3-D Wire Cloth Electrode for Higher Throughput DielectrophoreticSeparation of Bacterial Cell

Dielectrophoresis (DEP) is one of an alternative way for cell separation. It has mainly been limited to processing small volumes due to constrain in fabrication of microelectrode over large surface areas. This work incorporated the wire cloth electrode fabricated using textile technology into a high throughput chamber experiment. The plain-weave wire cloth consists of 71µm stainless steel wires as the microelectrode arrays hold together by polyester yarn warp. This work determines the cell separation yield with parameters on applied voltage, flow rate and cell concentration as well as its optimized variables on the chamber width of 1.2cm and 2.5cm. The optimum voltage achieved was 30Vpk-pk, with flow rate of 3.5 ml/min and maximum cell concentration of 2.08x107 cells/ml. In chamber width comparison, 1.2cm width chamber gives better total percentage yield of 96% than the 2.5cm width chamber of 85% total percentage yield.

The instability of gyrotactically trapped cell layers

Several metres below the coastal ocean surface there are areas of high ecological activity that contain thin layers of concentrated motile phytoplankton. Gyrotactic trapping has been proposed as a potential mechanism for layer formation of bottom-heavy swimming algae cells, especially in flows where the vorticity varies linearly with depth (Durham et al., Science, vol. 323(5917), 2009, pp. 1067–1070). Using a continuum model for dilute microswimmer suspensions, we report that an instability of a gyrotactically trapped cell layer can arise in a pressure-driven plane channel flow. The linear stability analysis reveals that the equilibrium cell-layer solution is hydrodynamically unstable due to negative microswimmer buoyancy (i.e. a gravitational instability) over a range of biologically relevant parameter values. The critical cell concentration for this instability is found to be $N_{c}\simeq 10^{4}~\text{cells}~\text{cm}^{-3}$, a value comparable to the typical maximum cell concentration observed in thin layers. This result indicates that the instability may be a potential mechanism for limiting the layer’s maximum cell concentration, especially in regions where turbulence is weak, and motivates the study of its nonlinear evolution, perhaps, in the presence of turbulence.

FEATURES OF SPIRULINA PLATENSIS CULTIVATED UNDER AUTOTROPHIC AND MIXOTROPHIC CONDITIONS

Cultivation of Spirulina platensis in Zarrouk media containing 0–20 g l-1 glucose was studied in a photobioreactor for 30 days using a light intensity of 3 klux. Various parameters were measured to evaluate the enhancement of cell performance with glucose such as cell number, osmolarity, membrane stability, biomass productivity, doubling time, stress intensity, stress tolerance, chlorophyll, protein, carbohydrates, and lipid contents. Based on the results, we concluded that S. platensis is able to grow and produce some ingredients in Zarrouk media containing up to 20 g l-1 of glucose which is the first to be reported. The cell concentration of the mixotrophic cultures (80 cells per mm2) corresponded well to the sum of the autotrophic cell concentrations (50 cells per mm2), showing that the addition of carbohydrate positively effects on the microalgae growth. The continuous operation supplemented with 0.5 g l-1 of glucose (G0.5) led to the maximum cell concentration about 9.06 g l-1 wet and 1.32 g l-1 dry weights. The highest tolerance index, specific growth rate, biomass productivity, cell division, osmolarity and membrane stability index were respectively 102.5%, 0.15 d-1, 0.04 g l-1d-1, 0.26 div d-1, 0.87 osmol kg-1 and 93.8%, obtained in the same treatment. Chlorophyll (6.7 % in G0; 0.046 g l-1 in G0.5), protein (79.9 % and 0.884 g l-1 in G0.5), carbohydrates (55.5% in G20; 0.492 g l-1 in G6) and lipid (53.3% in G10; 0.636 g l-1 in G0) percentages and yields were mostly enhanced in the mixotrophic condition. This study indicated that mixotrophic growth of S. platensis is useful for commercial biomass production.

PRODUCTION OF DOCOSAHEXAENOIC ACID (DHA) FROM Thraustochytrium sp. ATCC 26185 USING DIFFERENTS NITROGEN CONCENTRATIONS

Polyunsaturated fatty acids (PUFAs) of the type ω3 and ω6 play important physiological functions in human organism, since they are components of cell membranes and brain cells; they decrease the levels of triglycerides and can prevent the incidence of coronary heart disease. Various parameters, including concentration of the nitrogen source in the cultivation of oleaginous microorganisms have been reported to be essential in the biosynthesis and accumulation of PUFAs. The objective of this work is to study the effect of different concentrations of total nitrogen (TN) in the production of PUFAs, especially DHA, from Thraustochytrium sp. ATCC 26185. The concentrations of TN evaluated were 2.4 and 0.8 g/L (batch) and 0.009 g/L (hourly) under fed-batch process. The content of cell biomass, glucose consumption, TN and production of PUFAs was determined. The major composition of the PUFAs in Thraustochytrium sp. ATCC 26185 cell biomass were DPA ω6 (21-25%) and DHA (69-73%), regardless of the type and time of culture. The maximum cell concentration obtained (30.2 g/L) was using 2.4 g/L TN in 168 h of culture. With this same concentration of TN it was possible to produce the highest concentration of DHA (1.16 g/L) in 120 h of culture, demonstrating that the growth of Thraustochytrium sp. ATCC 26185 and yield of PUFAs are dependent on the concentration TN source available for consumption of this oleaginous microorganism, as well as culture time.

PRODUCTION OF DOCOSAHEXAENOIC ACID (DHA) FROM Thraustochytrium sp. ATCC 26185 USING DIFFERENTS NITROGEN CONCENTRATIONS

In this work it was studied the polyunsaturated fatty acids (PUFAs) production, especially DHA, from Thraustochytrium sp. ATCC 26185, under different total nitrogen (TN) availability. Three different TN conditions were evaluated: two with initial concentrations of 2.4 g/L and 0.8 g/L, and the third in a fed-batch process with a rate of 0.009 g/L.h. For each experiment the biomass, glucose, TN and PUFAs were determined. The major composition of the PUFAs in Thraustochytrium sp. ATCC 26185 cell biomass were DPA ω6 (21-25 %) and DHA (69-73 %), regardless of the type and time of culture. The maximum cell concentration (30.2 g/L) was obtained using 2.4 g/L TN in 168 h of culture. With this same concentration of TN it was possible to produce the highest concentration of DHA (1.16 g/L) in 120 h of culture, demonstrating that the growth of Thraustochytrium sp. ATCC 26185 and yield of PUFAs are dependent on the source concentration of TN available for consumption of this oleaginous microorganism, as well as culture time.

Rheological Properties of Cell-Hydrogel Composites Extruding Through Small-Diameter Tips

To form 3D structures with composites of living cells and hydrogel is now becoming an attractive technology in both bioengineering and manufacturing areas. Variant processes have been presented, most of which are based on the cell composite extrusion and deposition technique. So, the design of cell extrusion nozzle turns into an important phase, and the rheological properties of cell-hydrogel composites act as a key parameter and must be well investigated. In this paper, an entrance pressure detection device and adaptor for small-diameter tips were developed. With the hypothesis of power law fluid type, the power law constant and coefficient were calculated. As derived from the experimental results, the cell-hydrogel composites (with the maximum cell concentration of 107∕ml) show no difference from hydrogel materials in rheological properties. Based on these, the manufacturability of cell-hydrogel composites was discussed. Also, the cell viability, which is another important issue in cell assembly technology, was discussed. The peak percentage (97%) was achieved at the flow rate of 0.5mm3∕s. The extruding duration and the maximum shear stress loaded on cells were considered as two influence mechanisms.