parallel milliliter-scale stirred tank reactors (V = 12 mL). The device consists of an autoclavable fluidic section to dispense up to three liquids individually per reactor. The fluidic section contains 144 membrane pumps, which are magnetically driven by a clamped-on actuator section. The micropumps are designed to dose 1.6 μL per pump lift. Each micropump enables a continuous addition of liquid with a flow rate of up to 3 mL h(-1) . Viscous liquids up to a viscosity of 8.2 mPa s (corresponds to a 60% v/v glycerine solution) can be pumped without changes in the flow rates. Thus, nearly all feeding solutions can be delivered, which are commonly used in bioprocesses. The functionality of the first prototype of this microfluidic device was demonstrated by double-sided pH-controlled cultivations of Saccharomyces cerevisiae based on signals of fluorimetric sensors embedded at the bottom of the bioreactors. Furthermore, fed-batch cultivations with constant and exponential feeding profiles were successfully performed. Thus, the presented novel microfluidic device will be a useful tool for parallel and, thus, efficient optimization of controlled fed-batch bioprocesses in small-scale stirred tank bioreactors. This can help to reduce bioprocess development times drastically.

  8. [Technological characteristics of bioreactor landfill with aeration in the upper layer].

    Science.gov (United States)

    Tian, Ying; Wang, Shen; Xu, Qi-Yong

    2014-11-01

    In order to study the effects of upper-layer aerobic pretreatment in bioreactors on refuse degradation, leachate condition and methane production, two simulated columns were constructed, including traditional anaerobic bioreactor A1 and hybrid bioreactor C1 with aeration pretreatment in the upper layer. Results indicated that A1 was seriously inhibited by the accumulation of volatile fatty acids (VFA) with nearly no methane production and slower settlements. At the end of operations, refuse in A1 only deposited 5.4 cm which was less than half of that in C1. And up to 70 000 mg x L(-1) COD and 30 000 mg x L(-1) VFA could be monitored in the leachate. On the contrary, aerobic pretreatment effectively improved the removal of high VFA concentrations and remarkably accelerated the degradation rate. In bioreactor C1, COD and VFA concentrations were reduced to less than 14000 mg x L(-1) and 8900 mg x L(-1) at the end of the experiment, respectively. And about 61 976 mL methane gases were produced since aeration ceased on day 60 with its methane recovery efficiency rising to over 95%. However, the performance of hybrid bioreactors was still closely related to its operation conditions, such as aeration supply and leachate recirculation. Therefore, in order to guarantee better performance, appropriate aeration and leachate operations need to be provided.

  9. 甘草细胞在搅拌式生物反应器中的放大培养%Ampliifcation Culture of Glycyrrhiza uralensis Cell in Stirring Bioreactor

    Institute of Scientific and Technical Information of China (English)

    李雅丽; 孟婷婷; 王毛毛; 李晓雪; 李蓉蓉; 郭晓强

    2015-01-01

    Based on the establishment of a stable Glycyrrhiza uralensis (licorice) cells ampliifcation culture system in stirring bioreactor, the growth characteristics of cell suspension culture in stirring bioreactor were studied, including the cell growth, cell membrane permeability, the pH change of the culture system and licorice lfavonoids synthesis, and compared with the shake lfask culture. The results showed that cell biomass accumu-lation in the bioreactor was less than that in shake lfask under the same conditions, and the whole cultivation period was shortened. At the same time in the culture period, pH was slightly lower than that in the shake lfask, the H2O2 concentration in cells was 1.8 times of that in shake lfask, and the production of licorice lfavonoids was 1.5 times of that in the shake lfask. These indicated that the cultivation environment with mechanical agita-tion and lfuid shear in the bioreactor inhibited the cells growth at a certain level, but stimulated secondary me-tabolites licorice lfavonoids synthesis cell in higher levels.%在建立了稳定的甘草细胞搅拌式生物反应器放大培养体系的基础上,本文研究了甘草细胞在搅拌式反应器中悬浮培养的生长特性,包括细胞生长、细胞膜的透性、培养体系的pH变化及甘草黄酮合成情况等,并与摇瓶培养作比较。结果发现,同等条件下,反应器中培养细胞生物量的积累低于摇瓶培养,整个培养周期较摇瓶培养缩短。培养过程中同一时间段反应器中的pH值略低于摇瓶中的pH,细胞中H2O2的浓度是摇瓶中的1.8倍,甘草黄酮的产量是摇瓶培养的1.5倍,表明反应器中机械搅拌与流体剪切的培养环境对细胞生长起到一定程度的抑制作用,但刺激了细胞次生代谢产物甘草黄酮较高水平的合成。

  10. Proteins causing membrane fouling in membrane bioreactors.

    Science.gov (United States)

    Miyoshi, Taro; Nagai, Yuhei; Aizawa, Tomoyasu; Kimura, Katsuki; Watanabe, Yoshimasa

    2015-01-01

    In this study, the details of proteins causing membrane fouling in membrane bioreactors (MBRs) treating real municipal wastewater were investigated. Two separate pilot-scale MBRs were continuously operated under significantly different operating conditions; one MBR was a submerged type whereas the other was a side-stream type. The submerged and side-stream MBRs were operated for 20 and 10 days, respectively. At the end of continuous operation, the foulants were extracted from the fouled membranes. The proteins contained in the extracted foulants were enriched by using the combination of crude concentration with an ultrafiltration membrane and trichloroacetic acid precipitation, and then separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The N-terminal amino acid sequencing analysis of the proteins which formed intensive spots on the 2D-PAGE gels allowed us to partially identify one protein (OmpA family protein originated from genus Brevundimonas or Riemerella anatipestifer) from the foulant obtained from the submerged MBR, and two proteins (OprD and OprF originated from genus Pseudomonas) from that obtained from the side-stream MBR. Despite the significant difference in operating conditions of the two MBRs, all proteins identified in this study belong to β-barrel protein. These findings strongly suggest the importance of β-barrel proteins in developing membrane fouling in MBRs.

  11. Efficient Production Process for Food Grade Acetic Acid by Acetobacter aceti in Shake Flask and in Bioreactor Cultures

    Directory of Open Access Journals (Sweden)

    Hassan M. Awad

    2012-01-01

    Full Text Available Acetic acid is one of the important weak acids which had long history in chemical industries. This weak organic acid has been widely used as one of the key intermediate for many chemical, detergent, wood and food industries. The production of this acid is mainly carried out using submerged fermentation system and the standard strain Acetobacter aceti. In the present work, six different media were chosen from the literatures and tested for acetic acid production. The highest acetic acid production was produced in medium composed of glucose, yeast extract and peptone. The composition of this medium was optimized by changing the concentration of medium components. The optimized medium was composed of (g/L: glucose, 100; yeast extract, 12 and peptone 5 and yielded 53 g/L acetic acid in shake flask after 144 h fermentation. Further optimization in the production process was achieved by transferring the process to semi-industrial scale 16-L stirred tank bioreactor and cultivation under controlled pH condition. Under fully aerobic conditions, the production of acetic acid reached maximal concentration of about 76 g/L and 51 g/L for uncontrolled and controlled pH cultures, respectively.

  12. The carbon starvation response of Aspergillus niger during submerged cultivation: Insights from the transcriptome and secretome

    Directory of Open Access Journals (Sweden)

    Nitsche Benjamin M

    2012-08-01

    Full Text Available Abstract Background Filamentous fungi are confronted with changes and limitations of their carbon source during growth in their natural habitats and during industrial applications. To survive life-threatening starvation conditions, carbon from endogenous resources becomes mobilized to fuel maintenance and self-propagation. Key to understand the underlying cellular processes is the system-wide analysis of fungal starvation responses in a temporal and spatial resolution. The knowledge deduced is important for the development of optimized industrial production processes. Results This study describes the physiological, morphological and genome-wide transcriptional changes caused by prolonged carbon starvation during submerged batch cultivation of the filamentous fungus Aspergillus niger. Bioreactor cultivation supported highly reproducible growth conditions and monitoring of physiological parameters. Changes in hyphal growth and morphology were analyzed at distinct cultivation phases using automated image analysis. The Affymetrix GeneChip platform was used to establish genome-wide transcriptional profiles for three selected time points during prolonged carbon starvation. Compared to the exponential growth transcriptome, about 50% (7,292 of all genes displayed differential gene expression during at least one of the starvation time points. Enrichment analysis of Gene Ontology, Pfam domain and KEGG pathway annotations uncovered autophagy and asexual reproduction as major global transcriptional trends. Induced transcription of genes encoding hydrolytic enzymes was accompanied by increased secretion of hydrolases including chitinases, glucanases, proteases and phospholipases as identified by mass spectrometry. Conclusions This study is the first system-wide analysis of the carbon starvation response in a filamentous fungus. Morphological, transcriptomic and secretomic analyses identified key events important for fungal survival and their chronology. The

  13. In vivo bioreactors for mandibular reconstruction.

    Science.gov (United States)

    Tatara, A M; Wong, M E; Mikos, A G

    2014-12-01

    Large mandibular defects are difficult to reconstruct with good functional and aesthetic outcomes because of the complex geometry of craniofacial bone. While the current gold standard is free tissue flap transfer, this treatment is limited in fidelity by the shape of the harvested tissue and can result in significant donor site morbidity. To address these problems, in vivo bioreactors have been explored as an approach to generate autologous prefabricated tissue flaps. These bioreactors are implanted in an ectopic site in the body, where ossified tissue grows into the bioreactor in predefined geometries and local vessels are recruited to vascularize the developing construct. The prefabricated flap can then be harvested with vessels and transferred to a mandibular defect for optimal reconstruction. The objective of this review article is to introduce the concept of the in vivo bioreactor, describe important preclinical models in the field, summarize the human cases that have been reported through this strategy, and offer future directions for this exciting approach.

  14. Impact of stirred suspension bioreactor culture on the differentiation of murine embryonic stem cells into cardiomyocytes

    Directory of Open Access Journals (Sweden)

    Shafa Mehdi

    2011-12-01

    Full Text Available Abstract Background Embryonic stem cells (ESCs can proliferate endlessly and are able to differentiate into all cell lineages that make up the adult organism. Under particular in vitro culture conditions, ESCs can be expanded and induced to differentiate into cardiomyocytes in stirred suspension bioreactors (SSBs. However, in using these systems we must be cognizant of the mechanical forces acting upon the cells. The effect of mechanical forces and shear stress on ESC pluripotency and differentiation has yet to be clarified. The purpose of this study was to investigate the impact of the suspension culture environment on ESC pluripotency during cardiomyocyte differentiation. Results Murine D3-MHC-neor ESCs formed embyroid bodies (EBs and differentiated into cardiomyocytes over 25 days in static culture and suspension bioreactors. G418 (Geneticin was used in both systems from day 10 to enrich for cardiomyocytes by eliminating non-resistant, undifferentiated cells. Treatment of EBs with 1 mM ascorbic acid and 0.5% dimethyl sulfoxide from day 3 markedly increased the number of beating EBs, which displayed spontaneous and cadenced contractile beating on day 11 in the bioreactor. Our results showed that the bioreactor differentiated cells displayed the characteristics of fully functional cardiomyocytes. Remarkably, however, our results demonstrated that the bioreactor differentiated ESCs retained their ability to express pluripotency markers, to form ESC-like colonies, and to generate teratomas upon transplantation, whereas the cells differentiated in adherent culture lost these characteristics. Conclusions This study demonstrates that although cardiomyocyte differentiation can be achieved in stirred suspension bioreactors, the addition of medium enhancers is not adequate to force complete differentiation as fluid shear forces appear to maintain a subpopulation of cells in a transient pluripotent state. The development of successful ESC

  15. Plant growth and cultivation.

    Science.gov (United States)

    Podar, Dorina

    2013-01-01

    There is a variety of methods used for growing plants indoor for laboratory research. In most cases plant research requires germination and growth of plants. Often, people have adapted plant cultivation protocols to the conditions and materials at hand in their own laboratory and growth facilities. Here I will provide a guide for growing some of the most frequently used plant species for research, i.e., Arabidopsis thaliana, barley (Hordeum vulgare) and rice (Oryza sativa). However, the methods presented can be used for other plant species as well, especially if they are related to the above-mentioned species. The presented methods include growing plants in soil, hydroponics, and in vitro on plates. This guide is intended as a starting point for those who are just beginning to work on any of the above-mentioned plant species. Methods presented are to be taken as suggestive and modification can be made according to the conditions existing in the host laboratory.

  16. Pilot scale land-based cultivation of Saccharina latissima Linnaeus at southern European climate conditions: Growth and nutrient uptake at high temperatures

    DEFF Research Database (Denmark)

    Azevedo, Isabel C.; Silva Marinho, Goncalo; Silva, Diogo M.

    2016-01-01

    of integrated multi-trophic aquaculture (IMTA) systems, presenting good results considering both growth and biofiltration performance. In the present work, the cultivation of S. latissima in a pilot land-based system was performed in order to assess the efficiency of two different methods: tumbling in the water...... during high temperature periods. Densities around 8 kg m− 3 were effective in keeping epiphytes development low. This system may be used for seaweed monoculture or as a biofilter component of IMTA systems.......Saccharina latissima is a cold water seaweed species with commercial potential. The northern Portuguese coast is the southern distribution limit of the species, where some dispersed populations can be found. S. latissima has been identified as being a potential candidate for monoculture or as part...

  17. Trace Gas Emission from in-Situ Denitrifying Bioreactors

    Science.gov (United States)

    Pluer, W.; Walter, M. T.; Geohring, L.

    2014-12-01

    Despite decades of concerted effort to mitigate nonpoint source nitrate (NO3-) pollution from agricultural lands, these efforts have not been sufficient to arrest eutrophication. A primary process for removing excess NO3- from water is denitrification, where denitrifying bacteria use NO3- for respiration in the absence of oxygen. Denitrification results in reduced forms of nitrogen, often dinitrogen gas (N2) but also nitrous oxide (N2O), an aggressive greenhouse gas. A promising solution to NO3- pollution is to intercept agricultural discharges with denitrifying bioreactors (DNBRs). DNBRs provide conditions ideal for denitrifiers: an anaerobic environment, sufficient organic matter, and excess NO3-. These conditions are also ideal for methanogens, which produce methane (CH4), another harmful trace gas. While initial results from bioreactor studies show that they can cost-effectively remove NO3-, trace gas emissions are an unintended consequence. This study's goal was to determine how bioreactor design promotes denitrification while limiting trace gas production. Reactor inflow and outflow water samples were tested for nutrients, including NO3-, and dissolved inflow and outflow gas samples were tested for N2O and CH4. NO3- reduction and trace gas production were evaluated at various residence times, pHs, and inflow NO3- concentrations in field and lab-scale reactors. Low NO3- reduction indicated conditions that stressed denitrifying bacteria while high reductions indicated designs that optimized pollutant treatment for water quality. Several factors influenced high N2O, suggesting non-ideal conditions for the final step of complete denitrification. High CH4 emissions pointed to reactor media choice for discouraging methanogens, which may remove competition with denitrifiers. It is critical to understand all of potential impacts that DNBRs may have, which means identifying processes and design specifications that may affect them.

  18. Classification of cultivated plants.

    NARCIS (Netherlands)

    Brandenburg, W.A.

    1986-01-01

    Agricultural practice demands principles for classification, starting from the basal entity in cultivated plants: the cultivar. In establishing biosystematic relationships between wild, weedy and cultivated plants, the species concept needs re-examination. Combining of botanic classification, based

  19. THE USE OF CONTINUOUS, TEMPORARY IMMERSION BIOREACTOR SYSTEM AND SEMISOLID CULTURE MEDIUM FOR THE PRODUCTION OF Eucalyptus camaldulensis CLONES

    Directory of Open Access Journals (Sweden)

    Evânia Galvão Mendonça

    2016-01-01

    Full Text Available The plant micro-propagation in bioreactor systems is regarded as one way to reduce cost by automation and production scheduling. This research was carried out in order to obtain an efficient procedure for clone production of Eucalyptus camaldulensis on different types of bioreactor including continuous and temporary immersion bioreactor. To do so, the apical meristems (1 mm and the apical meristems with adjacent tissue (2,5 mm were used as initial explants. These tissues were cultured, for 60 days, in semisolid culture medium supplemented with 1 mg L -1 indole acetic acid (IAA and 0.32 mg L -1 benzylaminopurine (BA. After 60 days, the meristems with adjacent tissue were transferred to a continuous immersion bioreactor and maintained in dark or light conditions. In order to verify the effect of the explant source on bioreactor multiplication, the explants subcultured from meristems multiplied in semisolid culture medium and the meristems multiplied in continuous immersion bioreactor were tested and maintained in dark conditions. After establishing this parameters, the multiplication experiments were carried out in continuous and temporary immersion and the multiplied explants were then rooted in MS medium supplemented with 0, 2, 4, 8 and 20 mg L -1 indole butyric acid (IBA and kept in the dark or under controlled lighting conditions. After that, the rooting the plants were acclimatized in mist chamber. The meristem with adjacent tissue favored a greater number of buds/explants. The continuous immersion bioreactor in the dark provided higher shoots number and multiplication rate. The rooting was better on culture medium without auxin and kept in the dark for 15 days or the culture medium supplemented with auxin and maintained under light with 100% plantlet rooting. The Eucalyptus camaldulensis acclimatization was efficient, with high survival rate (76%. It was possible to establish the procedure for bioreactor micro-propagation of Eucalyptus

  20. Rhizosphere effect on survival of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium in manure-amended soil during cabbage (Brassica oleracea) cultivation under tropical field conditions in Sub-Saharan Africa.

    Science.gov (United States)

    Ongeng, Duncan; Muyanja, Charles; Ryckeboer, Jaak; Geeraerd, Annemie H; Springael, Dirk

    2011-09-15

    The effect of cabbage (Brassica oleracea) rhizosphere on survival of Escherichia coli O157:H7 and Salmonella Typhimurium in manure-amended soils under tropical field conditions was investigated in the Central Agro-Ecological Zone of Uganda. Three-week old cabbage seedlings were transplanted and cultivated for 120 days on manure-amended soil inoculated with 4 or 7 log CFU/g non-virulent E. coli O157:H7 and S. Typhimurium. Cabbage rhizosphere did not affect survival of the 4log CFU/g inocula in manure-amended soil and the two enteric bacteria were not detected on/in cabbage leaves at harvest. The 7 log CFU/g E. coli O157:H7 and S. Typhimurium survived in bulk soil for a maximum of 80 and 96 days, respectively, but the organisms remained culturable in cabbage rhizosphere up to the time of harvest. At 7 log CFU/g inoculum, E. coli O157:H7 and S. Typhimurium contamination on cabbage leaves occurred throughout the cultivation period. Leaf surface sterilisation with 1% AgNO(3) indicated that the organisms were present superficially and in protected locations on the leaves. These results demonstrate that under tropical field conditions, cabbage rhizosphere enhances the persistence of E. coli O157:H7 and S. Typhimurium in manure-amended soil at high inoculum density and is associated with long-term contamination of the leaves.

  1. Chip-based Three-dimensional Cell Culture in Perfused Micro-bioreactors

    Science.gov (United States)

    Gottwald, Eric; Lahni, Brigitte; Thiele, David; Giselbrecht, Stefan; Welle, Alexander; Weibezahn, Karl-Friedrich

    2008-01-01

    We have developed a chip-based cell culture system for the three-dimensional cultivation of cells. The chip is typically manufactured from non-biodegradable polymers, e.g., polycarbonate or polymethyl methacrylate by micro injection molding, micro hot embossing or micro thermoforming. But, it can also be manufactured from bio-degradable polymers. Its overall dimensions are 0.7 1 x 20 x 20 x 0.7 1 mm (h x w x l). The main features of the chips used are either a grid of up to 1156 cubic micro-containers (cf-chip) each the size of 120-300 x 300 x 300 μ (h x w x l) or round recesses with diameters of 300 μ and a depth of 300 μ (r-chip). The scaffold can house 10 Mio. cells in a three-dimensional configuration. For an optimal nutrient and gas supply, the chip is inserted in a bioreactor housing. The bioreactor is part of a closed steril circulation loop that, in the simplest configuration, is additionaly comprised of a roller pump and a medium reservoir with a gas supply. The bioreactor can be run in perfusion, superfusion, or even a mixed operation mode. We have successfully cultivated cell lines as well as primary cells over periods of several weeks. For rat primary liver cells we could show a preservation of organotypic functions for more than 2 weeks. For hepatocellular carcinoma cell lines we could show the induction of liver specific genes not or only slightly expressed in standard monolayer culture. The system might also be useful as a stem cell cultivation system since first differentiation experiments with stem cell lines were promising. PMID:19066592

  2. Effects of salinity build-up on the performance and bacterial community structure of a membrane bioreactor.

    Science.gov (United States)

    Luo, Wenhai; Phan, Hop V; Hai, Faisal I; Price, William E; Guo, Wenshan; Ngo, Hao H; Yamamoto, Kazuo; Nghiem, Long D

    2016-01-01

    This study investigated the effects of salinity increase on bacterial community structure in a membrane bioreactor (MBR) for wastewater treatment. The influent salt loading was increased gradually to simulate salinity build-up in the bioreactor during the operation of a high retention-membrane bioreactor (HR-MBR). Bacterial community diversity and structure were analyzed using 454 pyrosequencing of 16S rRNA genes of MBR mixed liquor samples. Results show that salinity increase reduced biological performance but did not affect microbial diversity in the bioreactor. Unweighted UniFrac and taxonomic analyses were conducted to relate the reduced biological performance to the change of bacterial community structure. In response to the elevated salinity condition, the succession of halophobic bacteria by halotolerant/halophilic microbes occurred and thereby the biological performance of MBR was recovered. These results suggest that salinity build-up during HR-MBR operation could be managed by allowing for the proliferation of halotolerant/halophilic bacteria.

  3. An integrated system for synchronous culture of animal cells under controlled conditions.

    Science.gov (United States)

    Mendoza-Pérez, Elena; Hernández, Vanessa; Palomares, Laura A; Serrato, José A

    2016-01-01

    The cell cycle has fundamental effects on cell cultures and their products. Tools to synchronize cultured cells allow the study of cellular physiology and metabolism at particular cell cycle phases. However, cells are most often arrested by methods that alter their homeostasis and are then cultivated in poorly controlled environments. Cell behavior could then be affected by the synchronization method and culture conditions used, and not just by the particular cell cycle phase under study. Moreover, only a few viable cells are recovered. Here, we designed an integrated system where a large number of cells from a controlled bioreactor culture is separated by centrifugal elutriation at high viabilities. In contrast to current elutriation methods, cells are injected directly from a bioreactor into an injection loop, allowing the introduction of a large number of cells into the separation chamber without stressful centrifugation. A low pulsation peristaltic pump increases the stability of the elutriation chamber. Using this approach, a large number of healthy cells at each cell cycle phase were obtained, allowing their direct inoculation into fully instrumented bioreactors. Hybridoma cells synchronized and cultured in this system behaved as expected for a synchronous culture.

  4. X-ray phase contrast imaging of calcified tissue and biomaterial structure in bioreactor engineered tissues.

    Science.gov (United States)

    Appel, Alyssa A; Larson, Jeffery C; Garson, Alfred B; Guan, Huifeng; Zhong, Zhong; Nguyen, Bao-Ngoc B; Fisher, John P; Anastasio, Mark A; Brey, Eric M

    2015-03-01

    Tissues engineered in bioreactor systems have been used clinically to replace damaged tissues and organs. In addition, these systems are under continued development for many tissue engineering applications. The ability to quantitatively assess material structure and tissue formation is critical for evaluating bioreactor efficacy and for preimplantation assessment of tissue quality. Techniques that allow for the nondestructive and longitudinal monitoring of large engineered tissues within the bioreactor systems will be essential for the translation of these strategies to viable clinical therapies. X-ray Phase Contrast (XPC) imaging techniques have shown tremendous promise for a number of biomedical applications owing to their ability to provide image contrast based on multiple X-ray properties, including absorption, refraction, and scatter. In this research, mesenchymal stem cell-seeded alginate hydrogels were prepared and cultured under osteogenic conditions in a perfusion bioreactor. The constructs were imaged at various time points using XPC microcomputed tomography (µCT). Imaging was performed with systems using both synchrotron- and tube-based X-ray sources. XPC µCT allowed for simultaneous three-dimensional (3D) quantification of hydrogel size and mineralization, as well as spatial information on hydrogel structure and mineralization. Samples were processed for histological evaluation and XPC showed similar features to histology and quantitative analysis consistent with the histomorphometry. These results provide evidence of the significant potential of techniques based on XPC for noninvasive 3D imaging engineered tissues grown in bioreactors.

