Quantitative determination of dietary lectin activities by enzyme-linked immunosorbent assay using specific glycoproteins immobilized on microtiter plates

An immunoenzymatic method for the quantitative determination of dietary lectin activities employing immobilized glycoproteins was studied. Lectins from wheat germ (WGA), peanut (PNA), and jack bean (ConA) were added to microtiter plates coated with ovalbumin or asialofetuin and quantified by enzyme-linked immunosorbent assay (ELISA) with lectin-specific antibodies. ELISA responses for lectin activity were dose-dependent in the concentration range 30-1000 ng/mL for WGA and 80-1000 ng/mL for both PNA and ConA. Inhibition assays carried out with different saccharides confirmed that the binding of lectins to immobilized glycoproteins was specific. The proposed method is specific and sensitive, allowing the quantitative determination of lectin activities on raw samples by simple dilution of the extracts. Examples of application to wheat germ and roasted peanut extracts are reported.     

Boron content in freshwaters of Northern Italy

The B concentration was determined in bulk deposition and in surface freshwaters (lakes and rivers) of the river Po watershed in Northern Italy. The curcumin photometric method was used to determine B content for all analyses. The B concentrations were under detection limits (0.06 mg B L^?1) in bulk deposition and below 0.09 mg B L^?1 in lake waters. Approximately 65 % of river samples measured had B concentrations close to natural background levels for natural waters (0.1 mg B L^?1). There was a strong correlation (r < 0.85) between B concentration and those of both total dissolved P and anionic detergents. The elevated B concentrations may be related to anthropogenic sources.     

Using Monte Carlo simulations to estimate relative fire ignition danger in a low-to-medium fire-prone region

A comprehensive assessment of fire ignition danger is nowadays a basic step towards the prioritization of fire management measures. In this study we propose performing a fire selectivity analysis using Monte Carlo simulations to statistically estimate the relative fire ignition danger in a low-to-intermediate fire-prone region such as Canton Ticino, Switzerland. We define fire ignition danger as the likelihood that at a given place a fire will be ignited. For each 25 m × 25 m pixel of the study area, landscape characteristics that may be related to the probability of fire ignition such as vegetation type, elevation, aspect, slope, urban-forest interface were first split into 9-12 categories. The selectivity of each category with respect to fire ignition was then statistically tested by means of Monte Carlo simulations. Finally, we proposed two different approaches for calculating the ignition danger index: cumulating the scores of the Monte Carlo simulations to a final index or producing synthetic scores by performing a principal component analysis of the Monte Carlo results. The validation of the resulting fire danger indices highlights the suitability of both proposed approaches. The PCA-option allows a slightly better discrimination between ignition and non-ignition points and may be of more general application.     

Chromosomal localisation and genetic variation of the SLC11A1 gene in goats (Capra hircus)

The solute carrier family 11 member A1 (SLC11A1) gene is associated with resistance to infectious diseases. Chromosomal localisation, genomic regions corresponding to functional domains and the genetic variability of microsatellites in the 3' untranslated region (3'-UTR) of this gene were investigated in 427 goats (Capra hircus) of six breeds. Using dual colour fluorescence in situ hybridisation, SLC11A1 was localised to goat chromosome 2. Single strand conformation polymorphism was used to screen for polymorphisms in SLC11A1 exons 2, 10 and 15. There was no variation among goat breeds in the sarcoma homology 3 (SH3) binding motif, the protein kinase C phosphorylation site or the two N-linked glycosylation sites. Exon 15 exhibited variability due to the presence of two polymorphic microsatellites. Genotyping of the upstream guanine-thymine repeat (GTn) at 3'-UTR revealed eight alleles (GT11, GT12, GT14-GT19) in goats, whereas GT13 (present in cattle) was absent. Most goats carried the GT16 allele and no allele was found to be exclusive to only one breed. The coefficient of genetic differentiation value (G(ST)) was 0.084. This microsatellite appears to be an informative DNA marker for genetic linkage analysis in goats.     

