Application of Experimental Design Technique to the Optimization of Bench-Scale Composting Conditions of Municipal Raw Sludge

Evaluation of the parameters affecting the compostability of dewatered raw sludge from a wastewater treatment plant has been carried out by means of a full factorial composite experimental design. The factors and their correspondent levels considered in the experimental design were: bulking agent:sludge ratio (from 1:1 to 4:1 by volume), bulking agent particle size (from 0-5 mm to 10-20 mm) and composting volume (in the range of typical laboratory-scale, from 1.5 to 25 L). Experimental design has permitted obtaining a polynomial second-order function that quantitatively describes the influence of the considered factors on the compostability of this waste in laboratory reactors. The function has been numerically optimized to find the optimal values for composting raw sludge resulting in a values of: 0-5 mm for bulking agent size, 1:1 for ratio bulking agent:sludge and a composting volume around 10 L. The use of this technique can be generalized and applied to the composting of other organic wastes and will permit comparison of composting performance of different wastes.     

Effect of beta-glycosidase activity of Oenococcus oeni on the glycosylated flavor precursors of Tannat wine during malolactic fermentation

Under traditional wine-making conditions, this work examines the beta-glycosidic activity of Oenococcus oeni on glycosylated aroma compounds of Tannat wines during malolactic fermentation (MLF) by comparing the changes on selected aglycones liberated. MLF diminished the content of all the glycosylated compounds. The level of the free aroma components was slightly modified by the action of the malolactic fermentation so that the cleavage of the glycosidic linkage by the beta-glycosidic activity of O. oeni did not appear to increase significatively the aglycone contents. The consequences of further chemical rearrangements of the algycones under wine conditions were explored using synthesized glycoconjugates on synthetic medium. Bacteria could also be responsible for the cleavage of aroma glycosylated compounds, being the aglycone adsorbed on polysaccharides or peptidoglycans and was released into the external medium. This hypothesis was studied through the evaluation of a stable arrangement of aroma compounds with polysaccharides produced by lactic acid bacteria. A possible retaining of free-made aroma compounds into the whole cells of O. oeni was also investigated through cell culture analysis. Through the results obtained, we assume stable linkage of aroma compounds with bacterial polysaccharides.     

Biological nitrogen fixation in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) by <Superscript>15</Superscript>N isotope-dilution and identification of associated culturable diazotrophs

The nitrogen-fixing capacity of a range of commercial cultivars of maize (Zea mays L.) was evaluated by the ¹⁵N isotope-dilution method. Biological nitrogen fixation (BNF) expressed as percent nitrogen derived from air (Ndfa) ranged from 12 to 33 regardless of nitrogen fertilization. BNF was not affected by mineral nitrogen fertilization except on cultivar Topacio and PAU-871 cultivars. Subsequently, culturable bacterial diazotrophs were isolated from endophytic tissue of maize: seed, root, stem, and leaf. All isolates were able to grow on N-free semisolid medium. Eleven bacteria isolates showed nitrogen-fixing capacity by the reduction of acetylene to ethylene and confirmed by PCR the presence of nifH gene in their genome. Identification of the 11 isolates was performed by bacteriological methods, 16S rRNA gene sequences, and phylogenetic analysis, which indicated that the bacteria isolated were closely related to Pantoea, Pseudomonas, Rhanella, Herbaspirillum, Azospirillum, Rhizobium (Agrobacterium), and Brevundimonas. This study demonstrated that maize cultivars obtain significant nitrogen from BNF, varying by maize cultivar and nitrogen fertilization level. The endophytic diazotrophic bacteria isolated from root, stem, and leaf tissues of maize cultivars may contribute to BNF in these plants.     

Photoprotection of vitamins in skimmed milk by an aqueous soluble lycopene-gum Arabic microcapsule

Riboflavin (Rf)-mediated photosensitized degradation of vitamins A and D3 in skimmed milk under illumination with a white fluorescence lamp was studied by using the HPLC technique. The photosensitized degradation of both vitamins followed first-order kinetics, and the temperature effect on the observed photodegradation rate constant allowed the determination of the activation energy Ea as being 4 and 16 kcal/mol for vitamins A and D3, respectively. The addition of lycopene microencapsulated by spray-drying with a gum arabic-sucrose (8:2) mixture (MIC) produced a reduction of ca. 45% in the photosensitized degradation rate of both vitamins. Front-face fluorescence experiments showed the same photoprotection factor in the degradation of Rf itself, indicating that the photodegradation mechanism involved Rf-mediated reactive species, such as the excited triplet state of Rf, 3Rf*, and/or singlet molecular oxygen, 1O2. The interaction of both 3Rf* and 1O2 with MIC was evaluated in aqueous solutions by using laser-induced time-resolved absorption or emission spectroscopy, and the contribution of an inner-filter effect in the presence of MIC in skimmed milk was evaluated by diffuse reflectance spectroscopy. The main operating mechanism of photoprotection is due to the deactivation of 3Rf* by the proteic component of gum arabic; thus, gum arabic based microcapsules could be used to improve the photostability of milk during its storage and/or processing under light.     