  5. Design considerations and challenges for mechanical stretch bioreactors in tissue engineering.

    Science.gov (United States)

    Lei, Ying; Ferdous, Zannatul

    2016-05-01

    With the increase in average life expectancy and growing aging population, lack of functional grafts for replacement surgeries has become a severe problem. Engineered tissues are a promising alternative to this problem because they can mimic the physiological function of the native tissues and be cultured on demand. Cyclic stretch is important for developing many engineered tissues such as hearts, heart valves, muscles, and bones. Thus a variety of stretch bioreactors and corresponding scaffolds have been designed and tested to study the underlying mechanism of tissue formation and to optimize the mechanical conditions applied to the engineered tissues. In this review, we look at various designs of stretch bioreactors and common scaffolds and offer insights for future improvements in tissue engineering applications. First, we summarize the requirements and common configuration of stretch bioreactors. Next, we present the features of different actuating and motion transforming systems and their applications. Since most bioreactors must measure detailed distributions of loads and deformations on engineered tissues, techniques with high accuracy, precision, and frequency have been developed. We also cover the key points in designing culture chambers, nutrition exchanging systems, and regimens used for specific tissues. Since scaffolds are essential for providing biophysical microenvironments for residing cells, we discuss materials and technologies used in fabricating scaffolds to mimic anisotropic native tissues, including decellularized tissues, hydrogels, biocompatible polymers, electrospinning, and 3D bioprinting techniques. Finally, we present the potential future directions for improving stretch bioreactors and scaffolds. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:543-553, 2016.

  6. X-ray Phase Contrast Imaging of Calcified Tissue and Biomaterial Structure in Bioreactor Engineered Tissues

    Energy Technology Data Exchange (ETDEWEB)

    Appel, Alyssa A. [Illinois Inst. of Technology, Chicago, IL (United States); Edward Hines Jr. VA Hospital, IL (United States); Larson, Jeffery C. [Illinois Inst. of Technology, Chicago, IL (United States); Edward Hines Jr. VA Hospital, IL (United States); Garson, III, Alfred B. [George Washington Univ., Washington, DC (United States); Guan, Huifeng [George Washington Univ., Washington, DC (United States); Zhong, Zhong [Brookhaven National Lab. (BNL), Upton, NY (United States); Nguyen, Bao-Ngoc [Univ. of Maryland, College Park, MD (United States); Fisher, John P. [Univ. of Maryland, College Park, MD (United States); Anastasio, Mark A. [George Washington Univ., Washington, DC (United States); Brey, Eric M. [Illinois Inst. of Technology, Chicago, IL (United States); Edward Hines Jr. VA Hospital, IL (United States)

    2014-11-04

    Tissues engineered in bioreactor systems have been used clinically to replace damaged tissues and organs. In addition, these systems are under continued development for many tissue engineering applications. The ability to quantitatively assess material structure and tissue formation is critical for evaluating bioreactor efficacy and for preimplantation assessment of tissue quality. These techniques allow for the nondestructive and longitudinal monitoring of large engineered tissues within the bioreactor systems and will be essential for the translation of these strategies to viable clinical therapies. X-ray Phase Contrast (XPC) imaging techniques have shown tremendous promise for a number of biomedical applications owing to their ability to provide image contrast based on multiple X-ray properties, including absorption, refraction, and scatter. In this research, mesenchymal stem cell-seeded alginate hydrogels were prepared and cultured under osteogenic conditions in a perfusion bioreactor. The constructs were imaged at various time points using XPC microcomputed tomography (µCT). Imaging was performed with systems using both synchrotron- and tube-based X-ray sources. XPC µCT allowed for simultaneous three-dimensional (3D) quantification of hydrogel size and mineralization, as well as spatial information on hydrogel structure and mineralization. Samples were processed for histological evaluation and XPC showed similar features to histology and quantitative analysis consistent with the histomorphometry. Furthermore, these results provide evidence of the significant potential of techniques based on XPC for noninvasive 3D imaging engineered tissues grown in bioreactors.

  7. Advanced microscale bioreactor system: a representative scale-down model for bench-top bioreactors.

    Science.gov (United States)

    Hsu, Wei-Ting; Aulakh, Rigzen P S; Traul, Donald L; Yuk, Inn H

    2012-12-01

    In recent years, several automated scale-down bioreactor systems have been developed to increase efficiency in cell culture process development. ambr™ is an automated workstation that provides individual monitoring and control of culture dissolved oxygen and pH in single-use, stirred-tank bioreactors at a working volume of 10-15 mL. To evaluate the ambr™ system, we compared the performance of four recombinant Chinese hamster ovary cell lines in a fed-batch process in parallel ambr™, 2-L bench-top bioreactors, and shake flasks. Cultures in ambr™ matched 2-L bioreactors in controlling the environment (temperature, dissolved oxygen, and pH) and in culture performance (growth, viability, glucose, lactate, Na(+), osmolality, titer, and product quality). However, cultures in shake flasks did not show comparable performance to the ambr™ and 2-L bioreactors.

  8. The culture of human embryonic stem cells in microchannel perfusion bioreactors

    Science.gov (United States)

    Korin, Natanel; Bransky, Avishay; Dinnar, Uri; Levenberg, Shulamit

    2007-12-01

    The culture of human Embryonic Stem (ES) cells in microchannel bioreactors can be highly beneficial for ES cell biology studies and ES tissue engineering applications. In the present study we examine the use of Human Foreskin Fibroblasts (HFF) cells as feeder cells for human ES culture in a microchannel perfusion bioreactor. PDMS microchannels (depth:130 micron) were fabricated using conventional soft-lithography techniques. The channels were sterilized, coated with a human fibronectin solution and seeded with cells. Following a period of static incubation, culture medium was perfused through the channels at various flow rates and cell growth was monitored throughout the culture process. Mass transport and fluid mechanics models were used to evaluate the culture conditions (shear stress, oxygen levels within the micro-bioreactor as a function of the medium flow rate. The conditions for successful long-term culture (>7 days) of HFF under flow were established. Experiments with human embryonic stem cells cultured in microchannels show that the conditions essential to co-culture human ES cell on HFF cells under perfusion differ from the conditions necessary for HFF cell culture. Human ES cells were found to be highly sensitive to flow and culture conditions and did not grow under flow rates which were suitable for HFF long-term culture. Successful culture of undifferentiated human ES cell colonies in a perfusion micro-bioreactor is a basic step towards utilizing microfluidic techniques to explore stem cell biology.

  9. Novel Sensor-Enabled Ex Vivo Bioreactor: A New Approach towards Physiological Parameters and Porcine Artery Viability.

    Science.gov (United States)

    Mundargi, Raghavendra; Venkataraman, Divya; Kumar, Saranya; Mogal, Vishal; Ortiz, Raphael; Loo, Joachim; Venkatraman, Subbu; Steele, Terry

    2015-01-01

    The aim of the present work is to design and construct an ex vivo bioreactor system to assess the real time viability of vascular tissue. Porcine carotid artery as a model tissue was used in the ex vivo bioreactor setup to monitor its viability under physiological conditions such as oxygen, pressure, temperature, and flow. The real time tissue viability was evaluated by monitoring tissue metabolism through a fluorescent indicator "resorufin." Our ex vivo bioreactor allows real time monitoring of tissue responses along with physiological conditions. These ex vivo parameters were vital in determining the tissue viability in sensor-enabled bioreactor and our initial investigations suggest that, porcine tissue viability is considerably affected by high shear forces and low oxygen levels. Histological evaluations with hematoxylin and eosin and Masson's trichrome staining show intact endothelium with fresh porcine tissue whereas tissues after incubation in ex vivo bioreactor studies indicate denuded endothelium supporting the viability results from real time measurements. Hence, this novel viability sensor-enabled ex vivo bioreactor acts as model to mimic in vivo system and record vascular responses to biopharmaceutical molecules and biomedical devices.

  10. EXPERIMENTAL STUDY ON THE GAS-LIQUID FLOW IN THE MEMBRANE MICROPORE AERATION BIOREACTOR

    Directory of Open Access Journals (Sweden)

    DONG LIU

    2008-12-01

    Full Text Available Particle Image Velocimetry (PIV has been developed to measure the typical two-phase flow of various work conditions in Membrane Micropore Aeration Bioreactor (MMAB. The fluid phase is separated out using image processing techniques, which provides accurate measurements for the Bioreactor’s flow field, and makes it possible for quantitative analysis of the momentum exchange, heat exchange and the process of micro-admixture. The experimental method PIV used in this paper can preferably measure the complex flow in the reactor and initiates a new approach for the bioreactor design which mainly depends on experience at present.

  11. Preparation of [11C]formaldehyde using a hollow fiber membrane bioreactor.

    Science.gov (United States)

    Hughes, J A; Jay, M

    1995-01-01

    A bioreactor consisting of the enzymes alcohol oxidase and catalase immobilized onto a hollow fiber membrane was used to convert [11C]methanol to [11C]formaldehyde. Using an alcohol oxidase:catalase ratio of 1:500 U, conversion yields of 90-95% were obtained allowing the production of up to 7400 MBq (200 mCi) of [11C]formaldehyde in 5 min. The hollow fiber bioreactor allowed for a convenient, rapid synthesis with yields significantly higher than the standard chemical procedures, has demonstrable advantages over glass bead immobilized systems (primarily due to convective flow), and was amenable to hot cell conditions.

  12. Optimal homogenization of perfusion flows in microfluidic bio-reactors; a numerical study

    CERN Document Server

    Okkels, Fridolin; Bruus, Henrik

    2009-01-01

    To ensure homogeneous conditions within the complete area of perfused microfluidic bio-reactors, we develop a general design of a continuously feed bio-reactor with uniform perfusion flow. This is achieved by introducing a specific type of perfusion inlet to the reaction area. The geometry of these inlets are found using the methods of topology optimization and shape optimization. The results are compared with two different analytic models, from which a general parametric description of the design is obtained and tested numerically. Such a parametric description will generally be beneficial for the design of a broad range of microfluidic bioreactors used for e.g. cell culturing and analysis, and in feeding bio-arrays.

  13. An Experimental Study on the Treating River Sewage with New Bioreactor

    Institute of Scientific and Technical Information of China (English)

    Jiang Fan; Chen Weiping; Zhang Tao

    2007-01-01

    A new bioreactor on the basis of a dynamic fluidized bed was designed, which combines advantages of the fluidized bed and a biological contactor. The experiments of start-up, normal operation and parameter adjustment are carried out. The results show that the bioreactor can be quickly started up in the condition that the fill is 50%, the hydraulic retention time is 72 min, aerate speed is 2.5 m3/h, rotation-cage rotated speed is 1.5 r/min, and the removal rates of chemical oxygen demand (CODCr)and Ammonia nitrogen (NH3-N) are 75.34% and 80.98% respectively. The influence of the operation parameter on removal rates of the bioreactor is analyzed, and an appropriate operation parameter is provided.

  14. Molecular analysis of methanogens involved in methanogenic degradation of tetramethylammonium hydroxide in full-scale bioreactors.

    Science.gov (United States)

    Whang, Liang-Ming; Hu, Tai-Ho; Liu, Pao-Wen Grace; Hung, Yu-Ching; Fukushima, Toshikazu; Wu, Yi-Ju; Chang, Shao-Hsiung

    2015-02-01

    This study investigated methanogenic communities involved in degradation of tetramethylammonium hydroxide (TMAH) in three full-scale bioreactors treating TMAH-containing wastewater. Based on the results of terminal-restriction fragment-length polymorphism (T-RFLP) and quantitative PCR analyses targeting the methyl-coenzyme M reductase alpha subunit (mcrA) genes retrieved from three bioreactors, Methanomethylovorans and Methanosarcina were the dominant methanogens involved in the methanogenic degradation of TMAH in the bioreactors. Furthermore, batch experiments were conducted to evaluate mcrA messenger RNA (mRNA) expression during methanogenic TMAH degradation, and the results indicated that a higher level of TMAH favored mcrA mRNA expression by Methansarcina, while Methanomethylovorans could only express considerable amount of mcrA mRNA at a lower level of TMAH. These results suggest that Methansarcina is responsible for methanogenic TMAH degradation at higher TMAH concentrations, while Methanomethylovorans may be important at a lower TMAH condition.

  15. Tubular bioreactor and its application; Tubular bioreactor to sono tekiyo

    Energy Technology Data Exchange (ETDEWEB)

    Endo, I.; Nagamune, T. [The University of Tokyo, Tokyo (Japan). Faculty of Engineering; Yuki, K. [Nikka Whisky Distilling Co. Ltd. Tokyo (Japan); Inaba, H. [Sumitomo Heavy Industries, Ltd., Tokyo (Japan)

    1994-09-05

    The loop type tubular bioreactor (TBR) was developed where biocatalysts are trapped in the reactor by membrane module. A UF membrane or MF membrane and crossflow filtration were adopted for the membrane module, and the reactor loop was composed of four membrane modules. The reactor was operated at 2-4 m/s in membrane surface velocity and 300-400 kPa in filtration pressure. As the result of the high-density culture of lactic acid bacteria and yeast, a biomass concentration was more than 10 times that in batch culture, suggesting the remarkable enhancement of a production efficiency. As the result of the continuous fermentation of cider, the fast fermentation more than 60 times that in conventional ones was obtained together with the same quality as conventional ones. Such a fast fermentation was probably achieved by yeast suspended in the fermenter of TBR, by yeast hardly affected physico-chemically as compared with immobilized reactors, and by small effect of mass transfer on reaction systems. 4 refs., 6 figs.

  16. Effect of Mixing on Microorganism Growth in Loop Bioreactors

    Directory of Open Access Journals (Sweden)

    A. M. Al Taweel

    2012-01-01

    Full Text Available The impact of mixing on the promotion of microorganism growth rate has been analyzed using a multiphase forced-circulation pipe-loop reactor model capable of identifying conditions under which it is possible to convert natural gas into Single-Cell Protein. The impact of mixing in the interphase mass transfer was found to exert a critical role in determining the overall productivity of the bioreactor, particularly at the high cell loadings needed to reduce the capital costs associated with the large-scale production needed for the production of relatively low-value SCP in a sustainable manner.

  17. Transient Behavior of Ethanol Fermentation in Immobilized Cell Bioreactors*

    OpenAIRE

    Tohru, KANNO; Yoshinori, FUJISHIGE; Hiroyuki, Ito; koichi, yamazaki; Masayoshi, KOBAYASHI

    1990-01-01

    The dynamic behavior of ethanol fermentation catalysed by an immobilized cell has been studied in batch and continuous stirred tank bioreactors, changing the operating conditions in a stepwise fashion. The rate of ethanol fermentation in the flow reactor reaches a new steady state within 60 min for the stepwise change in temperature or flow rate at 15〜30℃ and the residence time t_R=40 hr. The rate of fermentation obeys the Lineweaven-Burk plot and the Michaelis constant is calculated

  18. Composting of a solid olive-mill by-product ("alperujo") and the potential of the resulting compost for cultivating pepper under commercial conditions.

    Science.gov (United States)

    Alburquerque, J A; Gonzálvez, J; García, D; Cegarra, J

    2006-01-01

    A pollutant solid material called "alperujo" (AL), which is the main by-product from the Spanish olive oil industry, was composted with a cotton waste as bulking agent, and the compost obtained (ALC) was compared with a cattle manure (CM) and a sewage sludge compost (SSC) for use as organic amendment on a calcareous soil. The experiment was conducted with a commercial pepper crop in a greenhouse using fertigation. Composting AL involved a relatively low level of organic matter biodegradation, an increase in pH and clear decreases in the C/N and the fat, water-soluble organic carbon and phenol contents. The resulting compost, which was rich in organic matter and free of phytotoxicity, had a high potassium and organic nitrogen content but was low in phosphorus and micronutrients. The marketable yields of pepper obtained with all three organic amendments were similar, thus confirming the composting performance of the raw AL. When CM and SSC were used for soil amendment, the soil organic matter content was significantly reduced after cultivation, while it remained almost unchanged in the ALC-amended plots.

  19. Non-disruptive measurement system of cell viability in bioreactors

    Science.gov (United States)

    Rudek, F.; Nelsen, B. L.; Baselt, T.; Berger, T.; Wiele, M.; Prade, I.; Hartmann, P.

    2016-04-01

    Nutrient and oxygen transport, as well as the removal of metabolic waste are essential processes to support and maintain viable tissue. Current bioreactor technology used to grow tissue cultures in vitro has a fundamental limit to the thickness of tissues. Based on the low diffusion limit of oxygen a maximum tissue thickness of 200 μm is possible. The efficiency of those systems is currently under investigation. During the cultivation process of the artificial tissue in bioreactors, which lasts 28 days or longer, there are no possibilities to investigate the viability of cells. This work is designed to determine the influence of a non-disruptive cell viability measuring system on cellular activity. The measuring system uses a natural cellular marker produced during normal metabolic activity. Nicotinamide adenine dinucleotide (NADH) is a coenzyme naturally consumed and produced during cellular metabolic processes and has thoroughly been studied to determine the metabolic state of a cell. Measuring the fluorescence of NADH within the cell represents a non-disruptive marker for cell viability. Since the measurement process is optical in nature, NADH fluorescence also provides a pathway for sampling at different measurement depths within a given tissue sample. The measurement system we are using utilizes a special UV light source, to excite the NADH fluorescence state. However, the high energy potentially alters or harms the cells. To investigate the influence of the excitation signal, the cells were irradiated with a laser operating at a wavelength of 355 nm and examined for cytotoxic effects. The aim of this study was to develop a non-cytotoxic system that is applicable for large-scale operations during drug-tissue interaction testing.

  20. Design and Use of a Novel Bioreactor for Regeneration of Biaxially Stretched Tissue-Engineered Vessels.

    Science.gov (United States)

    Huang, Angela Hai; Lee, Yong-Ung; Calle, Elizabeth A; Boyle, Michael; Starcher, Barry C; Humphrey, Jay D; Niklason, Laura E

    2015-08-01

    Conventional bioreactors are used to enhance extracellular matrix (ECM) production and mechanical strength of tissue-engineered vessels (TEVs) by applying circumferential strain, which is uniaxial stretching. However, the resulting TEVs still suffer from inadequate mechanical properties, where rupture strengths and compliance values are still very different from native arteries. The biomechanical milieu of native arteries consists of both circumferential and axial loading. Therefore, to better simulate the physiological stresses acting on native arteries, we built a novel bioreactor system to enable biaxial stretching of engineered arteries during culture. This new bioreactor system allows for independent control of circumferential and axial stretching parameters, such as displacement and beat rate. The assembly and setup processes for this biaxial bioreactor system are reliable with a success rate greater than 75% for completion of long-term sterile culture. This bioreactor also supports side-by-side assessments of TEVs that are cultured under three types of mechanical conditions (static, uniaxial, and biaxial), all within the same biochemical environment. Using this bioreactor, we examined the impact of biaxial stretching on arterial wall remodeling of TEVs. Biaxial TEVs developed the greatest wall thickness compared with static and uniaxial TEVs. Unlike uniaxial loading, biaxial loading led to undulated collagen fibers that are commonly found in native arteries. More importantly, the biaxial TEVs developed the most mature elastin in the ECM, both qualitatively and quantitatively. The presence of mature extracellular elastin along with the undulated collagen fibers may contribute to the observed vascular compliance in the biaxial TEVs. The current work shows that biaxial stretching is a novel and promising means to improve TEV generation. Furthermore, this novel system allows us to optimize biomechanical conditioning by unraveling the interrelationships among the

  1. The modeling of ethanol production by Kluyveromyces marxianus using whey as substrate in continuous A-Stat bioreactors.

    Science.gov (United States)

    Gabardo, Sabrina; Pereira, Gabriela Feix; Rech, Rosane; Ayub, Marco Antônio Záchia

    2015-09-01

    We investigated the kinetics of whey bioconversion into ethanol by Kluyveromyces marxianus in continuous bioreactors using the "accelerostat technique" (A-stat). Cultivations using free and Ca-alginate immobilized cells were evaluated using two different acceleration rates (a). The kinetic profiles of these systems were modeled using four different unstructured models, differing in the expressions for the specific growth (μ) and substrate consumption rates (r s), taking into account substrate limitation and product inhibition. Experimental data showed that the dilution rate (D) directly affected cell physiology and metabolism. The specific growth rate followed the dilution rate (μ≈D) for the lowest acceleration rate (a = 0.0015 h(-2)), condition in which the highest ethanol yield (0.52 g g(-1)) was obtained. The highest acceleration rate (a = 0.00667 h(-2)) led to a lower ethanol yield (0.40 g g(-1)) in the system where free cells were used, whereas with immobilized cells ethanol yields increased by 23 % (0.49 g g(-1)). Among the evaluated models, Monod and Levenspiel combined with Ghose and Tyagi models were found to be more appropriate for describing the kinetics of whey bioconversion into ethanol. These results may be useful in scaling up the process for ethanol production from whey.

  2. Production of Biomass-Degrading Multienzyme Complexes under Solid-State Fermentation of Soybean Meal Using a Bioreactor.

    Science.gov (United States)

    Vitcosque, Gabriela L; Fonseca, Rafael F; Rodríguez-Zúñiga, Ursula Fabiola; Bertucci Neto, Victor; Couri, Sonia; Farinas, Cristiane S

    2012-01-01

    Biomass-degrading enzymes are one of the most costly inputs affecting the economic viability of the biochemical route for biomass conversion into biofuels. This work evaluates the effects of operational conditions on biomass-degrading multienzyme production by a selected strain of Aspergillus niger. The fungus was cultivated under solid-state fermentation (SSF) of soybean meal, using an instrumented lab-scale bioreactor equipped with an on-line automated monitoring and control system. The effects of air flow rate, inlet air relative humidity, and initial substrate moisture content on multienzyme (FPase, endoglucanase, and xylanase) production were evaluated using a statistical design methodology. Highest production of FPase (0.55 IU/g), endoglucanase (35.1 IU/g), and xylanase (47.7 IU/g) was achieved using an initial substrate moisture content of 84%, an inlet air humidity of 70%, and a flow rate of 24 mL/min. The enzymatic complex was then used to hydrolyze a lignocellulosic biomass, releasing 4.4 g/L of glucose after 36 hours of saccharification of 50 g/L pretreated sugar cane bagasse. These results demonstrate the potential application of enzymes produced under SSF, thus contributing to generate the necessary technological advances to increase the efficiency of the use of biomass as a renewable energy source.

  3. Woodchip denitrification bioreactors: Impact of temperature and hydraulic retention time on nitrate removal

    Science.gov (United States)

    Woodchip denitrification bioreactors, a relatively new technology for edge-of-field treatment of subsurface agricultural drainage water, have shown potential for nitrate removal. However, very few studies have evaluated the performance of these reactors under controlled conditions similar to the fie...

  4. Biological conversion of synthesis gas. Topical report: Bioreactor studies

    Energy Technology Data Exchange (ETDEWEB)

    Basu, R.; Klasson, K.T.; Clausen, E.C.; Gaddy, J.L.

    1993-09-01

    The purpose of the proposed research is to develop a technically and economically feasible process for biologically producing H{sub 2} from synthesis gas while, at the same time, removing harmful sulfur gas compounds. Six major tasks are being studied: culture development, where the best cultures are selected and conditions optimized for simultaneous hydrogen production and sulfur gas removal; mass transfer and kinetic studies in which equations necessary for process design are developed; bioreactor design studies, where the cultures chosen in Task 1 are utilized in continuous reaction vessels to demonstrate process feasibility and define operating conditions; evaluation of biological synthesis gas conversion under limiting conditions in preparation for industrial demonstration studies; process scale-up where laboratory data are scaled to larger-size units in preparation for process demonstration in a pilot-scale unit; and economic evaluation, where process simulations are used to project process economics and identify high cost areas during sensitivity analyses. The purpose of this report is to present results from bioreactor studies involving H{sub 2} production by water gas shift and H{sub 2}S removal to produce elemental sulfur. Many of the results for H{sub 2} production by Rhodospirillum rubrum have been presented during earlier contracts. Thus, this report concentrates mainly on H{sub 2}S conversion to elemental sulfur by R. rubrum.

  5. Mixotrophic Cultivation of Tetraselmis sp.-1 in Airlift Photobioreactor

    Institute of Scientific and Technical Information of China (English)

    Shen Jihong(沈继红); Yu Junhong; Lin Xuezheng; Li Guangyou; Liu Fayi

    2003-01-01

    Tetraselmis sp.-1 is a new microalgae strain constructed by cell fusion technique. In this paper, the mixotrophic cultivation of Tetraselmis sp.-1 in airlift photobioreactor is investigated. Firstly, the paper calculates the light attenuation in the mixotrophic medium, and sets the light attenuation model. Secondly, it uses the same dissolved oxygen coefficient (Kd) of flask culture to select the aeration of bioreactor. Finally, it sets the growth kinetic model, production (chlorophyll-a and total lipid) kinetic models and substrate (glucose) consumption kinetic model of Tetraselmis sp.-1 in airlift photobioreactor.

