Factors affecting the extraction and the relative proportions of multiple forms of plant α-galactosidases

The extractability of α-galactosidase activity from mature Vicia faba seeds and the conversion of the low molecular weight form II to the larger oligomer I, was examined over a range of salt concentrations. Specific and total activities of the preparations were high when strong salt solutions were used. Extraction of α-galactosidase I, in comparison with II, requires solutions with a high ionic strength (e.g. 0.5 M NaCl). Interpretation of gel filtration patterns are, however, complicated by conversion of II to I which occurs under these conditions. This conversion is also enhanced by routine procedures used for enzyme purification, such as citric acid precipitation.     

Efficacy of Metarhizium anisopliae in controlling the two‐spotted spider mite Tetranychus urticae on common bean in screenhouse and field experiments

The efficacy of aqueous and emulsifiable formulations of the fungus Metarhizium anisopliae isolate ICIPE78 was evaluated on the population density of Tetranychus urticae infesting common bean plants under screenhouse and field conditions. Synthetic acaricide abamectin was included as a check. Bean plants were artificially infested with T. urticae and allowed to multiply. Three treatments were applied in the screenhouse and 1 treatment in field trials. Mite density was recorded 2 d before spraying and weekly postspraying. The number of pods per plant, number of seeds per pod, and the dry weight of seeds per plant were recorded only in the screenhouse trials. In both screenhouse and field trials, fungal formulations applied at the concentration of 10⁸ conidia/mL and the acaricide reduced the population density of mites as compared to the controls. There were significant differences in T. urticae population densities between the treatments at the various post‐spraying sampling dates. In the screenhouse, the mite densities were near zero from 3‐week postspraying in the treated leaves. At 4‐week postspraying, there were no more leaves in the untreated control (T1) and in the control water + Silwet‐L77 (T2). Fungal formulations were as effective as abamectin in reducing mite densities in both screenhouse and field experiments. There were significant differences in the production parameters during the 2 screenhouse trials, with fungal and abamectin treatments generally having the highest yield. Results of this study underline the potential of the M. anisopliae isolate ICIPE78 as an alternative to acaricides for T. urticae management.     

Antifungal properties of selected lactic acid bacteria and application in the biological control of ochratoxin A producing fungi during cocoa fermentation

Thirteen Lactic acid bacteria strains isolated from fermenting cocoa and seven reference strains were used in order to assess their antifungal properties towards three ochratoxin A (OTA) producing fungi (Aspergillus carbonarius, Aspergillus niger and Aspergillus ochraceus). Furthermore, two of the isolates strains (A19 and A21) identified as belonging to the genus of Pediococcus as well as Lactobacillus plantarum B4496, Lactobacillus brevis 207 and Lactobacillus sanfranciscensis BB12 showed interesting in vitro broad antifungal activities towards the three ochratoxin-producing fungi with inhibition percentages ranging from 15% to 66.7%. Treatment of cell-free supernatant at 100°C affected antifungal activity suggesting that the main compounds responsible for this activity were of proteic nature, and hence could be bacteriocins. Application of isolate A19 in cocoa fermentation as starter inhibited the growth of each of the OTA-producing species. At the end of fermentation in boxes inoculated with A19, A. niger was not detectable while A. carbonarius concentration was found to be 2 Log CFU/g of wet beans. The assessment of the ochratoxin produced during fermentation of cocoa inoculated with A. carbonarius indicated that the use of isolate A19 as starter could reduce their level of growth so as to have only a toxin production of 0.0012 ± 0.0005 μg/kg after 40 days of storage, while this was 2.45 ± 0.35 μg/kg of fermented and dried cocoa beans in the absence of A19. This work is a contribution for the application of biological control of OTA-producing fungi during cocoa production.     

Sister Chromatid Exchanges Induced by Heavy Metals in Vicia Faba

The induction of sister chromatid exchanges (SCE) by chloride and nitrate salts of nickel, cobalt, cadmium and zinc were studied in meristematic root cells of Vicia faba. Salts of nickel, cobalt and cadmium significantly increased the frequency of SCE, whereas chloride and nitrate salts of zinc did not increase the frequency of SCE significantly above the spontaneous level. The reported data demonstrate that the induction of SCE in Vicia faba may represent a valuable bioindicator for detecting the cytogenetic damage of heavy metals.     

