Effect of differently post-treated dewatered sewage sludge on β-glucosidase activity,microbial biomass carbon,basal respiration and carbohydrates contents of soils from limestone quarries

This work has evaluated the effects of thermally dried (TDS) or composted (CDS) dewatered sewage sludge on β-glucosidase activity, total (TCH) and extractable (ECH) carbohydrate content, microbial biomass carbon and basal respiration of soils from limestone quarries under laboratory conditions. Two doses (low and high) of the dewatered sludge (DS) or of the respective TDS or CDS were applied to a clayey and a sandy soil, both coming from working quarries. The soil mixtures and the controls (soils with no added sludge) were incubated for 9 months at 25°C and 30% of field capacity. The addition of sludge increased all the studied soil parameters, and the increase depended on the amount of sludge. Except in the case of TCH and ECH, the enhancing effect decreased with time, but at the end of incubation, parameters of the treated soils were higher than those of the control. The rank order of the initial stimulating effect was soil–TDS ≥ soil–DS ≥ soil–CDS, and probably, this order depended on the proportion of stable organic matter, which was the lowest in the TDS. Values of metabolic quotient (qCO₂) were higher at the lower dose, and they did not change during incubation in the CDS-treated soils. Both TCH and ECH were the parameters with the greatest significant sludge and dose effects. Basal respiration, microbial biomass carbon and β-glucosidase activity were the best measured parameters in distinguishing the long-term effects of the three sludge types over the soils.     

Diversity in White Lupin (<Emphasis Type="Italic">Lupinus albus</Emphasis> L.) Landraces from Northwest Iberianplateau

All the accessions (35) of white lupin (Lupinus albus L.) landraces collected from northwest Iberian plateau and maintained at the Spanish germplasm collection (CRF of INIA), were studied with the general objective of assessing the biodiversity of these landraces and to ascertain their value as genetic resources for the development of germplasm adapted to the areas where they were collected with long and chilly winters. The characterization study comprised 51 qualitative and 50 quantitative characters. Quantitative parameters were analyzed by Principal Components Analysis (PCA). The 2-dimensional plot (49.3% of cumulative variability) formed a main group of accessions and 4 outliers (#1, #9, #27 and #28) separated in the first PC. The characters with a higher contribution to the first PC were inflorescence length, leaf petiole length and leaf central foliole area. The presence of alkaloids, the percentage of plants killed by soil borne fungal diseases, the phenology, the yield per plant and yield components were also assessed. Twenty six accessions (the main group in the PCA plot) showed high homogeneity, with the following highlights: bitterness, indeterminate growth habit, early ripeness and spring sowing with the exception of #18, in which seeding date is unknown and it had a later phenology. By contrast accessions #9 and #27 were bitter of autumn sowing with determinate growing habit and they had the longest values of primary inflorescence length and a later phenology when they were sown in spring. Average values for yield and yield components showed a broad range of variation between accessions. The main yield component was the number of pods per plant. Accession #22 had the highest mean seed mass, although this had not significant influence in yield. Accession #17 showed the highest yield and #1 the lowest. This last also had the lowest values of inflorescence length, leaf petiole length and leaf central foliole area. No significant differences of resistance to soil-borne fungi were found between accessions. The year had a significant effect in the phenology, yield and yield components except for number of seeds per pod. The studied material might be of interest for the development of spring sowing germplasm adapted to north Iberian peninsula.     

A calorimetric study of thermal denaturation of recombinant human lactoferrin from rice

Thermal denaturation of recombinant human lactoferrin from transgenic rice with different degrees of iron saturation has been studied by differential scanning calorimetry. The maximum temperature, enthalpy change, and activation energy of denaturation were higher when recombinant lactoferrin was more saturated with iron, indicating an increase in the stability of the protein structure. Maximum temperature and activation energy values for apo- and holo-lactoferrins were practically identical to those reported for the same forms of lactoferrin from human milk, which indicates a similar thermal stability. However, the value of enthalpy change for denaturation of the recombinant lactoferrin was 2.5-3-fold lower than that found for the human milk protein. This finding may reflect the influence that the different glycosylation pattern may have in the relationship between lactoferrin domains. Denaturation of recombinant lactoferrin in milk was compared with denaturation in phosphate buffer, and results indicated that the protein was more heat-sensitive when treated in milk than in buffer.     