DNA content,morphometric and molecular marker analyses of Citrus limonimedica, C. limon and C. medica for the determination of their variability and genetic relationships within the genus Citrus

This work investigated the fingerprinting and phenotyping of Citrus germplasm; species selected were of historical importance belonging to Citrus limonimedica Lush. and its supposed ancestors, along with some other species of the Citrus genus. An integrated approach based on the exploitation of nuclear DNA content, morphological traits and molecular markers, such as RAPD fingerprints and ITS-based SNPs, was employed. We studied a core collection of 54 distinct accessions, including 43 genotypes of the Citrus species (18 species or supposed species) and 11 genotypes of the Poncirus genus, which was used as the reference outgroup. Morphological trait analysis and statistical analysis of DNA content and markers were useful for reconstructing a Citrus phylogeny. In particular, our experiments aimed at estimating the genetic variation within and the genetic relatedness among C limon (L.) Burm., C. limonimedica and C. medica L. to shed light on the hybrid origin hypothesis of C. limonimedica. The results of the multidisciplinary analyses allowed us to confirm a remarkable differentiation between Poncirus and Citrus genera and to highlight a close relationship among the three investigated Citrus species but a distinct difference between these three species and other species in the Citrus genus. RAPD fingerprints and ITS polymorphisms enabled us to point out a variation gradient between lemon and citron, with C. limonimedica as a possible intermediate species. Some accessions of C. medica and C. limonimedica that deviate from such a trend suggest recurrent introgression and/or hybridisation with other species of Citrus.     

Impact of Agro-industrial Byproducts on Bioconversion,Chemical Composition,in vitro Digestibility,and Microbiota of the Black Soldier Fly (Diptera: Stratiomyidae) Larvae

The interest in using byproducts from agro-food industries as a rearing substrate for insects is increasing rapidly. We investigated the influence of byproducts of vegetal origin (okara—a byproduct of soy milk production, maize distillers with solubles, brewer’s grains), used as rearing diet for black soldier fly larvae (BSFL), on the following parameters: biomass production, substrate reduction (SR), nutritional profile and in vitro digestibility, and larval gut microbiota. Hen diet was used as a control substrate. The highest larval biomass was collected on maize distillers, whereas the highest SR was observed on okara. The rearing substrate affected ash, ether extract, and chitin larval content. The BSFL reared on okara were characterized by a lower lauric acid content (17.6% of total fatty acids). Diets also influenced in vitro crude protein digestibility (%) for monogastrics, with the highest values for BSFL reared on maize distillers (87.8), intermediate for brewer’s grains and okara BSFL, and the lowest for hen BSFL (82.7). The nutritive value for ruminants showed a lower Net Energy for lactation for BSFL reared on hen diet than okara and dried maize distillers BSFL. The different byproducts showed an influence on the larval gut microbiota, with a major bacterial complexity observed on larvae fed with the hen diet. The neutral detergent fiber concentration of dietary substrate was negatively correlated with Firmicutes and Actinobacteria relative abundance. Insects valorized byproducts converting them into high-value larval biomass to be used for feed production. The results evidenced the effects of the tested byproducts on the measured parameters, underling the chemical composition importance on the final insect meal quality.     

Response of rainbow trout gill (Na++K+)-ATPase and chloride cells to T3 and NaCl administration

