Evaluation of various techniques for microalgal biomass quantification.

Biomass concentration is one of the most important parameters in the biotechnology processes. Its measurement relies on the physical, chemical or biological properties of the cells. Several techniques were applied in this work to measure the cell concentration of four microalgae: Botryococcussp., Botryococcusbraunii, Chlorella vulgaris, and Ettlia sp. The experiments were performed using samples taken from a chemostat for each strain to provide microalgal cell suspensions in a stable physiological state and concentration. The dry cell weight (DCW) was used as the reference method for the evaluation of other methods. The two commercial sensors used to determine optical density and dielectric permittivity showed a broad effective measurement range up to more than 20gl(-1). A Red-Green-Blue model analysis of microalgal digital images in combination with Fourier equation significantly extended the measurements range up to 6gl(-1). Cell count using a flow cytometer showed a broad range of linearity to DCW in washed samples, but other counting methods using hemocytometer and microscopic automated count were limited. Finally, the oxygen production rate, representing the photosynthetic activity, showed a linear regression with DCW at cell concentrations lower than 1gl(-1).

Aeration effects on metabolic events during sporulation of Bacillus thuringiensis.

The metabolism of Bacillus thuringiensis during its sporulation process was investigated under different concentrations of oxygen. At the beginning of sporulation, the aeration conditions were regulated to obtain different oxygen transfer rates (OTR) in four separate fermentations, representing interrupted, limited, non-limited, and saturated oxygenation, respectively. A higher OTR resulted in a higher pH, up to about 9 in the case of saturated oxygenation, while the interrupted oxygenation resulted in a significantly acidic culture. In contrast, the absence of oxygen resulted in rapid sporangia lysis and caused acidification of the medium, indicating a distinctly different sporangia composition and different metabolism. The bacterium also showed different CO2 production rates during sporulation, although a maximum point was observed in every case.With a higher OTR, the maximal value was observed after a longer time and at a lower value (40, 26, and 13 mmol/L/h for limited, non-limited, and saturated cases, respectively). Despite the exhaustion of glucose prior to the sporulation phase, the interrupted oxygenation resulted in acetate, lactate, and citrate in the medium with a maximum concentration of 4.8, 1.3, and 5.0 g/L, respectively. Notwithstanding, while the metabolic events differed visibly in the absence of oxygen, once sporulation was triggered, it was completed, even in the case of an interrupted oxygen supply.

Growth, sporulation, delta-endotoxins synthesis, and toxicity during culture of bacillus thuringiensis H14.

Growth, sporulation, synthesis of delta-endotoxins, and toxicity against the larvae of Aedes aegypti and Culex pipiens were studied during fermentation of Bacillus thuringiensis H14 in a 20-L fermentor. Measurements of optical density and dielectric permittivity for biomass determination suggest a highly promising technique for on-line evaluation of sporulation. The synthesis of 65-, 25- and 130-kDa proteins started at 16, 18, and 23 h, respectively. These proteins were enriched in different ways until the end of culture (48 h). Toxicity in the course of sporulation was significantly different for the larvae of both mosquito species. Maximal activity against Ae. aegypti was obtained at the end of culture, whereas for Cx. pipiens, the sample at 38 h was the most active.