  6. Nitrogen removal and spatial distribution of denitrifier and anammox communities in a bioreactor for mine drainage treatment.

    Science.gov (United States)

    Herbert, Roger B; Winbjörk, Harry; Hellman, Maria; Hallin, Sara

    2014-12-01

    Mine drainage water may contain high levels of nitrate (NO3(-)) due to undetonated nitrogen-based explosives. The removal of NO3(-) and nitrite (NO2(-)) in cold climates through the microbial process of denitrification was evaluated using a pilot-scale fixed-bed bioreactor (27 m(3)). Surface water was diverted into the above-ground bioreactor filled with sawdust, crushed rock, and sewage sludge. At hydraulic residence times of ca.15 h and with the addition of acetate, NO3(-) and NO2(-) were removed to below detection levels at a NO3(-) removal rate of 5-10 g N m(-3) (bioreactor material) d(-1). The functional groups contributing to nitrogen removal in the bioreactor were studied by quantifying nirS and nirK present in denitrifying bacteria, nosZI and nosZII genes from the nitrous oxide - reducing community, and a taxa-specific part of the16S rRNA gene for the anammox community. The abundances of nirS and nirK were almost 2 orders of magnitude greater than the anammox specific 16S rRNA gene, indicating that denitrification was the main process involved in nitrogen removal. The spatial distribution of the quantified genes was heterogeneous in the bioreactor, with trends observed in gene abundance as a function of depth, distance from the bioreactor inlet, and along specific flowpaths. There was a significant relationship between the abundance of nirS, nirK, and nosZI genes and depth in the bioreactor, such that the abundance of organisms containing these genes may be controlled by oxygen diffusion and substrate supply in the partially or completely water-saturated material. Among the investigated microbial functional groups, nirS and anammox bacterial 16S rRNA genes exhibited a systematic trend of decreasing and increasing abundance, respectively, with distance from the inlet, which suggested that the functional groups respond differently to changing environmental conditions. The greater abundance of nirK along central flowpaths may indicate that the bioreactor

  7. Dependence of fungal characteristics on seed morphology and shear stress in bioreactors.

    Science.gov (United States)

    Lu, Hongzhong; Li, Chao; Tang, Wenjun; Wang, Zejian; Xia, Jianye; Zhang, Siliang; Zhuang, Yingping; Chu, Ju; Noorman, Henk

    2015-05-01

    The fungal morphology during submerged cultivations has a profound influence on the overall performance of bioreactors. In this research, glucoamylase production by Aspergillus niger has been taken as a model to improve more insights. The morphology engineering could be conducted effectively by changing the seed morphology, as well as specific power input. During the fed-batch cultivations, pellet formation under milder shear stress field helped to reduce the broth viscosity, thus relieving oxygen limitation and promoting the enzyme production. Furthermore, we found that the relation between the shear stress field, which was characterized by energy dissipation rate/circulation function (EDCF), and enzyme activity was consistent with quadratic parabola, which threw light on the process optimization and scale-up for industrial enzyme production.

  8. Hydrodynamics research of wastewater treatment bioreactors

    Institute of Scientific and Technical Information of China (English)

    REN Nan-qi; ZHANG Bing; ZHOU Xue-fei

    2009-01-01

    To optimize the design and improve the performance of wastewater treatment bioreactors, the review concerning the hydrodynamics explored by theoretical equations, process experiments, modeling of the hydrody-namics and flow field measurement is presented. Results of different kinds of experiments show that the hydro-dynamic characteristics can affect sludge characteristics, mass transfer and reactor performance significantly. A-long with the development of theoretical equations, turbulence models including large eddy simulation models and Reynolds-averaged Navier-Stokes (RANS) models are widely used at present. Standard and modified k-ε models are the most widely used eddy viscosity turbulence models for simulating flows in bioreactors. Numericalsimulation of hydrodynamics is proved to be efficient for optimizing design and operation. The development of measurement techniques with high accuracy and low intrusion enables the flow filed in the bioreactors to be transparent. Integration of both numerical simulation and experimental measurement can describe the hydrody-namics very well.

  9. Palladium nanoparticles produced by fermentatively cultivated bacteria as catalyst for diatrizoate removal with biogenic hydrogen

    Energy Technology Data Exchange (ETDEWEB)

    Hennebel, T.; Fitts, J.; Nevel, S. V.; Verschuere, S.; De Corte, S.; De Gusseme, B.; Cuvelier, C.; van der Lelie, D.; Boon, N.; Verstraete, W.

    2011-05-17

    A new biological inspired method to produce nanopalladium is the precipitation of Pd on a bacterium, i.e., bio-Pd. This bio-Pd can be applied as catalyst in dehalogenation reactions. However, large amounts of hydrogen are required as electron donor in these reactions resulting in considerable costs. This study demonstrates how bacteria, cultivated under fermentative conditions, can be used to reductively precipitate bio-Pd catalysts and generate the electron donor hydrogen. In this way, one could avoid the costs coupled to hydrogen supply. The catalytic activities of Pd(0) nanoparticles produced by different strains of bacteria (bio-Pd) cultivated under fermentative conditions were compared in terms of their ability to dehalogenate the recalcitrant aqueous pollutants diatrizoate and trichloroethylene. While all of the fermentative bio-Pd preparations followed first order kinetics in the dehalogenation of diatrizoate, the catalytic activity differed systematically according to hydrogen production and starting Pd(II) concentration in solution. Batch reactors with nanoparticles formed by Citrobacter braakii showed the highest diatrizoate dehalogenation activity with first order constants of 0.45 {+-} 0.02 h{sup -1} and 5.58 {+-} 0.6 h{sup -1} in batches with initial concentrations of 10 and 50 mg L{sup -1} Pd, respectively. Nanoparticles on C. braakii, used in a membrane bioreactor treating influent containing 20 mg L{sup -1} diatrizoate, were capable of dehalogenating 22 mg diatrizoate mg{sup -1} Pd over a period of 19 days before bio-Pd catalytic activity was exhausted. This study demonstrates the possibility to use the combination of Pd(II), a carbon source and bacteria under fermentative conditions for the abatement of environmental halogenated contaminants.

  10. Mathematical modeling of ultrasound in tissue engineering: From bioreactors to the cellular scale

    Science.gov (United States)

    Louw, Tobias M.

    predicted resonance. It appears that ultrasound applied close to the cell's resonant frequency effectively recreates the mechanical stimulation experienced by cells during natural movement. Ultrasonic bioreactors may therefore reproduce physiological conditions just as well as more complex bioreactors.

  11. Insect cell entrapment, growth and recovering using a single-use fixed-bed bioreactor. Scaling up and recombinant protein production.

    Science.gov (United States)

    Ventini-Monteiro, D; Dubois, S; Astray, R M; Castillo, J; Pereira, C A

    2015-12-20

    Insect cells are largely used for industrial production of vaccines, viral vectors and recombinant proteins as well as in research and development as an important tool for biology and bioprocess studies. They grow in suspension and are semi-adherent cells. Among the cell culture systems enabling scalable bioprocess the single-use fixed-bed iCELLis(®) bioreactors offer great advantages. We have established the conditions for Drosophila melanogaster Schneider 2 (S2) and Spodoptera frugiperda (Sf9) cells entrapment into the fixed-bed, cell growth and recover from the fixed-bed once high cell densities were attained. Our established protocol allowed these cells, at a cell seeding of 2×1E5 cells/microfiber carriers (MC) (3.5×1E6cells/mL; 1.7×1E4cells/cm(2)), to grow inside a 4m(2)/200mL fixed-bed attaining a concentration of 5.3×1E6 cells/MC (9.5×1E7cells/mL; 4.7×1E5 cells/cm(2)) for S2 cells or 4.6×1E6 cells/MC (8×1E7cells/mL; 4.1×1E5cells/cm(2)) for Sf9 cells. By washing the fixed-bed, entrapped cells could then be recovered from the fixed-bed at a high rate (>85%) with high viability (>95%) by increasing the agitation to 1200/1500rpm. Although the cell yields in the fixed-bed bioreactor were comparable to those obtained in a stirred tank (respectively, 1.3×1E10 and 2.5×1E10 total cells), S2 cells stably transfected with a cDNA coding for the rabies virus glycoprotein (RVGP) showed a 30% higher preserved rRVGP production (2.5±0.1 and 1.9±0.1μg/1E7 cells), as evidenced by a conformational ELISA evaluation. These findings demonstrate not only the possibility to entrap, cultivate to high densities and recover insect cells using a single-use fixed-bed bioreactor, but also that this system provides suitable physiological conditions for the entrapped cells to produce a cell membrane associated recombinant protein with higher specific biological activity as compared to classical suspension cell cultures.

  12. Transfer and internalisation of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium in cabbage cultivated on contaminated manure-amended soil under tropical field conditions in Sub-Saharan Africa.

    Science.gov (United States)

    Ongeng, D; Vasquez, G A; Muyanja, C; Ryckeboer, J; Geeraerd, A H; Springael, D

    2011-01-31

    Surface contamination and internalisation of Escherichia coli O157:H7 and Salmonella Typhimurium in cabbage leaf tissues at harvest (120 days post-transplantation) following amendment of contaminated bovine manure to soil at different times during crop cultivation were investigated under tropical field conditions in the Central Agro-Ecological Zone of Uganda. Fresh bovine manure inoculated with rifampicin-resistant derivatives of non-virulent strains of E. coli O157:H7 and S. Typhimurium was incorporated into the soil to achieve inoculum concentrations of 4 and 7 log CFU/g at the point of transplantation, 56 or 105 days post-transplantation of cabbage seedlings. Frequent sampling of the soil enabled the accurate identification of the survival kinetics in soil, which could be described by the Double Weibull model in all but one of the cases. The persistence of 4 log CFU/g E. coli O157:H7 and S. Typhimurium in the soil was limited, i.e. only inocula applied 105 days post-transplantation were still present at harvest. Moreover, no internalisation in cabbage leaf tissues was observed. In contrast, at the 7 log CFU/g inoculum level, E. coli O157:H7 and S. Typhimurium survived in the soil throughout the cultivation period. All plants (18/18) examined for leaf contamination were positive for E. coli O157:H7 at harvest irrespective of the time of manure application. A similar incidence of leaf contamination was found for S. Typhimurium. On the other hand, only plants (18/18) cultivated on soil amended with contaminated manure at the point of transplantation showed internalised E. coli O157:H7 and S. Typhimurium at harvest. These results demonstrate that under tropical field conditions, the risk of surface contamination and internalisation of E. coli O157:H7 and S. Typhimurium in cabbage leaf tissues at harvest depend on the inoculum concentration and the time of manure application. Moreover, the internalisation of E. coli O157:H7 and S. Typhimurium in cabbage leaf tissues

  13. Production of tropane alkaloids in Hyoscyamus niger (black henbane) hairy roots grown in bubble-column and spray bioreactors.

    Science.gov (United States)

    Jaremicz, Zbigniew; Luczkiewicz, Maria; Kokotkiewicz, Adam; Krolicka, Aleksandra; Sowinski, Pawel

    2014-04-01

    Hairy root cultures of Hyoscyamus niger were cultivated in shake-flasks, a bubble-column bioreactor and a hybrid bubble-column/spray bioreactor and evaluated for alkaloid production. The latter gave the highest anisodamine content (0.67 mg/g dry wt) whereas scopolamine, hyoscyamine and cuscohygrine concentrations were highest in the bubble-column reactor (5.3, 1.6 and 26.5 mg/g dry wt, respectively). Both bioreactors gave similar productivities of scopolamine (1 and 0.98 mg/l day) and cuscohygrine (5 and 5.4 mg/l day), but anisodamine productivity was 3.5-fold higher in the hybrid bioreactor (HB) (0.02 and 0.07 mg/l day, respectively). Elicitation with methyl jasmonate increased scopolamine productivity by 146 % in roots grown in the HB whereas their permeabilization with DMSO caused 4-, 5-, 25- and 28-fold increase in scopolamine, hyoscyamine, anisodamine and cuscohygrine concentrations in the growth medium. In situ extraction with Amberlite XAD-2 doubled scopolamine productivity in the hybrid reactor after 50 days.

  14. Bioreactor and methods for producing synchronous cells

    Science.gov (United States)

    Helmstetter, Charles E. (Inventor); Thornton, Maureen (Inventor); Gonda, Steve (Inventor)

    2005-01-01

    Apparatus and methods are directed to a perfusion culture system in which a rotating bioreactor is used to grow cells in a liquid culture medium, while these cells are attached to an adhesive-treated porous surface. As a result of this arrangement and its rotation, the attached cells divide, with one cell remaining attached to the substrate, while the other cell, a newborn cell is released. These newborn cells are of approximately the same age, that are collected upon leaving the bioreactor. The populations of newborn cells collected are of synchronous and are minimally, if at all, disturbed metabolically.

  15. Mass transport in a microchannel bioreactor with a porous wall.

    Science.gov (United States)

    Chen, Xiao Bing; Sui, Yi; Lee, Heow Pueh; Bai, Hui Xing; Yu, Peng; Winoto, S H; Low, Hong Tong

    2010-06-01

    A two-dimensional flow model has been developed to simulate mass transport in a microchannel bioreactor with a porous wall. A two-domain approach, based on the finite volume method, was implemented. For the fluid part, the governing equation used was the Navier-Stokes equation; for the porous medium region, the generalized Darcy-Brinkman-Forchheimer extended model was used. For the porous-fluid interface, a stress jump condition was enforced with a continuity of normal stress, and the mass interfacial conditions were continuities of mass and mass flux. Two parameters were defined to characterize the mass transports in the fluid and porous regions. The porous Damkohler number is the ratio of consumption to diffusion of the substrates in the porous medium. The fluid Damkohler number is the ratio of the substrate consumption in the porous medium to the substrate convection in the fluid region. The concentration results were found to be well correlated by the use of a reaction-convection distance parameter, which incorporated the effects of axial distance, substrate consumption, and convection. The reactor efficiency reduced with reaction-convection distance parameter because of reduced reaction (or flux), and smaller local effectiveness factor due to the lower concentration in Michaelis-Menten type reactions. The reactor was more effective, and hence, more efficient with the smaller porous Damkohler number. The generalized results could find applications for the design of bioreactors with a porous wall.

  16. Bioremediation of heavy metals using biostimulation in laboratory bioreactor.

    Science.gov (United States)

    Fulekar, M H; Sharma, Jaya; Tendulkar, Akalpita

    2012-12-01

    The present research study investigates bioremediation potential of biostimulated microbial culture isolated from heavy metals waste disposal contaminated site located at Bhayander (east), Mumbai, India. The physicochemical and microbial characterization including heavy metal contaminants have been studied at waste disposal site. The microorganisms adapted at heavy metal-contaminated environment were isolated, cultured, and biostimulated in minimal salt medium under aerobic conditions in a designed and developed laboratory bioreactor. Heavy metals such as Fe, Cu, and Cd at a selected concentration of 25, 50, and 100 μg/ml were taken in bioreactor wherein biostimulated microbial culture was added for bioremediation of heavy metals under aerobic conditions. The remediation of heavy metals was studied at an interval of 24 h for a period of 21 days. The biostimulated microbial consortium has been found effective for remediation of Cd, Cu, and Fe at higher concentration, i.e., 100 mg/l up to 98.5%, 99.6%, and 100%, respectively. Fe being a micronutrient was remediated completely compared to Cu and Cd. During the bioaccumulation of heavy metals by microorganisms, environmental parameters such as pH, total alkalinity, electronic conductivity, biological oxygen demand, chemical oxygen demand, etc. were monitored and assessed. The pilot scale study would be applicable to remediate heavy metals from waste disposal contaminated site to clean up the environment.

  17. Effects of Cultivation Conditions on the Activity of POD, SOD and CAT in the Leaf of Pinellia ternate Breit%栽培条件对半夏叶片POD、SOD、CAT活性的影响

    Institute of Scientific and Technical Information of China (English)

    章艳玲; 程纪伦; 李关荣

    2012-01-01

    To screen out a cultivation condition that can increase the Activity of POD, SOD and CAT in the leaf of Pinellia ternate Breit, measurements of POD, SOD and CAT activity in the leaf of Pinellia ternate Breit cultivated under different treatment were done with gaiacol, NBT and Ultraviolet spectrophotometry. The results showrd that the difference in leaf POD activity of Pinellia ternate Breit cultivated in the two media was insignificant; but different amount of fertilizers had very significant difference on POD activity, with 1/2MS+K+ P (KH2PO4100 mg/L) being the highest; the difference in leaf SOD and CAT activity of Pinellia ternate Breit cultivated in the two media was significant, activity of the two enzymes in perlite was very significantly higher than that in soil; different amount of fertilizers had some influence on SOD and CAT activity too, with 1/2MS+ K + P (KH2PO4100 mg/L) being the highest in SOD activity. The best cultivation that can enhance the POD, SOD, CAT activity in the leaf of Pinellia ternate Breit was fertilizing with 1/2MS+K+P (KH2PO4 100 mg/L) in perlite.%为了筛选能提高半夏叶片POD、SOD、CAT活性的栽培条件,用愈创木酚法、NBT光化还原抑制法和紫外分光光度法分别测定了不同基质不同施肥条件下半夏叶片中过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性.结果表明:半夏叶片POD酶活性受基质影响不明显,但不同施肥处理对其活性影响极显著,其中施用1/2MS+K+P(KH2PO4 100 mg/L)的处理半夏叶片POD酶的活性最强;半夏叶片中SOD和CAT酶的活性受不同基质的影响比较明显,其中以珍珠岩为基质栽培的半夏叶片中2种酶的活性均显著高于以土壤为基质栽培的半夏;不同施肥处理对这两种酶的活性也有影响,其中施用1/2MS+K+P(KH2PO4 100 mg/L)的处理半夏叶片SOD酶的活性最强.栽培半夏以珍珠岩为基质、施用1/2MS+K+P(KH2PO4 100 mg/L)的处理对于

  18. Hydrofocusing Bioreactor Produces Anti-Cancer Alkaloids

    Science.gov (United States)

    Gonda, Steve R.; Valluri, Jagan V.

    2011-01-01

    A methodology for growing three-dimensional plant tissue models in a hydrodynamic focusing bioreactor (HFB) has been developed. The methodology is expected to be widely applicable, both on Earth and in outer space, as a means of growing plant cells and aggregates thereof under controlled conditions for diverse purposes, including research on effects of gravitation and other environmental factors upon plant growth and utilization of plant tissue cultures to produce drugs in quantities greater and at costs lower than those of conventional methodologies. The HFB was described in Hydro focus - ing Bioreactor for Three-Dimensional Cell Culture (MSC-22358), NASA Tech Briefs, Vol. 27, No. 3 (March 2003), page 66. To recapitulate: The HFB offers a unique hydrofocusing capability that enables the creation of a low-shear liquid culture environment simultaneously with the herding of suspended cells and tissue assemblies and removal of unwanted air bubbles. The HFB includes a rotating cell-culture vessel with a centrally located sampling port and an internal rotating viscous spinner attached to a rotating base. The vessel and viscous spinner can be made to rotate at the same speed and direction or different speeds and directions to tailor the flow field and the associated hydrodynamic forces in the vessel in order to obtain low-shear suspension of cells and control of the locations of cells and air bubbles. For research and pharmaceutical-production applications, the HFB offers two major benefits: low shear stress, which promotes the assembly of cells into tissue-like three-dimensional constructs; and randomization of gravitational vectors relative to cells, which affects production of medicinal compounds. Presumably, apposition of plant cells in the absence of shear forces promotes cell-cell contacts, cell aggregation, and cell differentiation. Only gentle mixing is necessary for distributing nutrients and oxygen. It has been postulated that inasmuch as cells in the simulated

  19. 氨基酸硒对桃设施栽培环境适应性的影响%Effects of selenoamino acids addition on the adaptability of peach ( Prunus persica) to the condition of protected culti-vation

    Institute of Scientific and Technical Information of China (English)

    王孝娣; 刘凤之; 史祥宾; 王宝亮; 郑晓翠; 王海波

    2015-01-01

    探讨了硒提高桃适应设施栽培环境的效果, 为提高设施桃的环境适应能力和改善果实品质调控技术提供依据. 以4年生桃设施栽培的主栽品种中油4号油桃和春雪毛桃为试验材料, 研究氨基酸硒对桃光合作用光响应曲线、二氧化碳响应曲线、 温度响应曲线和蒸腾速率的影响. 喷施氨基酸硒使设施桃的光补偿点和二氧化碳补偿点显著降低, 表观量子效率和羧化效率显著提高, 使设施桃净光合速率在高温条件下保持较高数值且呈平稳状态, 显著促进了设施桃的蒸腾作用. 喷施氨基酸硒显著增强了桃对设施弱光、 二氧化碳浓度低、 高温和高湿环境的适应能力, 为设施桃品质的改善奠定了良好基础, 因此, 喷施氨基酸硒是一项切实可行的提高桃对设施环境适应能力的重要农艺措施.%To study the effects of application of selenoamino acids ( Se-aa) on the physiological parameters of peach tree and its adaptability to the protected cultivation condition, an experiment was carried using two 4-year-old peach ( Prunus persica) cultivars ('No. 4 Zhongyou' nectarine and'Springsnow' peach) which were cultivated in greenhouse as experimental materi-als. The results showed that Se-aa addition significantly decreased light compensation point and CO2 compensation point of peach plant, but significantly increased apparent quantum efficiency and carboxylation efficiency. Meanwhile, Se-aa addition improved net photosynthetic rate of protected peach plant under the condition of high temperature, and significantly improved the peach transpiration. These results fully showed that Se-aa addition significantly enhanced the adaptability of peach to the weak light condition with low concentration of CO2 , high temperature and high humidity under protected cultivation, which could be helpful for improveing the quality of peach fruits. It could be concluded that Se-aa addition could be taken as a

  20. RWPV bioreactor mass transport: earth-based and in microgravity

    Science.gov (United States)

    Begley, Cynthia M.; Kleis, Stanley J.

    2002-01-01

    Mass transport and mixing of perfused scalar quantities in the NASA Rotating Wall Perfused Vessel bioreactor are studied using numerical models of the flow field and scalar concentration field. Operating conditions typical of both microgravity and ground-based cell cultures are studied to determine the expected vessel performance for both flight and ground-based control experiments. Results are presented for the transport of oxygen with cell densities and consumption rates typical of colon cancer cells cultured in the RWPV. The transport and mixing characteristics are first investigated with a step change in the perfusion inlet concentration by computing the time histories of the time to exceed 10% inlet concentration. The effects of a uniform cell utilization rate are then investigated with time histories of the outlet concentration, volume average concentration, and volume fraction starved. It is found that the operating conditions used in microgravity produce results that are quite different then those for ground-based conditions. Mixing times for microgravity conditions are significantly shorter than those for ground-based operation. Increasing the differential rotation rates (microgravity) increases the mixing and transport, while increasing the mean rotation rate (ground-based) suppresses both. Increasing perfusion rates enhances mass transport for both microgravity and ground-based cases, however, for the present range of operating conditions, above 5-10 cc/min there are diminishing returns as much of the inlet fluid is transported directly to the perfusion exit. The results show that exit concentration is not a good indicator of the concentration distributions in the vessel. In microgravity conditions, the NASA RWPV bioreactor with the viscous pump has been shown to provide an environment that is well mixed. Even when operated near the theoretical minimum perfusion rates, only a small fraction of the volume provides less than the required oxygen levels

  1. Improved poliovirus d-antigen yields by application of different Vero cell cultivation methods

    OpenAIRE

    Thomassen, Y.E.; Rubingh, O.; Wijffels, R.H.; Pol, van de, F.C.M.

    2014-01-01

    Vero cells were grown adherent to microcarriers (Cytodex 1; 3 g L-1) using animal component free media in stirred-tank type bioreactors. Different strategies for media refreshment, daily media replacement (semi-batch), continuous media replacement (perfusion) and recirculation of media, were compared with batch cultivation. Cell densities increased using a feed strategy from 1 × 106 cells mL-1 during batch cultivation to 1.8, 2.7 and 5.0 × 106 cells mL-1 during semi-batch, perfusion and recir...

