Purification and biochemical properties of dipeptidyl peptidase I from porcine skeletal muscle

Dipeptidyl peptidase I (DPP I; EC 3.4.14.1) was purified from porcine skeletal muscle after several steps such as heat treatment, ammonium sulfate fractionation, gel filtration chromatography, and HPLC anion exchange chromatography. The purified enzyme showed a native molecular mass of approximately 200 kDa on Sephacryl S-200 column chromatography. Two protein bands of 65 and 42 kDa were obtained by SDS-PAGE, indicating its oligomeric nature. Maximum activity was reached at pH 5.5 and 55 degrees C. DPP I shared some common substrate specificities, both on synthetic derivatives and on real peptides, with porcine muscle DPP III. The enzyme required reducing agents for full activation, although the halide requirement was not proved. DPP I was inhibited by the assayed cysteine peptidase inhibitors except p-CMB. The serine peptidase inhibitor 3, 4-DCI also inhibited the enzyme as did the divalent cations Co(2+), Mn(2+), and Zn(2+). On the basis of its properties, DPP I may contribute to the generation of dipeptides during the processing of meat and/or meat products, including cooked ham.     

Recent Developments in Microbiotesting and Early Millennium Prospects

Small-scale toxicity testing with microbiotests is a rapidly-expanding component of the field of aquatic toxicology which contributes diverse contamination assessment tools and approaches for a variety of environmental (liquid and solid) media. In this short review on microbiotesting, some of the recent developments conducted under the second St. Lawrence River Action Plan (1993–1998) at the St. Lawrence Centre (Environment Canada, Quebec Region, Montreal) are recalled. These include 1) employing the SOS Chromotest to determine the genotoxic status of major industrial effluents discharging to the St. Lawrence River and their potential impact on downstream biota, 2) developing an algal solid phase assay to predict the toxic potential of freshwater sediments, 3) developing a microplate-based cnidarian assay to screen for toxicity of chemicals and environmental samples, 4) developing an alternative assay to whole fish acute (sub)lethal toxicity testing with the help of rainbow trout primary hepatocytes, 5) developing a microplate-based phagocytosis assay to check for immunocompetence of feral bivalve shellfish and 6) conducting a major investigation to develop a cost-effective multitrophic bioanalytical battery to assess the (geno)toxicity of freshwater sediments. In addition, integrative tools with specific microbiotests were respectively constructed to determine the toxic potential of industrial effluents (PEEP: Potential Ecotoxic Effects Probe) and that of sediments (SED-TOX). Such examples illustrate the diversity of on-going endeavors in the field of small-scale toxicity testing internationally, as further corroborated by recent books entirely dedicated to the subject. It is undeniable that many important challenges still lie ahead for this field early into the third millennium and likely well beyond.     

Determination of total dietary fiber in foods and food products: collaborative study

A collaborative study was conducted to determine the total dietary fiber (TDF) content of food and food products, using a combination of enzymatic and gravimetric procedures. The method was basically the same as published earlier (J. Assoc. Off. Anal. Chem. (1984) 67, 1044-1052), with changes in the concentration of alcohol and buffers, time of incubation, sample preparation, and some explanatory notes, all with the intent of decreasing the coefficient of variation (CV) of the method. Duplicate blind samples of soy isolate, white wheat flour, rye bread, potatoes, rice, wheat bran, oats, corn bran, and whole wheat flour were analyzed by 9 collaborators. TDF was calculated as the weight of the residue minus the weight of protein and ash. CV values of the data from all laboratories for 7 of the samples ranged from 1.56 to 9.80%. The rice and soy isolate samples had CV values of 53.71% and 66.25%, respectively; however, each sample contained only about 1% TDF. The enzymatic-gravimetric method for determining TDF has been adopted official first action.     

