Search results for: stirred airlift bioreactor

Authors: Prangya Ranjan Rout, Puspendu Bhunia, Rajesh Roshan Dash

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In a developing country like India, providing reliable and affordable wastewater treatment facilities in rural areas is a huge challenge. With the aim of enhancing the nutrient removal from rural domestic wastewater while reducing the cost of treatment process, a novel, integrated treatment system consisting of a multistage bio-filter with drop aeration and a post positioned attached growth carbonaceous denitrifying-bioreactor was designed and developed in this work. The bio-filter was packed with ‘dolochar’, a sponge iron industry waste, as an adsorbent mainly for phosphate removal through physiochemical approach. The Denitrifying bio-reactor was packed with many waste organic solid substances (WOSS) as carbon sources and substrates for biomass attachment, mainly to remove nitrate in biological denitrification process. The performance of the modular system, treating real domestic wastewater was monitored for a period of about 60 days and the average removal efficiencies during the period were as follows: phosphate, 97.37%; nitrate, 85.91%, ammonia, 87.85%, with mean final effluent concentration of 0.73, 9.86, and 9.46 mg/L, respectively. The multistage bio-filter played an important role in ammonium oxidation and phosphate adsorption. The multilevel drop aeration with increasing oxygenation, and the special media used, consisting of certain oxides were likely beneficial for nitrification and phosphorus removal, respectively, whereas the nitrate was effectively reduced by biological denitrification in the carbonaceous bioreactor. This treatment system would allow multipurpose reuse of the final effluent. Moreover, the saturated dolochar can be used as nutrient suppliers in agricultural practices and the partially degraded carbonaceous substances can be subjected to composting, and subsequently used as an organic fertilizer. Thus, the system displays immense potential for treating domestic wastewater significantly decreasing the concentrations of nutrients and more importantly, facilitating the conversion of the waste materials into usable ones.

Keywords: nutrient removal, denitrifying bioreactor, multi-stage bio-filter, dolochar, waste organic solid substances

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Authors: Karan Bhandari

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Almost quarter region of Indian terrain is covered by grasslands. Grass being a low maintenance perennial crop is in abundance. Farmers are well acquainted with its nature, yield and storage. The aim of this paper is to study and identify the applicability of grass as a source of bio fuel. Anaerobic break down is a well-recognized technology. This process is vital for harnessing bio fuel from grass. Grass is a lignocellulosic material which is fibrous and can readily cause problems with parts in motion. Further, it also has a tendency to float. This paper also deals with the ideal digester configuration for biogas generation from grass. Intensive analysis of the literature is studied on the optimum production of grass storage in accordance with bio digester specifications. Subsequent to this two different digester systems were designed, fabricated, analyzed. The first setup was a double stage wet continuous arrangement usually known as a Continuously Stirred Tank Reactor (CSTR). The next was a double stage, double phase system implementing Sequentially Fed Leach Beds using an Upflow Anaerobic Sludge Blanket (SLBR-UASB). The above methodologies were carried for the same feedstock acquired from the same field. Examination of grass silage was undertaken using Biomethane Potential values. The outcomes portrayed that the Continuously Stirred Tank Reactor system produced about 450 liters of methane per Kg of volatile solids, at a detention period of 48 days. The second method involving Leach Beds produced about 340 liters of methane per Kg of volatile solids with a detention period of 28 days. The results showcased that CSTR when designed exclusively for grass proved to be extremely efficient in methane production. The SLBR-UASB has significant potential to allow for lower detention times with significant levels of methane production. This technology has immense future for research and development in India in terms utilizing of grass crop as a non-conventional source of fuel.

Keywords: biomethane potential values, bio digester specifications, continuously stirred tank reactor, upflow anaerobic sludge blanket

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Authors: Ehsan Arabian, Thomas Sattelmayer

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The formation of nitrogen dioxide NO2 in the exhaust duct of a MAN dual fuel test engine has been investigated numerically. The dual fuel engine concept with premixed lean methane combustion ignited through diesel pilot flames reveals high potential for the abatement of the NOx formation. The drawback of this combustion method, however, is the high NO2 formation due to the increasing concentration of unburned hydrocarbons. This promotes the conversion of NO to NO2, which is toxic and characterized through its yellow color. The results presented in this paper cover a wide range of engine operation points from full load to part load for different air to fuel ratios. The effects of temperature, pressure and concentrations of unburned methane and nitric oxide on NO2 formation in the exhaust duct has been investigated on the basis of a zero-dimensional well stirred reactor model implemented in Cantera, which calculates the steady state of a uniform composition for a certain residence time. It can be shown that the simulated conversion of NO to NO2 match the experimental results fairly well. The partial oxidation of methane followed by CO production can be predicted as well. It can also be concluded that the lower temperature limit for which no conversion takes place, depends mainly on the concentration of the unburned hydrocarbons in the exhaust.

Keywords: cantera, dual fuel engines, exhaust tract, numerical modeling of NO2 formation, well stirred reactor

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Authors: F. A. Ogundolie, F. Okogue, D. V. Adegunloye

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Greywater samples were obtained from three restaurants in the Federal University of Technology; Akure coded SSR, MGR and GGR. Fungi isolates obtained include Rhizopus stolonifer, Aspergillus niger, Mucor mucedo, Aspergillus flavus, Saccharomyces cerevisiae. Of these fungi isolates obtained, R. stolonifer, A. niger and A. flavus showed significant degradation ability on grey water and was used for this research. A simple bioreactor was constructed using biodegradation process in purification of waste water samples. Waste water undergoes primary treatment; secondary treatment involves the introduction of the isolated organisms into the waste water sample and the tertiary treatment which involved the use of filter candle and the sand bed filtration process to achieve the end product without the use of chemicals. A. niger brought about significant reduction in both the bacterial load and the fungi load of the greywater samples of the three respective restaurants with a reduction of (1.29 × 108 to 1.57 × 102 cfu/ml; 1.04 × 108 to 1.12 × 102 cfu/ml and 1.72 × 108 to 1.60 × 102 cfu/ml) for bacterial load in SSR, MGR and GGR respectively. Reduction of 2.01 × 104 to 1.2 × 101; 1.72 × 104 to 1.1 × 101, and 2.50 × 104 to 1.5 × 101 in fungi load from SSR, MGR and GGR respectively. Result of degradation of these selected waste water by the fungi showed that A. niger was probably more potent in the degradation of organic matter and hence, A. niger could be used in the treatment of wastewater.