Lectin from Phaseolus acutifolius var. escumite: chemical characterization, sugar specificity, and effect on human T-lymphocytes

Purification of the lectin from Phaseolus acutifolius var. escumite was achieved by affinity chromatography on a column containing glutaraldehyzed membranes from blood group O erythrocytes. The lectin is a tetrameric glycoprotein of 121 kDa with 10% of sugar by weight composed by four subunits of 30 kDa as determined by SDS-PAGE. The lectin is composed of four isolectins as determined by ion-exchange chromatography on a mono-S column. The lectin and its isolectins showed identical NH2 terminal residues (ANDLSFNFQR FNETN) with homology to the PHA leucoagglutinin-precursor. Peptide mass fingerprint from each lectin isoform determined from tryptic peptides by MALDI-TOF (matrix assisted laser desorption ionization-time-of-flight) showed differences among subunits, thus suggesting microheterogeneity in their amino acid sequences or different glycosylation patterns. The lectin and its four isolectins agglutinated erythrocytes without serological specificity and showed mitogenic activity on human leukocytes; moreover, the main effect was rather toward CD8+ than to CD4+ human peripheral lymphocytes. The lectin from escumite was not inhibitable by simple sugars; however, the specificity of the lectin and its isoforms was mainly addressed toward galactose residues present in bi- or triantennary N-acetyllactosamine-type glycans.     

Proteolytic digestive enzymes and peritrophic membranes during the development of Plodia interpunctella (Lepidoptera: Piralidae): targets for the action of soybean trypsin inhibitor (SBTI) and chitin-binding vicilin (EvV)

The digestive system of P. interpunctella was characterized during its larval development to determine possible targets for the action of proteinaceous enzyme inhibitors and chitin-binding proteins. High proteolytic activities using azocasein at pH 9.5 as substrate were found. These specific enzymatic activities (AU/mg protein) showed an increase in the homogenate of third instar larvae, and when analyzed by individual larvae (AU/gut), the increase was in sixth instar larvae. Zymograms showed two bands corresponding to those enzymatic activities, which were inhibited by TLCK and SBTI, indicating that the larvae mainly used serine proteinases at pH 9.5 in their digestive process. The presence of a peritrophic membrane in the larvae was confirmed by chemical testing and light microscopy. In a bioassay, P. interpunctella was not susceptible to the soybean trypsin inhibitor, which did not affect larval mass and mortality, likely due to the weak association with its target digestive enzyme. EvV (Erythrina velutina vicilin), when added to the diet, affected mortality (LD50 0.23%) and larval mass (ED50 0.27%). This effect was associated with EvV-binding to the peritrophic membrane, as seen by immunolocalization. EvV was susceptible to gut enzymes and after the digestion process, released an immunoreactive fragment that was bound to the peritrophic matrix, which probably was responsible for the action of EvV.     

Photodegradation of the acaricide abamectin: a kinetic study

The acaricide abamectin is a mixture of two colorless homologues in a molar ratio of at least 4:1 with the same structure of macrocyclic lactone. The kinetics of its degradation under direct (254 nm) and dye-sensitized (>400 nm) photoirradiation in methanol solution has been studied by UV-vis spectrophotometry, potentiometric detection of dissolved oxygen, stationary fluorescence, laser flash photolysis, and time-resolved detection of singlet molecular oxygen (O2((1)Delta(g))) phosphorescence. The results indicate that the degradation is very efficient under direct irradiation with UV light (254 nm), with a quantum yield of 0.23. On the contrary, under visible-light irradiation, using the natural pigment riboflavin or the synthetic dye rose bengal as sensitizers, the degradation is very inefficient and proceeds through a O2((1)Delta(g))-mediated mechanism, with a bimolecular rate constant for the overall O2((1)Delta(g)) quenching (the sum of physical and chemical quenching) of 5.5 x 10(5) M(-1) s(-1). This value is similar to those reported for the rate constants of the reactions of O2((1)Delta(g)) with isolated double bonds or conjugated dienes and points to similar processes in the case of abamectin.     

Survival and growth of hatchery-produced postlarvae and spat of the Caribbean scallops Argopecten nucleus and Nodipecten nodosus

A study was made of the growth and survival of postlarvae and spat of Argopecten nucleus and Nodipecten nodosus produced in the laboratory and exposed to different conditions of settlement and culture. An evaluation was made of the different types of collection substrates (onion-bags, artificial-grass, fibreglass-plates), two types of settlement inducers (epinephrine, low temperature), and two culture environments (sea, laboratory). The results showed that larger numbers of postlarvae were obtained in artificial-grass collectors than on fibreglass-plates or in the onion-bags. The growth of the A. nucleus postlarvae was not affected by the collector type, while that of N. nodosus was higher on fibreglass-plates, and lesser in the onion-bags. Both thermal and chemical stimulation of the competent larvae resulted in recovery of a greater percentage of postlarvae. The growth of postlarvae of A. nucleus was better in groups exposed to the two methods of settlement induction, while those of N. nodosus remained unaffected by this treatment. The survival of the postlarvae and spat was greater in the laboratory than at sea except in the case of A. nucleus postlarvae attached to collectors, whose survival was similar in both environments. The growth of postlarvae and spat was greater in the sea than in the laboratory.