Differences in common bean rhizobial populations associated with soil tillage management in southern Brazil

Progressive adoption of no-tillage (NT) agriculture in the tropics is finally reversing physical, chemical, and biological erosion of soil and in Brazil, an estimated 19 Mha are now devoted to NT. Common bean (Phaseolus vulgaris L.) is a main component of Brazilian agriculture, and enhancement of yields has been achieved under NT as a result of mitigation of environmental stresses, resulting in higher N₂ fixation. However, the effects of NT on rhizobial diversity are poorly understood. This study evaluated rhizobial diversity in soils planted to common bean under NT or conventional tillage (CT) systems that were compared with natural grassland used for grazing, in the State of Santa Catarina, southern Brazil. Genetic diversity was assessed by the amplification of the DNA by PCR with specific primers (BOX-PCR) and by RFLP-PCR analyses of the 16S rDNA region. A high level of diversity was observed among strains from all three systems, such that the similarity in the clustering analysis of BOX-PCR products ranged from 36% under natural grassland to only 23% for CT strains. High polymorphism was confirmed in the RFLP-PCR analysis; forty-seven different profiles were obtained, none sharing high similarity with the profiles of reference species of common bean rhizobia. These results indicate that other tropical rhizobial species remain to be described. Genetic diversity was higher among the NT than the CT rhizobial strains, especially when the RFLP-PCR profiles were considered. Genetic diversity in the natural grassland was lower than in the cropped systems, possibly due to absence of the host plant and stubble burning in winter. Average yields in the area under NT (e.g. common bean, approximately 1500 kg ha⁻¹) have been about 30% higher than under CT, therefore high rhizobial diversity may be a parameter indicative of superior soil quality.     

Effects of tillage systems on soil characteristics, glomalin and mycorrhizal propagules in a Chilean Ultisol

Tillage affects the soil physical and chemical environment in which soil microorganisms live, thereby affecting their number, diversity and activity. However, soil disturbance generally has the greatest impact on biological properties, including both free and symbiotic fungal populations. Interest in more ecologically sustainable agricultural systems is rising with increasing recognition that agricultural intensification can adversely affect environmental quality. This paper discusses the effect of tillage system on some soil characteristics, such as pH, C, N and S levels, total and Olsen-P contents including some P forms associated with organic matter, glomalin contents and arbuscular mycorrhizae (AM) parameters, such as root colonization, spore number and total and active hyphal length. Measurements were in the sixth year of an on-going tillage-rotation experiment conducted on an Ultisol under no-till (NT), reduced tillage (RT) and conventional tillage with stubble mixed into the soil (CTS) or stubble burnt (CTB). Soil was sampled at two dates; after wheat (Triticum aestivum) harvest (autumn) and 6 months after subsequent grassland seeding (spring). Higher C, N, S, total P and fulvic acid-P concentrations and pH occurred under NT and RT than under CTS and CTB after wheat harvest. However, results at the second sampling were not consistent. AM spore number and active hyphal length were highest under NT having the greatest incidence on AM root colonization and P concentration in shoots of the pasture. Glomalin concentration was higher under NT and RT than under CTS and CTB but no differences in calculated glomalin to total C (ca. 5%) were found. It is concluded that a less disruptive effect of NT influences positively all soil characteristics and also increases P acquisition by the following crop in the rotation system.     

Assessing air-drying and rewetting pre-treatment effect on some soil enzyme activities under Mediterranean conditions

Soil enzyme activities are useful indicators of soil quality as they are very sensitive to disturbance. Sample storage or pre-treatments could affect the results in these assays, which are normally determined in fresh samples, kept cold or frozen. The objectives of this study were to (i) evaluate the effect of air-drying or air-drying and rewetting on β-glucosidase, acid phosphatase and urease activities in soils from different locations, degradation status and sampling seasons, and (ii) assess if air-drying or air-drying and rewetting is an accurate sample storage and pre-treatment procedure for enzyme activities in soil quality evaluations under semiarid Mediterranean conditions. Our results showed that urease, phosphatase and β-glucosidase activities were hardly affected by air-drying of degraded and non-degraded soils from the two locations studied in all seasons. Short incubations (4, 8 and 12 d at 23 °C) of rewetted air-dried soil at 55% of water-holding capacity showed different patterns depending on the enzyme studied. Urease and β-glucosidase activities were relatively stable during incubation, with several significant (P<0.05) shifts up and down in some soils and samplings. However, acid phosphatase showed an increase in activity with incubation, of between 5% and 50% relative to air-dried samples. These increases followed no pattern and were unrelated to soil characteristics or sampling date. Hence, urease, phosphatase and β-glucosidase activities determined in air-dried soil samples seem to be representative of those obtained under field-moist conditions. In contrast, short incubations of rewetted soil samples can produce fluctuations in these enzyme activities, mainly of acid phosphatase, and estimations in these conditions are not so representative of field-moist soil values.     

