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The present study used bench scale columns filled with biochar for phosphorous (P) recovery from real ultrafiltered wastewater. No studies are available about the potentiality of biochar using ultrafiltered real wastewater. Therefore, this study aimed to assess phosphate (PO4 3-) recovery by biochar-packed columns employing real treated wastewater from an ultrafiltration process. Three flow rates were tested, specifically 0.7, 1.7 and 2.3 L h-1, to gain insights into the optimal working conditions. Results revealed that the maximum amount of PO4 3- recovery (namely, 3.43 mg g-1 biochar) can be achieved after 7 h by employing the highest tested flow rate. Furthermore, the phosphorus exchange capacity (PEC) was inversely correlated with the feeding flow rate (FFR), with PEC values equal to 35, 25 and 9 % for FFR of 0.67, 1.7 and 2.3 L h-1, respectively. The pseudo-first order model best approximated the adsorption kinetics, thus suggesting that the adsorption of phosphate by biochar depends on its concentrations (i.e. physiosorption mechanism).

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Therapeutic oligonucleotides (ONs) are typically manufactured via solid-phase synthesis, characterized by limited scalability and huge environmental footprint, limiting their availability. Biomanufactured ONs have the potential to reduce the immunogenic side-effects, and to improve the sustainability of their chemical counterparts. Rhodovulum sulfidophilum was demonstrated a valuable host for the extracellular production of recombinant ONs. However, low viable cell densities and product titer were reported so far. In this work, perfusion cell cultures were established for the intensification of ON biomanufacturing. First, the perfusion conditions were simulated in 50 mL spin tubes, selected as a scale-down model of the process, with the aim of optimizing the medium composition and process parameters. This optimization stage led to an increase in the cell density by 44 % compared to the reference medium formulation. In addition, tests at increasing perfusion rates were conducted until achieving the maximum viable cell density (VCDmax), allowing the determination of the minimum cell-specific perfusion rate (CSPRmin) required to sustain the cell culture. Intriguingly, we discovered in this system also a maximum CSPR, above which growth inhibition starts. By leveraging this process optimization, we show for the first time the conduction of perfusion cultures of R. sulfidophilum in bench-scale bioreactors. This process development pipeline allowed stable cultures for more than 20 days and the continuous biomanufacturing of ONs, testifying the great potential of perfusion processes.

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Most pharmaceutical biotechnology companies use stirred-tank bioreactors (STR) for recombinant protein manufacturing. These bioreactors are used at a variety of different scales ranging from bench to production scales, with working volumes from 10 mL to 25,000 L. Bench-scale STRs are commonly used to culture mammalian cells for process development, to troubleshoot production scale bioreactors using scale-down models (SDM), or to conduct fundamental research. In this chapter, we describe the operations of a bench-scale STR for the production of recombinant proteins with suspension-adapted Chinese hamster ovary (CHO) cells. These operations include bioreactor setup and configuration, batching media, inoculation of the seed cell culture, production phase, and harvest of cell-free fluids.

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Powder segregation can cause severe issues in processes of pharmaceutical drugs for control of content uniformity if the powder is likely to be free or easy flowing. Assessing segregation intensity of formulated powders in a process is challenging at the formulation stage because of the limited availability of samples. An advanced segregation evaluation using small bench-scale testers can be useful for formulation decisions and suggestions of operation conditions in the process, which has not been practically investigated before. In this study, eight formulations (two co-processed excipients blended with one active pharmaceutical ingredient at different ratios) were used for the segregation study on two types of bench-scale testers (air-induced and surface rolling segregation tester), and a pilot simulation process rig as a comparative study. The results show that segregation measured on the bench-scale testers can give a good indication of the segregation intensity of a blend if the segregation intensity is not more than 20%. The comparison also shows that both the bench-scale testers have a good correlation to the process rig, respectively, which means either segregation tester can be used independently for the evaluation. A linear regression model was explored for prediction of segregation in the process.

