The exponential fed-batch cultivation of Bacillus sphaericus UPMB10 in 2 l stirred tank fermenter was performed by feeding the initial batch culture with 14 g l-1 of glycerol according to the algorithm aimed at controlling the specific growth rate (μ) of the bacterium. Very high viable cell count (1.14 x 1010 cfu ml-1), which was four times higher as compared to batch cultivation, was achieved in the fed-batch with a controlled μ at 0.4 h-1. In repeated exponential fed-batch cultivation, consisting of four cycles of harvesting and recharging, a final cell concentration of 1.9 x 1011 cfu ml-1 was obtained at the end of the fourth cycle (46 h). Meanwhile, acetylene reduction of cell samples collected from repeated fed-batch cultivation remained unchanged and was maintained at around 20 nmol C2H2 h-1 ml-1 after prolonged cultivation period, and was comparable to those obtained in batch and exponential fed-batch cultivation. Glycerol could be used as a carbon source for high performance cultivation of B. sphaericus, a nitrogen fixing bacterium, in repeated fed-batch cultivation with high cell yield and cell productivity. The productivity (0.68 g l-1 h-1) for repeated fed-batch cultivation increased about 6 times compared to that obtained in conventional batch cultivation (0.11 g l1 h-1). A innovative method in utilizing glycerol for efficient cultivation of nitrogen fixing bacterium could be beneficial to get more understanding and reference in manipulating the integrated plans for sustainable and profitable biodiesel industry.
The aim of this study is to prepare a model for the production of Newcastle disease virus (NDV) lentogenic F strain using cell culture in bioreactor for live attenuated vaccine preparation. In this study, firstly we investigated the growth of Vero cells in several culture media. The maximum cell number was yielded by culture of Vero cells in Dulbecco's Modified Eagle Medium (DMEM) which was 1.93 x 10(6) cells/ml. Secondly Vero cells were grown in two-litre stirred tank bioreactor by using several commercial microcarriers. We achieved the maximum cell concentration about 7.95 x 10(5) cells/ml when using Cytodex 1. Later we produced Newcastle Disease virus in stirred tank bioreactor based on the design developed using Taguchi L4 method. Results reveal that higher multiplicity of infection (MOI) and size of cell inoculums can yield higher virus titer. Finally, virus samples were purified using high-speed centrifugation based on 3( * *)(3-1) Fractional Factorial Design. Statistical analysis showed that the maximum virus titer can be achieved at virus sample concentration of 58.45% (v/v), centrifugation speed of 13729 rpm, and centrifugation time of 4 hours. As a conclusion, high yield of virus titer could be achieved through optimization of cell culture in bioreactor and separation by high-speed centrifugation.
In this study, a novel continuous reactor has been developed to produce high quality methyl esters (biodiesel) from palm oil. A microporous TiO2/Al2O3 membrane was packed with potassium hydroxide catalyst supported on palm shell activated carbon. The central composite design (CCD) of response surface methodology (RSM) was employed to investigate the effects of reaction temperature, catalyst amount and cross flow circulation velocity on the production of biodiesel in the packed bed membrane reactor. The highest conversion of palm oil to biodiesel in the reactor was obtained at 70 °C employing 157.04 g catalyst per unit volume of the reactor and 0.21 cm/s cross flow circulation velocity. The physical and chemical properties of the produced biodiesel were determined and compared with the standard specifications. High quality palm oil biodiesel was produced by combination of heterogeneous alkali transesterification and separation processes in the packed bed membrane reactor.