  2. Comparison of microalgae cultivation in photobioreactor, open raceway pond, and a two-stage hybrid system

    Directory of Open Access Journals (Sweden)

    Rakesh R Narala

    2016-08-01

    Full Text Available In the wake of intensive fossil fuel usage and CO2 accumulation in the environment, research is targeted towards sustainable alternate bioenergy that can suffice the growing need for fuel and also that leaves a minimal carbon footprint. Oil production from microalgae can potentially be carried out more efficiently, leaving a smaller footprint and without competing for arable land or biodiverse landscapes. However, current algae cultivation systems and lipid induction processes must be significantly improved and are threatened by contamination with other algae or algal grazers. To address this issue, we have developed an efficient two-stage cultivation system using the marine microalga Tetraselmis sp. M8. This hybrid system combines exponential biomass production in positive pressure air lift-driven bioreactors with a separate synchronized high-lipid induction phase in nutrient deplete open raceway ponds. A comparison to either bioreactor or open raceway pond cultivation system suggests that this process potentially leads to significantly higher productivity of algal lipids. Nutrients are only added to the closed bioreactors while open raceway ponds have turnovers of only a few days, thus reducing the issue of microalgal grazers.

  3. Spirulina cultivation in China

    Science.gov (United States)

    Wu, Bo-Tang; Xiang, Wen-Zhou; Zeng, Cheng-Kui

    1998-03-01

    This paper reviews and discusses the development and many problems of Spirulina cultivation in China, points out the advantages and disadvantages of open photobioreactor system, and predicts that seawater Spirulina cultivation will be a new trend to be strengthened and emphasized due to its special physiological characteristics, easier management, lower fertilizer cost, and higher resistance to contaminants and rare pollution of chemicals.

  4. Engineering skeletal muscle tissue in bioreactor systems

    Institute of Scientific and Technical Information of China (English)

    An Yang; Li Dong

    2014-01-01

    Objective To give a concise review of the current state of the art in tissue engineering (TE) related to skeletal muscle and kinds of bioreactor environment.Data sources The review was based on data obtained from the published articles and guidelines.Study selection A total of 106 articles were selected from several hundred original articles or reviews.The content of selected articles is in accordance with our purpose and the authors are authorized scientists in the study of engineered muscle tissue in bioreactor.Results Skeletal muscle TE is a promising interdisciplinary field which aims at the reconstruction of skeletal muscle loss.Although numerous studies have indicated that engineering skeletal muscle tissue may be of great importance in medicine in the near future,this technique still represents a limited degree of success.Since tissue-engineered muscle constructs require an adequate connection to the vascular system for efficient transport of oxygen,carbon dioxide,nutrients and waste products.Moreover,functional and clinically applicable muscle constructs depend on adequate neuromuscular junctions with neural calls.Third,in order to engineer muscle tissue successfully,it may be beneficial to mimic the in vivo environment of muscle through association with adequate stimuli from bioreactors.Conclusion Vascular system and bioreactors are necessary for development and maintenance of engineered muscle in order to provide circulation within the construct.

  5. LANDFILL BIOREACTOR PERFORMANCE, SECOND INTERIM REPORT

    Science.gov (United States)

    A bioreactor landfill is a landfill that is operated in a manner that is expected to increase the rate and extent of waste decomposition, gas generation, and settlement compared to a traditional landfill. This Second Interim Report was prepared to provide an interpretation of fie...

  6. MONITORING APPROACHES FOR BIOREACTOR LANDFILLS - Report

    Science.gov (United States)

    Experimental bioreactor landfill operations at operating Municipal Solid Waste (MSW) landfills can be approved under the research development and demonstration (RD&D) provisions of 30CFR 258.4. To provide a basis for consistent data collection for future decision-making in suppor...

  7. Sulfate-reducing bacteria in anaerobic bioreactors.

    NARCIS (Netherlands)

    Oude Elferink, S.J.W.H.

    1998-01-01

    The treatment of industrial wastewaters containing high amounts of easily degradable organic compounds in anaerobic bioreactors is a well-established process. Similarly, wastewaters which in addition to organic compounds also contain sulfate can be treated in this way. For a long time, the occurrenc

  8. Bioreactor Studies and Computational Fluid Dynamics

    Science.gov (United States)

    Singh, H.; Hutmacher, D. W.

    The hydrodynamic environment “created” by bioreactors for the culture of a tissue engineered construct (TEC) is known to influence cell migration, proliferation and extra cellular matrix production. However, tissue engineers have looked at bioreactors as black boxes within which TECs are cultured mainly by trial and error, as the complex relationship between the hydrodynamic environment and tissue properties remains elusive, yet is critical to the production of clinically useful tissues. It is well known in the chemical and biotechnology field that a more detailed description of fluid mechanics and nutrient transport within process equipment can be achieved via the use of computational fluid dynamics (CFD) technology. Hence, the coupling of experimental methods and computational simulations forms a synergistic relationship that can potentially yield greater and yet, more cohesive data sets for bioreactor studies. This review aims at discussing the rationale of using CFD in bioreactor studies related to tissue engineering, as fluid flow processes and phenomena have direct implications on cellular response such as migration and/or proliferation. We conclude that CFD should be seen by tissue engineers as an invaluable tool allowing us to analyze and visualize the impact of fluidic forces and stresses on cells and TECs.

  9. Vortex breakdown in a truncated conical bioreactor

    DEFF Research Database (Denmark)

    Balci, Adnan; Brøns, Morten; Herrada, Miguel A.;

    2015-01-01

    This numerical study explains the eddy formation and disappearance in a slow steady axisymmetric air–water flow in a vertical truncated conical container, driven by the rotating top disk. Numerous topological metamorphoses occur as the water height, Hw, and the bottom-sidewall angle, α, vary. It ...... are of fundamental interest and can be relevant for aerial bioreactors....

  10. Continuous-Flow Gas-Phase Bioreactors

    Science.gov (United States)

    Wise, Donald L.; Trantolo, Debra J.

    1994-01-01

    Continuous-flow gas-phase bioreactors proposed for biochemical, food-processing, and related industries. Reactor contains one or more selected enzymes dehydrated or otherwise immobilized on solid carrier. Selected reactant gases fed into reactor, wherein chemical reactions catalyzed by enzyme(s) yield product biochemicals. Concept based on discovery that enzymes not necessarily placed in traditional aqueous environments to function as biocatalysts.

  11. Anaerobic membrane bioreactors: Are membranes really necessary?

    NARCIS (Netherlands)

    Davila, M.; Kassab, G.; Klapwijk, A.; Lier, van J.B.

    2008-01-01

    Membranes themselves represent a significant cost for the full scale application of anaerobic membrane bioreactors (AnMBR). The possibility of operating an AnMBR with a self-forming dynamic membrane generated by the substances present in the reactor liquor would translate into an important saving. A

  12. Denitrifying bioreactor clogging potential during wastewater treatment

    Science.gov (United States)

    Chemoheterotrophic denitrification technologies using woodchips as a solid carbon source (i.e., woodchip bioreactors) have been widely trialed for treatment of diffuse-source agricultural nitrogen pollution. There is growing interest in the use of this simple, relatively low-cost biological wastewat...

  13. Scalable cultivation of human pluripotent stem cells on chemically-defined surfaces

    Science.gov (United States)

    Hsiung, Michael Chi-Wei

    Human stem cells (SCs) are classified as self-renewing cells possessing great ability in therapeutic applications due of their ability to differentiate along any major cell lineage in the human body. Despite their restorative potential, widespread use of SCs is hampered by strenuous control issues. Along with the need for strict xeno-free environments to sustain growth in culture, current methods for growing human pluripotent stem cells (hPSCs) rely on platforms which impede large-scale cultivation and therapeutic delivery. Hence, any progress towards development of large-scale culture systems is severely hindered. In a concentrated effort to develop a scheme that can serve as a model precursor for large scale SC propagation in clinical use, we have explored methods for cultivating hPSCs on completely defined surfaces. We discuss novel approaches with the potential to go beyond the limitations presented by current methods. In particular, we studied the cultivation of human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) on surface which underwent synthetic or chemical modification. Current methods for hPSCs rely on animal-based extracellular matrices (ECMs) such as mouse embryonic fibroblasts (MEFs) or feeders and murine sacoma cell-derived substrates to facilitate their growth. While these layers or coatings can be used to maximize the output of hPSC production, they cannot be considered for clinical use because they risk introducing foreign pathogens into culture. We have identified and developed conditions for a completely defined xeno-free substrate used for culturing hPSCs. By utilizing coupling chemistry, we can functionalize ester groups on a given surface and conjugate synthetic peptides containing the arginine-glycine-aspartic acid (RGD) motif, known for their role in cell adhesion. This method offers advantages over traditional hPSC culture by keeping the modified substrata free of xenogenic response and can be scaled up in

  14. NASA-approved rotary bioreactor enhances proliferation of human epidermal stem cells and supports formation of 3D epidermis-like structure.

    Directory of Open Access Journals (Sweden)

    Xiao-hua Lei

    Full Text Available The skin is susceptible to different injuries and diseases. One major obstacle in skin tissue engineering is how to develop functional three-dimensional (3D substitute for damaged skin. Previous studies have proved a 3D dynamic simulated microgravity (SMG culture system as a "stimulatory" environment for the proliferation and differentiation of stem cells. Here, we employed the NASA-approved rotary bioreactor to investigate the proliferation and differentiation of human epidermal stem cells (hEpSCs. hEpSCs were isolated from children foreskins and enriched by collecting epidermal stem cell colonies. Cytodex-3 micro-carriers and hEpSCs were co-cultured in the rotary bioreactor and 6-well dish for 15 days. The result showed that hEpSCs cultured in rotary bioreactor exhibited enhanced proliferation and viability surpassing those cultured in static conditions. Additionally, immunostaining analysis confirmed higher percentage of ki67 positive cells in rotary bioreactor compared with the static culture. In contrast, comparing with static culture, cells in the rotary bioreactor displayed a low expression of involucrin at day 10. Histological analysis revealed that cells cultured in rotary bioreactor aggregated on the micro-carriers and formed multilayer 3D epidermis structures. In conclusion, our research suggests that NASA-approved rotary bioreactor can support the proliferation of hEpSCs and provide a strategy to form multilayer epidermis structure.

  15. Effects of hydraulic retention time on anaerobic hydrogenation performance and microbial ecology of bioreactors fed with glucose-peptone and starch-peptone

    Energy Technology Data Exchange (ETDEWEB)

    Li, Shiue-Lin; Chao, Yu-Chieh; Wang, Yu-Hsuan; Hsiao, Chia-Jung; Bai, Ming-Der [Department of Environmental Engineering, National Cheng-Kung University, No. 1, University Road, Tainan 701 (China); Whang, Liang-Ming; Wang, Yung-Fu; Cheng, Sheng-Shung [Department of Environmental Engineering, National Cheng-Kung University, No. 1, University Road, Tainan 701 (China); Sustainable Environment Research Center (SERC), National Cheng-Kung University, No. 1, University Road, Tainan 701 (China); Tseng, I.-Cheng [Sustainable Environment Research Center (SERC), National Cheng-Kung University, No. 1, University Road, Tainan 701 (China); Department of Life Science, National Cheng-Kung University, No. 1, University Road, Tainan 701 (China)

    2010-01-15

    This study evaluated anaerobic hydrogenation performance and microbial ecology in bioreactors operated at different hydraulic retention time (HRT) conditions and fed with glucose-peptone (GP) and starch-peptone (SP). The maximum hydrogen production rates for GP- and SP-fed bioreactors were found to be 1247 and 412 mmol-H{sub 2}/L/d at HRT of 2 and 3 h, respectively. At HRT > 8 h, hydrogen consumption due to peptone fermentation could occur and thus reduced hydrogen yield from carbohydrate fermentation. Results of cloning/sequencing and denaturant gradient gel electrophoresis (DGGE) indicated that Clostridium sporogenes and Clostridium celerecrescens were dominant hydrogen-producing bacteria in the GP-fed bioreactor, presumably due to their capability on protein hydrolysis. In the SP-fed bioreactor, Lactobacillus plantarum, Propionispira arboris, and Clostridium butyricum were found to be dominant populations, but the presence of P. arboris at HRT > 3 h might be responsible for a lower hydrogen yield from starch fermentation. As a result, optimizing HRT operation for bioreactors was considered an important asset in order to minimize hydrogen-consuming activities and thus maximize net hydrogen production. The limitation of simple parameters such as butyrate to acetate ratio (B/A ratio) in predicting hydrogen production was recognized in this study for bioreactors fed with multiple substrates. It is suggested that microbial ecology analysis, in addition to chemical analysis, should be performed when complex substrates and mixed cultures are used in hydrogen-producing bioreactors. (author)

  16. Osmotic membrane bioreactor for phenol biodegradation under continuous operation.

    Science.gov (United States)

    Praveen, Prashant; Loh, Kai-Chee

    2016-03-15

    Continuous phenol biodegradation was accomplished in a two-phase partitioning osmotic membrane bioreactor (TPPOMBR) system, using extractant impregnated membranes (EIM) as the partitioning phase. The EIMs alleviated substrate inhibition during prolonged operation at influent phenol concentrations of 600-2000mg/L, and also at spiked concentrations of 2500mg/L phenol restricted to 2 days. Filtration of the effluent through forward osmosis maintained high biomass concentration in the bioreactor and improved effluent quality. Steady state was reached in 5-6 days at removal rates varying between 2000 and 5500mg/L-day under various conditions. Due to biofouling and salt accumulation, the permeate flux varied from 1.2-7.2 LMH during 54 days of operation, while maintaining an average hydraulic retention time of 7.4h. A washing cycle, comprising 1h osmotic backwashing using 0.5M NaCl and 2h washing with water, facilitated biofilm removal from the membranes. Characterization of the extracellular polymeric substances (EPS) through FTIR showed peaks between 1700 and 1500cm(-1), 1450-1450cm(-1) and 1200-1000cm(-1), indicating the presence of proteins, phenols and polysaccharides, respectively. The carbohydrate to protein ratio in the EPS was estimated to be 0.3. These results indicate that TPPOMBR can be promising in continuous treatment of phenolic wastewater.

  17. Thiosulphate conversion in a methane and acetate fed membrane bioreactor.

    Science.gov (United States)

    Suarez-Zuluaga, Diego A; Timmers, Peer H A; Plugge, Caroline M; Stams, Alfons J M; Buisman, Cees J N; Weijma, Jan

    2016-02-01

    The use of methane and acetate as electron donors for biological reduction of thiosulphate in a 5-L laboratory membrane bioreactor was studied and compared to disproportionation of thiosulphate as competing biological reaction. The reactor was operated for 454 days in semi-batch mode; 30 % of its liquid phase was removed and periodically replenished (days 77, 119, 166, 258, 312 and 385). Although the reactor was operated under conditions favourable to promote thiosulphate reduction coupled to methane oxidation, thiosulphate disproportionation was the dominant microbial process. Pyrosequencing analysis showed that the most abundant microorganisms in the bioreactor were phototrophic green sulphur bacteria (GSB) belonging to the family Chlorobiaceae and thiosulphate-disproportionating bacteria belonging to the genus Desulfocapsa. Even though the reactor system was surrounded with opaque plastic capable of filtering most of the light, the GSB used it to oxidize the hydrogen sulphide produced from thiosulphate disproportionation to elemental sulphur. Interrupting methane and acetate supply did not have any effect on the microbial processes taking place. The ultimate goal of our research was to develop a process that could be applied for thiosulphate and sulphate removal and biogenic sulphide formation for metal precipitation. Even though the system achieved in this study did not accomplish the targeted conversion using methane as electron donor, it does perform microbial conversions which allow to directly obtain elemental sulphur from thiosulphate.

  18. Applicability of dynamic membrane technology in anaerobic membrane bioreactors.

    Science.gov (United States)

    Ersahin, Mustafa Evren; Ozgun, Hale; Tao, Yu; van Lier, Jules B

    2014-01-01

    This study investigated the applicability of dynamic membrane technology in anaerobic membrane bioreactors for the treatment of high strength wastewaters. A monofilament woven fabric was used as support material for dynamic membrane formation. An anaerobic dynamic membrane bioreactor (AnDMBR) was operated under a variety of operational conditions, including different sludge retention times (SRTs) of 20 and 40 days in order to determine the effect of SRT on both biological performance and dynamic membrane filtration characteristics. High COD removal efficiencies exceeding 99% were achieved during the operation at both SRTs. Higher filtration resistances were measured during the operation at SRT of 40 days in comparison to SRT of 20 days, applying a stable flux of 2.6 L/m(2) h. The higher filtration resistances coincided with lower extracellular polymeric substances concentration in the bulk sludge at SRT of 40 days, likely resulting in a decreased particle flocculation. Results showed that dynamic membrane technology achieved a stable and high quality permeate and AnDMBRs can be used as a reliable and satisfactory technology for treatment of high strength wastewaters.

  19. 乌鲁木齐南山杏树种植气候条件分析%Analysis on Cultivation Climate Conditions of Apricot Trees in Nanshan Mountains

    Institute of Scientific and Technical Information of China (English)

    郑玉萍

    2012-01-01

    Based on the climate data from Urumqi Nanshan mountains, the climate conditions for planting apricot trees were analyzed. The results showed that heat, frost-free period, precipitation, sunshine intensity and other weather conditions in Urumqi Nanshan mountains are fit for the growth of apricots. In order to avoid freezing disaster in winter, apricot trees should be planted in a strong temperature inversion area. Frost Damage rate was 19.6% which was within mild risk degree during the flowering and fruiting period in spring. This study provides reference for the reasonable adjustment of cropping industry in local area.%利用乌鲁木齐南山山区历史气象资料,进行了杏树种植的气候条件分析。结果表明:乌鲁木齐南山的热量、无霜期、降水、光照等气候条件基本可以满足杏树的生长需求;为了避免冬季冻害,应选择在逆温强的地带发展杏树;春季花果期霜冻灾损率为19.6%,处于中轻度风险区。

  20. Bioreactors in tissue engineering - principles, applications and commercial constraints.

    Science.gov (United States)

    Hansmann, Jan; Groeber, Florian; Kahlig, Alexander; Kleinhans, Claudia; Walles, Heike

    2013-03-01

    Bioreactor technology is vital for tissue engineering. Usually, bioreactors are used to provide a tissue-specific physiological in vitro environment during tissue maturation. In addition to this most obvious application, bioreactors have the potential to improve the efficiency of the overall tissue-engineering concept. To date, a variety of bioreactor systems for tissue-specific applications have been developed. Of these, some systems are already commercially available. With bioreactor technology, various functional tissues of different types were generated and cultured in vitro. Nevertheless, these efforts and achievements alone have not yet led to many clinically successful tissue-engineered implants. We review possible applications for bioreactor systems within a tissue-engineering process and present basic principles and requirements for bioreactor development. Moreover, the use of bioreactor systems for the expansion of clinically relevant cell types is addressed. In contrast to cell expansion, for the generation of functional three-dimensional tissue equivalents, additional physical cues must be provided. Therefore, bioreactors for musculoskeletal tissue engineering are discussed. Finally, bioreactor technology is reviewed in the context of commercial constraints.

  1. Organic Matter Effect on Glomus Intrarradices in Beans (Phaseolus Vulgaris L. Growth Cultivated in Soils with Two Sources of Water under Greenhouse Conditions

    Directory of Open Access Journals (Sweden)

    A. K. Gardezi

    2014-02-01

    Full Text Available The objective of this research was to evaluate the effect of organic matter on the association with Glomus intrarradices and soil contamination on beans (Phaseolus vulgaris L.. The study was done under greenhouse conditions at the Montecillo Campus of the Postgraduate College, Mexico. Two soils were used, one irrigated with sewage water and the other one with clean water from a well. Half of the plants were inoculated with Glomus intrarradices. Vermicompost was used as a source of organic matter. There were highly significant increases (p≤0.05 in all the variables recorded due to the application of organic matter, and to the inoculation with Glomus intarradices. The irrigation source of the soils used for this experiment only had a significant effect (p≤0.05 on pod number and nitrogen fixation. The best growth and grain yield occurred with inoculated plants and supplementary organic matter.

  2. Expansion of activated lymphocytes obtained from renal cell carcinoma in an automated hollow fiber bioreactor.

    Science.gov (United States)

    Hillman, G G; Wolf, M L; Montecillo, E; Younes, E; Ali, E; Pontes, J E; Haas, G P

    1994-01-01

    Immunotherapy using IL-2 alone or combined with activated lymphocytes has been promising for metastatic renal cell carcinoma. Cytotoxic lymphocytes can be isolated from tumors, expanded in vitro with IL-2, and adoptively transferred back into the tumor-bearing host. These cells can also be transduced with the genes coding for cytokines for local delivery to tumor sites. A major drawback in adoptive immunotherapy is the cumbersome and expensive culture technology associated with the growth of large numbers of cells required for their therapeutic effect. To reduce the cost, resources, and manpower, we have developed the methodology for lymphocyte activation and expansion in the automated hollow fiber bioreactor IMMUNO*STAR Cell Expander (ACT BIOMEDICAL, INC). Tumor Infiltrating Lymphocytes (TIL) isolated from human renal cell carcinoma tumor specimens were inoculated at a number of 10(8) cells in a small bioreactor of 30 ml extracapillary space volume. We have determined the medium flow rates and culture conditions to obtain a significant and repeated expansion of TIL at weekly intervals. The lymphocytes cultured in the bioreactor demonstrated the same phenotype and cytotoxic activity as those expanded in parallel in tissue culture plates. Lymphocyte expansion in the hollow fiber bioreactor required lower volumes of medium, human serum, IL-2 and minimal labor. This technology may facilitate the use of adoptive immunotherapy for the treatment of refractory malignancies.

  3. Carbon dioxide capture using Escherichia coli expressing carbonic anhydrase in a foam bioreactor.

    Science.gov (United States)

    Watson, Stuart K; Han, Zhenlin; Su, Wei Wen; Deshusses, Marc A; Kan, Eunsung

    2016-12-01

    The present study reports CO2 capture and conversion to bicarbonate using Escherichia coli expressing carbonic anhydrase (CA) on its cell surface in a novel foam bioreactor. The very large gas-liquid interfacial area in the foam bioreactor promoted rapid CO2 absorption while the CO2 in the aqueous phase was subsequently converted to bicarbonate ions by the CA. CO2 gas removal in air was investigated at various conditions such as gas velocity, cell density and CO2 inlet concentration. Regimes for kinetic and mass transfer limitations were defined. Very high removal rates of CO2 were observed: 9570 g CO2 m(-3) bioreactor h(-1) and a CO2 removal efficiency of 93% at 4% inlet CO2 when the gas retention time was 24 s, and cell concentration was 4 gdw L(-1). These performances are superior to earlier reports of experimental bioreactors using CA for CO2 capture. Overall, this bioreactor system has significant potential as an alternative CO2 capture technology.

  4. Antioxidant compounds in Salvia officinalis L. shoot and hairy root cultures in the nutrient sprinkle bioreactor

    Directory of Open Access Journals (Sweden)

    Izabela Grzegorczyk

    2011-01-01

    Full Text Available The study focused on the production of compounds with antioxidant activity in hairy root and shoot cultures of Salvia officinalis grown in laboratory-scale sprinkle nutrient bioreactors. HPLC analysis showed that production of rosmarinic acid in transformed roots (34.65 ±1.07 mg l-1 was higher that in shoot culture (26.24 ±0.48 mg l-1. In the latter diterpenoids: carnosic acid (1.74 ±0.02 mg l-1 and carnosol (1.34 ±0.01 mg l-1 were also found. Biomass accumulation after a growth period in the bioreactor was also studied. An 18-fold increase in hairy root biomass was recorded after 40 days of culture. In sage shoot culture, biomass increased 43 times after 21 days of bioreactor run. The current operating conditions of the bioreactor were not suitable for the propagation of Salvia officinalis mainly due to the hyperhydricity problem of leaves and stems.

  5. Perchlorate and Nitrate Remediation Efficiency and Microbial Diversity in a Containerized Wetland Bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Jr., B D; Dibley, V; Pinkart, H; Legler, T

    2004-06-09

    We have developed a method to remove perchlorate (14 to 27 {micro}g/L) and nitrate (48 mg/L) from contaminated groundwater using a wetland bioreactor. The bioreactor has operated continuously in a remote field location for more than two years with a stable ecosystem of indigenous organisms. This study assesses the bioreactor for long-term perchlorate and nitrate remediation by evaluating influent and effluent groundwater for reduction-oxidation conditions and nitrate and perchlorate concentrations. Total community DNA was extracted and purified from 10-g sediment samples retrieved from vertical coring of the bioreactor during winter. Analysis by denaturing gradient gel electrophoresis of short, 16S rDNA, polymerase-chain-reaction products was used to identify dominant microorganisms. Bacteria genera identified were closely affiliated with bacteria widely distributed in soils, mud layers, and fresh water. Of the 17 dominant bands sequenced, most were gram negative and capable of aerobic or anaerobic respiration with nitrate as the terminal electron acceptor (Pseudomonas, Acinetobacter, Halomonas, and Nitrospira). Several identified genera (Rhizobium, Acinetobactor, and Xanthomonas) are capable of fixing atmospheric nitrogen into a combined form (ammonia) usable by host plants. Isolates were identified from the Proteobacteria class, known for the ability to reduce perchlorate. Initial bacterial assessments of sediments confirm the prevalence of facultative anaerobic bacteria capable of reducing perchlorate and nitrate in situ.