Keywords: Aspergillus niger, greywater, bacterial, fungi, microbial load, bioreactor, biodegradation, purification, organic matter and filtration

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Authors: Simon Grossemy, Peggy P. Y. Chan, Pauline M. Doran

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Nerve tissue engineering is the main field of research aimed at finding an alternative to autografts as a treatment for nerve injuries. Scaffolds are used as a support to enhance nerve regeneration. In order to successfully design novel scaffolds and in vitro cell culture systems, a deep understanding of the factors affecting nerve regeneration processes is needed. Physical and biological parameters associated with the culture environment have been identified as potentially influential in nerve cell differentiation, including electrical stimulation, exposure to extracellular-matrix (ECM) proteins, dynamic medium conditions and co-culture with glial cells. The mechanisms involved in driving the cell to differentiation in the presence of these factors are poorly understood; the complexity of each of them raises the possibility that they may strongly influence each other. Some questions that arise in investigating nerve regeneration include: What are the best protein coatings to promote neural cell attachment? Is the scaffold design suitable for providing all the required factors combined? What is the influence of dynamic stimulation on cell viability and differentiation? In order to study these effects, scaffolds adaptable to bioreactor culture conditions were designed to allow electrical stimulation of cells exposed to ECM proteins, all within a dynamic medium environment. Gold coatings were used to make the surface of viscose rayon microfiber scaffolds (VRMS) conductive, and poly-L-lysine (PLL) and laminin (LN) surface coatings were used to mimic the ECM environment and allow the attachment of rat PC12 neural cells. The robustness of the coatings was analyzed by surface resistivity measurements, scanning electron microscope (SEM) observation and immunocytochemistry. Cell attachment to protein coatings of PLL, LN and PLL+LN was studied using DNA quantification with Hoechst. The double coating of PLL+LN was selected based on high levels of PC12 cell attachment and the reported advantages of laminin for neural differentiation. The underlying gold coatings were shown to be biocompatible using cell proliferation and live/dead staining assays. Coatings exhibiting stable properties over time under dynamic fluid conditions were developed; indeed, cell attachment and the conductive power of the scaffolds were maintained over 2 weeks of bioreactor operation. These scaffolds are promising research tools for understanding complex neural cell behavior. They have been used to investigate major factors in the physical culture environment that affect nerve cell viability and differentiation, including electrical stimulation, bioreactor hydrodynamic conditions, and combinations of these parameters. The cell and tissue differentiation response was evaluated using DNA quantification, immunocytochemistry, RT-qPCR and functional analyses.

Keywords: bioreactor, electrical stimulation, nerve differentiation, PC12 cells, scaffold

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Authors: Payal T. Patel, Ramakant Panchal, Ketankumar G. Patel

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Gear in stirred vessel is one of the most critical components in machinery which has power transmission system and it is rotating machinery cost and redesign being the major constraints, there is always a great scope for a mechanical engineer to apply skills to improve the design. Gear will be most effective means of transmitting power in future machinery due to their high degree of compactness. The Galliard moved in the industry from heavy industries such as textile machinery and shipbuilding to industries such as automobile manufacture tools will necessitate the affable application of gear technology. The two-stage planetary reduction gear unit is designed to meet the output specifications. In industries, where the bevel gears are used in turret vessel to transmit the power, that unit is replaced by this planetary gearbox. Use of this type of gearbox is to get better efficiency and also the manufacturing of the bevel gear is more complex than the spur gears. Design a gearbox with the epicyclic gear train. In industries, the power transmission from gearbox to vessel is done through the bevel gears, which transmit the power at a right angle. In this work, the power is to be transmitted vertically from gearbox to vessel, which will increase the efficiency and life of gears. The arrangement of the gears is quite difficult as well as it needs high manufacturing cost and maintenance cost. The design is replaced by the planetary gearbox to reduce the difficulties, and same output is achieved but with a different arrangement of the planetary gearbox.

Keywords: planetary gearbox, epicyclic gear, optimization, dynamic balancing

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Authors: Elahe Najafizade, Ahmad Mohammad Alkhateeb, Seyed Ali Hossein Zahraei, Iman Dianat

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Introduction: Hangnails are short, torn, down parts of the skin surrounding the nails. At times they are very painful. The usual treatment advised is cutting the excess skin with clippers or scissors. To provide instant relief to the patients, we describe a simpler and more effective way to use surgical glue to paste them back into their original position. Method: The cream should not be on the heat; it is on the bain-marie. To achieve the desired emulsifier, 1 gram of borax was mixed in 10 grams of distilled water in a bain-marie until it melted, then stirred oserin, beeswax, and oil in the bain-marie until it melted. After that, 32 grams of distilled water was added little by little. We add and stir and gradually add the borax dissolved in 10 grams of distilled water. The bowl of cream was placed in a bowl of cold water and stirred until the cream was smooth. After that, we add gasoline, alcohol, or methylparaben preservatives. It should be noted that this amount of ingredients is enough for a 350-gram can (when we prepare the cream, we also add the extract). Result: The patient was a 40-year-old female with a hangnail problem that had been used several different creams and Vaseline, but the treatment was not useful, but after this cream was applied for treatment; the hangnail started to cure within one week, and complete treatment achieved after two weeks. Conclusion: Traditional methods with modification without using chemical substances somehow work better and safer, so research programs on them will be useful for less risky treatment procedures.

Keywords: nail, cream, formula, traditional medicine

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Authors: Alexandra Tucaliuc, Alexandra C. Blaga, Anca I. Galaction, Lenuta Kloetzer, Dan Cascaval

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Ergosterol (ergosta-5,7,22-trien-3β-ol), also known as provitamin D2, is the precursor of vitamin D2 (ergocalciferol), because it is converted under UV radiation to this vitamin. The natural sources of ergosterol are mainly the yeasts (Saccharomyces sp., Candida sp.), but it can be also found in fungus (Claviceps sp.) or plants (orchids). In the yeasts cells, ergosterol is accumulated in membranes, especially in free form in the plasma membrane, but also as esters with fatty acids in membrane lipids. The chemical synthesis of ergosterol does not represent an efficient method for its production, in these circumstances, the most attractive alternative for producing ergosterol at larger-scale remains the aerobic fermentation using S. cerevisiae on glucose or by-products from agriculture of food industry as substrates, in batch or fed-batch operating systems. The aim of this work is to analyze comparatively the influence of aeration efficiency on ergosterol production by S. cerevisiae in batch and fed-batch fermentations, by considering different levels of mixing intensity, aeration rate, and n-dodecane concentration. The effects of the studied factors are quantitatively described by means of the mathematical correlations proposed for each of the two fermentation systems, valid both for the absence and presence of oxygen-vector inside the broth. The experiments were carried out in a laboratory stirred bioreactor, provided with computer-controlled and recorded parameters. n-Dodecane was used as oxygen-vector and the ergosterol content inside the yeasts cells has been considered at the fermentation moment related to the maximum concentration of ergosterol, 9 hrs for batch process and 20 hrs for fed-batch one. Ergosterol biosynthesis is strongly dependent on the dissolved oxygen concentration. The hydrocarbon concentration exhibits a significant influence on ergosterol production mainly by accelerating the oxygen transfer rate. Regardless of n-dodecane addition, by maintaining the glucose concentration at a constant level in the fed-batch process, the amount of ergosterol accumulated into the yeasts cells has been almost tripled. In the presence of hydrocarbon, the ergosterol concentration increased by over 50%. The value of oxygen-vector concentration corresponding to the maximum level of ergosterol depends mainly on biomass concentration, due to its negative influences on broth viscosity and interfacial phenomena of air bubbles blockage through the adsorption of hydrocarbon droplets–yeast cells associations. Therefore, for the batch process, the maximum ergosterol amount was reached for 5% vol. n-dodecane, while for the fed-batch process for 10% vol. hydrocarbon.