Hydrocarbon pollution in marine ecosystems occurs mainly by accidental oil spills, deliberate discharge of ballast waters from oil tankers and bilge waste discharges; causing site pollution and serious adverse effects on aquatic environments as well as human health. A large number of petroleum hydrocarbons are biodegradable, thus bioremediation has become an important method for the restoration of oil polluted areas. In this research, a series of natural attenuation, crude oil (CO) and dispersed crude oil (DCO) bioremediation experiments of artificially crude oil contaminated seawater was carried out. Bacterial consortiums were identified as Acinetobacter, Alcaligenes, Bacillus, Pseudomonas and Vibrio. First order kinetics described the biodegradation of crude oil. Under abiotic conditions, oil removal was 19.9% while a maximum of 31.8% total petroleum hydrocarbons (TPH) removal was obtained in natural attenuation experiment. All DCO bioreactors demonstrated higher and faster removal than CO bioreactors. Half life times were 28, 32, 38 and 58 days for DCO and 31, 40, 50 and 75 days for CO with oil concentrations of 100, 500, 1000 and 2000 mg/L, respectively. The effectiveness of Corexit 9500 dispersant was monitored in the 45 day study; the results indicated that it improved the crude oil biodegradation rate.
The main objective of this work was to determine the effectiveness of various biofouling reducers (BFRs) to operational condition in hybrid membrane bioreactor (MBR) of palm oil mill effluent (POME). A series of tests involving three bench scale (100 L) hybrid MBR were operated at sludge retention times (SRTs) of 30 days with biofouling reducer (BFR). Three different biofouling reducers (BFRs) were powdered actived carbon (PAC), zeolite (Ze), and Moringa oleifera (Mo) with doses of 4, 8 and 12 g L(-1) respectively were used. Short-term filtration trials and critical flux tests were conducted. Results showed that, all BFRs successfully removed soluble microbial products (SMP), for PAC, Ze, and Mo at 58%, 42%, and 48%, respectively. At their optimum dosages, PAC provided above 70% reductions and 85% in fouling rates during the short-term filtration and critical flux tests.
Bacillus strain NS 8, a lipase-producing bacterium isolated from a Malaysian hot spring, is able to tolerate a broad range of temperature and pH, which makes it beneficial for this study. It generated PCR products with molecular weight of 1,532 bp, and the 16S rRNA sequence analysis identified it as Bacillus subtilis with accession number AB110598. It showed a 71% similarity index with B. subtilis using Biolog Microstation System. Its lipase production was optimized using a shake flask system by changing the physical (agitation speed, pH and temperature) and nutritional (nitrogen, carbon and minerals) factors. The most suitable combination of the basal medium for lipase production was 2.5% olive oil (carbon), 1.5% peptone (nitrogen), 0.1% MgSO(4) (mineral) at an optimum temperature of 50°C, pH 7.5 and 150 rpm agitation, giving an enzyme yield of 4.23 U/ml. Statistical optimization using response surface methodology was carried out. An optimum lipase production of 5.67 U/ml was achieved when olive oil concentration of 3%, peptone 2%, MgSO(4)·7H(2)O 0.2% and an agitation rate of 200 rpm were combined. Lipase production was further carried out inside a 2-liter bioreactor, which yielded an enzyme activity of 14.5 U/ml after 15 h of incubation.
Solid-state fermentation (SSF) was employed to enhance the nutritive values of palm kernel cake (PKC) for poultry feeding. Aspergillus flavus was isolated from local PKC and utilized to increase the mannose content of PKC via the degradation of β-mannan in PKC; evaluation was done for batch SSF in Erlenmeyer flasks and in a novel laterally aerated moving bed (LAMB) bioreactor. The optimum condition for batch SSF in flasks was 110% initial moisture content, initial pH 6.0, 30 °C, 855 μm particle size, and 120 h of fermentation, yielding 90.91 mg mannose g⁻¹ dry PKC (5.9-fold increase). Batch SSF in the LAMB at the optimum condition yielded 79.61 mg mannose g⁻¹ dry PKC (5.5-fold increase) within just 96 h due to better heat and mass transfer when humidified air flowed radially across the PKC bed. In spite of a compromise of 12% reduction in mannose content when compared with the flasks, the LAMB facilitated good heat and mass transfer, and improved the mannose content of PKC in a shorter fermentation period. These attributes are useful for batch production of fermented PKC feed in an industrial scale.
