Production, purification, and properties of an endoglucanase produced by the hyphomycete Chalara (Syn. Thielaviopsis) paradoxa CH32

The hyphomycete Chalara (syn. Thielaviopsis) paradoxa produces endoglucanase activity during the late trophophase. The low molecular mass (35 kDa) endoglucanase purified from cultured broths works optimally at 37 degrees C and pH 5.0. The enzyme inactivates at pH below 3.0 and also at temperatures of 50 degrees C or higher, but it is stable at lower temperatures, including refrigeration temperature and freezing. The enzyme is inhibited by detergents, by EDTA, and by the divalent cations Hg(2+) and Ag(2+). It is also inhibited to some extent by 10 mM Zn(2+), Fe(2+), and Mg(2+), but it is stimulated by Mn(2+). Enzyme activity is not affected by reducing agents. In the presence of low concentrations of water miscible organic solvents (20%) endoglucanase activity is inhibited by 7% (for methanol) to 50% (for acetonitrile), and it is totally inhibited at higher solvent concentrations (50%). Enzyme activity is not affected by the water immiscible solvent ethyl acetate. Carboxymethylcellulose is the preferred substrate (K(m(app)) = 8.3 g/L; V(max(app)) = 1.1 microM/min). Hydrolysis of crystalline cellulosic substrates is very limited, but it is greatly enhanced by phosphoric acid swelling. The purified enzyme shows no activity toward disaccharides or aryl-glucosides. Its activity is inhibited by cellobiose.     

Significance of soil properties in the adsorption and mobility of the fungicide metalaxyl in vineyard soils

Adsorption and mobility of the fungicide metalaxyl were studied in 16 vineyard soils from the La Rioja region (Spain), with organic matter (OM) contents in the 0.31--1.37% range, and in 7 natural soils with OM contents in the 3.30--8.24% range. Adsorption isotherms were obtained using the batch equilibrium technique, and mobility was studied by soil thin-layer chromatography (soil-TLC). In all cases, the adsorption isotherms fit the Freundlich equation. The values of the K(f) constants were low in the vineyard soils (0.01--0.64) and increased in the natural soils (1.05--2.83). The n(f) values were in general lower than unity. K(f) constants were significantly correlated (p < 0.001) with the OM content when all of the soils were considered. According to the determination coefficient, r(2), OM would account for 88% of the variance in adsorption. When the vineyard soils alone were considered, a significant correlation was seen between K(f) and the OM and clay contents; both parameters, varying simultaneously, explain 80% of the variance in adsorption. Study of the mobility of metalaxyl with soil-TLC indicated that in vineyard soils the fungicide has the potential for being highly mobile in 19% of the soils and mobile in 81% of them. In natural soils, the fungicide has the potential for being moderately mobile or mobile in 86 and 14% of the soils, respectively. This type of behavior of metalaxyl indicates that in vineyards soils of the La Rioja region (Spain) with low OM contents, where application of the compound is continuous, a leaching of the fungicide from the soil to groundwaters could potentially occur. These results should be borne in mind when metalaxyl is to be used in the soils of this region.     

Determination of phytohormones of environmental impact by capillary zone electrophoresis

A test mixture of five phytohormones [naphthaleneacetic acid (NAA), naphthoxyacetic acid (NOA), indoleacetic acid (IAA), indolebutyric acid (IBA), and indolepropionic acid (IPA)] was investigated. These compounds were cleanly separated with good resolution by capillary zone electrophoresis with a UV diode array detector using 20 mM sodium phosphate buffer (pH 7.25). The lowest detection limit was obtained for IPA (0.45 mg L(-)(1) or 0.005 mg kg(-)(1)) and the highest for NAA (1.04 mg L(-)(1) or 0.014 mg kg(-)(1)). The method has been applied for tomato samples fortified with the five phytohormones using a liquid-liquid extraction procedure, obtaining recovery percentages ranging from 91 to 109.0%.     