Keywords: bioreactors, ergosterol, fermentation, oxygen-vector

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Authors: Nikolaos Katsoulas, Sofia Faliagka, George Kountrias, Eleni Dimitriou, Eleftheria Pechlivani

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The challenges to feed the world in 2050 are becoming more and more apparent. This calls for producing more with fewer inputs (most of them under scarcity), higher resource efficiency, minimum or zero effect on the environment, and higher sustainability. Therefore, increasing the circularity of production systems is highly significant for their sustainability. Protected horticulture offers opportunities for maximum resource efficiency across various levels within and between farms and at the regional level), high-quality production, and contributes significantly to the nutrition security as part of the world food production. In greenhouses, closed soilless cultivation systems give the opportunity to increase the water and nutrient use efficiency and reduce the environmental impact of the cultivation system by the reuse of the drained water and nutrients. However, due to the low quality of the water used in the Mediterranean countries, a completely closed system is not feasible. Partial discharge of the drainage nutrient solution when the levels of electrical conductivity (EC) or of the toxic ions in the system are reached is still a necessity. Thus, in the frame of the circular economy concept, this work presents the utilisation of the drainage solution of soilless cultivation systems for microalgae and biofertilisers production. The system includes a greenhouse equipped with a soilless cultivation system, a drainage solution collection tank, a closed bioreactor for microalgae production, and a biocatalysis tank. The bioreactor tested in the frame of this work includes two closed tube loops of a capacity of 1000 L each where, after the initial inoculation, the microalgae is developed using as a growth medium the drainage solution collected from the greenhouse crops. The bioreactor includes light and temperature control while pH is still manually regulated. As soon as the microalgae culture reaches a certain density level, 20% of the culture is harvested, and the culture system is refiled by a drainage nutrient solution. The microalgae produced goes through a biocatalysis process, which leads to the production of a rich aminoacids (and nitrogen) biofertiliser. The produced biofertiliser is then used for the fertilisation of greenhouse crops. The complete production cycle along with the effects of the biofertiliser produced on crop growth and yield are presented and discussed in this manuscript. Acknowledgment: This work was carried out under the PestNu project that has received funding from the European Union’s Horizon 2020 research and innovation programme under the Green Deal grant agreement No. 101037128 — PestNu.

Keywords: soilless, water use efficiency, nutrients use efficiency, biostimulant

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Authors: Andres Diaz Garcia

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The successful technological development of any bioproduct, including those of the biostimulant type, requires to adequately completion of a series of stages allied to different disciplines that are related to microbiological, engineering, pharmaceutical chemistry, legal and market components, among others. Engineering as a discipline has a key contribution in different aspects of fermentation processes such as the design and optimization of culture media, the standardization of operating conditions within the bioreactor and the scaling of the production process of the active ingredient that it will be used in unit operations downstream. However, all aspects mentioned must take into account many biological factors of the microorganism such as the growth rate, the level of assimilation to various organic and inorganic sources and the mechanisms of action associated with its biological activity. This paper focuses on the practical experience within the Colombian Corporation for Agricultural Research (Agrosavia), which led to the development of a biostimulant bioproduct based on native rhizobacteria Bacillus amyloliquefaciens, oriented mainly to plant growth promotion in cape gooseberry nurseries and fruit crops in Colombia, and the challenges that were overcome from the expertise in the area of engineering. Through the application of strategies and engineering tools, a culture medium was optimized to obtain concentrations higher than 1E09 CFU (colony form units)/ml in liquid fermentation, the process of biomass production was standardized and a scale-up strategy was generated based on geometric (H/D of bioreactor relationships), and operational criteria based on a minimum dissolved oxygen concentration and that took into account the differences in the capacity of control of the process in the laboratory and pilot scales. Currently, the bioproduct obtained through this technological process is in stages of registration in Colombia for cape gooseberry fruits for export.

Keywords: biochemical engineering, liquid fermentation, plant growth promoting, scale-up process

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Authors: Ahmed Tawfik

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Oxidation of 1,4 dioxane produces metabolites by-products involving glycolaldehyde and acids that have geno- and cytotoxicity impact on microbial degradation. Thereby, the incorporation of algae with bacteria in the treatment system would eliminate and overcome the accumulation of metabolites that are utilized as a carbon source for the build-up of biomass. Therefore, the aim of the present study is to assess the potential of algae/bacteria-based membrane bioreactor (AB-MBR) for biodegradation of 1,4 dioxane-rich wastewater at a high imposed loading rate. Three identical reactors, i.e., AB-MBR1, AB-MBR2, and AB-MBR3, were operated in parallel at 1,4 dioxane loading rates of 641.7, 320.9, and 160.4 mg/L. d., and HRTs of 6.0, 12 and 24 h. respectively. The AB-MBR1 achieved 1,4 dioxane removal rate of 263.7 mg/L.d., where the residual value in the treated effluent amounted to 94.4±22.9 mg/L. Reducing the 1,4 dioxane loading rate (LR) to 320.9 mg/L.d in the AB-MBR2 maximized the removal rate efficiency of 265.9 mg/L.d., with a removal efficiency of 82.8±3.2%. The minimum value of 1,4 dioxane of 17.3±1.8 mg/L in the treated effluent of AB-MBR3 was obtained at an HRT of 24.0 h and loading rate of 160.4 mg/L.d. The mechanism of 1,4 dioxane degradation in AB-MBR was a combination of volatilization (8.03±0.6%), UV oxidation (14.1±0.9%), microbial biodegradation (49.1±3.9%) and absorption/uptake and assimilation by algae (28.8±2.%). Further, the Thioclava, Afipia, and Mycobacterium genera oxidized and produced the required enzymes for hydrolysis and cleavage of the dioxane ring into 2-hydroxy-1,4 dioxane. Moreover, the fungi, i.e., Basidiomycota and Cryptomycota, played a big role in the degradation of the 1,4 dioxane into 2-hydroxy-1,4 dioxane. Xanthobacter and Mesorhizobium were involved in the metabolism process by secreting alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), and glycolate oxidase. Bacteria and fungi produced dehydrogenase (DH) for the transformation of 2-hydroxy-1,4 dioxane into 2-hydroxy-ethoxyacetaldehyde. The latter is converted into Ethylene glycol by Aldehyde hydrogenase (ALDH). Ethylene glycol is oxidized into acids using Alcohol hydrogenase (ADH). The Diatomea, Chlorophyta, and Streptophyta utilize the metabolites for biomass assimilation and produce the required oxygen for further oxidation of the dioxane and its metabolites by-products of bacteria and fungi. The major portion of metabolites (ethylene glycol, glycolic acid, and oxalic acid were removed due to uptake and absorption by algae (43±4.3%), followed by adsorption (18.4±0.9%). The volatilization and UV oxidation contribution for the degradation of metabolites were 8.7±0.7% and 12.3±0.8%, respectively. The capabilities of genera Defluviimonas, Thioclava, Luteolibacter, and Afipia. The genera of Defluviimonas, Thioclava, Luteolibacter, and Mycobacterium were grown under a high 1,4 dioxane LR of 641.7 mg/L.d. The Chlorophyta (4.1-43.6%), Streptophyta (2.5-21.7%), and Diatomea (0.8-1.4%) phyla were dominant for degradation of 1,4 dioxane. The results of this study strongly demonstrated that the bioremediation and bioaugmentation process can safely remove 1,4 dioxane from industrial wastewater while minimizing environmental concerns and reducing economic costs.