A mechanical-conditioning bioreactor has been developed to provide bi-axial loading to three-dimensional (3D) tissue constructs within a highly controlled environment. The computer-controlled bioreactor is capable of applying axial compressive and shear deformations, individually or simultaneously at various regimes of strain and frequency. The reliability and reproducibility of the system were verified through validation of the spatial and temporal accuracy of platen movement, which was maintained over the operating length of the system. In the presence of actual specimens, the system was verified to be able to deliver precise bi-axial load to the specimens, in which the deformation of every specimen was observed to be relatively homogeneous. The primary use of the bioreactor is in the culture of chondrocytes seeded within an agarose hydrogel while subjected to physiological compressive and shear deformation. The system has been designed specifically to permit the repeatable quantification and characterisation of the biosynthetic activity of cells in response to a wide range of short and long term multi-dimensional loading regimes.
High cell density cultivation of microalgae via heterotrophic growth mechanism could effectively address the issues of low productivity and operational constraints presently affecting the solar driven biodiesel production. This paper reviews the progress made so far in the development of commercial-scale heterotrophic microalgae cultivation processes. The review also discusses on patentable concepts and innovations disclosed in the past four years with regards to new approaches to microalgal cultivation technique, improvisation on the process flow designs to economically produced biodiesel and genetic manipulation to confer desirable traits leading to much valued high lipid-bearing microalgae strains.
The influence of water activity and water content was investigated with farnesyl laurate synthesis catalyzed by Lipozyme RM IM. Lipozyme RM IM activity depended strongly on initial water activity value. The best results were achieved for a reaction medium with an initial water activity of 0.11 since it gives the best conversion value of 96.80%. The rate constants obtained in the kinetics study using Ping-Pong-Bi-Bi and Ordered-Bi-Bi mechanisms with dead-end complex inhibition of lauric acid were compared. The corresponding parameters were found to obey the Ordered-Bi-Bi mechanism with dead-end complex inhibition of lauric acid. Kinetic parameters were calculated based on this model as follows: V (max) = 5.80 mmol l(-1) min(-1) g enzyme(-1), K (m,A) = 0.70 mmol l(-1) g enzyme(-1), K (m,B) = 115.48 mmol l(-1) g enzyme(-1), K (i) = 11.25 mmol l(-1) g enzyme(-1). The optimum conditions for the esterification of farnesol with lauric acid in a continuous packed bed reactor were found as the following: 18.18 cm packed bed height and 0.9 ml/min substrate flow rate. The optimum molar conversion of lauric acid to farnesyl laurate was 98.07 ± 0.82%. The effect of mass transfer in the packed bed reactor has also been studied using two models for cases of reaction limited and mass transfer limited. A very good agreement between the mass transfer limited model and the experimental data obtained indicating that the esterification in a packed bed reactor was mass transfer limited.
A membrane sequencing batch reactor (MSBR) treating hypersaline oily wastewater was modeled by artificial neural network (ANN). The MSBR operated at different total dissolved solids (TDSs) (35,000; 50,000; 100,000; 150,000; 200,000; 250,000mg/L), various organic loading rates (OLRs) (0.281, 0.563, 1.124, 2.248, and 3.372kg COD/(m(3)day)) and cyclic time (12, 24, and 48h). A feed-forward neural network trained by batch back propagation algorithm was employed to model the MSBR. A set of 193 operational data from the wastewater treatment with the MSBR was used to train the network. The training, validating and testing procedures for the effluent COD, total organic carbon (TOC) and oil and grease (O&G) concentrations were successful and a good correlation was observed between the measured and predicted values. The results showed that at OLR of 2.44kg COD/(m(3)day), TDS of 78,000mg/L and reaction time (RT) of 40h, the average removal rate of COD was 98%. In these conditions, the average effluent COD concentration was less than 100mg/L and met the discharge limits.