Further studies on the analysis of DSP toxin profiles in galician mussels

Further studies on mussel samples from Galicia, Spain, have revealed the presence of okadaic acid (OA), dinophysistoxin 2 (DTX2), and the fatty acid acyl esters of both of these toxins as the "DTX3" complex. Measurements were performed with an improved in situ method for the formation of 9-anthryldiazomethane (ADAM) derivatives followed by liquid chromatography with fluorescence detection. Base hydrolysis of DTX3 toxins gave free OA and DTX2, which were determined following ADAM derivatization. Results were confirmed by liquid chromatography/mass spectrometry analyses, and in most of the samples, free DTX2 was the most abundant toxin. However, the OA/DTX2 ratio in the DTX3 conjugated form was different, with OA being the most abundant in all cases. This difference could be due to different rates of metabolism of OA and DTX2 to the acyl esters or due to contamination of the shellfish by the two toxins at different points in time, resulting in less acyl ester formation for one toxin versus the other. The second possibility would be reasonable if two different source organisms were producing the toxins.     

Intensive beach management as an improved sea turtle conservation strategy in Mexico

We evaluate the role of intensive beach management, meaning intense patrolling and nest reburial to a central hatchery, as a strategy for improving the success of sea turtle conservation at nesting sites in Mexico. We report the results of an experimental program at Playa Cuixmala, Jalisco, western Mexico. Sea turtle conservation efforts in Mexico have, in general, poor results because of lack of funds, which leads to insufficient beach protection and severe negative effects of nest removal on hatching success and sex ratios. Alternative strategies are needed to optimize limited resources. We predicted that intensive beach management, which included intense patrolling and careful nest reburial, could be an effective way to maximize nest survival and hatchling release under limited financial and human resources. The results of our 9-year study were very positive. Survival rate increased several fold during the study period. Hatching success and sex ratios were not significantly different between in-situ and removed nests. Survival in removed nests was, however, much higher that in-situ nests, because of predation and beach erosion. In total, the small (3 km length) Playa Cuixmala became the second most productive sea turtle nesting beach in the region because of these concentrated efforts. Intense beach management can be an important technique for sea turtle conservation, and can be properly applied to small beaches or the most productive sections of large beaches.     

The hydrophobic character of peanut (Arachis hypogaea) isoagglutinins

Peanut seed lectin (PNA) is widely used to identify tumor-specific antigens on the eukaryotic cell surface. In this work PNA was purified by affinity chromatography, using a column containing glutaraldehyde-treated human erythrocytes, whereas PNA isoforms were purified by hydrophobic interaction chromatography using Phenyl-Sepharose. The affinity-purified PNA and its isoforms consist of four equal subunits of 24.5 kDa each, all of which agglutinated human sialidase-treated erythrocytes equally well; however, differences in their relative thermostabilities and sugar specificities for lactose were observed. Fractions PNA-I and PNA-II possess higher affinity for lactose residues than the more hydrophobic isoforms III and IV. These findings suggest that the differences observed in PNA isoagglutinins are due to hydrophobic regions of the protein that influence the three-dimensional organization of the molecule as well as its thermal stability and sugar specificity.     

Hydrological properties of a Mediterranean soil burned with different fire intensities

The influence of vegetation cover on soil hydrological properties and its response to the impact of different fire intensities, in a Mediterranean forest environment, has been evaluated. The study was carried out in the Permanent Experimental Field Station of La Concordia (Llíria–Valencia, Spain), on a set of nine erosion plots (4 × 20 m²). The Station is located on a calcareous hillside S–SE oriented, with soils of Rendzic Leptosol type and supporting Mediterranean shrubland vegetation. All runoff generated and sediment produced in every rain event was collected from each plot. The set up includes a system of sensors for the continuous monitoring of climatic parameters (air temperature and humidity, rain volume, intensity, etc.).In June 1995, a set of experimental fires was carried out to the Station. Three of the plots were burned with high intensity fire, three with moderate intensity and the remaining were left unaltered. Soil water content and water retention capacity (WRC) were measured in the different plots and in two different vegetation covers: under canopy (UC) and in bare soil (BS). The pF curves were also obtained for each fire treatment.A year after the fires (June 1995–June 1996), great differences, reaching 77.15%, in runoff generation between fire treatments and the control plots were observed.No significant differences were detected on water retention capacity between soils UC and BS in the burned plots. However, these differences appeared in the control plots, giving UC and BS values of 13% and 18%, respectively. Plots corresponding to the high intensity fire treatment showed values of WRC significantly higher than those of the moderate intensity and of the control treatments.The pF curves show that the values of water volume, at the different pressure points studied, were slightly greater on UC soil. Values obtained for BS samples are higher in the fire treatments, showing significant differences in respect to the control plots at pF 1 and 2. These differences were also observed for UC soil, but in this case at pF 2, 2.5 and 4.2.