Isolation,purification and characterization of phytase from germinating mung beans

Phytase isolated from mung bean cotyledons was purified about 80-fold with a recovery of 28%. The enzyme is stable at 0°, has a pH optimum at 7·5 and optimal temperature of 57°. The energy of activation is approximately 8500 cal/mole between 37° and 57°. Inhibition by P_i has been found to be competitive, the K_i value being 0·40–0·43 × 10⁻³ M; the K_m value with phytate is 0·65 × 10⁻³ M. Divalent cations are not required for activity. Lower members of inositol phosphates are better substrates, as shown by their V_max and K_m values. When subjected to polyacrylamide gel electrophoresis two bands have been resolved; one (major) corresponds to phytase and the other (minor) to phosphatase and pyrophosphatase activity. Filtration through Biogel P-200 partially resolves phytase from phosphatase and pyrophosphatase. The molecular weight of phytase is approximately 160,000.     

Influence of extremely low-frequency electric fields on the growth of Vigna radiata seedlings

The biological effects of extremely low-frequency electric fields (ELF) on living organisms have been explored in many studies, but the results are controversial and only a few studies investigated the influence of the intensity of the applied field on seedling growth. Here we assess the effects of a 50 Hz sinusoidal electric field on the early growth of Vigna radiata seedlings while varying the field intensity. Experiments performed in a dark, constant-climate chamber on several thousands of seedlings show that the field produces an inhibitory effect at a low field intensity and an enhancing one at a higher intensity. The maximum negative effect occurs at about 450 V/m, which is an intensity much lower than the exposure limits currently in force in the safety regulations.     

大鼠压杆行为的运动皮层局部场电位研究

目的：提取与大鼠右前肢运动相关联的初级运动皮层场电位的信号特征,并探讨依据局部场电位（LFP）识别前肢运动行为的可行性。方法：4只SD大鼠通过训练习得压杆取水操作,然后在左右脑初级运动皮层M1区分别植入多通道束状微电极,术后恢复后进行大鼠压杆行为实验,以2 kHz/s速率记录深部脑电信号及压杆状态信号,同步记录行为过程的视频信号。以通道间的差分信号作为局部场电位信号,分析局部场电位信号的时域特征,进行聚类分析。以压杆状态信号和视频分析为判定依据,对聚类结果进行分析。结果：局部场电位信号在大鼠压杆动作时明显增强,不同通道的局部场电位信号幅值、波形有差异,表明与前肢运动相关联的M1区局部场电位信号有空间分布特征;依据阈值准则从局部场电位信号检测压杆行为的检出率为80%。结论：依据局部场电位信号特征对大鼠前肢运动进行检测具有可行性。     

Determination of the triacylglycerol composition of coffee beans by reverse-phase high-performance liquid chromatography

Reverse-phase HPLC with refractive index and light scattering detectors in isocratic and gradient elution modes, respectively, was applied for the separation of the major triacylglycerols (TAG) in coffee lipids. Twelve TAG species could be identified and determined using a linear gradient of acetonitrile in dichloromethane: dichloroethane. The quantitative evaluation was based on the relative area percentages derived directly from a data-station. The procedure was applied to determine the TAG composition of three types of coffee beans harvested in two coffee producing areas in Brazil and dried by two commonly used procedures. No significant differences in the TAG compositions due to the type, origin and drying procedure were found.     

Mycoflora and Potential for Mycotoxin Production of Freshly Harvested Black Bean from the Argentinean Main Production Area

A mycological survey was carried out, for the first time, on black bean samples from the northwestern Argentinean province of Salta in the 1999 harvest season. Ten varieties of black beans were evaluated at three locations. Species of the genus Alternaria were the most prevalent component of the black bean mycoflora. Species of Fusarium, Sclerotinia, Rhizoctonia and Acremonium were also recorded. The predominant species of the genera isolated were Alternaria alternata, Sclerotinia sclerotiorum, Rhizoctonia solani, Fusarium semitectum and Acremonium strictum. An analysis of variance was applied to determine possible differences between black bean varieties. Variety FT88/519 was the most susceptible to Sclerotinia sclerotinia infection, while the variety DOR 604 was the least susceptible. As toxigenic species were recovered, Alternaria toxins, zearalenone and trichothecenes may pose a contamination risk for black bean.