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Bioethanol is a renewable fuel widely used in road transportation and is generally regarded as a clean energy source. Although fermentation is one of the major processes in bioethanol production, studies on improving its efficiency through operational design are limited, especially compared to other steps (pretreatment and hydrolysis/saccharification). In this study, two adapted feeding strategies, in which feed medium addition (sugar delivery) was adjusted to increase the supply of fermentable sugar, were developed to improve ethanol productivity in 5-L fed-batch fermentation by Saccharomyces cerevisiae. Specifically, a linear adapted feeding strategy was established based on changes in cell biomass, and an exponential adapted feeding strategy was developed based on cell biomass accumulation. By implementing these two feeding strategies, the overall ethanol productivity reached 0.88±0.04 and 0.87±0.06 g/L/h, respectively. This corresponded to ~20% increases in ethanol productivity compared to fixed pulsed feeding operations. Additionally, there was no residual glucose at the end of fermentation, and final ethanol content reached 95±3 g/L under the linear adapted operation and 104±3 g/L under the exponential adapted feeding strategy. No statistical difference was observed in the overall ethanol yield (ethanol-to-sugar ratio) between fixed and adapted feeding strategies (~91%). These results demonstrate that sugar delivery controlled by adapted feeding strategies was more efficient than fixed feeding operations, leading to higher ethanol productivity. Overall, this study provides novel adapted feeding strategies to improve sugar delivery and ethanol productivity. Integration into the current practices of the ethanol industry could improve productivity and reduce production costs of fermentation processes.

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With the emergence of multiple side effects on the usage of commercial L-asparaginase formulations, keen interest is provoked to investigate new sources of L-asparaginases that possess antileukemic properties with minimal side effects. The present study reports the cost-effective bench-scale production, homogeneity purification and apoptosis induction potential of a new L-asparaginase preparation from Bacillus indicus against human leukemia cells. The enzyme is highly specific toward the natural substrate L-asparagine. The study initiated with the enzyme production using cost-effective substrates in which a 3.28-fold enhancement of enzyme activity was achieved in comparison with an unoptimized medium using the central composite experimental design approach. The scale-up of the process in a 3.7-L batch bioreactor resulted in 16.42 ± 0.17 IU/mL of L-asparaginase activity in 24 h. The crude extracellular enzyme was purified to homogeneity using anion exchange chromatography followed by gel filtration chromatography. A single band of approximately 35 kDa molecular weight was obtained on SDS-PAGE, while native PAGE analysis confirmed it to be a tetramer of four identical subunits. The circular dichroism spectroscopic study revealed the α + ß mixed type of secondary structure with 38.7% α-helices and 27.4% ß pleated sheets. The antitumor toxicity exhibited on the MOLT-4 leukemia cells by the new L-asparaginase was revealed using the MTT assay and acridine orange/propidium iodide dual staining for live/dead cells. The flow cytometry analysis established the potential of the purified L-asparaginase to induce the apoptotic cell death mechanism in MOLT-4 leukemia cells. Conclusively, the L-asparaginase of Bacillus indicus is a highly promising candidate that can be introduced as a new enzyme therapeutic against various leukemia disorders.

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Bacterial L-asparaginase (LA) is a chemotherapeutic drug that has remained mainstay of cancer treatment for several decades. LA has been extensively used worldwide for the treatment of acute lymphoblastic leukemia (ALL). A halotolerant bacterial strain Bacillus licheniformis sp. isolated from marine environment was used for LA production. The enzyme produced was subjected to purification and physico-chemical characterisation. Purified LA was thermotolerant and demonstrated more than 90% enzyme activity after 1 h of incubation at 80 °C. LA has also proved to be resistant against pH gradient and retained activity at pH ranging from 3.0 to 10. The enzyme also had high salinity tolerance with 90% LA activity at 10% NaCl concentration. Detergents like Triton X-100 and Tween-80 were observed to inhibit LA activity while more than 70% catalytic activity was maintained in the presence of metals. Electrophoretic analysis revealed that LA is a heterodimer (~ 63 and ~ 65 kDa) and has molecular mass of around 130 kDa in native form. The kinetic parameters of LA were tested with LA having low Km value of 1.518 µM and Vmax value of 6.94 µM/min/mL. Purified LA has also exhibited noteworthy antiproliferative activity against cancer cell lines-HeLa, SiHa, A549, and SH-SY-5Y. In addition, bench-scale LA production was conducted in a 5-L bioreactor using moringa leaves as cost-effective substrate.