Biohydrogen is regarded as an attractive future clean energy carrier due to its high energy content and environmental-friendly conversion. It has the potential for renewable biofuel to replace current hydrogen production which rely heavily on fossil fuels. While biohydrogen production is still in the early stage of development, there have been a variety of laboratory- and pilot-scale systems developed with promising potential. This work presents a review of advances in bioreactor and bioprocess design for biohydrogen production. The state-of-the art of biohydrogen production is discussed emphasizing on production pathways, factors affecting biohydrogen production, as well as bioreactor configuration and operation. Challenges and prospects of biohydrogen production are also outlined.
The effects of different hydraulic retention time (HRT) on (RS)-MCPP utilisation was investigated by decreasing the feed flow rate in an anaerobic membrane bioreactor (AnMBR). Results showed an average COD removal efficiency of 91.4%, 96.9% and 94.4% when the reactor was operated at HRT 3, 7 and 17 d, respectively. However, when the HRT was reduced to 1d, the COD removal efficiency declined to just only 60%, confirming the AnMBR is stable to a large transient hydraulic shock loads. The (RS)-MCPP removal efficiency fluctuated from 6% to 39% at HRT 3 d, however when it was increased to 7 and 17 d, the removal efficiency increased to an average of 60% and 74.5%. In addition, (RS)-MCPP specific utilisation rates (SUR) were dependent on the HRT and gradually improved from 18 to 43 μg mg VSS(-1) d(-1) as flow rate increased.
The present work aims to address the gas-phase biotransformation of geraniol into citronellol using growing cells of Saccharomyces cerevisiae (baker's yeast) in a continuous-closed-gas-loop bioreactor (CCGLB). This study revealed that the gaseous geraniol had a severe effect on the production of biomass during the growing cell biotransformation resulting in the decrease in the specific growth rate from 0.07 to 0.05 h⁻¹. The rate of reaction of the growing cell biotransformation was strongly affected by agitation and substrate flow rates. The highest citronellol concentration of 1.18 g/L and initial rate of reaction of 7.06 × 10⁻⁴ g/min g(cell) were obtained at 500 rpm and 8 L/min, respectively.
The UASB process among other treatment methods has been recognized as a core method of an advanced technology for environmental protection. This paper highlights the treatment of seven types of wastewaters i.e. palm oil mill effluent (POME), distillery wastewater, slaughterhouse wastewater, piggery wastewater, dairy wastewater, fishery wastewater and municipal wastewater (black and gray) by UASB process. The purpose of this study is to explore the pollution load of these wastewaters and their treatment potential use in upflow anaerobic sludge blanket process. The general characterization of wastewater, treatment in UASB reactor with operational parameters and reactor performance in terms of COD removal and biogas production are thoroughly discussed in the paper. The concrete data illustrates the reactor configuration, thus giving maximum awareness about upflow anaerobic sludge blanket reactor for further research. The future aspects for research needs are also outlined.
The performance of moving bed sequencing batch reactors (MBSBRs) added with 8 % (v/v) of polyurethane (PU) foam cubes as carrier media in nitrogen removal was investigated in treating low COD/N wastewater. The results indicate that MBSBR with 8-mL cubes achieved the highest total nitrogen (TN) removal efficiency of 37% during the aeration period, followed by 31%, 24% and 19 % for MBSBRs with 27-, 64- and 125-mL cubes, respectively. The increased TN removal in MBSBRs was mainly due to simultaneous nitrification and denitrification (SND) process which was verified by batch studies. The relatively lower TN removal in MBSBR with larger PU foam cubes was attributed to the observation that larger PU foam cubes were not fully attached by biomass. Higher concentrations of 8-mL PU foam cubes in batch reactors yielded higher TN removal.