Broccoli processing wastes as a source of peroxidase

A peroxidase isozyme (BP) was purified to homogeneity from broccoli stems ( Brassica oleraceae var. maraton) discarded from industrial processing wastes. BP specific activity was 1216 ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)] units/mg, representing 466-fold that of crude extract. BP is a monomeric glycoprotein containing 16% carbohydrates, with a molecular mass of 49 kDa and an isoelectric point close to 4.2. From kinetic data it showed a two-substrate ping-pong mechanism, and the catalytic efficiency measured as the rate-limiting step of free BP regeneration was 3.4 x 10(6) M(-1) s(-1). The ABTS K m value was 0.2 mM, which was about 20 times lower than that reported for acidic commercial horseradish peroxidase (HRP). Assessment of BP secondary structure showed 30% helical character, similar to HRP and cytochrome c peroxidase. BP lost only 25% activity after 10 min of heating at 55 degrees C and pH 6; it was stable in the pH range from 4 to 9 and showed an optimum pH of 4.6 using ABTS as substrate. BP was active on substrates normally involved in lignin biosynthesis, such as caffeic and ferulic acids, and also displayed good catechol oxidation activity in the presence of hydrogen peroxide. Reverse micellar extraction was successfully used as potential large-scale prepurification of broccoli peroxidase, achieving a purification factor of 7, with 60% activity yield. Stems from the broccoli processing industry have a high potential as an alternative for peroxidase purification.     

Enzyme activities and microbial community structure in semiarid agricultural soils

This study investigated the effect of management on -glucosidase, -glucosaminidase, alkaline phosphatase, and arylsulfatase activities and the microbial community structure in semiarid soils from West Texas, USA. Surface samples (0–5 cm) were taken from a fine sandy loam, sandy clay loam, and loam that were under continuous cotton ( Gossypium hirsutum L.) or in cotton rotated with peanut ( Arachis hypogaea L.), sorghum ( Sorghum bicolor L.), rye ( Secale cereale) or wheat ( Triticum aestivum L.), and had different water management (irrigated or dryland), and tillage (conservation or conventional). The enzyme activities were higher in the loam and sandy clay loam than in the fine sandy loam. Soil pH was not affected by management, but the soil organic C and total N contents were generally affected by the different crop rotations and tillage practices studied. The trends of the enzyme activities as affected by management depended on the soil, but in general crop rotations and conservation tillage increased the enzyme activities in comparison to continuous cotton and conventional tillage. The soil enzyme activities were significantly correlated with the soil organic C ( r -values up to 0.90, P< 0.001), and were correlated among each other ( r -values up to 0.90, P <0.001). There were differences in the fatty acid methyl ester profiles between the fine sandy loam and the sandy clay loam and loam, and they reflected the differences in the enzyme activities found among the soils. For example, a 15:0 ranged from 1.61±0.25% in cotton-peanut/irrigated/no-till in the fine sandy loam to 3.86±0.48% in cotton-sorghum/dryland/conservation tillage in the sandy clay loam. There were no differences due to management within the same soil.Trade names and company names are included for the benefit of the reader and do not infer any endorsement or preferential treatment of the product by USDA-ARS     

Effect of germination on the carbohydrate composition of the dietary fiber of peas (Pisum sativum L.)

The effect of different conditions of pea germination on dietary fiber (DF) composition was studied. Insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) were subjected to acid hydrolysis, and the resultant neutral sugars, uronic acids, and Klason lignin were quantified. Germinated peas exhibited significantly higher contents of total dietary fiber (TDF) than the raw sample, due to the increases of both DF fractions. Under darkness conditions, germination exhibited the highest contents of IDF and SDF. Decreasing IDF/SDF ratios showed that the carbohydrate changes did not take place to the same extent during germination, the SDF fraction being the most affected. The detailed chemical composition of fiber fractions reveals increases of cellulose in the IDF of germinated samples, whereas SDF exhibits a decrease of pectic polysaccharides and also increases of polysaccharides rich in glucose and mannose. The DF results were corroborated by a comparative examination of the cell wall carbohydrate composition.     