Keywords: wastewater, membrane bioreactor, bacterial community, algal community

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Authors: Deybith Venegas-Rojas, Jens Budde, Dominik Nörz, Manfred Jücker, Hoc Khiem Trieu

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Microfluidics is a promising approach for biomedicine cell culture experiments with microfluidic bioreactors (MBR), which can provide high precision in volume and time control over mass transport and microenvironments in small-scale studies. Nevertheless, shear stress and oxygen concentration are important factors that affect the microenvironment and then the cell culture. It is presented a novel MBR design in which differences in geometry, shear stress, and oxygen concentration were studied and optimized for cell culture. The aim is to mimic the in vivo condition with biocompatible materials and continuous perfusion of nutrients, a healthy shear stress, and oxygen concentration. The design consists of a capture system of PDMS micropillars which keep cells in place, so it is not necessary any hydrogel or complicated scaffolds for cells immobilization. Besides, the design allows continuous supply with nutrients or even any other chemical for cell experimentation. Finite element method simulations were used to study and optimize the effect of parameters such as flow rate, shear stress, oxygen concentration, micropillars shape, and dimensions. The micropillars device was fabricated with microsystem technology such as soft-lithography, deep reactive ion etching, self-assembled monolayer, replica molding, and oxygen plasma bonding. Eight different geometries were fabricated and tested, with different flow rates according to the simulations. During the experiments, it was observed the effect of micropillars size, shape, and configuration for stability and shear stress control when increasing flow rate. The device was tested with several successful HepG2 3D cell cultures. With this MBR, the aforementioned parameters can be controlled in order to keep a healthy microenvironment according to specific necessities of different cell types, with no need of hydrogels and can be used for a wide range of experiments with cells.

Keywords: cell culture, micro-bioreactor, microfluidics, micropillars, oxygen concentration, shear stress

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Authors: C. Müller, T. Ortmann, S. Scholl, H. J. Jördening

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Multienzymatic catalysis can be used as an alternative to chemical synthesis or hydrolysis of polysaccharides for the production of high value oligosaccharides from cheap resources such as sucrose. However, development of multienzymatic processes is complex, especially with respect to suitable conditions for enzymes originating from different organisms. Furthermore, an optimal configuration of the catalysts in a reaction cascade has to be found. These challenges can be approached by design of experiments. The system investigated in this study is a trienzymatic catalyzed reaction which results in laminaribiose production from sucrose and comprises covalently immobilized sucrose phosphorylase (SP), glucose isomerase (GI) and laminaribiose phosphorylase (LP). Operational windows determined with design of experiments and kinetic data of the enzymes were used to optimize the enzyme ratio for maximum product formation and minimal production of byproducts. After adjustment of the enzyme activity ratio to 1: 1.74: 2.23 (SP: LP: GI), different process options were investigated in silico. The considered options included substrate dependency, the use of glucose as co-substrate and substitution of glucose isomerase by glucose addition. Modeling of batch operation in a stirred tank reactor led to yields of 44.4% whereas operation in a continuous stirred tank reactor resulted in product yields of 22.5%. The maximum yield in a bienzymatic system comprised of sucrose phosphorylase and laminaribiose phosphorylase was 67.7% with sucrose and different amounts of glucose as substrate. The experimental data was in good compliance with the process model for batch operation. The continuous operation will be investigated in further studies. Simulation of operational process possibilities enabled us to compare various operational modes regarding different aspects such as cost efficiency, with the minimum amount of expensive and time-consuming practical experiments. This gives us more flexibility in process implementation and allows us, for example, to change the production goal from laminaribiose to higher oligosaccharides.

Keywords: design of experiments, enzyme kinetics, multi-enzymatic system, in silico process development

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Authors: Vytautas Galvanauskas, Rimvydas Simutis, Donatas Levisauskas, Vykantas Grincas, Renaldas Urniezius

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The paper deals with model-based development and implementation of efficient control strategies for recombinant protein synthesis in fed-batch E.coli cultivation processes. Based on experimental data, a kinetic dynamic model for cultivation process was developed. This model was used to determine substrate feeding strategies during the cultivation. The proposed feeding strategy consists of two phases – biomass growth phase and recombinant protein production phase. In the first process phase, substrate-limited process is recommended when the specific growth rate of biomass is about 90-95% of its maximum value. This ensures reduction of glucose concentration in the medium, improves process repeatability, reduces the development of secondary metabolites and other unwanted by-products. The substrate limitation can be enhanced to satisfy restriction on maximum oxygen transfer rate in the bioreactor and to guarantee necessary dissolved carbon dioxide concentration in culture media. In the recombinant protein production phase, the level of substrate limitation and specific growth rate are selected within the range to enable optimal target protein synthesis rate. To account for complex process dynamics, to efficiently exploit the oxygen transfer capability of the bioreactor, and to maintain the required dissolved oxygen concentration, adaptive control algorithms for dissolved oxygen control have been proposed. The developed model-based control strategies are useful in scale-up of cultivation processes and accelerate implementation of innovative biotechnological processes for industrial applications.

Keywords: adaptive algorithms, model-based control, recombinant E. coli, scale-up of bioprocesses

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Authors: W. Baumgartner, M. Welti, N. Hild, S. C. Hess, W. J. Stark, G. Meier Bürgisser, P. Giovanoli, J. Buschmann

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Perfusion bioreactors are used to solve problems in tissue engineering in terms of sufficient nutrient and oxygen supply. Such problems especially occur in critical size grafts because vascularization is often too slow after implantation ending up in necrotic cores. Biominerizable and biocompatible nanocomposite materials are attractive and suitable scaffold materials for bone tissue engineering because they offer mineral components in organic carriers – mimicking natural bone tissue. In addition, human adipose derived stem cells (ASCs) can potentially be used to increase bone healing as they are capable of differentiating towards osteoblasts or endothelial cells among others. In the present study, electrospun nanocomposite disks of poly-lactic-co-glycolic acid and amorphous calcium phosphate nanoparticles (PLGA/a-CaP) were seeded with human ASCs and eight disks were stacked in a bioreactor running with normal culture medium (no differentiation supplements). Under continuous perfusion and uniaxial cyclic compression, load-displacement curves as a function of time were assessed. Stiffness and energy dissipation were recorded. Moreover, stem cell densities in the layers of the piled scaffold were determined as well as their morphologies and differentiation status (endothelial cell differentiation, chondrogenesis and osteogenesis). While the stiffness of the cell free constructs increased over time caused by the transformation of the a-CaP nanoparticles into flake-like apatite, ASC-seeded constructs showed a constant stiffness. Stem cell density gradients were histologically determined with a linear increase in the flow direction from the bottom to the top of the 3.5 mm high pile (r2 > 0.95). Cell morphology was influenced by the flow rate, with stem cells getting more roundish at higher flow rates. Less than 1 % osteogenesis was found upon osteopontin immunostaining at the end of the experiment (9 days), while no endothelial cell differentiation and no chondrogenesis was triggered under these conditions. All ASCs had mainly remained in their original pluripotent status within this time frame. In summary, we have fabricated a critical size bone graft based on a biominerizable bone-biomimetic nanocomposite with preserved stiffness when seeded with human ASCs. The special feature of this bone graft was that ASC densities inside the piled construct varied with a linear gradient, which is a good starting point for tissue engineering interfaces such as bone-cartilage where the bone tissue is cell rich while the cartilage exhibits low cell densities. As such, this tissue-engineered graft may act as a bone-cartilage interface after the corresponding differentiation of the ASCs.