  6. Mathematical modelling of cell layer growth in a hollow fibre bioreactor.

    Science.gov (United States)

    Chapman, Lloyd A C; Whiteley, Jonathan P; Byrne, Helen M; Waters, Sarah L; Shipley, Rebecca J

    2017-04-07

    Generating autologous tissue grafts of a clinically useful volume requires efficient and controlled expansion of cell populations harvested from patients. Hollow fibre bioreactors show promise as cell expansion devices, owing to their potential for scale-up. However, further research is required to establish how to specify appropriate hollow fibre bioreactor operating conditions for expanding different cell types. In this study we develop a simple model for the growth of a cell layer seeded on the outer surface of a single fibre in a perfused hollow fibre bioreactor. Nutrient-rich culture medium is pumped through the fibre lumen and leaves the bioreactor via the lumen outlet or passes through the porous fibre walls and cell layer, and out via ports on the outer wall of the extra-capillary space. Stokes and Darcy equations for fluid flow in the fibre lumen, fibre wall, cell layer and extra-capillary space are coupled to reaction-advection-diffusion equations for oxygen and lactate transport through the bioreactor, and to a simple growth law for the evolution of the free boundary of the cell layer. Cells at the free boundary are assumed to proliferate at a rate that increases with the local oxygen concentration, and to die and detach from the layer if the local fluid shear stress or lactate concentration exceed critical thresholds. We use the model to predict operating conditions that maximise the cell layer growth for different cell types. In particular, we predict the optimal flow rate of culture medium into the fibre lumen and fluid pressure imposed at the lumen outlet for cell types with different oxygen demands and fluid shear stress tolerances, and compare the growth of the cell layer when the exit ports on the outside of the bioreactor are open with that when they are closed. Model simulations reveal that increasing the inlet flow rate and outlet fluid pressure increases oxygen delivery to the cell layer and, therefore, the growth rate of cells that are

  7. Good manufacturing practice-compliant animal-free expansion of human bone marrow derived mesenchymal stroma cells in a closed hollow-fiber-based bioreactor.

    Science.gov (United States)

    Nold, Philipp; Brendel, Cornelia; Neubauer, Andreas; Bein, Gregor; Hackstein, Holger

    2013-01-01

    Mesenchymal stroma cells (MSC) are increasingly recognized for various applications of cell-based therapies such as regenerative medicine or immunomodulatory treatment strategies. Standardized large-scale expansions of MSC under good manufacturing practice (GMP)-compliant conditions avoiding animal derived components are mandatory for further evaluation of these novel therapeutic approaches in clinical trials. We applied a novel automated hollow fiber cell expansion system (CES) for in vitro expansion of human bone marrow derived MSC employing a GMP-compliant culture medium with human platelet lysate (HPL). Between 8 and 32 ml primary bone marrow aspirate were loaded into the hollow fiber CES and cultured for 15-27 days. 2-58 million MSC were harvested after primary culture. Further GMP-compliant cultivation of second passage MSC for 13 days led to further 10-20-fold enrichment. Viability, surface antigen expression, differentiation capacity and immunosuppressive function of MSC cultured in the hollow fiber CES were in line with standard criteria for MSC definition. We conclude that MSC can be enriched from primary bone marrow aspirate in a GMP-conform manner within a closed hollow fiber bioreactor and maintain their T lymphocyte inhibitory capacity. Standardized and reliable conditions for large scale MSC expansion pave the way for safe applications in humans in different therapeutic approaches.

  8. Endomycorrhizal Inoculation Effect On Beans (Phaseolus vulgaris L., OAT (Avena sativa L., And Wheat (Triticum aestivum L. Growth Cultivated In Two Soil Types Under Greenhouse Conditions

    Directory of Open Access Journals (Sweden)

    Abdul Khalil Gardezi

    2012-10-01

    Full Text Available The objective of this research was to evaluate the effect of inoculation with Glomus intraradices on oat, beans, and wheat. The study was done under greenhouse conditions at the Montecillo Campus of the Postgraduate College, Mexico. Two soil types from San Luis Potosí State were used, one was red (Xerosol, and the other one was grey (Litosol. With and without Glomus intraradices inoculation. Three bean cultivars: Pinto Saltillo, Bayo comercial and Flor de Mayo; Chihuahua (oat variety; and Tlaxcala wheat genotype were planted. The experimental design was factorial complete randomized block and three replications. The result showed that bean yield (average 3.7 g plant-1, pod number and dry weight, leaf area, plant height, stem diameter, and aerial part dry weight were positively affected by the inoculation with Glomus intraradices, but not by soil type. A similar trend was observed in root length, volume and dry weight, and in the nodule number. In relation with the species studied, Phaseolus vulgaris varieties had higher values than wheat and oats in growth and yield variables evaluated. It is concluded that endomycorrhiza inoculation (Glomus intraradices gave better growth and yield, especially in beans. The soil types studied did not affect significantly plant responses in this study.

  9. STUDIES ON THE EFFECT OF GENOTYPE ON GROWTH AND SEED YIELD IN SOME Camelina sativa L. VARIETIES CULTIVATED UNDER CONTROLLED ENVIRONMENTAL CONDITIONS

    Directory of Open Access Journals (Sweden)

    Emanuela PODGOREANU

    2015-10-01

    Full Text Available The recently emerged interest in using renewable vegetable sources as feedstock for biofuel production requires the identification of effective solutions aimed to follow the principles of sustainable development. Biofuels from the first generation were produced from sugarcane, starch and oils derived from agricultural products, but they may not be sustainable because are competing with the agricultural production. Currently, studies are focused on the obtaining of a second generation of biofuels that originate from plants other then food crops, from agricultural residues and from municipal wastes. Being economically attractive and having certain agro-technical advantages, the Camelina sativa seed oil is considered a sustainable source for obtaining second generation biofuels. The aim of the present research was to record the effect of genotype in 3 foreign varieties of Camelina sativa (CALENA, GP 204 and GP 202 and in 3 autochthonous ones (CAMELIA, one local population Fundulea and one hybrid line as regards both the growth characteristics and the yield components in a greenhouse experiment, in order to evaluate their potential performance and adaptability in field conditions. Periodic phenological observations were performed on the occurrence of flowering, on the pod maturation and genotypic differences of the biometric measurements such as plant height, number of branches, number of pods per plant, number of seeds in pods and thousand seed weight are discussed.

  10. In silico multi-scale model of transport and dynamic seeding in a bone tissue engineering perfusion bioreactor.

    Science.gov (United States)

    Spencer, T J; Hidalgo-Bastida, L A; Cartmell, S H; Halliday, I; Care, C M

    2013-04-01

    Computer simulations can potentially be used to design, predict, and inform properties for tissue engineering perfusion bioreactors. In this work, we investigate the flow properties that result from a particular poly-L-lactide porous scaffold and a particular choice of perfusion bioreactor vessel design used in bone tissue engineering. We also propose a model to investigate the dynamic seeding properties such as the homogeneity (or lack of) of the cellular distribution within the scaffold of the perfusion bioreactor: a pre-requisite for the subsequent successful uniform growth of a viable bone tissue engineered construct. Flows inside geometrically complex scaffolds have been investigated previously and results shown at these pore scales. Here, it is our aim to show accurately that through the use of modern high performance computers that the bioreactor device scale that encloses a scaffold can affect the flows and stresses within the pores throughout the scaffold which has implications for bioreactor design, control, and use. Central to this work is that the boundary conditions are derived from micro computed tomography scans of both a device chamber and scaffold in order to avoid generalizations and uncertainties. Dynamic seeding methods have also been shown to provide certain advantages over static seeding methods. We propose here a novel coupled model for dynamic seeding accounting for flow, species mass transport and cell advection-diffusion-attachment tuned for bone tissue engineering. The model highlights the timescale differences between different species suggesting that traditional homogeneous porous flow models of transport must be applied with caution to perfusion bioreactors. Our in silico data illustrate the extent to which these experiments have the potential to contribute to future design and development of large-scale bioreactors.

  11. Very high cell density perfusion of CHO cells anchored in a non-woven matrix-based bioreactor.

    Science.gov (United States)

    Zhang, Ye; Stobbe, Per; Silvander, Christian Orrego; Chotteau, Véronique

    2015-11-10

    Recombinant Chinese Hamster Ovary (CHO) cells producing IgG monoclonal antibody were cultivated in a novel perfusion culture system CellTank, integrating the bioreactor and the cell retention function. In this system, the cells were harbored in a non-woven polyester matrix perfused by the culture medium and immersed in a reservoir. Although adapted to suspension, the CHO cells stayed entrapped in the matrix. The cell-free medium was efficiently circulated from the reservoir into- and through the matrix by a centrifugal pump placed at the bottom of the bioreactor resulting in highly homogenous concentrations of the nutrients and metabolites in the whole system as confirmed by measurements from different sampling locations. A real-time biomass sensor using the dielectric properties of living cells was used to measure the cell density. The performances of the CellTank were studied in three perfusion runs. A very high cell density measured as 200 pF/cm (where 1 pF/cm is equivalent to 1 × 10(6)viable cells/mL) was achieved at a perfusion rate of 10 reactor volumes per day (RV/day) in the first run. In the second run, the effect of cell growth arrest by hypothermia at temperatures lowered gradually from 37 °C to 29 °C was studied during 13 days at cell densities above 100 pF/cm. Finally a production run was performed at high cell densities, where a temperature shift to 31 °C was applied at cell density 100 pF/cm during a production period of 14 days in minimized feeding conditions. The IgG concentrations were comparable in the matrix and in the harvest line in all the runs, indicating no retention of the product of interest. The cell specific productivity was comparable or higher than in Erlenmeyer flask batch culture. During the production run, the final harvested IgG production was 35 times higher in the CellTank compared to a repeated batch culture in the same vessel volume during the same time period.

  12. Novel approach of high cell density recombinant bioprocess development: Optimisation and scale-up from microlitre to pilot scales while maintaining the fed-batch cultivation mode of E. coli cultures

    Directory of Open Access Journals (Sweden)

    Rimšeliene Renata

    2010-05-01

    Full Text Available Abstract Background Bioprocess development of recombinant proteins is time consuming and laborious as many factors influence the accumulation of the product in the soluble and active form. Currently, in most cases the developmental line is characterised by a screening stage which is performed under batch conditions followed by the development of the fed-batch process. Performing the screening already under fed-batch conditions would limit the amount of work and guarantee that the selected favoured conditions also work in the production scale. Results Here, for the first time, high throughput multifactorial screening of a cloning library is combined with the fed-batch technique in 96-well plates, and a strategy is directly derived for scaling to bioreactor scale. At the example of a difficult to express protein, an RNase inhibitor, it is demonstrated that screening of various vector constructs and growth conditions can be performed in a coherent line by (i applying a vector library with promoters and ribosome binding sites of different strength and various fusion partners together with (ii an early stage use of the fed-batch technology. It is shown that the EnBase® technology provides an easy solution for controlled cultivation conditions in the microwell scale. Additionally the high cell densities obtained provide material for various analyses from the small culture volumes. Crucial factors for a high yield of the target protein in the actual case were (i the fusion partner, (ii the use of of a mineral salt medium together with the fed-batch technique, and (iii the preinduction growth rate. Finally, it is shown that the favorable conditions selected in the microwell plate and shake flask scales also work in the bioreactor. Conclusions Cultivation media and culture conditions have a major impact on the success of a screening procedure. Therefore the application of controlled cultivation conditions is pivotal. The consequent use of fed

  13. conditions

    Directory of Open Access Journals (Sweden)

    M. Venkatesulu

    1996-01-01

    Full Text Available Solutions of initial value problems associated with a pair of ordinary differential systems (L1,L2 defined on two adjacent intervals I1 and I2 and satisfying certain interface-spatial conditions at the common end (interface point are studied.

  14. Hatchery cultivation of the common cockle (

    NARCIS (Netherlands)

    Pronker, A.E.; Peene, F.; Donner, S.; Wijnhoven, S.; Geijsen, P.; Bossier, P.; Nevejan, N.M.

    2015-01-01

    This study describes for the first time the cultivation of Cerastoderma edule on a commercial scale. A protocol to grow F2 generation cockles was developed, which led to fine-tuning experiments for broodstock conditioning and spat growth.Broodstock animals were conditioned with die

  15. Biological hydrogen production using a membrane bioreactor.

    Science.gov (United States)

    Oh, Sang-Eun; Iyer, Prabha; Bruns, Mary Ann; Logan, Bruce E

    2004-07-01

    A cross-flow membrane was coupled to a chemostat to create an anaerobic membrane bioreactor (MBR) for biological hydrogen production. The reactor was fed glucose (10,000 mg/L) and inoculated with a soil inoculum heat-treated to kill non-spore-forming methanogens. Hydrogen gas was consistently produced at a concentration of 57-60% in the headspace under all conditions. When operated in chemostat mode (no flow through the membrane) at a hydraulic retention time (HRT) of 3.3 h, 90% of the glucose was removed, producing 2200 mg/L of cells and 500 mL/h of biogas. When operated in MBR mode, the solids retention time (SRT) was increased to SRT = 12 h producing a solids concentration in the reactor of 5800 mg/L. This SRT increased the overall glucose utilization (98%), the biogas production rate (640 mL/h), and the conversion efficiency of glucose-to-hydrogen from 22% (no MBR) to 25% (based on a maximum of 4 mol-H(2)/mol-glucose). When the SRT was increased from 5 h to 48 h, glucose utilization (99%) and biomass concentrations (8,800 +/- 600 mg/L) both increased. However, the biogas production decreased (310 +/- 40 mL/h) and the glucose-to-hydrogen conversion efficiency decreased from 37 +/- 4% to 18 +/- 3%. Sustained permeate flows through the membrane were in the range of 57 to 60 L/m(2) h for three different membrane pore sizes (0.3, 0.5, and 0.8 microm). Most (93.7% to 99.3%) of the membrane resistance was due to internal fouling and the reversible cake resistance, and not the membrane itself. Regular backpulsing was essential for maintaining permeate flux through the membrane. Analysis of DNA sequences using ribosomal intergenic spacer analysis indicated bacteria were most closely related to members of Clostridiaceae and Flexibacteraceae, including Clostridium acidisoli CAC237756 (97%), Linmingia china AF481148 (97%), and Cytophaga sp. MDA2507 AF238333 (99%). No PCR amplification of 16s rRNA genes was obtained when archaea-specific primers were used.

  16. Biotreatment of chlorpyrifos in a bench scale bioreactor using Psychrobacter alimentarius T14.

    Science.gov (United States)

    Khalid, Saira; Hashmi, Imran

    2016-01-01

    Bacteria tolerant to high pesticide concentration could be used for designing an efficient treatment technology. Bacterial strains T14 was isolated from pesticide-contaminated soil in mineral salt medium (MSM) and identified as Psychrobacter alimentarius T14 using 16S rRNA gene sequence analysis. Bench scale bioreactor was evaluated for biotreatment of high Chlorpyrifos (CP) concentration using P. alimentarius T14. Effect of various parameters on bioreactor performance was examined and optimum removal was observed at optical density (OD600 nm): 0.8; pH: 7.2; CP concentration: 300 mg L(-1) and hydraulic retention time: 48 h. At optimum conditions, 70.3/79% of CP/chemical oxygen demand (COD) removal was achieved in batch bioreactors. In addition, P. alimentarius T14 achieved 95/91, 62.3/75, 69.8/64% CP/COD removal efficiency with addition of CS (co-substrates), CS1 (yeast extract + synthetic wastewater), CS2 (glucose + synthetic wastewater) and CS3 (yeast extract), respectively. Addition of CS1 to bioreactor could accelerate CP removal rate up to many cycles with considerable efficiency. However, accumulation of 3, 5, 6-trichloro-2-pyridinol affects reactor performance in cyclic mode. First-order rate constant k1 0.062 h(-1) and t1/2 11.1 h demonstrates fast degradation. Change in concentration of total chlorine and nitrogen could be the result of complete mineralization. Photodegradation of CP in commercial product was more than its pure form. Commercial formulation accelerated photodegradation process; however no effect on biodegradation process was observed. After bio-photodegradation, negligible toxicity for seeds of Triticum aestivum was observed. Study suggests an efficient treatment of wastewater containing CP and its metabolites in batch bioreactors could be achieved using P. alimentarius.

  17. Production of rhEPO with a serum-free medium in the packed bed bioreactor.

    Science.gov (United States)

    Deng, J; Yang, Q; Cheng, X; Li, L; Zhou, J

    1997-01-01

    Recombinant CHO (C2) cells producing human erythropoietin (rhEPO) were cultured with DMEM:F12 media containing 5% FBS for 8-10 days in a packed bed bioreactor, then rhEPO was produced with a serum-free medium (SFM-p) which was prepared in our laboratory. The SFM-p medium can support the growth of C2 cells and the production of rhEPO, and furthermore, it easily separates rhEPO from the culture supernatant. The cell culture in a packed bed bioreactor system using SFM-p was maintained in a stable condition for 20-25 days. The expression level of rhEPO was 12-28.4 mg/L. The bioreactor productivity was 71.0 mg/L.d and increased by 12-14 fold over that of the roller bottle. The glucose consumption rate was 21 g/L.d. At the end of 30 days of perfusion circulation, a final cell density of over 3.0 x 10(7)/ml of culture volume was achieved. Since the cells were entrapped in the polyester disk, the culture supernatant contained only a few detachment cells. Variations in lactate and ammonia production in the reactor were observed, and results showed that the productions of lactate and ammonia by the bioreactor were 3.5 g/L and 5 mmol/L, respectively, and did not affect the expression of interest protein. This experiment demonstrates that SFM-p is suitable for the growth and rhEPO production of recombinant C2 in the packed bed bioreactor.

  18. Bioreactor and process design for biohydrogen production.

    Science.gov (United States)

    Show, Kuan-Yeow; Lee, Duu-Jong; Chang, Jo-Shu

    2011-09-01

    Biohydrogen is regarded as an attractive future clean energy carrier due to its high energy content and environmental-friendly conversion. It has the potential for renewable biofuel to replace current hydrogen production which rely heavily on fossil fuels. While biohydrogen production is still in the early stage of development, there have been a variety of laboratory- and pilot-scale systems developed with promising potential. This work presents a review of advances in bioreactor and bioprocess design for biohydrogen production. The state-of-the art of biohydrogen production is discussed emphasizing on production pathways, factors affecting biohydrogen production, as well as bioreactor configuration and operation. Challenges and prospects of biohydrogen production are also outlined.

  19. Hydrostatic pressure and shear stress affect endothelin-1 and nitric oxide release by endothelial cells in bioreactors.

    Science.gov (United States)

    Vozzi, Federico; Bianchi, Francesca; Ahluwalia, Arti; Domenici, Claudio

    2014-01-01

    Abundant experimental evidence demonstrates that endothelial cells are sensitive to flow; however, the effect of fluid pressure or pressure gradients that are used to drive viscous flow is not well understood. There are two principal physical forces exerted on the blood vessel wall by the passage of intra-luminal blood: pressure and shear. To analyze the effects of pressure and shear independently, these two stresses were applied to cultured cells in two different types of bioreactors: a pressure-controlled bioreactor and a laminar flow bioreactor, in which controlled levels of pressure or shear stress, respectively, can be generated. Using these bioreactor systems, endothelin-1 (ET-1) and nitric oxide (NO) release from human umbilical vein endothelial cells were measured under various shear stress and pressure conditions. Compared to the controls, a decrease of ET-1 production by the cells cultured in both bioreactors was observed, whereas NO synthesis was up-regulated in cells under shear stress, but was not modulated by hydrostatic pressure. These results show that the two hemodynamic forces acting on blood vessels affect endothelial cell function in different ways, and that both should be considered when planning in vitro experiments in the presence of flow. Understanding the individual and synergic effects of the two forces could provide important insights into physiological and pathological processes involved in vascular remodeling and adaptation.

  20. 鲤鱼冻血细胞培养及染色体制备条件优化研究%Optimum Conditions Research on Frozen Blood Cell Cultivation and Chromosome Preparation of Common Carp( Cyprinus carpio L.)

    Institute of Scientific and Technical Information of China (English)

    安晶; 张桂贤; 李运东; 刘青; 陈维伟

    2012-01-01

    鱼类染色体制备方法中,外周血淋巴细胞培养法远远优于PHA体内注射法,使用冻血省力省功。由于外周血培养需要一定的条件和技术。该文就鲤鱼血液保存方法、细胞培养温度、秋水仙素处理浓度及滴加时间、低渗温度、固定处理次数等条件进行分析,得到较好的鲤鱼全血细胞培养及染色体标本制备方法,可为鲤鱼分子细胞遗传的后续研究奠定基础。%Among fish chromosomes preparation methods ,the peripheral blood lymphocyte cuhure method is far superior to the PHA shots in the body.Frozen blood is easier to get ,but peripheral blood cultivation need some conditions and techniques. This experiment analyzed the common carp blood conservation techniques,cell culture temperature,concentrations of colchicine and adding-time,hypotonic treatment temperature,fixed processing times and so on.It got a good carp complete blood incubation and chromosome preparation methods,which laid the foundation of the follow-up molecular cytogenetics research of carp.

  1. Effect of co-substrate on production of poly-β- hydroxybutyrate (PHB and copolymer PHBV from newly identified mutant Rhodobacter sphaeroides U7 cultivated under aerobic-dark condition

    Directory of Open Access Journals (Sweden)

    Kemarajt Kemavongse

    2007-07-01

    Full Text Available Photosynthetic bacterial mutant strain U7 was identified using both classical and molecular (16S rDNA techniques to be Rhodobacter sphaeroides. The glutamate-acetate (GA medium containing sodium acetate and sodium glutamate as carbon and nitrogen sources was used for production of poly-β-hydroxybutyrate (PHB from R. sphaeroides U7 cultivated under aerobic-dark condition (200 rpm at 37oC. Effect of auxiliary carbon sources (propionate and valerate and concentrations (molar ratio of 40/0, 40/20, 40/40 and 40/80 on copolymer production were studied. Both combinations of acetate with valerate and acetate with propionate were found to induce the accumulation of poly-β-hydroxybutyrate-co-β-hydroxyvalerate (PHBV within the cell. Acetate with propionate in the molar ratio of 40/40 gave the highest poly-β-hydroxyalkanoates (PHA content (77.68%, followed by acetate with valerate at the same molar ratio (77.42%. Although their polymer contents were similar, the presence of 40 mM valerate gave more than 4 times higher hydroxyvalerate (HV fraction (84.77% than in the presence of 40 mM propionate (19.12% HV fraction.

  2. 基于GRNN和粒子群算法的酱油种曲培养条件优化%Optimization of koji cultivation conditions in soy sauce production base on GRNN and PSO

    Institute of Scientific and Technical Information of China (English)

    张如意; 王学雷

    2012-01-01

    The spores number is an important index of soy sauce's koji quality. In order to improve the quality of soy sauce koji, Based on process data, A General Regression Neural Network (GRNN) with cross validation for optimizing model parameter was proposed to predict the final spores number, then particle swarm optimization (PSO) combined with GRNN was applied to optimize cultivation conditions. Simulation shows the algorithm runs fast and the optimization result is practical to guide koji production.%孢子数是酱油种曲质量的一个重要指标.为提高种曲质量,基于培养过程数据,给出一种广义回归神经网络和粒子群算法相结合的优化种曲培养条件方法.首先建立预测种曲孢子数的广义回归神经网络模型,并利用交叉验证确定GRNN模型的最优参数,而后使用粒子群算法寻优种曲培养条件.仿真实验显示优化结果可应用于实践指导生产,且算法易于实施、运行速度快.