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Municipal solid waste incineration (MSWI) fly ash could be used as supplementary cementitious material in cement-based materials. However, heavy metal leaching, such as Cd, Cr, Cu, Pb and Zn, both from the MSWI fly ash and cement-based materials containing MSWI fly ash, remains a persistent obstacle. Here, an up-scaled electrodialytic treatment was used as a pre-treatment to remove heavy metals from MSWI fly ash before using the fly ash in mortar. Mortar samples with 10 wt% replacement of cement with either raw or elecrtodialytically treated MSWI fly ash were subjected to monolithic (in-use scenario) and crushed mortar (end-of-life scenario) leaching tests. The environmental conditions (e.g., exposure to chlorides or sulfates) at the surface of cement-based materials can affect leaching. Acidified H2O, NaCl or Na2SO4 solutions were, therefore, used for the leaching tests. Up to 80% heavy metal removal by the up-scaled electrodialytic pre-treatment was feasible. Regulatory limits for disposing of the MSWI fly ash in non-hazardous waste landfills were exceeded, even if the electrodialytic treatment removed heavy metals. However, leaching from monolithic mortar samples complied with the regulatory limits, while Cr leaching exceeded the regulatory limits for all crushed mortar samples when using NaCl or Na2SO4. Both NaCl and Na2SO4 generally increased the heavy metal leaching yield from fly ash and mortar compared to leaching with acidified H2O. The results of the study suggest that environmental conditions should be taken into account when assessing leaching from cement-based materials with MSWI fly ash.

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One of the expected outcomes of global warming is increased algal and cyanobacterial blooms. Based on its ability to separate algal particles, dissolved air flotation (DAF) is considered as a climate change adaptation technology for water treatment. The feasibility of DAF treatment is often assessed using DAF jar tests; however, they are not particularly good at predicting a full-scale DAF system's turbidity removals. Therefore, our group has developed a more reliable larger-diameter/larger-volume batch apparatus (LB-DAF), which was optimized by comparison with a full-scale DAF plant treating a low turbidity, highly coloured river water (SUVA ∼ 4.3). The objective of this study was to verify that the LB-DAF was capable of simulating full-scale DAF systems treating two significantly different waters. One was water from a large eutrophic bay in Lake Ontario (SUVA ∼2.6) and the second was a river water (SUVA ∼3.5). The turbidity removals achieved by the full-scale DAF systems treating these waters were compared with those for the LB-DAF tests conducted using different flocculation velocity gradients, saturated water pressures, recycle ratios and water depth to diameter ratios. The LB-DAF tests are good predictors of the full-scale DAF turbidity removals, the average difference for the two waters tested were 2% and 6%. The LB-DAF natural organic matter (NOM) removals for both waters differed by less than 1% from that measured at the corresponding treatment plants. In addition, as in our previous LB-DAF study, varying the different LB-DAF operational variables did not have a significant impact on turbidity and NOM removals.

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Accurate determination of evaporative flux from water surfaces and liquid containing porous media is critical for geotechnical and geoenvironmental applications. Laboratory simulations can isolate the various parameters influencing evaporative fluxes. However, most simulators capture selected surface and atmospheric conditions, and published literature generally provide limited information on the development and operation of the instruments. The new simulator adequately captures a wide range of relevant field parameters, maintains controlled conditions over the required testing time, utilizes readily available components for modular fabrication, and facilitates operational efficiency between individual modules.•This paper presents the modified Bench-Scale Atmosphere Simulator (BAS2).•This paper summarizes various atmosphere simulators developed over the last 25 years.•This paper describes the design, fabrication, operation, calibration, and validation of BAS2.

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The effectiveness and the failure mechanism of fire barriers in residential upholstered furniture were investigated by full-scale flaming tests on upholstered chair mock-ups. Six commercial fire barriers were tested in this study. Fire barriers were screened for (1) the presence of elements that are typically used in fire retardants and, (2) the presence of targeted fire retardants. For each fire barrier, triplicate flammability tests were run on chair mock-ups where polyurethane foam and polyester fiber fill were used as the padding materials, and each chair component was fully wrapped with the fire barrier of choice and a polypropylene cover fabric. The ignition source was an 18 kW square propane burner, impinging on the top surface of the seat cushion for 80 s. Results showed all six fire barriers reduced the peak heat release rate (as much as ≈ 64 %) and delayed its occurrence (up to ≈ 19 min) as compared to the control chair mock-ups. The heat release rate remained at a relatively low plateau level until liquid products (generated by either melting or pyrolysis of the padding material) percolated through the fire barrier at the bottom of the seat cushion and ignited, while the fire barrier was presumably intact. The flaming liquid products dripped and quickly formed a pool fire under the chair and the peak heat release rate occurred shortly thereafter. Ultimately, the ignition of the percolating liquid products at the bottom of the seat cushion was identified as the mechanism triggering the failure of the fire barrier.