Keywords: bioreactor, bone, cartilage, nanocomposite, stem cell gradient

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Authors: Nassima Hamzaoui, Mostefa Ghamnia

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Nanoscale thin films of pure and Mn-doped Nickel oxide (NiO) were prepared by dissolving nickel chloride hexahydrate (NiCl2, 6H2O) and manganese chloride tetrahydrate (MnCl2,4H2O) under experimental conditions. The resulting solution was stirred at room temperature for 30 OC minutes in order to obtain homogeneity. The solution was sprayed onto heated glass substrates. The films obtained were characterized by X-ray diffraction to verify crystallinity. Atomic force microscopy (AFM) reveals surface topographical structure. UV-visible spectroscopy shows good transparency of the NiO layers.

Keywords: films, NiO, AFM, X-ray diffraction

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Authors: Anubha Kaushik, Raman Preet

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Distillery wastewater in the form of spent wash is a complex and strong industrial effluent, with high load of organic pollutants that may deplete dissolved oxygen on being discharged into aquatic systems and contaminate groundwater by leaching of pollutants, while untreated spent wash disposed on land acidifies the soil. Stringent legislative measures have therefore been framed in different countries for discharge standards of distillery effluent. Utilising the organic pollutants present in various types of wastes as food by mixed microbial populations is emerging as an eco-friendly approach in the recent years, in which complex organic matter is converted into simpler forms, and simultaneously useful gases are produced as renewable and clean energy sources. In the present study, wastewater from a rice bran based distillery has been used as the substrate in a dark fermenter, and native microbial consortium from the digester sludge has been used as the inoculum to treat the wastewater and produce hydrogen. After optimising the operational conditions in batch reactors, sequential batch mode and continuous flow stirred tank reactors were used to study the best operational conditions for enhanced and sustained hydrogen production and removal of pollutants. Since the rate of hydrogen production by the microbial consortium during dark fermentation is influenced by concentration of organic matter, pH and temperature, these operational conditions were optimised in batch mode studies. Maximum hydrogen production rate (347.87ml/L/d) was attained in 32h dark fermentation while a good proportion of COD also got removed from the wastewater. Slightly acidic initial pH seemed to favor biohydrogen production. In continuous stirred tank reactor, high H₂ production from distillery wastewater was obtained from a relatively shorter substrate retention time (SRT) of 48h and a moderate organic loading rate (OLR) of 172 g/l/d COD.

Keywords: distillery wastewater, hydrogen, microbial consortium, organic pollution, sludge

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Authors: Akram Abi, Clarissa Müller, Hans-Joachim Jördening

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Laminaribiose is a beta-1,3-glycoside which is used in the medical field for the treatment of dermatitis and also can be used as a building block for new pharmaceutics. The conventional process of laminaribiose production is the uneconomical process of hydrolysis of laminarin extracted from natural polysaccharides of plant origin. A more economical approach however is attainable by enzymatically synthesis of laminaribiose via a reverse phosphorylase reaction catalyzed by laminaribiose phosphorylase (LP) from Euglena gracilis. Different cultivation methods of Euglena gracilis and the effect on LP production have been investigated. Buffered/unbuffered heterotrophic and mixotrophic cultivations of Euglena gracilis has been carried out. Changes of biomass and LP production, glucose level and pH, cell count and shape has been monitored in the course of time. The results obtained from experiments each in three repetitions, show that in the heterotrophic cultivation of Euglena gracilis not only more biomass is produced compared to mixotrophic cultivation, but also higher specific protein concentration is achieved. Furthermore, the LP activity test showed that the protein extracted from heterotrophically cultured cells has a higher LP activity. It was also observed that the cells develop in a distinctive different shape between these two cultures and have different length to width ratios. Taking the heterotrophic culture as the more efficient cultivation method in LP production, another comparative experiment between buffered and unbuffered heterothrophic culture was carried out that showed the unbuffered culture has advantages over the other one in respect of both LP production and resulting activity. A hetrotrophic cultivation of Euglena gracilis in a 5L bioreactor with controlled operating conditions showed a distinctive improvement of all the aspects of culture compared to the shaking flask cultivations. Biomass production was improved from 5 to more than 8 g/l (dry weight) which resulted in a specific protein concentration of 45 g/l in the heterotrophic cultivation in the bioreactor. In further attempts to improve LP production, different purification methods were tested and each method was checks through an activity assay. A laminaribiose yield of 35% was achieved which was by far the highest amount amongst different methods tested.

Keywords: euglena gracilis, heterotrophic culture, laminaribiose production, mixotrophic culture

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Authors: Aadila Cayenne, Hinrich Uellendahl

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Optimization of the anaerobic digestion (AD) process of halophytic plants is essential as the biomass contains a high salt content that can inhibit the AD process. Anaerobic co-digestion, together with manure, can resolve the inhibitory effects of saline biomass in order to dilute the salt concentration and establish favorable conditions for the microbial consortia of the AD process. The present laboratory study investigated the co-digestion of S. ramosissima (Sram), and pig manure (PM) in batch and semi-continuous stirred tank reactors (CSTR) under mesophilic (38oC) conditions. The 0.5L batch reactor experiments were in mono- and co-digestion of Sram: PM using different percent volatile solid (VS) based ratios (0:100, 15:85, 25:75, 35:65, 50:50, 100:0) with an inoculum to substate (I/R) ratio of 2. Two 5L CSTR systems (R1 and R2) were operated for 133 days with a feed of PM in a control reactor (R1) and with a co-digestion feed in an increasing Sram VS ratio of Sram: PM of 15:85, 25:75, 35:65 in reactor R2 at an organic loading rate (OLR) of 2 gVS/L/d and hydraulic retention time (HRT) of 20 days. After a start-up phase of 8 weeks for both reactors R1 and R2 with PM feed alone, the halophyte biomass Sram was added to the feed of R2 in an increasing ratio of 15 – 35 %VS Sram over an 11-week period. The process performance was monitored by pH, total solid (TS), VS, total nitrogen (TN), ammonium-nitrogen (NH4 – N), volatile fatty acids (VFA), and biomethane production. In the batch experiments, biomethane yields of 423, 418, 392, 365, 315, and 214 mL-CH4/gVS were achieved for mixtures of 0:100, 15:85, 25:75, 35:65, 50:50, 100:0 %VS Sram: PM, respectively. In the semi-continuous reactor processes, the average biomethane yields were 235, 387, and 365 mL-CH4/gVS for the phase of a co-digestion feed ratio in R2 of 15:85, 25:75, and 35:65 %VS Sram: PM, respectively. The methane yield of PM alone in R1 was in the corresponding phases on average 260, 388, and 446 mL-CH4/gVS. Accordingly, in the continuous AD process, the methane yield of the halophyte Sram was highest at 386 mL-CH4/gVS in the co-digestion ratio of 25:75%VS Sram: PM and significantly lower at 15:85 %VS Sram: PM (100 mL-CH4/gVS) and at 35:65 %VS Sram (214 mL-CH4/gVS). The co-digestion process showed no signs of inhibition at 2 – 4 g/L NH4 – N, 3.5 – 4.5 g/L TN, and total VFA of 0.45 – 2.6 g/L (based on Acetic, Propionic, Butyric and Valeric acid). This study demonstrates that a stable co-digestion process of S. ramosissima and pig manure can be achieved with a feed of 25%VS Sram at HRT of 20 d and OLR of 2 gVS/L/d.

