Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata.

Pseudokirchneriella subcapitata is a unicellular green algae widely distributed in freshwater and soils. Due to its cosmopolitan characteristic, its use is recommended by national and international protocols in ecotoxicity studies. The alteration of phosphatase activities by agriculture pollutants like heavy metals has been extensively used as a biomarker in risk assessment and biomonitoring. In this study, we compared the extraction of acid phosphatase from P. subcapitata by different procedures and we studied the stability, substrates specificity, kinetics and the effect of Hg2+ in the crude extract. The freezing and thawing technique associated with probe sonication was the most suitable method of extraction. The enzyme was stable when frozen at -20ºC for at least six months, showed an optimum pH of 5 and a Km value of 0.27 mM for p-nitrophenylphosphate (pNPP) as substrate. Some natural organic substrates were cleaved by a similar extent as the synthetic substrate pNPP. Short term exposure (24 hours) to Hg2+ had little effect but inhibition of the specific activity was observed after 7 days with EC50 (concentration of Hg2+ that promotes 50% decrease of specific activity) value of 12.63 μM Hg2+ .

An algebraic pedotransfer function to calculate standardized in situ determined field capacity.

Despite the large applicability of the field capacity (FC) concept in hydrology and engineering, it presents various ambiguities and inconsistencies due to a lack of methodological procedure standardization. Experimental field and laboratory protocols taken from the literature were used in this study to determine the value of FC for different depths in 29 soil profiles, totaling 209 soil samples. The volumetric water content (θ) values were also determined at three suction values (6 kPa, 10 kPa, 33 kPa), along with bulk density (BD), texture (T) and organic matter content (OM). The protocols were devised based on the water processes involved in the FC concept aiming at minimizing hydraulic inconsistencies and procedural difficulty while maintaining the practical meaning of the concept. A high correlation between FC and θ(6 kPa) allowed the development of a pedotransfer function (Equation 3) quadratic for θ(6 kPa), resulting in an accurate and nearly bias-free calculation of FC for the four database geographic areas, with a global root mean squared residue (RMSR) of 0.026 m3·m-3. At the individual soil profile scale, the maximum RMSR was only 0.040 m3·m-3. The BD, T and OM data were generally of a low predicting quality regarding FC when not accompanied by the moisture variables. As all the FC values were obtained by the same experimental protocol and as the predicting quality of Equation 3 was clearly better than that of the classical method, which considers FC equal to θ(6), θ(10) or θ(33), we recommend using Equation 3 rather than the classical method, as well as the protocol presented here, to determine in-situ FC.