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The main sugarcane wastes from the non-centrifugal cane sugar (NCS) agro-industry, agricultural crop residue (ACR) and sugarcane scum (SCS), were used to produce biogas in a bench-scale semi-continuous anaerobic tubular digester. A two-stage strategy was proposed to achieve the appropriate operability and stability of the digester. In the first stage, the operability of the digester was achieved with ACR mono-digestion. In the second stage, the digester feed was changed until it reached an ACR:SCS ratio (co-digestion) of 75:25, based on volatile solids, and until stability was achieved. The strategy was successful, and specific biogas production of 0.132 m3 kg-1VS with a methane content of 50.4% was achieved, confirming the technical feasibility of the process. Economic viability was established through a case study at a typical NCS mill. Therefore, anaerobic co-digestion can be consolidated as a technological alternative for the treatment of ACR + SCS and the sustainable benefit of the NCS agro-industry.

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An integrated process for bioethanol production from Miscanthus sacchariflorus was used to construct a bench-scale plant constructed and an economic analysis was carried out to investigate the feasibility of its application to a commercial plant. The bench-scale plant was operated for 1 month and an economic analysis and sensitivity analysis was performed on the data acquired. In this study, 100,000 kL of bioethanol could be produced annually from 606,061 tons of M. sacchariflorus and the production cost was calculated to be US$1.76/L. However, the by-products of this process such as xylose molasses and lignin can be sold or used as a heat source, which can decrease the ethanol production costs. Therefore, the final ethanol production cost was calculated to be US$1.31/L, and is considerably influenced by the enzyme cost. The results and data obtained should contribute to the development of a commercial-scale lignocellulosic bioethanol plant.

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Lignocellulosic biomass is an abundant renewable energy source that can be converted into various liquid fuels via thermochemical processes such as pyrolysis. Pyrolysis is a thermal decomposition method, in which solid biomass are thermally depolymerized to liquid fuel called bio-oil or pyrolysis oil. However, the low quality of pyrolysis oil caused by its high oxygen content necessitates further catalytic upgrading to increase the content of oxygen-free compounds, such as aromatic hydrocarbons. Among the three different types of lignocellulosic biomass components (hemicellulose, lignin, and cellulose), lignin is the most difficult fraction to be pyrolyzed because of its highly recalcitrant structure for depolymerization, forming a char as a main product. The catalytic conversion of lignin-derived pyrolyzates is also more difficult than that of furans and levoglucosan which are the main pyrolysis products of hemicellulose and cellulose. Hence, the main purpose of this study was to develop a bench-scale catalytic pyrolysis process using a tandem catalyst (both in-situ and ex-situ catalysis mode) for an efficient pyrolysis and subsequent upgrading of lignin components. While HZSM-5 was employed as an ex-situ catalyst for its excellent aromatization efficiency, the potential of the low-cost additives of bentonite, olivine, and spent FCC as in-situ catalysts in the Kraft lignin pyrolysis at 500 °C was investigated. The effects of these in-situ catalysts on the product selectivity were studied; bentonite resulted in higher selectivity to aromatic hydrocarbons compared to olivine and spent FCC. The reusability of HZSM-5 (with and without regeneration) was examined in the pyrolysis of lignin mixed with the in-situ catalysts of bentonite, olivine, and spent FCC. In the case of using bentonite and spent FCC as in-situ catalysts, there were no obvious changes in the activity of HZSM-5 after regeneration, whereas using olivine as in-situ catalyst resulted in a remarkable decrease in the activity of HZSM-5 after regeneration.

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To provide information for the design and improvement of full-scale biofilters, pilot-scale biofiltration studies are the current industry standard because they utilize the same filter media size and loading rate as the full-scale biofilters. In the current study, bench-scale biofilters were designed according to a biofilter scaling model from the literature, and the ability of the bench-scale biofilters to accurately represent the organics removal of pilot-scale biofilters was tested. To ensure similarity in effluent water quality between bench- and pilot- or full-scale biofilters at the same influent substrate concentration, the tested model requires that either mass transport resistance or biofilm shear loss takes primacy over the other. The potential primacy of mass transport resistance or biofilm shear loss was evaluated via water quality testing (dissolved organic carbon, specific ultraviolet absorbance, liquid chromatography - organic carbon detection, trihalomethane formation potential, and haloacetic acid formation potential). The biofilters also were characterized for adenosine triphosphate (ATP) content, enzyme activity, extracellular polymeric substances, and microbial community structure. The results of this study indicate that biofilm shear loss takes primacy over mass transport resistance for bench-scale biofilter design in this system; thus, bench-scale biofilters designed in this manner accurately represent organics removal in pilot-scale biofilters. Applying this scaling procedure can reduce filter media requirements from many kilograms to just a few grams and daily water requirements from thousands of liters to less than 10 L. This scaling procedure will allow future researchers to test alternative treatment designs and operating conditions without the need for expensive pilot-scale studies.