Keywords: anaerobic co-digestion, biomethane production, halophytes, pig manure, salicornia ramosissima

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Authors: Sylvana A. Vega, Cesar E. Huilinir, Carlos J. Gonzalez

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Co-digestion in anaerobic biodigesters is a technology improving hydrolysis by increasing methane generation. In the present study, the dimensional computational fluid dynamics (CFD) is numerically analyzed using Ansys Fluent software for agitation in a full-scale Continuous Stirred Tank Reactor (CSTR) biodigester during the co-digestion process. For this, a rheological study of the substrate is carried out, establishing rotation speeds of the stirrers depending on the microbial activity and energy ranges. The substrate is organic waste from industrial sources of sanitary water, butcher, fishmonger, and dairy. Once the rheological behavior curves have been obtained, it is obtained that it is a non-Newtonian fluid of the pseudoplastic type, with a solids rate of 12%. In the simulation, the rheological results of the fluid are considered, and the full-scale CSTR biodigester is modeled. It was coupling the second-order continuity differential equations, the three-dimensional Navier Stokes, the power-law model for non-Newtonian fluids, and three turbulence models: k-ε RNG, k-ε Realizable, and RMS (Reynolds Stress Model), for a 45° tilt vane impeller. It is simulated for three minutes since it is desired to study an intermittent mixture with a saving benefit of energy consumed. The results show that the absolute errors of the power number associated with the k-ε RNG, k-ε Realizable, and RMS models were 7.62%, 1.85%, and 5.05%, respectively, the numbers of power obtained from the analytical-experimental equation of Nagata. The results of the generalized Reynolds number show that the fluid dynamics have a transition-turbulent flow regime. Concerning the Froude number, the result indicates there is no need to implement baffles in the biodigester design, and the power number provides a steady trend close to 1.5. It is observed that the levels of design speeds within the biodigester are approximately 0.1 m/s, which are speeds suitable for the microbial community, where they can coexist and feed on the substrate in co-digestion. It is concluded that the model that more accurately predicts the behavior of fluid dynamics within the reactor is the k-ε Realizable model. The flow paths obtained are consistent with what is stated in the referenced literature, where the 45° inclination PBT impeller is the right type of agitator to keep particles in suspension and, in turn, increase the dispersion of gas in the liquid phase. If a 24/7 complete mix is considered under stirred agitation, with a plant factor of 80%, 51,840 kWh/year are estimated. On the contrary, if intermittent agitations of 3 min every 15 min are used under the same design conditions, reduce almost 80% of energy costs. It is a feasible solution to predict the energy expenditure of an anaerobic biodigester CSTR. It is recommended to use high mixing intensities, at the beginning and end of the joint phase acetogenesis/methanogenesis. This high intensity of mixing, in the beginning, produces the activation of the bacteria, and once reaching the end of the Hydraulic Retention Time period, it produces another increase in the mixing agitations, favoring the final dispersion of the biogas that may be trapped in the biodigester bottom.

Keywords: anaerobic co-digestion, computational fluid dynamics, CFD, net power, organic waste

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Authors: Ilze Dimanta, Zane Rutkovska, Vizma Nikolajeva, Janis Kleperis, Indrikis Muiznieks

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Majority of word’s steadily increasing energy consumption is provided by non-renewable fossil resources. Need to find an alternative energy resource is essential for further socio-economic development. Hydrogen is renewable, clean energy carrier with high energy density (142 MJ/kg, accordingly – oil has 42 MJ/kg). Biological hydrogen production is an alternative way to produce hydrogen from renewable resources, e.g. using organic waste material resource fermentation that facilitate recycling of sewage and are environmentally benign. Hydrogen gas is produced during the fermentation process of bacteria in anaerobic conditions. Bacteria are producing hydrogen in the liquid phase and when thermodynamic equilibrium is reached, hydrogen is diffusing from liquid to gaseous phase. Because of large quantities of available crude glycerol and the highly reduced nature of carbon in glycerol per se, microbial conversion of it seems to be economically and environmentally viable possibility. Such industrial organic waste product as crude glycerol is perspective for usage in feedstock for hydrogen producing bacteria. The process of biodiesel production results in 41% (w/w) of crude glycerol. The developed lab-scale test system (experimental bioreactor) with hydrogen micro-electrode (Unisense, Denmark) was used to determine hydrogen production yield and rate in the liquid phase. For hydrogen analysis in the gas phase the RGAPro-100 mass-spectrometer connected to the experimental test-system was used. Fermentative bacteria strains were tested for hydrogen gas production rates. The presence of hydrogen in gaseous phase was measured using mass spectrometer but registered concentrations were comparatively small. To decrease the hydrogen partial pressure in liquid phase reactor with a system for continuous bubbling with inert gas was developed. H2 production rate for the best producer in liquid phase reached 0,40 mmol H2/l, in gaseous phase - 1,32 mmol H2/l. Hydrogen production rate is time dependent – higher rate of hydrogen production is at the fermentation process beginning when concentration increases, but after three hours of fermentation, it decreases.

Keywords: bio-hydrogen, fermentation, experimental bioreactor, crude glycerol

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Authors: Ahmed AlNouss, Salim Ahmed

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Process control refers to the techniques to control the variables in a process in order to maintain them at their desired values. Advanced process control (APC) is a broad term within the domain of control where it refers to different kinds of process control and control related tools, for example, model predictive control (MPC), statistical process control (SPC), fault detection and classification (FDC) and performance assessment. APC is often used for solving multivariable control problems and model predictive control (MPC) is one of only a few advanced control methods used successfully in industrial control applications. Advanced control is expected to bring many benefits to the plant operation; however, the extent of the benefits is plant specific and the application needs a large investment. This requires an analysis of the expected benefits before the implementation of the control. In a real plant simulation studies are carried out along with some experimentation to determine the improvement in the performance of the plant due to advanced control. In this research, such an exercise is undertaken to realize the needs of APC application. The main objectives of the paper are as follows: (1) To apply MPC to a number of simulations set up to realize the need of MPC by comparing its performance with that of proportional integral derivatives (PID) controllers. (2) To study the effect of controller parameters on control performance. (3) To develop appropriate performance index (PI) to compare the performance of different controller and develop novel idea to present tuning map of a controller. These objectives were achieved by applying PID controller and a special type of MPC which is dynamic matrix control (DMC) on the multi-tanks process simulated in loop-pro. Then the controller performance has been evaluated by changing the controller parameters. This performance was based on special indices related to the difference between set point and process variable in order to compare the both controllers. The same principle was applied for continuous stirred tank heater (CSTH) and continuous stirred tank reactor (CSTR) processes simulated in Matlab. However, in these processes some developed programs were written to evaluate the performance of the PID and MPC controllers. Finally these performance indices along with their controller parameters were plotted using special program called Sigmaplot. As a result, the improvement in the performance of the control loops was quantified using relevant indices to justify the need and importance of advanced process control. Also, it has been approved that, by using appropriate indices, predictive controller can improve the performance of the control loop significantly.

Keywords: advanced process control (APC), control loop, model predictive control (MPC), proportional integral derivatives (PID), performance indices (PI)

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Authors: Azila Adnan, Giridhar R. Nair, Mark C. Lay, Janis E. Swan

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The effect of aeration on bacterial cellulose (BC) production by Gluconacetobacter xylinus DSM46604 was studied in 5-L bioreactor. Four aeration rates were applied (0.3, 0.6, 1.0 and 1.5 vvm) in the fermentation media at constant agitation rate, 150 rpm. One vvm enhanced BC concentration (4.4 g/L) and productivity (0.44 g/L/day) while greater agitation rate (1.5 vvm) decreased BC concentration (4.0 g/L) and productivity (0.40 g/L/day).