  3. Culturing and applications of rotating wall vessel bioreactor derived 3D epithelial cell models.

    Science.gov (United States)

    Radtke, Andrea L; Herbst-Kralovetz, Melissa M

    2012-04-03

    Cells and tissues in the body experience environmental conditions that influence their architecture, intercellular communications, and overall functions. For in vitro cell culture models to accurately mimic the tissue of interest, the growth environment of the culture is a critical aspect to consider. Commonly used conventional cell culture systems propagate epithelial cells on flat two-dimensional (2-D) impermeable surfaces. Although much has been learned from conventional cell culture systems, many findings are not reproducible in human clinical trials or tissue explants, potentially as a result of the lack of a physiologically relevant microenvironment. Here, we describe a culture system that overcomes many of the culture condition boundaries of 2-D cell cultures, by using the innovative rotating wall vessel (RWV) bioreactor technology. We and others have shown that organotypic RWV-derived models can recapitulate structure, function, and authentic human responses to external stimuli similarly to human explant tissues (1-6). The RWV bioreactor is a suspension culture system that allows for the growth of epithelial cells under low physiological fluid shear conditions. The bioreactors come in two different formats, a high-aspect rotating vessel (HARV) or a slow-turning lateral vessel (STLV), in which they differ by their aeration source. Epithelial cells are added to the bioreactor of choice in combination with porous, collagen-coated microcarrier beads (Figure 1A). The cells utilize the beads as a growth scaffold during the constant free fall in the bioreactor (Figure 1B). The microenvironment provided by the bioreactor allows the cells to form three-dimensional (3-D) aggregates displaying in vivo-like characteristics often not observed under standard 2-D culture conditions (Figure 1D). These characteristics include tight junctions, mucus production, apical/basal orientation, in vivo protein localization, and additional epithelial cell-type specific properties

  4. Aujeszky's disease virus production in disposable bioreactor

    Indian Academy of Sciences (India)

    I Slivac; V Gaurina Srček; K Radošević; I Kmetič; Z Kniewald

    2006-09-01

    A novel, disposable-bag bioreactor system that uses wave action for mixing and transferring oxygen was evaluated for BHK 21 C13 cell line growth and Aujeszky’s disease virus (ADV) production. Growth kinetics of BHK 21 C13 cells in the wave bioreactor during 3-day period were determined. At the end of the 3-day culture period and cell density of 1.82 × 106 cells ml–1, the reactor was inoculated with 9 ml of gE- Bartha K-61 strain ADV suspension (105.9 TCID50) with multiplicity of infection (MOI) of 0.01. After a 144 h incubation period, 400 ml of ADV harvest was obtained with titre of 107.0 TCID50 ml–1, which corresponds to 40,000 doses of vaccine against AD. In conclusion, the results obtained with the wave bioreactor using BHK 21 C13 cells showed that this system can be considered as suitable for ADV or BHK 21 C13 cell biomass production.

  5. Membrane bioreactors and their uses in wastewater treatments

    Energy Technology Data Exchange (ETDEWEB)

    Le-Clech, Pierre [New South Wales Univ., Sydney (Australia). UNESCO Centre for Membrane Science and Technology

    2010-12-15

    With the current need for more efficient and reliable processes for municipal and industrial wastewaters treatment, membrane bioreactor (MBR) technology has received considerable attention. After just a couple of decades of existence, MBR can now be considered as an established wastewater treatment system, competing directly with conventional processes like activated sludge treatment plant. However, MBR processes still suffer from major drawbacks, including high operational costs due to the use of anti-fouling strategies applied to the system to maintain sustainable filtration conditions. Moreover, this specific use of membranes has not reached full maturity yet, as MBR suppliers and users still lack experience regarding the long-term performances of the system. Still, major improvements of the MBR design and operation have been witnessed over the recent years, making MBR an option of choice for wastewater treatment and reuse. This mini-review reports recent developments and current research trends in the field. (orig.)

  6. Membrane bioreactors and their uses in wastewater treatments.

    Science.gov (United States)

    Le-Clech, Pierre

    2010-12-01

    With the current need for more efficient and reliable processes for municipal and industrial wastewaters treatment, membrane bioreactor (MBR) technology has received considerable attention. After just a couple of decades of existence, MBR can now be considered as an established wastewater treatment system, competing directly with conventional processes like activated sludge treatment plant. However, MBR processes still suffer from major drawbacks, including high operational costs due to the use of anti-fouling strategies applied to the system to maintain sustainable filtration conditions. Moreover, this specific use of membranes has not reached full maturity yet, as MBR suppliers and users still lack experience regarding the long-term performances of the system. Still, major improvements of the MBR design and operation have been witnessed over the recent years, making MBR an option of choice for wastewater treatment and reuse. This mini-review reports recent developments and current research trends in the field.

  7. Hydrodynamic effects on cell growth in agitated microcarrier bioreactors

    Science.gov (United States)

    Cherry, Robert S.; Papoutsakis, E. Terry

    1988-01-01

    The net growth rate of bovine embryonic kidney cells in microcarrier bioreactor is the result of a variable death rate imposed on a cell culture trying to grow at a constant intrinsic growth rate. The death rate is a function of the agitation conditions in the system, and increases at higher agitation because of increasingly energetic interactions of the cell covered microcarriers with turbulent eddies in the fluid. At very low agitation rates bead-bead bridging becomes important; the large clumps formed by bridging can interact with larger eddies than single beads, leading to a higher death rate at low agitation. The growth and death rate were correlated with a dimensionless eddy number which compares eddy forces to the buoyant force on the bead.

  8. Decolourisation of textile wastewater in a submerged anaerobic membrane bioreactor.

    Science.gov (United States)

    Spagni, Alessandro; Casu, Stefania; Grilli, Selene

    2012-08-01

    Azo dye decolourisation can be easily achieved by biological reduction under anaerobic conditions. The aim of this study was to evaluate the applicability of submerged anaerobic membrane bioreactors (SAMBRs) for the decolourisation of dyeing wastewater containing azo dyes. The reactive orange 16 was used as model of an azo dye. The results demonstrated that very high decolourisation (higher than 99%) can be achieved by SAMBRs. Although decolourisation was not significantly influenced by the azo dye concentrations up to 3.2 g L(-1), methane production was greatly inhibited (up to 80-85%). Since volatile fatty acids accumulated in the treatment system with the azo dye concentration increase, methanogenes seem to be the most sensitive microbial populations of the anaerobic ecological community. The results demonstrated that anaerobic process combined with membrane filtration can deal with highly concentrated wastewaters that result from stream separation of industrial discharges.

  9. Membrane fouling control by ultrasound in an anaerobic membrane bioreactor

    Institute of Scientific and Technical Information of China (English)

    SUI Pengzhe; WEN Xianghua; HUANG Xia

    2007-01-01

    In this study,ultrasound was used to control the membrane fouling online in an anaerobic membrane bioreactor (AMBR).Short-term running experiments were carried out under different operating conditions to explore feasible ultrasonic parameters.The experimental results indicated that when the crossflow velocity was more than 1.0 m/s,membrane fouling could be controlled effectively only by hydrodynamic methods without ultrasound.When ultrasound was applied,an ultrasonic power range of 60-150 W was suitable for the membrane fouling control in the experimental system.The experimental results showed that the membrane fouling was controlled so well that membrane filtration resistance(ΣR) could stay at 5×1011 m-1 for more than a week with the crossflow velocity of 0.75 m/s,which equaled the effect of crossflow velocity of more than 1.0 m/s without ultrasound.

  10. Feasibility of using sodium chloride as a tracer for the characterization of the distribution of matter in complex multi-compartment 3D bioreactors for stem cell culture.

    Science.gov (United States)

    Gerlach, Jörg C; Witaschek, Tom; Strobel, Catrin; Brayfield, Candace A; Bornemann, Reinhard; Catapano, Gerardo; Zeilinger, Katrin

    2010-06-01

    The experimental characterization of the distribution of matter in complex multi-compartment three-dimensional membrane bioreactors for human cell culture is complicated by tracer interactions with the membranes and other bioreactor constituents. This is due to the fact that membranes with a high specific surface area often feature a hydrophobic chemical backbone that may adsorb tracers often used to this purpose, such as proteins and dyes. Membrane selectivity, and its worsening caused by protein adsorption, may also hinder tracer transfer across neighboring compartments, thus preventing effective characterization of the distribution of matter in the whole bioreactor. Tracer experiments with sodium chloride (NaCl) may overcome some of these limitations and be effectively used to characterize the distribution of matter in complex 3D multi-compartments membrane bioreactors for stem cell culture. NaCl freely permeates most used membranes, it does not adsorb on uncharged membranes, and its concentration may be accurately measured in terms of solution conductivity. In this preliminary study, the feasibility of complex multi-compartment membrane bioreactors was investigated with a NaCl concentration pulse challenge to characterize how their distribution of matter changes when they are operated under different conditions. In particular, bioreactors consisting of three different membrane types stacked on top of one another to form a 3D network were characterized under different feed conditions.

  11. Production of high-titer human influenza A virus with adherent and suspension MDCK cells cultured in a single-use hollow fiber bioreactor.

    Science.gov (United States)

    Tapia, Felipe; Vogel, Thomas; Genzel, Yvonne; Behrendt, Ilona; Hirschel, Mark; Gangemi, J David; Reichl, Udo

    2014-02-12

    Hollow fiber bioreactors (HFBRs) have been widely described as capable of supporting the production of highly concentrated monoclonal antibodies and recombinant proteins. Only recently HFBRs have been proposed as new single-use platforms for production of high-titer influenza A virus. These bioreactors contain multiple hollow fiber capillary tubes that separate the bioreactor in an intra- and an extra-capillary space. Cells are usually cultured in the extra-capillary space and can grow to a very high cell concentration. This work describes the evaluation of the single-use hollow fiber bioreactor PRIMER HF (Biovest International Inc., USA) for production of influenza A virus. The process was setup, characterized and optimized by running a total of 15 cultivations. The HFBRs were seeded with either adherent or suspension MDCK cells, and infected with influenza virus A/PR/8/34 (H1N1), and the pandemic strain A/Mexico/4108/2009 (H1N1). High HA titers and TCID₅₀ of up to 3.87 log₁₀(HA units/100 μL) and 1.8 × 10(10)virions/mL, respectively, were obtained for A/PR/8/34 influenza strain. Influenza virus was collected by performing multiple harvests of the extra-capillary space during a virus production time of up to 12 days. Cell-specific virus yields between 2,000 and 8,000 virions/cell were estimated for adherent MDCK cells, and between 11,000 and 19,000 virions/cell for suspension MDCK.SUS2 cells. These results do not only coincide with the cell-specific virus yields obtained with cultivations in stirred tank bioreactors and other high cell density systems, but also demonstrate that HFBRs are promising and competitive single-use platforms that can be considered for commercial production of influenza virus.

  12. Towards a Tissue-Engineered Ligament: Design and Preliminary Evaluation of a Dedicated Multi-Chamber Tension-Torsion Bioreactor

    Directory of Open Access Journals (Sweden)

    Cédric P. Laurent

    2014-02-01

    Full Text Available Tissue engineering may constitute a promising alternative to current strategies in ligament repair, providing that suitable scaffolds and culture conditions are proposed. The objective of the present contribution is to present the design and instrumentation of a novel multi-chamber tension-torsion bioreactor dedicated to ligament tissue engineering. A preliminary biological evaluation of a new braided scaffold within this bioreactor under dynamic loading is reported, starting with the development of a dedicated seeding protocol validated from static cultures. The results of these preliminary biological characterizations confirm that the present combination of scaffold, seeding protocol and bioreactor may enable us to head towards a suitable ligament tissue-engineered construct.

  13. Multi-objective optimization of an industrial penicillin V bioreactor train using non-dominated sorting genetic algorithm.

    Science.gov (United States)

    Lee, Fook Choon; Rangaiah, Gade Pandu; Ray, Ajay Kumar

    2007-10-15

    Bulk of the penicillin produced is used as raw material for semi-synthetic penicillin (such as amoxicillin and ampicillin) and semi-synthetic cephalosporins (such as cephalexin and cefadroxil). In the present paper, an industrial penicillin V bioreactor train is optimized for multiple objectives simultaneously. An industrial train, comprising a bank of identical bioreactors, is run semi-continuously in a synchronous fashion. The fermentation taking place in a bioreactor is modeled using a morphologically structured mechanism. For multi-objective optimization for two and three objectives, the elitist non-dominated sorting genetic algorithm (NSGA-II) is chosen. Instead of a single optimum as in the traditional optimization, a wide range of optimal design and operating conditions depicting trade-offs of key performance indicators such as batch cycle time, yield, profit and penicillin concentration, is successfully obtained. The effects of design and operating variables on the optimal solutions are discussed in detail.

  14. Optimization of complex medium composition for heterotrophic cultivation of Euglena gracilis and paramylon production.

    Science.gov (United States)

    Ivušić, Franjo; Šantek, Božidar

    2015-06-01

    Heterotrophic cultivation of Euglena gracilis was carried out on synthetic (Hutner medium) and complex cultivation media in order to optimize production of β-1,3-glucan (paramylon). For preparation of complex media, various industrial by-products (e.g., molasses, corn steep solid, yeast extract, and beef extract) were used with or without addition of pure compounds [glucose, galactose, fructose, lactose, maltose, sucrose, and (NH4)2HPO4]. Heterotrophic cultivation of E. gracilis was performed in Erlenmeyer flasks and additionally confirmed during research in the stirred tank bioreactor. The results clearly show that E. gracilis can easily metabolize glucose and fructose as carbon sources and corn steep solid as complex nitrogen and growth factors source for biomass growth and paramylon synthesis. Furthermore, it was also proved that addition of (NH4)2HPO4, beef extract, or gibberellic acid did not have positive effect on the biomass growth and paramylon synthesis. After optimization of complex medium composition and verification in the stirred tank bioreactor, it was concluded that medium composed of glucose (20 g/L) and corn steep solid (30 g/L) is the most suitable complex medium for industrial cultivation of E. gracilis and paramylon production.

  15. An experimental investigation of the multiphase flows in a photobioreactor for algae cultivation

    Science.gov (United States)

    Yang, Zifeng; Hu, Hui; Del Ninno, Matteo; Wen, Zhiyou

    2011-11-01

    Algal biomass is a promising feedstock for biofuels production, with photobioreactors being one of the major cultivation systems for algal cells. Light absorption, fluid dynamics, and algal metabolism are three key factors in determining the overall performance of a photobioreactor. The behavior of the multiphase flow (i.e., liquid phase - water, gas phase - CO2 and O2, and solid phase - algal cells) and turbulent mixing inside the reactor are the core connecting the three factors together. One of the major challenges in the optimal design of photobioreactors for algae cultivation is the lack of in-depth understanding of the characteristics of the multiphase flows and turbulent mixing. In this study, we present a comprehensive experimental study to investigate the effects of turbulent mixing in photobioreactors on the performance of a photobioreactor for algae cultivation. A high-resolution particle image velocity (PIV) system is used to achieve time-resolved, in-situ flow field measurements to quantify the turbulent mixing of the multiphase flows inside the bioreactor, while algal cultures are also grown in the same reactor with the same experimental settings. The mixing characteristics of the multiphase flow are correlated with the algal growth performance in the bioreactors to elucidate the underlying physics to explore/optimize design paradigms for the optimization of photobioreactor designs for algae cultivation.

  16. Producing bioethanol from cellulosic hydrolyzate via co-immobilized cultivation strategy.

    Science.gov (United States)

    Liu, Yu-Kuo; Yang, Chih-An; Chen, Wei-Chuan; Wei, Yu-Hong

    2012-08-01

    Lignocellulose was converted into reducing sugars by using saccharification enzymes from cocultivated Trichoderma reesei and Aspergillus niger and reducing sugars as nutrients for Zymomonas mobilis to produce bioethanol in an immobilization system. After 96 h of cultivation, cocultivated T. reesei and A. niger had enzymatical synergistic effects that enabled a reducing sugar production of 1.29 g/L and a cellulose conversion rate of 23.27%. An 18% total inoculum concentration and a 1/1 inoculation ratio of T. reesei to A. niger obtained a reducing sugar production rate and a cellulose conversion rate of 2.57 g/L and 46.27%, respectively. The co-immobilization cultivation results showed that using polyurethane as a carrier optimized total saccharification enzyme activity at an inoculum ratio of 1/1 and a total inoculum concentration of 6.5×10(6)spores/mL. Based on the experimental results, the bioreactor design was further modified to enhance bioethanol production. The three strains (A. niger, T. reesei and Z. mobilis) were cocultivated with a co-immobilization cultivation system. The experimental results showed that, after 24 h cultivation, bioethanol production reached 0.56 g/L, and reducing sugar conversion rate reached 11.2% when using carboxymethylcellulose (CMC) substrates. The experimental results confirmed that the modified bioreactor enhances bioethanol production. However, further experiments are needed to determine how to prevent multi-stage failure of reducing medium volume.

  17. 太子参不定根组织培养的研究%Study on tissue cultivation of adventitious roots of Pseudoxtellariae heterophylla

    Institute of Scientific and Technical Information of China (English)

    梁玉勇; 尹双双; 左北梅; 高文远

    2012-01-01

    Objective: To systematically optimize the cultivation conditions of adventitious roots of Pseudoxtellariae heterophylla. Method: Tissue cultivation technology and ultraviolet spectrophotometry were adopted to observe the effect of inoculum volume, sucrose concentration, inorganic salt concentration, number of cultivation days, gradual scale-up cultivation and bubble different angles of bioreactor on the growth of adventitious roots of P. heterophylla, and determine the content of constituents such as saponin, polysaccha-ride and amino acid. Result: The propagation multiple of adventitious roots reached the maximum when the inoculum was 6 g in a 1 L culture shake flask. With the increase in sucrose concentration, the dry weight propagation multiples of adventitious roots followed an up and down trend. The inorganic salt concentration in a cultivation dish had a greater effect on the growth of adventitious roots, particularly 3/4 MS was the most favorable for the growth of adventitious roots. The growth curve of P. heterophylla was " S" , with the bio-mass reaching the maximum at the 28th day. Conclusion: The inoculum volume, sucrose concentration, inorganic salt concentration, gradual scale-up cultivation and angles of bubble bioreactor had a significant effect on the growth of adventitious roots of P. heterophylla. The contents of saponin and amino acid in adventitious roots were higher than that in cultivated P. heterophylla, whereas the poly-saccharide content were lower than that in cultivated P. heterophylla.%目的:对太子参不定根的培养条件进行系统的优化.方法:利用组织培养技术结合紫外分光光度法,考察了接种量、蔗糖浓度、无机盐浓度、培养天数、逐级扩大培养以及不同角度鼓泡式反应器对太子参不定根生长的影响,并对不定根中皂苷、多糖和氨基酸等成分进行含量测定.结果:每1L培养基接种的不定根鲜重为6 g时,太子参不定根的干重增殖倍数达到

  18. New generation NMR bioreactor coupled with high-resolution NMR spectroscopy leads to novel discoveries in Moorella thermoaceticum metabolic profiles

    Energy Technology Data Exchange (ETDEWEB)

    Xue, Junfeng; Isern, Nancy G.; Ewing, R James; Liyu, Andrey V.; Sears, Jesse A.; Knapp, Harlan; Iversen, Jens; Sisk, Daniel R.; Ahring, Birgitte K.; Majors, Paul D.

    2014-06-20

    An in-situ nuclear magnetic resonance (NMR) bioreactor was developed and employed to monitor microbial metabolism under batch-growth conditions in real time. We selected Moorella thermoacetica ATCC 49707 as a test case. M. thermoacetica (formerly Clostridium thermoaceticum) is a strictly anaerobic, thermophilic, acetogenic, gram-positive bacterium with potential for industrial production of chemicals. The metabolic profiles of M. thermoacetica were characterized during growth in batch mode on xylose (a component of lignocellulosic biomass) using the new generation NMR bioreactor in combination with high-resolution, high sensitivity NMR (HR-NMR) spectroscopy. In-situ NMR measurements were performed using water-suppressed H-1 NMR spectroscopy at an NMR frequency of 500 MHz, and aliquots of the bioreactor contents were taken for 600 MHz HR-NMR spectroscopy at specific intervals to confirm metabolite identifications and expand metabolite coverage. M. thermoacetica demonstrated the metabolic potential to produce formate, ethanol and methanol from xylose, in addition to its known capability of producing acetic acid. Real-time monitoring of bioreactor conditions showed a temporary pH decrease, with a concomitant increase in formic acid during exponential growth. Fermentation experiments performed outside of the magnet showed that the strong magnetic field employed for NMR detection did not significantly affect cell metabolism. Use of the in-situ NMR bioreactor facilitated monitoring of the fermentation process in real time, enabling identification of intermediate and end-point metabolites and their correlation with pH and biomass produced during culture growth. Real-time monitoring of culture metabolism using the NMR bioreactor in combination with the HR-NMR spectroscopy will allow optimization of the metabolism of microorganisms producing valuable bioproducts.

  19. New generation NMR bioreactor coupled with high-resolution NMR spectroscopy leads to novel discoveries in Moorella thermoacetica metabolic profiles.

    Science.gov (United States)

    Xue, Junfeng; Isern, Nancy G; Ewing, R James; Liyu, Andrei V; Sears, Jesse A; Knapp, Harlan; Iversen, Jens; Sisk, Daniel R; Ahring, Birgitte K; Majors, Paul D

    2014-10-01

    An in situ nuclear magnetic resonance (NMR) bioreactor was developed and employed to monitor microbial metabolism under batch growth conditions in real time. We selected Moorella thermoacetica ATCC 49707 as a test case. M. thermoacetica (formerly Clostridium thermoaceticum) is a strictly anaerobic, thermophilic, acetogenic, gram-positive bacterium with potential for industrial production of chemicals. The metabolic profiles of M. thermoacetica were characterized during growth in batch mode on xylose (a component of lignocellulosic biomass) using the new generation NMR bioreactor in combination with high-resolution NMR (HR-NMR) spectroscopy. In situ NMR measurements were performed using water-suppressed H-1 NMR spectroscopy at 500 MHz, and aliquots of the bioreactor contents were taken for 600-MHz HR-NMR spectroscopy at specific intervals to confirm metabolite identifications and expand metabolite coverage. M. thermoacetica demonstrated the metabolic potential to produce formate, ethanol, and methanol from xylose, in addition to its known capability of producing acetic acid. Real-time monitoring of bioreactor conditions showed a temporary pH decrease, with a concomitant increase in formic acid during exponential growth. Fermentation experiments performed outside of the magnet showed that the strong magnetic field employed for NMR detection did not significantly affect cell metabolism. Use of the in situ NMR bioreactor facilitated monitoring of the fermentation process, enabling identification of intermediate and endpoint metabolites and their correlation with pH and biomass produced during culture growth. Real-time monitoring of culture metabolism using the NMR bioreactor in combination with HR-NMR spectroscopy will allow optimization of the metabolism of microorganisms producing valuable bioproducts.

  20. Effect of sucrose and potassium nitrate on biomass and saponin content of Talinum paniculatum Gaertn. hairy root in balloon-type bubble bioreactor

    Institute of Scientific and Technical Information of China (English)

    Yosephine Sri Wulan Manuhara; Alfinda Novi Kristanti; Edy Setiti Wida Utami; Arif Yachya

    2015-01-01

    Objective: To increase biomass and saponin production in hairy root culture of Talinum paniculatum Gaertn. (T. paniculatum) in balloon-type bubble bioreactor (BTBB). Methods: Hairy roots which were collected from leaf explants of T. paniculatum were infected by Agrobacterium rhizogenes strain LB510. The hairy roots were cultivated at 400 mL Murashige and Skoog liquid medium without growth regulator (MS0) in 1 000 mL BTBB. Each BTBB had 2 g hairy roots as initial inoculum and these cultures were treated with various concentrations of sucrose (3%, 4%, 5%, 6%w/v) and potassium nitrate (0.5, 1.0, 1.5 and 2.0 strength of MS medium). Cultures were maintained for 14 days. Fresh and dry weights of hairy roots at the end of culture were investigated. Results: Various concentrations of sucrose influenced the biomass accumulation of hairy roots. Maximum biomass was reached by MS medium supplemented with 6% sucrose and it was approximately threefold higher than control. Culture supplemented with po-tassium nitrate at 2.0 strength of MS0 could increase biomass accumulation of hairy roots until 0.14 g dry weight and it was almost threefold higher than control. However, the maximum saponin content was obtained by MS medium supplemented with 5%sucrose and 2.0 strength potassium nitrate of MS. Conclusions: Based on this research, those conditions can be used to produce biomass and saponin of hairy root of T. paniculatum in the large scale.

  1. Analysis of ammonia-oxidizing bacteria dominating in lab-scale bioreactors with high ammonium bicarbonate loading

    NARCIS (Netherlands)

    Vejmelkova, D.; Sorokin, D.Y.; Abbas, B.; Kovaleva, O.L.; Kleerebezem, R.; Kampschreur, M.J.; Muyzer, G.; Van Loosdrecht, M.C.M.