Purification and characterization of an N-acetylglucosamine-binding lectin from Koelreuteria paniculata seeds and its effect on the larval development of Callosobruchus maculatus (Coleoptera: Bruchidae) and Anagasta kuehniella (Lepidoptera: Pyralidae)

This study describes the purification of an N-acetylglucosamine-binding lectin from Koelreuteria paniculata seeds and its effects on the larval development of Callobruchus maculatus and Anagasta kuehniella. The lectin (KpLec) was characterized and isolated by gel filtration, affinity column, and reverse phase chromatography. SDS-PAGE indicated that this lectin is a dimer composed of subunits of 22 and 44 kDa. The N terminus exhibited 40% similarity with Urtiga dioica agglutinin. KpLec was tested for anti-insect activity against C. maculatus and A. kuehniella. With regard to C. maculatus, an artificial diet containing 0.7 and 1% KpLec produced LD(50) and ED(50) value, respectively. However, for A. kuenhiella, an artificial diet containing 0.65% KpLec produced an LD(50), whereas 0.2% KpLec produced an ED(50). The transformation of genes coding for this lectin could be useful in the development of insect resistance in important agricultural crops.     

Organophosphorus pesticide residues in Mexican commercial pasteurized milk

A study was conducted to measure residues of 13 organophosphorus (OP) pesticides, widely used as dairy cattle ectoparasiticides or in crops used for animal feed, in homogenized and pasteurized Mexican milk samples. Four different milk brands with high distribution were collected biweekly during a 12 month period (n = 96) in supermarkets. OP pesticide residues were measured by gas chromatography with a flame photometric detector. Approximately 39.6% of the samples contained detectable levels of OP pesticide residues. Eight samples contained residues exceeding established maximum residue limits (MRL), and the OP pesticides present in these samples were dichlorvos (five samples), phorate, chlorpyrifos, and chlorfenvinphos (one sample, respectively). Average residues of 13 OP pesticides measured were below established MRLs ranging between 0.0051 and 0.0203 ppm.     

Multiple domestications of the Mesoamerican gene pool of lima bean (Phaseolus lunatus L.): evidence from chloroplast DNA sequences

The Mesoamerican (MA) gene pool of lima bean (Phaseolus lunatus L.) is widely distributed from northern Mexico to northern Argentina in its wild forms and from the southern United States to the east coast of Brazil in its domesticated forms. This broad distribution and lack of wild accessions of the MA gene pool in many areas of its natural distribution has impeded determining its center of domestication and establishing whether it has a single or multiple centers. To answer these questions, we evaluated 262 accessions of P. lunatus using two intergenic spacers of chloroplast DNA: atpB-rbcL and trnL-trnF. The data were analyzed using a maximum likelihood tree (ML), a haplotype network and two estimators of genetic differentiation (N _ST and G _ST). Nucleotide diversity (π) and haplotype (Hd) were quantified to estimate the percentage of reduction in genetic diversity (%r) as a founder effect. The ML tree and haplotype network indicated the existence of three groups (AI, MI and MII), which was supported by the high values of N _ST (0.61–0.80). Values for %r were high (58.67–60.83 %). Existence of Mesoamerican and Andean gene pools was confirmed, with two genetically and geographically distinct groups (MI and MII) within the MA gene pool. We present the first evidence for multiple origins of domestication for the MA gene pool. For MI, we propose western central Mexico as the domestication area and between Guatemala and Costa Rica for MII. We observed a founder effect in the MA gene pool as a result of domestication.     

Usefulness of amino acid composition to discriminate between honeydew and floral honeys. Application to honeys from a small geographic area

With the aim of finding methods that could constitute a solid alternative to melissopalynological and physicochemical analyses to determine the botanical origin (floral or honeydew) of honeys, the free amino acid content of 46 honey samples has been determined. The honeys were collected in a small geographic area of approximately 2000 km(2) in central Spain. Twenty-seven honey samples were classified as floral and 19 as honeydew according to their palynological and physicochemical analyses. The resulting data have been subjected to different multivariant analysis techniques. One hundred percent of honey samples have been correctly classified into either the floral or the honeydew groups, according to their content in glutamic acid and tryptophan. It is concluded that free amino acids are good indicators of the botanical origin of honeys, saving time compared with more tedious analyses.