Keywords: aeration, bacterial cellulose, fermentation, gluconacetobacter xylinus

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Authors: Roman Matejka, Vaclav Prochazka, Tibor Izak, Jana Stepanovska, Martina Travnickova, Alexander Kromka

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Cell-based impedance spectroscopy is suitable method for electrical monitoring of cell activity especially on substrates that cannot be easily inspected by optical microscope (without fluorescent markers) like decellularized tissues, nano-fibrous scaffold etc. Special sensor for this measurement was developed. This sensor consists of corning glass substrate with gold interdigitated electrodes covered with diamond layer. This diamond layer provides biocompatible non-conductive surface for cells. Also, a special PPFC flow cultivation chamber was developed. This chamber is able to fix sensor in place. The spring contacts are connecting sensor pads with external measuring device. Construction allows real-time live cell imaging. Combining with perfusion system allows medium circulation and generating shear stress stimulation. Experimental evaluation consist of several setups, including pure sensor without any coating and also collagen and fibrin coating was done. The Adipose derived stem cells (ASC) and Human umbilical vein endothelial cells (HUVEC) were seeded onto sensor in cultivation chamber. Then the chamber was installed into microscope system for live-cell imaging. The impedance measurement was utilized by vector impedance analyzer. The measured range was from 10 Hz to 40 kHz. These impedance measurements were correlated with live-cell microscopic imaging and immunofluorescent staining. Data analysis of measured signals showed response to cell adhesion of substrates, their proliferation and also change after shear stress stimulation which are important parameters during cultivation. Further experiments plan to use decellularized tissue as scaffold fixed on sensor. This kind of impedance sensor can provide feedback about cell culture conditions on opaque surfaces and scaffolds that can be used in tissue engineering in development artificial prostheses. This work was supported by the Ministry of Health, grants No. 15-29153A and 15-33018A.

Keywords: bio-impedance measuring, bioreactor, cell cultivation, diamond layer, gold interdigitated electrodes, tissue engineering

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Authors: J. Ritonja

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The biochemical technology has been developing extremely fast since the middle of the last century. The main reason for such development represents a requirement for large production of high-quality biologically manufactured products such as pharmaceuticals, foods, and beverages. The impact of the biochemical industry on the world economy is enormous. The great importance of this industry also results in intensive development in scientific disciplines relevant to the development of biochemical technology. In addition to developments in the fields of biology and chemistry, which enable to understand complex biochemical processes, development in the field of control theory and applications is also very important. In the paper, the control for the biochemical reactor for the milk fermentation was studied. During the fermentation process, the biophysical quantities must be precisely controlled to obtain the high-quality product. To control these quantities, the bioreactor’s stirring drive and/or heating system can be used. Available commercial biochemical reactors are equipped with open loop or conventional linear closed loop control system. Due to the outstanding parameters variations and the partial nonlinearity of the biochemical process, the results obtained with these control systems are not satisfactory. To improve the fermentation process, the self-tuning adaptive control system was proposed. The use of the self-tuning adaptive control is suggested because the parameters’ variations of the studied biochemical process are very slow in most cases. To determine the linearized mathematical model of the fermentation process, the recursive least square identification method was used. Based on the obtained mathematical model the linear quadratic regulator was tuned. The parameters’ identification and the controller’s synthesis are executed on-line and adapt the controller’s parameters to the fermentation process’ dynamics during the operation. The use of the proposed combination represents the original solution for the control of the milk fermentation process. The purpose of the paper is to contribute to the progress of the control systems for the biochemical reactors. The proposed adaptive control system was tested thoroughly. From the obtained results it is obvious that the proposed adaptive control system assures much better following of the reference signal as a conventional linear control system with fixed control parameters.

Keywords: adaptive control, biochemical reactor, linear quadratic regulator, recursive least square identification

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Authors: Petros Gkotsis, Anastasios Zouboulis, Manasis Mitrakas, Dimitrios Zamboulis, E. Peleka

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The most serious drawback in wastewater treatment using membrane bioreactors (MBRs) is membrane fouling which gradually leads to membrane permeability decrease and efficiency deterioration. This work is part of a research project that aims to develop an integrated methodology for membrane fouling control, using specific chemicals which will enhance the coagulation and flocculation of compounds responsible for fouling, hence reducing biofilm formation on the membrane surface and limiting the fouling rate acting as a pre-treatment step. For this purpose, a pilot-scale plant with fully automatic operation achieved by means of programmable logic controller (PLC) has been constructed and tested. The experimental set-up consists of four units: wastewater feed unit, bioreactor, membrane (side-stream) filtration unit and permeate collection unit. Synthetic wastewater was fed as the substrate for the activated sludge. The dissolved oxygen (DO) concentration of the aerobic tank was maintained in the range of 2-3 mg/L during the entire operation by using an aerator below the membrane module. The membranes were operated at a flux of 18 LMH while membrane relaxation steps of 1 min were performed every 10 min. Both commercial and composite coagulants are added in different concentrations in the pilot-scale plant and their effect on the overall performance of the ΜΒR system is presented. Membrane fouling was assessed in terms of TMP, membrane permeability, sludge filterability tests, total resistance and the unified modified fouling index (UMFI). Preliminary tests showed that particular attention should be paid to the addition of the coagulant solution, indicating that pipe flocculation effectively increases hydraulic retention time and leads to voluminous sludge flocs. The most serious drawback in wastewater treatment using MBRs is membrane fouling, which gradually leads to membrane permeability decrease and efficiency deterioration. This results in increased treatment cost, due to high energy consumption and the need for frequent membrane cleaning and replacement. Due to the widespread application of MBR technology over the past few years, it becomes clear that the development of a methodology to mitigate membrane fouling is of paramount importance. The present work aims to develop an integrated technique for membrane fouling control in MBR systems and, thus, contribute to sustainable wastewater treatment.

Keywords: coagulation, membrane bioreactor, membrane fouling, pilot plant

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Authors: Ashish Pathak, Dong-Jin Kim, Byoung-Gon Kim

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Biological processes based on oxidation of sulfur compounds by chemolithotrophic microorganisms are emerging as an efficient and eco-friendly technique for removal of sulfur from the coal. In the present article, study was carried out to investigate the potential of biodesulfurization process in removing the sulfur from lignite coal sample collected from a Mongolian coal mine. The batch biodesulfurization experiments were conducted in 2.5 L borosilicate baffle type reactors at 35 ºC using Acidithiobacillus ferrooxidans. The effect of pulp density on efficiency of biodesulfurization was investigated at different solids concentration (1-10%) of coal. The results of the present study suggested that the rate of desulfurization was retarded at higher coal pulp density. The optimum pulp density found 5% at which about 48% of the total sulfur was removed from the coal.