    2011-01-01

    The ammonia-oxidizing bacterial community (AOB) was investigated in two types of laboratory-scale bioreactors performing partial oxidation of ammonia to nitrite or nitrate at high (80 mM) to extremely high (428 mM) concentrations of ammonium bicarbonate. At all conditions, the dominant AOB was affil

  2. Analysis of ammonia-oxidizing bacteria dominating in lab-scale bioreactors with high ammonium bicarbonate loading

    NARCIS (Netherlands)

    D. Vejmelkova; D.Y. Sorokin; B. Abbas; O.L. Kovaleva; R. Kleerebezem; M.J. Kampschreur; G. Muyzer; M.C.M. van Loosdrecht

    2012-01-01

    The ammonia-oxidizing bacterial community (AOB) was investigated in two types of laboratory-scale bioreactors performing partial oxidation of ammonia to nitrite or nitrate at high (80 mM) to extremely high (428 mM) concentrations of ammonium bicarbonate. At all conditions, the dominant AOB was affil

  3. Greenhouse Gas Emission from In-situ Denitrifying Bioreactors

    Science.gov (United States)

    Pluer, W.; Walter, M. T.; Geohring, L.

    2013-12-01

    Despite decades of concerted effort to mitigate nonpoint source nitrate (NO3-) pollution from agricultural lands, these efforts have not been sufficient to arrest eutrophication, which continues to be a serious and chronic problem. Two primary processes for removing excess NO3- from water are biological assimilation and denitrification. Denitrifying bacteria use NO3- as the electron acceptor for respiration in the absence of oxygen. Denitrification results in reduced forms of nitrogen, often dinitrogen gas (N2) but also nitrous oxide (N2O), an aggressive greenhouse gas (GHG). A promising solution to NO3- pollution is to intercept agricultural discharges with denitrifying bioreactors (DNBRs), though research has been limited to NO3- level reduction and omitted process mechanisms. DNBRs work by providing an anaerobic environment with plenty of organic matter (commonly woodchips) for denitrifying bacteria to flourish. While, initial results from bioreactor studies show that they can cost-effectively remove NO3-, GHG emission could be an unintended consequence. The study's goal is to determine how bioreactor design promotes microbial denitrification while limiting N2O production. It specifically focuses on expanding the body of knowledge concerning DNBRs in the areas of design implications and internal processes by measuring intermediate compounds and not solely NO3-. Nutrient samples are collected at inflow and outflow structures and tested for NO3- and nitrite (NO2-). Dissolved and headspace gas samples are collected and tested for N2O. Additional gas samples will be analyzed for naturally-occurring isotopic N2 to support proposed pathways. Designs will be analyzed both through the N2O/N2 production ratio and NO2- production caused by various residence times and inflow NO3- concentrations. High GHG ratios and NO2- production suggest non-ideal conditions or flow patterns for complete denitrification. NO3- reduction is used for comparison with previous studies. Few

  4. Production of asiaticoside from centella (Centella asiatica L. Urban) cells in bioreactor

    Institute of Scientific and Technical Information of China (English)

    Nguyen Hoang Loc; Nguyen Thi Duy Nhat

    2013-01-01

    Objective:To investigate the effects of some culture conditions on production of asiaticoside from centella (Centella asiatica L. Urban) cells cultured in 5-L bioreactor. Methods: The centell cell suspension culture was conducted in 5-L bioreactor to investigate the growth and asiaticoside accumulation under various conditions. Asiaticoside content was determined by HPLC analysis. Results:The results showed that the cell growth and asiaticoside accumulation peaked after 24 d of culture at an agitation speed of 150 r/min and aeration rate of 2.5 L/min. The cell biomass reached a maximum value of 302.45 g fresh weight (31.45 g dry weight) and growth index of 3.03 with inoculum size of 100 g. However, asiaticoside content was the highest (60.08 mg/g dry weight) when culture was initiated with an inoculum size of 50 g. Conclusions: The present study found the suitable conditions for growth of centella cells and their asiaticoside production in bioreactor.

  5. Teratoma formation of human embryonic stem cells in three-dimensional perfusion culture bioreactors.

    Science.gov (United States)

    Stachelscheid, H; Wulf-Goldenberg, A; Eckert, K; Jensen, J; Edsbagge, J; Björquist, P; Rivero, M; Strehl, R; Jozefczuk, J; Prigione, A; Adjaye, J; Urbaniak, T; Bussmann, P; Zeilinger, K; Gerlach, J C

    2013-09-01

    Teratoma formation in mice is today the most stringent test for pluripotency that is available for human pluripotent cells, as chimera formation and tetraploid complementation cannot be performed with human cells. The teratoma assay could also be applied for assessing the safety of human pluripotent cell-derived cell populations intended for therapeutic applications. In our study we examined the spontaneous differentiation behaviour of human embryonic stem cells (hESCs) in a perfused 3D multi-compartment bioreactor system and compared it with differentiation of hESCs and human induced pluripotent cells (hiPSCs) cultured in vitro as embryoid bodies and in vivo in an experimental mouse model of teratoma formation. Results from biochemical, histological/immunohistological and ultrastuctural analyses revealed that hESCs cultured in bioreactors formed tissue-like structures containing derivatives of all three germ layers. Comparison with embryoid bodies and the teratomas revealed a high degree of similarity of the tissues formed in the bioreactor to these in the teratomas at the histological as well as transcriptional level, as detected by comparative whole-genome RNA expression profiling. The 3D culture system represents a novel in vitro model that permits stable long-term cultivation, spontaneous multi-lineage differentiation and tissue formation of pluripotent cells that is comparable to in vivo differentiation. Such a model is of interest, e.g. for the development of novel cell differentiation strategies. In addition, the 3D in vitro model could be used for teratoma studies and pluripotency assays in a fully defined, controlled environment, alternatively to in vivo mouse models.

  6. Modulation of mesenchymal stromal cell characteristics by microcarrier culture in bioreactors.

    Science.gov (United States)

    Hupfeld, Julia; Gorr, Ingo H; Schwald, Christian; Beaucamp, Nicola; Wiechmann, Kornelius; Kuentzer, Karin; Huss, Ralf; Rieger, Bernhard; Neubauer, Markus; Wegmeyer, Heike

    2014-11-01

    Mesenchymal stromal cells (MSCs) are promising candidates for cell therapy. Their therapeutic use requires extensive expansion to obtain a sufficiently high number of cells for clinical applications. State-of-the-art expansion systems, that is, primarily culture flask-based systems, are limited regarding scale-up, automation, and reproducibility. To overcome this bottleneck, microcarrier (MC)-based expansion processes have been developed. For the first time, MSCs from the perinatal sources umbilical cord (UC) and amniotic membrane (AM) were expanded on MCs. This study focuses on the comparison of flask- and Cytodex 1 MC-expanded MSCs by evaluating the influence of the expansion process on biological MSC characteristics. Furthermore, we tested the hypothesis to obtain more homogeneous MSC preparations by expanding cells on MCs in controlled large-scale bioreactors. MSCs were extensively characterized determining morphology, cell growth, surface marker expression, and functional properties such as differentiation capacity, secretion of paracrine factors, and gene expression. Based on their gene expression profile MSCs from different donors and sources clearly clustered in distinct groups solely depending on the expansion process-MC or flask culture. MC- and flask-expanded MSCs significantly differed from each other regarding surface markers and both paracrine factors and gene expression profiles. Furthermore, based on gene expression analysis, MC cultivation of MSCs in controlled bioreactor systems resulted in less heterogeneity between cells from different donors. In conclusion, MC-based MSC expansion in controlled bioreactors has the potential to reliably produce MSCs with altered characteristics and functions as compared to flask-expanded MSCs. These findings may be useful for the generation of MSCs with tailored properties for clinical applications.

  7. An Optical Oxygen Sensor for Long-Term Continuous Monitoring of Dissolved Oxygen in Perfused Bioreactors

    Science.gov (United States)

    Gao, F. G.; Jeevarajan, A. S.; Anderson, M. M.

    2002-01-01

    For long-term growth of man1ITlalian cells in perfused bioreactors, it is essential to monitor the concentration of dissolved oxygen (DO) present in the culture medium to quantitate and control level of DO. Continuous measurement of the amount of DO in the cell culture medium in-line under sterile conditions in NASA's perfused bioreactor requires that the oxygen sensor provide increased sensitivity and be sterilizable and nontoxic. Additionally, long-term cell culture experiments require that the calibration be maintained several weeks or months. Although there are a number of sensors for dissolved oxygen on the market and under development elsewhere, very few meet these stringent conditions. An optical oxygen sensor (BOXY) based on dynamic fluorescent quenching and a pulsed blue LED light source was developed in our laboratory to address these requirements. Tris( 4,7 -diphenyl-l, 1 O-phenanthroline )ruthenium(II) chloride is employed as the fluorescent dye indicator. The sensing element consists of a glass capillary (OD 4.0 mm; ID 2.0 mm) coated internally with a thin layer of the fluorescent dye in silicone matrix and overlayed with a black shielding layer. Irradiation of the sensing element with blue light (blue LED with emission maximum at 475 nm) generates a red fluorescence centered at 626 nm. The fluorescence intensity is correlated to the concentration of DO present in the culture medium, following the modified non-linear Stern-Volmer equation. By using a pulsed irradiating light source, the problem of dye-bleaching, which is often encountered in long-term continuous measurements of tIns type, 'is minimized. To date we achieved sensor resolution of 0.3 mmHg at 50 mmHg p02, and 0.6 mmHg at 100 mmHg p02, with a response time of about one minute. Calibration was accomplished in sterile phosphate-buffered saline with a blood-gas analyzer (BGA) measurement as reference. Stand-alone software was also developed to control the sensor and bioreactor as well as to

  8. Denitrifying bioreactors for nitrate removal from tile drained cropland

    Science.gov (United States)

    Denitrification bioreactors are a promising technology for mitigation of nitrate-nitrogen (NO3-N) losses in subsurface drainage water. Bioreactors are constructed with carbon substrates, typically wood chips, to provide a substrate for denitrifying microorganisms. Researchers in Iowa found that for ...

  9. Microbial community analysis of a full-scale DEMON bioreactor.

    Science.gov (United States)

    Gonzalez-Martinez, Alejandro; Rodriguez-Sanchez, Alejandro; Muñoz-Palazon, Barbara; Garcia-Ruiz, Maria-Jesus; Osorio, Francisco; van Loosdrecht, Mark C M; Gonzalez-Lopez, Jesus

    2015-03-01

    Full-scale applications of autotrophic nitrogen removal technologies for the treatment of digested sludge liquor have proliferated during the last decade. Among these technologies, the aerobic/anoxic deammonification process (DEMON) is one of the major applied processes. This technology achieves nitrogen removal from wastewater through anammox metabolism inside a single bioreactor due to alternating cycles of aeration. To date, microbial community composition of full-scale DEMON bioreactors have never been reported. In this study, bacterial community structure of a full-scale DEMON bioreactor located at the Apeldoorn wastewater treatment plant was analyzed using pyrosequencing. This technique provided a higher-resolution study of the bacterial assemblage of the system compared to other techniques used in lab-scale DEMON bioreactors. Results showed that the DEMON bioreactor was a complex ecosystem where ammonium oxidizing bacteria, anammox bacteria and many other bacterial phylotypes coexist. The potential ecological role of all phylotypes found was discussed. Thus, metagenomic analysis through pyrosequencing offered new perspectives over the functioning of the DEMON bioreactor by exhaustive identification of microorganisms, which play a key role in the performance of bioreactors. In this way, pyrosequencing has been proven as a helpful tool for the in-depth investigation of the functioning of bioreactors at microbiological scale.

  10. Expression Systems and Species Used for Transgenic Animal Bioreactors

    OpenAIRE

    Yanli Wang; Sihai Zhao; Liang Bai; Jianglin Fan; Enqi Liu

    2013-01-01

    Transgenic animal bioreactors can produce therapeutic proteins with high value for pharmaceutical use. In this paper, we compared different systems capable of producing therapeutic proteins (bacteria, mammalian cells, transgenic plants, and transgenic animals) and found that transgenic animals were potentially ideal bioreactors for the synthesis of pharmaceutical protein complexes. Compared with other transgenic animal expression systems (egg white, blood, urine, seminal plasma, and silkworm ...

  11. Sulfur formation and recovery in a thiosulfateoxidizing bioreactor

    NARCIS (Netherlands)

    Gonzalez-Sanchez, A.; Meulepas, R.J.W.; Revah, S.

    2008-01-01

    This work describes the design and Performance of a thiosulfate-oxidizing bioreactor that allowed high elemental sulfur production and recovery efficiency. The reactor system, referred to as a Supernatant-Recycling Settler Bioreactor (SRSB), consisted of a cylindrical upflow reactor and a separate a

  12. Evaluation of woodchip bioreactors for improved water quality

    Science.gov (United States)

    Woodchip bioreactors are gaining popularity with farmers because of their edge-of-field nitrate removal capabilities, which do not require changes in land management practices. However, limited research has been conducted to study the potential of these bioreactors to also reduce downstream transpor...

  13. Production of Validamycins from Crude Substrates by Streptomyces hygroscopicus in an External-loop Airlift Bioreactor with a Low Height-to-Diameter Ratio%低高径比外循环气升式生物反应器带渣发酵生产有效霉素

    Institute of Scientific and Technical Information of China (English)

    郑裕国; 陈小龙; 汪钊; 沈寅初

    2004-01-01

    Fermentation experiments to produce validamycins from crude substrates by Streptomyces hygroscopicus were carried out in an external-loop airlift bioreactor (0.0115 m3 ) with a low ratio of height to diameter of the riser of 2.9 and a ratio of riser to downcomer diameter of 6.6. The influences of gas flow rate and liquid volume on fermentation of validamycins were investigated. Comparisons of validamycin fermentation were made among the external-loop airlift bioreactor, a mechanically stirred tank bioreactor (0.010m3 ) and shaking flasks. Under the same operation conditions including fermentation medium composition, inoculum ratio and culture temperature,the fermentation time in the external-loop airlift bioreactor (45 h) was shorter than that in the shaking flasks (100 h)and the same as that in the mechanically stirred tank bioreactor. After a total fermentation time of 45 h under optimized operation conditions, average validamycin concentration obtained in the external-loop airlift bioreactor was close to 19630μg.m1-1 validamycin concentration in the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the mechanically stirred tank bioreactor in production of validamycins from crude substrates with dregs by Streptomyces hygroscopicus.

  14. First use of the WAVE™ disposable rocking bioreactor for enhanced bioproduct synthesis by N2 -fixing cyanobacteria.

    Science.gov (United States)

    Cirés, Samuel; Alvarez-Roa, Carlos; Heimann, Kirsten

    2015-03-01

    WAVE™ rocking disposable bioreactors have been successfully utilized for bioproduct development from bacteria, yeast, microalgae, and animal and plant cells but not from cyanobacteria so far. N2 -fixing cyanobacteria represent a prolific bioproducts source with reduced cultivation costs. In this study, 1 L cultures of the N2 -fixing cyanobacterium Anabaena siamensis grown diazotrophically in the WAVE™ bioreactor exhibited increased phosphate consumption and 37-70% higher CO2 fixation rates than those grown in conventional bubbled suspension (BS) batch cultures. This generated 40-80% increased biomass productivities in the WAVE™ bioreactor reaching 60 mg L(-1)  day(-1) when supplemented with 10% CO2 . Consequently, WAVE™ generated 36-153% more protein, lipid, and carbohydrate than BS, including 47-100% increased productivity of phycocyanin and stearidonic acid (SA) with relevant biomedical applications. While the type of culture system (BS or WAVE(TM) ) did not affect the biochemical profile of cyanobacterial biomass, 10% CO2 supplementation induced a significant decrease in fatty acids and phycocyanin contents (mg g(-1)  DW). Therefore, for commercial applications, the CO2 supplementation of WAVE™ should be optimized for each targeted bioproduct separately. This study opens possibilities for upgrading the WAVE™ systems to photobioreactors (PBRs) for bioproduct development from cyanobacteria, with opportunities and challenges critically evaluated herein.

  15. STATE OF THE PRACTICE FOR BIOREACTOR LANDFILLS - SUMMARY OF USEPA WORKSHOP ON BIOREACTOR LANDFILLS: SUMMARY

    Science.gov (United States)

    This is a summary of the Workshop on Landfill Bioreactors, held 9/6-7/2000 in Arlington, VA. The purpose of the workshop was to provide a forum to EPA, state and local governments, solid waste industry, and academic research representatives to exchange information and ideas on b...

  16. Wastewater treatments by membrane bioreactors (MBR); Bioreactores de membrana (MBR) para la depuracion de aguas residuales

    Energy Technology Data Exchange (ETDEWEB)

    Guardino Ferre, R.

    2001-07-01

    Wastewater treatments by membrane bioreactors (MBR), are a good alternative of treatment to the conventional processes when wish to obtain very high quality of the treated water or to try high load contaminants in low flow. Simultaneously, the article explains the significant reduction of the wastewater treatment plant space, eliminating the secondary septic tank. (Author) 7 refs.

  17. Seasonal growth variation of peach palms cultivated in containers under subtropical conditions Variação estacional do crescimento em pupunheiras cultivadas em recipientes em condição subtropical

    Directory of Open Access Journals (Sweden)

    Maria Luiza Sant'Anna Tucci

    2007-01-01

    Full Text Available Peach palm (Bactris gasipaes Kunth is grown in the São Paulo State, Brazil, under climate seasonal variation conditions, mainly temperature and rainfal with possible effects on plant physiology. Recently, due to a higher interest in carrying out physiological experiments on the species, there has been a requirement for more controlled experimental conditions. Therefore, with the aim of studying the seasonal variation of peach palm growth for heart-of-palm production, as well as the possibility of growing them until harvest in pots, for future utilization in physiological experiments, this work was carried out in Campinas, SP, Brazil, with 40 spineless peach palms. One year after seed germination, seedlings were transplanted to 80 L plastic pots, spaced 2 x 1 m, arranged in four rows of ten plants. All plants had vegetative growth evaluated monthly by measurements of main stem height, number of functional leaves, number of offshoots and length of leaf raquis. Seasonal variations were observed in terms of height and diameter growth as well as raquis length of the youngest leaf and in the evolution of the number of leaves. After two years, plants had an average height of 230 cm, six functional leaves and 11.7 offshoots. Positive correlations (P A pupunheira (Bactris gasipaes Kunth é cultivada no Estado de São Paulo, Brasil, sob condições de variação estacional do clima, particularmente temperatura e precipitação pluviométrica, com possíveis efeitos na fisiologia das plantas. Recentemente, devido ao crescente interesse em se realizar experimentos sobre a fisiologia da espécie, tem havido necessidade de cultivá-la sob condições experimentais mais controladas. Com o objetivo de estudar a variação estacional do crescimento de pupunheiras, bem como a possibilidade de cultivá-las em recipientes até a colheita, para a realização de futuros experimentos fisiológicos, foi executado este trabalho, em Campinas, SP, com 40 pupunheiras

  18. Continuous anaerobic bioreactor with a fixed-structure bed (ABFSB) for wastewater treatment with low solids and low applied organic loading content.

    Science.gov (United States)

    Mockaitis, G; Pantoja, J L R; Rodrigues, J A D; Foresti, E; Zaiat, M

    2014-07-01

    This paper describes a new type of anaerobic bioreactor with a fixed-structure bed (ABFSB) in which the support for the biomass consists of polyurethane foam strips placed along the length of the bioreactor. This configuration prevents the accumulation of biomass or solids in the bed as well as clogging and channeling effects. In this study, complex synthetic wastewater with a chemical oxygen demand of 404.4 mg O(2) L(-1) is treated by the reactor. The ABFSB, which has a working volume of 4.77 L, was inoculated with anaerobic sludge obtained from an upflow anaerobic sludge blanket bioreactor. A removal efficiency of 78 % for organic matter and an effluent pH of 6.97 were achieved. An analysis of the organic volatile acids produced by the ABFSB indicated that it operated under stable conditions during an experimental run of 36 days. The stable and efficient operation of the bioreactor was compared with the configurations of other anaerobic bioreactors used for complex wastewater treatment. The results of the study indicate that the ABFSB is a technological alternative to packed-bed bioreactors.

  19. Disposable bioreactors for inoculum production and protein expression.

    Science.gov (United States)

    Eibl, Regine; Löffelholz, Christian; Eibl, Dieter

    2014-01-01

    Disposable bioreactors have been increasingly implemented over the past ten years. This relates to both R & D and commercial manufacture, in particular, in animal cell-based processes. Among the numerous disposable bioreactors which are available today, wave-mixed bag bioreactors and stirred bioreactors are predominant. Whereas wave-mixed bag bioreactors represent the system of choice for inoculum production, stirred systems are often preferred for protein expression. For this reason, the authors present protocols instructing the reader how to use the wave-mixed BIOSTAT CultiBag RM 20 L for inoculum production and the stirred UniVessel SU 2 L for recombinant protein production at benchtop scale. All methods described are based on a Chinese hamster ovary (CHO) suspension cell line expressing the human placental secreted alkaline phosphatase (SEAP).

  20. H2S removal from biogas using bioreactors: a review

    Directory of Open Access Journals (Sweden)

    E. Dumont

    2015-01-01

    Full Text Available This review aims to provide an overview of the bioprocesses used for the removal of H2S from biogas. The ability of aerobic and anoxic bioreactors (biotrickling filters, bioscrubbers, and a combination of chemical scrubbers and bioreactors to perform the degradation of H2S is considered. For each operating mode (aerobic and anoxic, the bioprocesses are presented, the operating conditions affecting performance are summarized, the state of the art of research studies is described and commercial applications are given. At laboratory-scale, whatever their operating mode, biological processes are effective for biogas cleaning and provide the same performance. The clogging of the packed bed due to the deposit of elemental sulfur S0 and biomass accumulation clearly represents the main drawback of bioprocesses. Although elimination capacities (EC determined at laboratory-scale can be very high, EC should not be higher than 90 g m-3 h-1 at industrial-scale in order to limit clogging effects. For aerobic processes, the need to control the oxygen mass transfer accurately remains a key issue for their development at full-scale. As a result, the aerobic processes alone are probably not the most suitable bioprocesses for the treatment of biogas highly loaded with H2S. For anaerobic bioprocesses using nitrate as an electron acceptor, the scale-up of the laboratory process to a full-size plant remains a challenge. However, the use of wastewater from treatment plants, which constitutes a cheap source of nitrates, represents an interesting opportunity for the development of innovative bioprocesses enabling the simultaneous removal of H2S and nitrates.

  1. L-乳酸调控乳酸产生菌产物光学纯度的分析%Effects of cultivation conditions on the optical purity of L(+)-lactic acid

    Institute of Scientific and Technical Information of China (English)

    孟武; 李十中; 封文涛; 张晗星; 王瑞明

    2009-01-01

    发酵初期在米根霉菌发酵培养基中添加L-乳酸可以调控发酵产物乳酸的光学纯度.随着L-乳酸添加量的增加,所产L-乳酸的光学纯度随之增加,当L-乳酸的添加量≥1.5 g/L时,D-乳酸不再产生.同时,L-乳酸的产量、生物量、糖转化率也随之降低.该调控方法对乳酸菌调控产L-乳酸光学纯度影响不大,对大肠杆菌发酵调控产D-乳酸光学纯度没有效果.%The effect of cultivation conditions on the optical purity of L(+)-lactic acid produced by Rhizopus oryzae HZS6 was investigated.The isomeric composition of lactic acid was influenced by the supplementation of L(+)-lactic acid to fermentation medium.L(+)-isomer increased with the dosage, no D(-)-lactic acid was observed when the concentration of supplemented L(+)-lactic acid in matrix was ≥ 1.5 g/L.However, the L(+)-lactic acid yield, biomass and glucose conversion rate decreased with the dosage.With the same method, the supplementation of L(+)-lactic to substrate had no influence on isomeric composition of lactic acid by Lactobacillus and Escherichia coli.