The objective of this study was to evaluate the effects ofNi(II) and Cr(VI) individually and in combination on the simultaneous removal of chemical oxygen demand (COD), nitrogen and metals under a sequencing batch reactor (SBR) operation. Three identical laboratory-scale SBRs were operated with FILL, REACT, SETTLE, DRAW and IDLE periods in a ratio of 1:12:1:2:8 for a cycle time of 24 h until the steady state was achieved. Nickel(II) at increasing concentrations up to 35 mg/L was added to one of the reactors; Cr(VI) at increasing concentrations up to 25 mg/L was added to a second reactor; while a combination of Ni(II) and Cr(VI) in equal concentrations up to 10 mg/L was added to a third reactor. The results demonstrate that both Ni(II) and Cr(VI) exerted a more pronounced inhibitory effect on the removal of ammonia nitrogen (AN) than on COD removal. Synergistic and antagonistic inhibitory effects on the rates of COD and AN removal, respectively, were observed for the 50% Ni(II) and 50% Cr(VI) (w/w) mixture in the concentration range between 10 and 20 mg/L. The simultaneous presence of 50% Ni(II) and 50% Cr(VI) at a concentration of 20 mg/L resulted in system failure.
Agitation speed was found to influence the tannase production and fungal growth of Aspergillus niger FETL FT3. The optimal agitation speed was at 200 rpm which produced 1.41 U/ml tannase and 3.75 g/l of fungal growth. Lower or higher agitation speeds than 200 rpm produced lower enzyme production and fungal growth. Based on the SEM and TEM micrograph observation, there was a significant correlation between agitation speed and the morphology of the fungal mycelia. The results revealed an increase of the enzyme production with the change of the fungal growth morphology from filamentous to pelleted growth forms. However, the exposure to higher shear stress with an increasing agitation speed of the shaker also resulted in lower biomass yields as well as enzyme production.
The granulation process in palm oil mill effluent using calcium oxide-cement kiln dust (CaO-CKD) provides an attractive and cost effective treatment option. In this study the efficiency of CaO-CKD at doses of 1.5-20 g/l was tested in batch experiments and found that 10 g of CaO/l caused the greatest degradation of VFA, butyrate and acetate. An upflow anaerobic sludge blanket (UASB) reactor was operated continuously at 35°C for 150 days to investigate the effect of CaO-CKD on sludge granulation and methanogenesis during start-up. The treatment of POME emphasized the influence of varying organic loading rates (OLR). Up to 94.9% of COD was removed when the reactor was fed with the 15.5-65.5 g-CODg/l at an OLR of 4.5-12.5 kg-COD/m(3)d, suggesting the feasibility of using CaO in an UASB process to treat POME. The ratio of volatile solids/total solids (VS/TS) and volatile fatty acids in the anaerobic sludge in the UASB reactor decreased significantly after long-term operation due to the precipitation of calcium carbonate in the granules. Granulation and methanogenesis decreased with an increase in the influent CaO-CKD concentration.
A study has been carried out to define the effect of drastic temperature changes on the performance of lab-scale hollow-fibre MBR in treating municipal wastewater at a flux of 10 L m(-2) h(-1) (LMH). The objectives of the study were to estimate the activated sludge properties, the removal efficiencies of COD and NH(3)-N and the membrane fouling tendency under critical conditions of drastic temperature changes (23, 33, 42 & 33 °C) and MLSS concentration ranged between 6,382 and 8,680 mg/L. The study exhibited that the biomass reduction, the low sludge settleability and the supernatant turbidity were results of temperature increase. The temperature increase led to increase in SMP carbohydrate and protein, and to decrease in EPS carbohydrate and protein. The BRE of COD dropped from 80% at 23 °C to 47% at 42 °C, while the FRE was relatively constant at about 90%. Both removal efficiencies of NH(3)-N trended from about 100% at 33 °C to less than 50% at 42 °C. TMP and BWP ascended critically with temperature increase up to 336 and 304 mbar respectively by the end of the experiment. The values of suspended solids (SS) and the turbidity in the final effluent were negligible. The DO in the mixed liquor was varying with temperature change, while the pH was within the range of 6.7-8.3.