Keywords: biodesulfurization, bioreactor, coal, pyrite

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Authors: masmoudi Jabri Khaoula, Zitouni Hana, Bousselmi Latifa, Akrout Hanen

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Mathematical modeling has become an essential tool for sustainable wastewater management, particularly for the simulation and the optimization of complex processes involved in activated sludge systems. In this context, the activated sludge model (ASM3h) was used for the simulation of a Biological Membrane Reactor (MBR) as it includes the integration of biological wastewater treatment and physical separation by membrane filtration. In this study, the MBR with a useful volume of 12.5 L was fed continuously with poultry slaughterhouse wastewater (PSWW) for 50 days at a feed rate of 2 L/h and for a hydraulic retention time (HRT) of 6.25h. Throughout its operation, High removal efficiency was observed for the removal of organic pollutants in terms of COD with 84% of efficiency. Moreover, the MBR has generated a treated effluent which fits with the limits of discharge into the public sewer according to the Tunisian standards which were set in March 2018. In fact, for the nitrogenous compounds, average concentrations of nitrate and nitrite in the permeat reached 0.26±0.3 mg. L-1 and 2.2±2.53 mg. L-1, respectively. The simulation of the MBR process was performed using SIMBA software v 5.0. The state variables employed in the steady state calibration of the ASM3h were determined using physical and respirometric methods. The model calibration was performed using experimental data obtained during the first 20 days of the MBR operation. Afterwards, kinetic parameters of the model were adjusted and the simulated values of COD, N-NH4+and N- NOx were compared with those reported from the experiment. A good prediction was observed for the COD, N-NH4+and N- NOx concentrations with 467 g COD/m³, 110.2 g N/m³, 3.2 g N/m³ compared to the experimental data which were 436.4 g COD/m³, 114.7 g N/m³ and 3 g N/m³, respectively. For the validation of the model under dynamic simulation, the results of the experiments obtained during the second treatment phase of 30 days were used. It was demonstrated that the model simulated the conditions accurately by yielding a similar pattern on the variation of the COD concentration. On the other hand, an underestimation of the N-NH4+ concentration was observed during the simulation compared to the experimental results and the measured N-NO3 concentrations were lower than the predicted ones, this difference could be explained by the fact that the ASM models were mainly designed for the simulation of biological processes in the activated sludge systems. In addition, more treatment time could be required by the autotrophic bacteria to achieve a complete and stable nitrification. Overall, this study demonstrated the effectiveness of mathematical modeling in the prediction of the performance of the MBR systems with respect to organic pollution, the model can be further improved for the simulation of nutrients removal for a longer treatment period.

Keywords: activated sludge model (ASM3h), membrane bioreactor (MBR), poultry slaughter wastewater (PSWW), reuse

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Authors: E. Nakamachi, K. Matsumoto, K. Yamamoto, Y. Morita, H. Sakamoto

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In recently, electromagnetic and mechanical stimulations have been recognized as the effective extracellular environment stimulation technique to enhance the defected peripheral nerve tissue regeneration. In this study, we developed a new hybrid bioreactor by adopting 50 Hz uniform alternative current (AC) magnetic stimulation and 4% strain mechanical stimulation. The guide tube for nerve regeneration is mesh structured tube made of biodegradable polymer, such as polylatic acid (PLA). However, when neural damage is large, there is a possibility that peripheral nerve undergoes necrosis. So it is quite important to accelerate the nerve tissue regeneration by achieving enhancement of nerve axonal extension rate. Therefore, we try to design and fabricate the system that can simultaneously load the uniform AC magnetic field stimulation and the stretch stimulation to cells for enhancement of nerve axonal extension. Next, we evaluated systems performance and the effectiveness of each stimulation for rat adrenal pheochromocytoma cells (PC12). First, we designed and fabricated the uniform AC magnetic field system and the stretch stimulation system. For the AC magnetic stimulation system, we focused on the use of pole piece structure to carry out in-situ microscopic observation. We designed an optimum pole piece structure using the magnetic field finite element analyses and the response surface methodology. We fabricated the uniform AC magnetic field stimulation system as a bio-reactor by adopting analytically determined design specifications. We measured magnetic flux density that is generated by the uniform AC magnetic field stimulation system. We confirmed that measurement values show good agreement with analytical results, where the uniform magnetic field was observed. Second, we fabricated the cyclic stretch stimulation device under the conditions of particular strains, where the chamber was made of polyoxymethylene (POM). We measured strains in the PC12 cell culture region to confirm the uniform strain. We found slightly different values from the target strain. Finally, we concluded that these differences were allowable in this mechanical stimulation system. We evaluated the effectiveness of each stimulation to enhance the nerve axonal extension using PC12. We confirmed that the average axonal extension length of PC12 under the uniform AC magnetic stimulation was increased by 16 % at 96 h in our bio-reactor. We could not confirm that the axonal extension enhancement under the stretch stimulation condition, where we found the exfoliating of cells. Further, the hybrid stimulation enhanced the axonal extension. Because the magnetic stimulation inhibits the exfoliating of cells. Finally, we concluded that the enhancement of PC12 axonal extension is due to the magnetic stimulation rather than the mechanical stimulation. Finally, we confirmed that the effectiveness of the uniform AC magnetic field stimulation for the nerve axonal extension using PC12 cells.

Keywords: nerve cell PC12, axonal extension, nerve regeneration, electromagnetic-mechanical stimulation, bioreactor

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Authors: Shin Woo Kim, Eui Ju Lee

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The recent development of biofuel production technology facilitates the use of bioethanol and biodiesel on automobile. Bioethanol, especially, can be used as a fuel for gasoline vehicles because the addition of ethanol has been known to increase octane number and reduce soot emissions. However, the wide application of biofuel has been still limited because of lack of detailed combustion properties such as auto-ignition temperature and pollutant emissions such as NOx and soot, which has been concerned mainly on the vehicle fire safety and environmental safety. In this study, the combustion characteristics of gasoline/ethanol fuel were investigated both numerically and experimentally. For auto-ignition temperature and NOx emission, the numerical simulation was performed on the well-stirred reactor (WSR) to simulate the homogeneous gasoline engine and to clarify the effect of ethanol addition in the gasoline fuel. Also, the response surface method (RSM) was introduced as a design of experiment (DOE), which enables the various combustion properties to be predicted and optimized systematically with respect to three independent variables, i.e., ethanol mole fraction, equivalence ratio and residence time. The results of stoichiometric gasoline surrogate show that the auto-ignition temperature increases but NOx yields decrease with increasing ethanol mole fraction. This implies that the bioethanol added gasoline is an eco-friendly fuel on engine running condition. However, unburned hydrocarbon is increased dramatically with increasing ethanol content, which results from the incomplete combustion and hence needs to adjust combustion itself rather than an after-treatment system. RSM results analyzed with three independent variables predict the auto-ignition temperature accurately. However, NOx emission had a big difference between the calculated values and the predicted values using conventional RSM because NOx emission varies very steeply and hence the obtained second order polynomial cannot follow the rates. To relax the increasing rate of dependent variable, NOx emission is taken as common logarithms and worked again with RSM. NOx emission predicted through logarithm transformation is in a fairly good agreement with the experimental results. For more tangible understanding of gasoline/ethanol fuel on pollutant emissions, experimental measurements of combustion products were performed in gasoline/ethanol pool fires, which is widely used as a fire source of laboratory scale experiments. Three measurement methods were introduced to clarify the pollutant emissions, i.e., various gas concentrations including NOx, gravimetric soot filter sampling for elements analysis and pyrolysis, thermophoretic soot sampling with transmission electron microscopy (TEM). Soot yield by gravimetric sampling was decreased dramatically as ethanol was added, but NOx emission was almost comparable regardless of ethanol mole fraction. The morphology of the soot particle was investigated to address the degree of soot maturing. The incipient soot such as a liquid like PAHs was observed clearly on the soot of higher ethanol containing gasoline, and the soot might be matured under the undiluted gasoline fuel.

Keywords: gasoline/ethanol fuel, NOx, pool fire, soot, well-stirred reactor (WSR)

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