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Removing protozoa from a water supply using coagulation, flocculation, dissolved air flotation (DAF) and filtration on a bench scale was evaluated. Calcium carbonate flocculation with and without immunomagnetic separation (IMS) was chosen to detect Giardia spp. cysts and Cryptosporidium spp. oocysts in the studied samples. The results indicated that DAF removed between 1.31 log and 1.79 log of cysts and between 1.08 log and 1.42 log of oocysts. The performance was lower in filtration, with the removal of 1.07 log-1.44 log for cysts and 0.82 log-0.98 log for oocysts. The coagulation, flocculation, DAF and filtration steps removed more than 2.2 log of cysts and oocysts from the water studied. However, protozoa were detected in the filtered water, even with turbidity values of 0.2 NTU. The recovery of the detection method met the international criteria and was higher when there was no IMS. Including the third acid dissociation in the IMS was critical to improve the performance of the protocol tested. However, there was an increase in the technical and analytical complexity and costs. It was also observed that the efficiency of the treatment was linked to the performance of the selected method of detecting protozoa.

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Most pharmaceutical biotechnology companies use stirred-tank bioreactors (STR) for recombinant protein manufacturing. These bioreactors are used at a variety of different scales ranging from bench to production scales, with working volumes from 10 mL to 25,000 L. Bench-scale STRs are commonly used to culture mammalian cells for process development, to troubleshoot production scale bioreactors using scale-down models (SDM), or to conduct fundamental research. In this chapter, we describe the operations of a bench-scale STR for the production of recombinant proteins with suspension-adapted Chinese hamster ovary (CHO-DG44) cells. These operations include bioreactor setup and configuration, batching media, inoculation of the seed cell culture, production phase, and harvest of cell-free fluids.

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A bench-scale facility was developed for the evaluation of thermal imaging cameras. Smoke obscuration conditions in the optical smoke cell were characterized by measuring laser light transmittance through the cell. Measurements showed that the laser transmittance along the axial direction of the optical smoke cell was relatively uniform in the upper and lower halves of the cell for various smoke obscuration conditions. The thermal sensitivity of thermal imagers was investigated using the Michelson Contrast (CM) as a performance metric for a bar target viewed through the smoke-filled cell for different background thermal conditions. The results of the study indicate that the optical smoke cell can be utilized as a well-controlled and effective bench-scale test apparatus to evaluate aspects of the performance of thermal imagers for fire service applications.

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L-asparaginase (LA), an enzyme with anticancer activities, produced by marine-derived Aspergillus niger was subjected to purification and characterization. The purified enzyme was observed to have molecular weight ∼90KDa. The enzyme retained activity over a wide range of pH, i.e. pH 4-10. The enzyme was quite stable in temperature range 20-40°C. Tween 80 and Triton X-100 were observed to enhance LA activity while inhibition of LA activity was observed in presence of heavy metals. The values for Km was found to be 0.8141 mM and Vmax was 6.228µM/mg/min. The enzyme exhibited noteworthy antiproliferative activity against various cancer cell lines tested. Successful bench scale production (in 5L bioreacator) of LA using groundnut oil cake as low cost substrate has also been carried out.

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Bench-scale systems are often used to evaluate pretreatment methods and operational conditions that can be applied in full-scale ultrafiltration (UF) systems. However, the membrane packing density is substantially different in bench and full-scale systems. Differences in concentration factor (CF) at the solution-membrane interface as a result of packing density may impact the mass transfer and fouling rate and the applicability of bench-scale systems. The present study compared membrane resistance when considering raw water (CF = 1) and reject water (also commonly referred to as concentrate water) (CF > 1) as feed in UF systems operated in deposition (dead-end) mode. A positive relationship was observed between the concentration of the organic matter in the solution being filtered and resistance. Bench-scale trials conducted with CF = 1 water were more representative of full-scale operation than trials conducted with elevated CFs when considering membrane resistance and permeate quality. As such, the results of this study indicate that the use of the same feed water as used at full-scale (CF = 1) is appropriate to evaluate fouling in UF systems operated in deposition mode.