  2. Mathematical modelling and optimization of hydrogen continuous production in a fixed bed bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Palazzi, E.; Perego, P.; Fabiano, B. [University of Genoa, Genova (Italy). Chemical and Process Engineering Department ' G.B. Bonino'

    2002-09-01

    The purpose of this paper is to investigate, both theoretically and experimentally, hydrogen production from agro-industrial by-products using a continuous bioreactor packed with a mixture of spongy and glass beads and inoculated with Enterobacter aerogenes. Replicated series of experimental runs were performed to study the effects of residence time on hydrogen evolution rate and to characterize the critical conditions for the wash out, as a function of the inlet glucose concentration and of the fluid superficial velocity. A further series of experimental runs was focused on the effects of both residence time and inlet glucose concentration over hydrogen productivity. A kinetic model of the process was developed and showed good agreement with experimental data, thus representing a potential tool to design a large-scale fermenter. In fact, the model was applied to the optimal design of a bioreactor suitable of feeding a phosphoric acid fuel cell of a target power. (author)

  3. Determination of Ammonia Oxidizing Bacteria and Nitrate Oxidizing Bacteria in Wastewater and Bioreactors

    Science.gov (United States)

    Francis, Somilez Asya

    2014-01-01

    The process of water purification has many different physical, chemical, and biological processes. One part of the biological process is the task of ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB). Both play critical roles in the treatment of wastewater by oxidizing toxic compounds. The broad term is nitrification, a naturally occurring process that is carried out by AOB and NOB by using oxidation to convert ammonia to nitrite and nitrite to nitrate. To monitor this biological activity, bacterial staining was performed on wastewater contained in inoculum tanks and biofilm samples from bioreactors. Using microscopy and qPCR, the purpose of this experiment was to determine if the population of AOB and NOB in wastewater and membrane bioreactors changed depending on temperature and hibernation conditions to determine the optimal parameters for AOB/NOB culture to effectively clean wastewater.

  4. Dynamic fouling behaviors of submerged nonwoven bioreactor for filtration of activated sludge with different SRT.

    Science.gov (United States)

    Chuang, Shun-Hsing; Lin, Po-Kuen; Chang, Wei-Chin

    2011-09-01

    The flux variations and resistances accumulated during filtration of activated sludge with sludge retention time (SRT) of 15, 30, and 60 days were analyzed to investigate the dynamic fouling behavior in a submerged nonwoven bioreactor. Different SRT values varied sludge condition and particle size distribution in the supernatants, which caused dissimilar fouling characteristics. Short-term fouling of the nonwoven bioreactor during filtration of activated sludge with SRT of 15 days was fully reversible, and the resistance percentages of solutes, colloids, and suspended solids were 6%, 27%, and 67%, respectively. On the other hand, significant increases of colloid resistance, such as with the filtration of activated sludge with SRT of 30 and 60 days, were related to the occurrence of irreversible fouling. The phenomenon of pore blocking by particles or colloids with size analogous to the pore of nonwoven fabric was a decisive factor leading to irreversible fouling in the large-pore materials.

  5. [Cultivation of pathogenic free-living amoebae].

    Science.gov (United States)

    Peng, Heng; Zhu, Huai-Min

    2009-08-01

    The isolation and culture of pathogenic free-living amoebae are useful in the diagnosis and research. This review focuses on the methods of isolation and cultivation of pathogenic free-living amoebae, including sample treatment, culture conditions, passage culture, pathogen detection, and maintenance.

  6. Experimental investigation on feasible bioreactor using mechanism of hydrogen oxidation of natural soil for detritiation system.

    Science.gov (United States)

    Edao, Yuki; Iwai, Yasunori; Sato, Katsumi; Hayashi, Takumi

    2016-08-01

    A passive reactor for tritium oxidation at room temperature has been widely studied in nuclear engineering especially for a detritiation system (DS) of a tritium process facility taking possible extraordinary situation severely into consideration. We have focused on bacterial oxidation of tritium by hydrogen-oxidizing bacteria in natural soil to realize the passive oxidation reactor. The purpose of this study was to examine the feasibility of a bioreactor with hydrogen-oxidizing bacteria in soil from a point of view of engineering. The efficiency of the bioreactor was evaluated by kinetics. The bioreactor packed with natural soil shows a relative high conversion rate of tritium under the saturated moisture condition at room temperature, which is obviously superior to that of a Pt/Al2O3 catalyst generally used for tritium oxidation in the existing tritium handling facilities. The order of reaction for tritium oxidation with soil was the pseudo-first order as assessed with Michaelis-Menten kinetics model. Our engineering suggestion to increase the reaction rate is the intentional addition of hydrogen at a small concentration in the feed gas on condition that the oxidation of tritium with soil is expressed by the Michaelis-Menten kinetics model.

  7. Componentes da radiação solar em cultivo de tomate sob condições de ambiente protegido Components of the solar radiation in tomato cultivated under greenhouse conditions

    Directory of Open Access Journals (Sweden)

    Ligia S. Reis

    2012-07-01

    Full Text Available Objetivou-se, com este trabalho, avaliar o saldo de radiação e a irradiação solar e fotossintética em condições de ambiente protegido cultivado com o tomateiro e suas relações com a irradiação solar do ambiente externo. O tomateiro foi cultivado em casa de vegetação não climatizada, com cobertura de polietileno de 0,12 mm de espessura. A irradiação solar (Rgi, o saldo de radiação (Rni e a densidade de fluxo de fótons fotossintéticos foram obtidos por radiômetros ligados a um datalogger instalado no interior do ambiente protegido. Os dados externos (irradiância solar global, Rg foram coletados na Estação Agrometeorológica do Centro de Ciências Agrárias da Universidade Federal de Alagoas. O ambiente protegido promoveu redução na irradiação solar expressa como a transmitância do polietileno em 62%. As relações entre os componentes de radiação do ambiente interno e externo foram expressas satisfatoriamente por regressões lineares, com coeficientes de determinação (R² superiores a 0,88 enquanto para a irradiação fotossintética (PARi os R² foram maiores que 0,52. As relações obtidas mostraram que Rni representa 0,60 da Rgi e a PARi é proporcional a 0,28 da Rg e 0,44 da Rgi. O albedo médio da cultura durante o ciclo foi de 0,15.The objective of this paper was to evaluate of net radiation, photosynthetic and solar irradiation in greenhouse conditions cultivated with tomato crop and its relationship with the global solar irradiation of the external environment. The tomato was cultivated in greenhouse (not acclimatized, with covering of polyethylene 0.12 mm of thickness. The global solar irradiation (Rgi, the net radiation (Rni and the flux density of photosynthetic photons were obtained by radiometers connected to a datalogger installed into the protected environment. The external data (global solar irradiation, Rg were collected in the Agrometeorological Station of the Agricultural Science Center of

  8. Desarrollo vegetativo de patrones cítricos cultivados en condiciones de invernadero bajo dos sistemas de riego Vegetative development of citrus seedlings cultivated at greenhouse conditions and submitted to two irrigations systems

    Directory of Open Access Journals (Sweden)

    Gilmar Schäfer

    2006-08-01

    Full Text Available En el presente estudio se evaluó el desarrollo vegetativo de patrones cítricos cultivados en invernadero bajo dos sistemas de riego. El experimento se realizó en la Estação Experimental Agronômica de la Universidade Federal do Rio Grande do Sul, ubicada en Eldorado do Sul, Rio Grande do Sul, Brasil, entre los meses de septiembre de 2003 y abril de 2004, totalizando 225 días de experimentación. El diseño experimental fue de parcelas subdivididas, en factorial 2 x 3, con 4 repeticiones de 22 contenedores cada. En las parcelas principales se evaluaron los sistemas de riego (microaspersión y capilaridad y en las subparcelas los patrones cítricos Poncirus trifoliata (L. Raf., citrangero 'C37' [P. trifoliata x Citrus sinensis (L. Osb. cv. Pêra] y lima 'Rangpur' (C. limonia Osb.. En condiciones de invernadero los patrones cítricos presentan un desarrollo vegetativo más rápido bajo riego por capilaridad respecto a la microaspersión. Los patrones cítricos evaluados presentan desarrollos vegetativos distintos, donde el citrangero 'C37' supera a los demás.The aim of the present work was to evaluate the vegetative development of citrus rootstock seedlings cultivated under greenhouse conditions with two irrigation systems. The experiment was conducted at the Estação Experimental Agronômica , Universidade Federal do Rio Grande do Sul, located in Eldorado do Sul, Rio Grande do Sul, Brazil, from September 2003 to April 2004, totalizing 225 days of experimentation. The experimental design was a split-plot, in a 2x3 factorial, with 4 replications of 22 pots each. In the main plot the irrigation systems was evaluated (micro sprinkler and capillarity and in the split-plot the citrus rootstocks [Trifoliate orange - Poncirus trifoliata (L. Raf., 'C37' citrange - P. trifoliata x Citrus sinensis (L. Osb. cv. Pêra and 'Rangpur' lime - C. limonia Osb.] were evaluated. The main result showed in conditions of greenhouse citrus rootstock seedlings

  9. Growth of oleaginous Rhodotorula glutinis in an internal-loop airlift bioreactor by using lignocellulosic biomass hydrolysate as the carbon source.

    Science.gov (United States)

    Yen, Hong-Wei; Chang, Jung-Tzu

    2015-05-01

    The conversion of abundant lignocellulosic biomass (LCB) to valuable compounds has become a very attractive idea recently. This study successfully used LCB (rice straw) hydrolysate as a carbon source for the cultivation of oleaginous yeast-Rhodotorula glutinis in an airlift bioreactor. The lipid content of 34.3 ± 0.6% was obtained in an airlift batch with 60 g reducing sugars/L of LCB hydrolysate at a 2 vvm aeration rate. While using LCB hydrolysate as the carbon source, oleic acid (C18:1) and linoleic acid (C18:2) were the predominant fatty acids of the microbial lipids. Using LCB hydrolysate in the airlift bioreactor at 2 vvm achieved the highest cell mass growth as compared to the agitation tank. Despite the low lipid content of the batch using LCB hydrolysate, this low cost feedstock has the potential of being adopted for the production of β-carotene instead of lipid accumulation in the airlift bioreactor for the cultivation of R. glutinis.

  10. Production of manganese peroxidase and laccase in a solid-state bioreactor and modeling of enzyme production kinetics.

    Science.gov (United States)

    Moilanen, Ulla; Winquist, Erika; Mattila, Tuomas; Hatakka, Annele; Eerikäinen, Tero

    2015-01-01

    Lignin-modifying enzymes have various promising applications such as biobleaching, biopulping, the functionalization of lignocellulosic materials, the modification of wood fibers, the remediation of contaminated soil and effluents, as well as improvement of the enzymatic hydrolysis of lignocellulosic substrates. In this study, the production of laccase and manganese peroxidase (MnP) in solid-state cultivation was examined. Oat husks were used as an inexpensive substrate for the white-rot fungus Cerrena unicolor PM170798 (FBCC 387). The addition of a fines fraction (consisting of oat flour and finely ground husks) enhanced MnP production fivefold and laccase production almost threefold. The enzyme production was studied first on a 100 g scale, and the cultivation experiments were then repeated at a larger laboratory-scale (4 kg) in a solid-state bioreactor. High enzyme activity levels were obtained (MnP: 340 nkat g(-1) DM, laccase: 470 nkat g(-1) DM). In addition, the correlation between the CO2 evolution rate and enzyme production was mathematically modeled from the bioreactor experimental data. The model parameters could be used to predict enzyme production.

  11. Recombinant Escherichia coli strains with inducible Campylobacter jejuni single domain hemoglobin CHb expression exhibited improved cell growth in bioreactor culture.

    Directory of Open Access Journals (Sweden)

    Li Xu

    Full Text Available Maintaining an appropriate concentration of dissolved oxygen in aqueous solution is critical for efficient operation of a bioreactor, requiring sophisticated engineering design and a system of regulation to maximize oxygen transfer from the injected air bubbles to the cells. Bacterial hemoglobins are oxygen-binding proteins that transfer oxygen from the environment to metabolic processes and allow bacteria to grow even under microaerophilic conditions. To improve the oxygen utilization efficiency of cells and overcome the oxygen shortage in bioreactors, the gene coding for the Campylobacter jejuni single domain hemoglobin (CHb gene was artificially synthesized and functionally expressed under the control of inducible expression promoters PT7 and Pvgh in Escherichia coli. The effects of the recombinants PT7-CHb and Pvgh-CHb on cell growth were evaluated in aerobic shake flasks, anaerobic capped bottles and a 5-L bioreactor, and a pronounced improvement in cell biomass was observed for CHb-expressing cells. To determine the growth curves, CHb gene expression, and CHb oxygen-binding capacity of specific recombinants with different promoters, we determined the time course of CHb gene expression in the two recombinants by semi-quantitative RT-PCR and CO differential spectrum assays. Based on the growth patterns of the two recombinants in the bioreactor, we proposed different recombinant types with optimal performance under specific culture conditions.

  12. Prospects for Sorghum cultivation in Poland

    Directory of Open Access Journals (Sweden)

    Roman Prażak

    2016-06-01

    Full Text Available The article presents the origin and cultivation history of sorghum (Sorghum spp., its biology, requirements, cultivation techniques, and utilization. Sorghum is a cereal of the Poaceae. It is one of the most important crop plants grown in warmer parts of the world. Sorghum comes from Africa and therefore has very high heat requirements. In comparison with other crop plants, it is characterized by more efficient nutrient and water utilization. Sorghum grain is used to produce porridge, flour, syrup, sugar, ethanol, vegetable oil, starch, wax, paints, and animal fodder (the grain and entire plant. Sorghum straw is used to produce fibres, paper, and building materials. Sorghum has high energy value and can be an excellent source of renewable energy. It is easy to cultivate, with low soil and nutrient requirements. Due to its content of allelopathic compounds, it inhibits weed growth and has a phytosanitary effect. It is also resistant to disease and pests. It is a short-day plant, and in Polish climate conditions, it does not form sufficiently mature seeds, but produces a very high yield of green matter that can be used for fodder. Cultivation of sorghum during periodic water shortages may be an alternative solution for obtaining fodder when maize cultivation is unreliable.

  13. [Dendrobium officinale stereoscopic cultivation method].

    Science.gov (United States)

    Si, Jin-Ping; Dong, Hong-Xiu; Liao, Xin-Yan; Zhu, Yu-Qiu; Li, Hui

    2014-12-01

    The study is aimed to make the most of available space of Dendrobium officinale cultivation facility, reveal the yield and functional components variation of stereoscopic cultivated D. officinale, and improve quality, yield and efficiency. The agronomic traits and yield variation of stereoscopic cultivated D. officinale were studied by operating field experiment. The content of polysaccharide and extractum were determined by using phenol-sulfuric acid method and 2010 edition of "Chinese Pharmacopoeia" Appendix X A. The results showed that the land utilization of stereoscopic cultivated D. officinale increased 2.74 times, the stems, leaves and their total fresh or dry weight in unit area of stereoscopic cultivated D. officinale were all heavier than those of the ground cultivated ones. There was no significant difference in polysaccharide content between stereoscopic cultivation and ground cultivation. But the extractum content and total content of polysaccharide and extractum were significantly higher than those of the ground cultivated ones. In additional, the polysaccharide content and total content of polysaccharide and extractum from the top two levels of stereoscopic culture matrix were significantly higher than that of the ones from the other levels and ground cultivation. Steroscopic cultivation can effectively improves the utilization of space and yield, while the total content of polysaccharides and extractum were significantly higher than that of the ground cultivated ones. The significant difference in Dendrobium polysaccharides among the plants from different height of stereo- scopic culture matrix may be associated with light factor.

  14. Disposable polymeric cryogel bioreactor matrix for therapeutic protein production.

    Science.gov (United States)

    Jain, Era; Kumar, Ashok

    2013-05-01

    Low cost and high efficiency make disposable bioreactors feasible for small-scale therapeutic development and initial clinical trials. We have developed a cryogel-based disposable bioreactor matrix, which has been used for production of protein therapeutics such as urokinase and monoclonal antibodies (mAbs). The protocol discusses the application of a cryogel bioreactor for mAb production. Cryogels composed of either polyacrylamide (PAAm) coupled to gelatin or semi-interpenetrating PAAm-chitosan are synthesized by free-radical polymerization at -12 °C. Hybridoma cells are immobilized over the cryogel bioreactor and incubated for 48 h. Medium is circulated thereafter at 0.2 ml min(-1) and bioreactors can be run continuously for 60 d. The cryogel-based packed-bed bioreactor can be formulated as a monolith or as beads; it also has an efficiency four times what can be obtained using a tissue-culture flask, a high surface-to-volume ratio and effective nutrient transport. After incubation, the bioreactor setup will take about 60 min using a pre-prepared sterilized cryogel.

  15. Disposable Bioreactors for Plant Micropropagation and Mass Plant Cell Culture

    Science.gov (United States)

    Ducos, Jean-Paul; Terrier, Bénédicte; Courtois, Didier

    Different types of bioreactors are used at Nestlé R&D Centre - Tours for mass propagation of selected plant varieties by somatic embryogenesis and for large scale culture of plants cells to produce metabolites or recombinant proteins. Recent studies have been directed to cut down the production costs of these two processes by developing disposable cell culture systems. Vegetative propagation of elite plant varieties is achieved through somatic embryogenesis in liquid medium. A pilot scale process has recently been set up for the industrial propagation of Coffea canephora (Robusta coffee). The current production capacity is 3.0 million embryos per year. The pre-germination of the embryos was previously conducted by temporary immersion in liquid medium in 10-L glass bioreactors. An improved process has been developed using a 10-L disposable bioreactor consisting of a bag containing a rigid plastic box ('Box-in-Bag' bioreactor), insuring, amongst other advantages, a higher light transmittance to the biomass due to its horizontal design. For large scale cell culture, two novel flexible plastic-based disposable bioreactors have been developed from 10 to 100 L working volumes, validated with several plant species ('Wave and Undertow' and 'Slug Bubble' bioreactors). The advantages and the limits of these new types of bioreactor are discussed, based mainly on our own experience on coffee somatic embryogenesis and mass cell culture of soya and tobacco.

  16. Dynamic global sensitivity analysis in bioreactor networks for bioethanol production.

    Science.gov (United States)

    Ochoa, M P; Estrada, V; Di Maggio, J; Hoch, P M

    2016-01-01

    Dynamic global sensitivity analysis (GSA) was performed for three different dynamic bioreactor models of increasing complexity: a fermenter for bioethanol production, a bioreactors network, where two types of bioreactors were considered: aerobic for biomass production and anaerobic for bioethanol production and a co-fermenter bioreactor, to identify the parameters that most contribute to uncertainty in model outputs. Sobol's method was used to calculate time profiles for sensitivity indices. Numerical results have shown the time-variant influence of uncertain parameters on model variables. Most influential model parameters have been determined. For the model of the bioethanol fermenter, μmax (maximum growth rate) and Ks (half-saturation constant) are the parameters with largest contribution to model variables uncertainty; in the bioreactors network, the most influential parameter is μmax,1 (maximum growth rate in bioreactor 1); whereas λ (glucose-to-total sugars concentration ratio in the feed) is the most influential parameter over all model variables in the co-fermentation bioreactor.

  17. Long term cultivation of larger benthic Foraminifera

    Science.gov (United States)

    Wöger, Julia; Eder, Wolfgang; Kinoshita, Shunichi; Antonino, Briguglio; Carles, Ferrandes-Cañadell; Hohenegger, Johann

    2015-04-01

    Benthic Foraminifera are used in a variety of applications employing numerous different methods, i.e. ecological monitoring, studying the effects of ocean acidification, reconstructing palaeo-bathymetry or investigating palaeo-salinity and palaeo-temperature to name only a few. To refine our understanding of ecological influences on larger benthic foraminiferal biology and to review inferences from field observations, culture experiments have become an indispensable tool. While culture experiments on smaller benthic foraminifera have become increasingly frequent in the past century, reports of the cultivation of symbiont bearing larger Foraminifera are rare. Generally, cultivation experiments can be divided into two groups: Culturing of populations and cultivation of single specimens allowing individual investigation. The latter differ form the former by several restrictions resulting from the need to limit individual motility without abridging microenvironmental conditions in the Foraminiferans artificial habitat, necessary to enable the individual to development as unfettered as possible. In this study we present first experiences and preliminary results of the long-term cultivation of larger benthic Foraminifera conducted at the 'Tropical Biosphere Research Station Sesoko Island, University of the Ryukyus', Japan, trying to reproduce natural conditions as closely as possible. Individuals of three species of larger benthic Foraminifera (Heterostegina depressa, Palaeonummulites venosus and Operculina complanata) have been cultured since April 2014. At the time of the general assembly the cultivation experiments will have been going on for more than one year, with the aim to investigate growth rates, longevities and reproduction strategies for comparison with results statistically inferred from application of the of the 'natural laboratory' method. The most important factor influencing foraminiferal health and development was found to be light intensity and light

  18. MEMBRANE BIOREACTOR FOR TREATMENT OF RECALCITRANT WASTEWATERS

    Directory of Open Access Journals (Sweden)

    Suprihatin Suprihatin

    2012-02-01

    Full Text Available The low biodegradable wastewaters remain a challenge in wastewater treatment technology. The performance of membrane bioreactor systems with submerged hollow fiber micro- and ultrafiltration membrane modules were examined for purifying recalcitrant wastewaters of leachate of a municipal solid waste open dumping site and effluent of pulp and paper mill. The use of MF and UF membrane bioreactor systems showed an efficient treatment for both types wastewaters with COD reduction of 80-90%. The membrane process achieved the desirable effects of maintaining reasonably high biomass concentration and long sludge retention time, while producing a colloid or particle free effluent. For pulp and paper mill effluent a specific sludge production of 0.11 kg MLSS/kg COD removed was achieved. A permeate flux of about 5 L/m²h could be achieved with the submerged microfiltration membrane. Experiments using ultrafiltration membrane produced relatively low permeate fluxes of 2 L/m²h. By applying periodical backwash, the flux could be improved significantly. It was indicated that the particle or colloid deposition on membrane surface was suppressed by backwash, but reformation of deposit was not effectively be prevented by shear-rate effect of aeration. Particle and colloid started to accumulate soon after backwash. Construction of membrane module and operation mode played a critical role in achieving the effectiveness of aeration in minimizing deposit formation on the membrane surface.

  19. Landfill leachate treatment in assisted landfill bioreactor

    Institute of Scientific and Technical Information of China (English)

    HE Pin-jing; QU Xian; SHAO Li-ming; LEE Duu-jong

    2006-01-01

    Landfill is the major disposal route of municipal solid waste(MSW) in most Asian countries. Leachate from landfill presents a strong wastewater that needs intensive treatment before discharge. Direct recycling was proposed as an effective alternative for leachate treatment by taking the landfill as a bioreactor. This process was proved not only considerably reducing the pollution potential of leachate, but also enhancing organic degradation in the landfill. However, as this paper shows, although direct leachate recycling was effective in landfilled MSW with low food waste fraction (3.5%, w/w), it failed in MSW containing 54% food waste, as normally noted in Asian countries. The initial acid stuck would inhibit methanogenesis to build up, hence strong leachate was yielded from landfill to threaten the quality of receiving water body. We demonstrated the feasibility to use an assisted bioreactor landfill, with a well-decomposed refuse layer as ex-situ anaerobic digester to reducing COD loading in leachate. By doing so, the refuse in simulated landfill column (2.3 m high) could be stabilized in 30 weeks while the COD in leachate reduced by 95%(61000 mg/L to 3000 mg/L). Meanwhile, the biogas production was considerably enhanced, signaling by the much greater amount and much higher methane content in the biogas.

  20. Novel Hydrogen Bioreactor and Detection Apparatus.

    Science.gov (United States)

    Rollin, Joseph A; Ye, Xinhao; Del Campo, Julia Martin; Adams, Michael W W; Zhang, Y-H Percival

    2016-01-01

    In vitro hydrogen generation represents a clear opportunity for novel bioreactor and system design. Hydrogen, already a globally important commodity chemical, has the potential to become the dominant transportation fuel of the future. Technologies such as in vitro synthetic pathway biotransformation (SyPaB)-the use of more than 10 purified enzymes to catalyze unnatural catabolic pathways-enable the storage of hydrogen in the form of carbohydrates. Biohydrogen production from local carbohydrate resources offers a solution to the most pressing challenges to vehicular and bioenergy uses: small-size distributed production, minimization of CO2 emissions, and potential low cost, driven by high yield and volumetric productivity. In this study, we introduce a novel bioreactor that provides the oxygen-free gas phase necessary for enzymatic hydrogen generation while regulating temperature and reactor volume. A variety of techniques are currently used for laboratory detection of biohydrogen, but the most information is provided by a continuous low-cost hydrogen sensor. Most such systems currently use electrolysis for calibration; here an alternative method, flow calibration, is introduced. This system is further demonstrated here with the conversion of glucose to hydrogen at a high rate, and the production of hydrogen from glucose 6-phosphate at a greatly increased reaction rate, 157 mmol/L/h at 60 °C.