Increased degradation of phenanthrene in soil by Pseudomonas sp. GF3 in the presence of wheat

A phenanthrene-degrading bacterial strain Pseudomonas sp. GF3 was examined for plant-growth promoting effects and phenanthrene removal in soil artificially contaminated with low and high levels of phenanthrene (0, 100 and 200 mg kg⁻¹) in pot experiments. Low and high phenanthrene treatments significantly decreased the growth of wheat. Inoculation with bacterial strain Pseudomonas sp. GF3 was found to increase root and shoot growth of wheat. Strain GF3 was able to degrade phenanthrene effectively in the unplanted and planted soils. Over a period of 80 days the concentration of phenanthrene in soil in which wheat was grown was significantly lower than in unplanted soil (p<0.05). At the end of the 80-d experiments, 62.2% and 42.3% of phenanthrene had disappeared from planted soils without Pseudomonas sp. GF3 when the phenanthrene was added at 100 and 200 mg kg⁻¹ soil, respectively, but 84.8% and 70.2% of phenanthrene had disappeared from planted soils with the bacterial inoculation. The presence of vegetation significantly enhances the dissipation of phenanthrene in the soil. There was no significant difference in soil polyphenol oxidase activities among the applications of 0, 100 and 200 mg kg⁻¹ of phenanthrene. However, the enzyme activities in planted and unplanted soils inoculated with the strain Pseudomonas sp. GF3 were significantly higher than those of non-inoculation controls. The bacterial isolate was also able to colonize and develop in the rhizosphere soil of wheat after inoculation.     

Nitrogen loading of Eucalyptus globulus seedlings: nutritional dynamics and influence on morphology and root growth potential

New Forests - Achieving successful outplanting of trees on increasingly harsher sites is a global concern. In Chile, for example, new Eucalyptus globulus plantations are being targeted to poorer,...     

Impact of tree coppicing on tree-crop competition in parkland and alley farming systems in semiarid Burkina Faso

The aim of this study was to evaluate the effects of tree coppicing on tree-crop competition in farmed parkland and in alley farming, both in semiarid Burkina Faso.Azadirachta indica A. Juss (neem) was studied in the parklands, while neem,Albizia lebbeck (L.) Benth. (albizia) andLeucaena leucocephala (Lam.) de Wit (leucaena) were investigated in the alley farming system. The crop was sorghum (Sorghum bicolor (L.) Moench) in both cases.Under non-coppiced neem trees in the parkland, the sorghum yield at 0–3 m distance from the tree (under the tree canopy) was 82% of open field yields at 6–9 m distance, but the difference was not significant (p=0.07). Close to trees (0–3 m distance) which had been coppiced before planting, the sorghum yield was 148% of open field yields. In alley farming, early coppicing was superior; sorghum yields were highest close to trees coppiced early and lowest close to trees coppiced late.

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Résumé La présente étude se propose d'évaluer les effets de la taille sur la concurrence entre arbres et cultures dans les parcs arborés et dans les cultures en couloir, en climat semiaride (Burkina Faso). Les espèces étudiées ont été:Azadirachta indica A. Juss (neem) dans les parcs et neem,Albizia lebbeck (L.) Benth. (albizia) ainsi queLeucaena leucocephala (Lam.) de Wit (leucaena) dans les cultures en couloir. Dans les deux cas la plante cultivée concerné a été le sorgho blanc (Sorghum bicolor (L.) Moench).Sous les neems non taillés des parcs, le rendement du sorgho cultivé de 0 à 3 m de l'arbre (sous le bouppier de l'arbre) a atteint 82% du rendement obtenu en plein champ, soit à une distance de 6 à 9 m de l'arbre (différence non significative,p=0.07). Une taille effectuée en début de campagne a entraîné un accroissement du rendement de 148% pour les cultures situées de 0 à 3 m de l'arbre, par rapport aux rendements en plein champ.Dans les cultures en couloir, on a constaté qu'une taille precoce est préférable, c'est à dire que les rendements de sorgho ont été supérieurs dans les lignes proches des arbres taillés tôt que dans celles proches des arbres taillés tard.