With the aim of comparing the effects of oral T₃ and NaCl administration on trout hypoosmoregulatory mechanisms, three groups of rainbow trout (Oncorhynchus mykiss Walbaum) held in freshwater (FW) were fed a basal diet (C), the same diet containing 8.83 ppm of 3,5,3-triiodo-L-thyronine (T₃) (T) or 10% (w/w) NaCl (N) respectively for 30 d. They were then transferred to brackish water (BW) for 22 d and fed on diet C. Gill (Na⁺+K⁺)-ATPase activity and its dependence on ATP, Na⁺ and pH, number of gill chloride cells (CC), serum T₃ level as well as fish growth, condition factor (K) and mortality were evaluated. During the FW phase, as compared to C trout, T trout showed a two fold higher serum T₃ level, had unchanged gill (Na⁺+K⁺)-ATPase activity and increased CC number, whereas N trout showed higher gill (Na⁺+K⁺)-ATPase activity and CC number. At the end of the experiment the enzyme activity was in the order T>N>C groups and all groups showed similar CC number. Both treatments changed the enzyme activation kinetics by ATP and Na⁺. A transient increase in K value occurred in N group during the period of salt administration. In BW, T and N groups had higher and lower survival than C group respectively. Other parameters were unaffected by the treatments. This trial suggests that T₃ administration promotes the development of hypoosmoregulatory mechanisms of trout but it leaves the (Na⁺+K⁺)-ATPase activity unaltered till the transfer to a hyperosmotic environment.     

Phagocytosis of Astaxanthin-Loaded Microparticles Modulates TGFβ Production and Intracellular ROS Levels in J774A.1 Macrophages

Radiation-induced fibrosis is a serious long-lasting side effect of radiation therapy. Central to this condition is the role of macrophages that, activated by radiation-induced reactive oxygen species and tissue cell damage, produce pro-inflammatory cytokines, such as transforming growth factor beta (TGFβ). This, in turn, recruits fibroblasts at the site of the lesion that initiates fibrosis. We investigated whether astaxanthin, an antioxidant molecule extracted from marine and freshwater organisms, could help control macrophage activation. To this purpose, we encapsulated food-grade astaxanthin from Haematococcus pluvialis into micrometer-sized whey protein particles to specifically target macrophages that can uptake material within this size range by phagocytosis. The data show that astaxanthin-loaded microparticles are resistant to radiation, are well-tolerated by J774A.1 macrophages, induce in these cells a significant reduction of intracellular reactive oxygen species and inhibit the release of active TGFβ as evaluated in a bioassay with transformed MFB-F11 fibroblasts. Micro-encapsulation of bioactive molecules is a promising strategy to specifically target phagocytic cells and modulate their own functions.     

Sulphur Isotopes, Trace Elements and Mineral Stability Diagrams of Waters from the Abandoned Fe–Cu Mines of Libiola and Vigonzano (Northern Apennines, Italy)

The geochemical characteristics of rills draining pyrite-chalcopyrite tailings impoundments and of bordering streams were investigated at the ophiolite-hosted Libiola and Vigonzano abandoned massive sulphide mines, northern Apennines Italy. Water samples were analysed for major and trace chemical composition, hydrogen and oxygen isotope composition, and sulphur isotope composition of aqueous sulphate. Sulphur isotope composition was determined also for some samples of ore sulphides. At Libiola, the newly acquired chemical results on waters corroborate those from previous investigations, thus providing additional support to existing geochemical models in terms of metal distribution, solid phases precipitation, reaction path modelling and mixing reaction paths, and environmental problems. At Vigonzano, the chemical characteristics of waters are similar to those at Libiola. In both localities, solution-secondary phase equilibria estimated using an updated thermodynamic dataset account for mineralogy in the field, including poorly crystalline phases like jurbanite and hydrowoodwardite. The hydrogen and oxygen isotope composition of waters at Libiola and Vigonzano agrees with their meteoric origin. Acid to neutral mine waters do not show any significant isotope shift with respect to the initial water, in spite of the oxidation of even large amounts of pyrite/chalcopyrite ore. The sulphur isotope composition of aqueous sulphate in mine rills at Libiola (δ ³⁴S?=?5.6 to 8.5‰; mean 6.5‰) matches that of massive sulphide ore (δ ³⁴S?=??0.5 to 6.7‰; mean 5.8‰), in keeping with the supergenic origin of the sulphate and related isotope effects in the sulphide oxidation process. Sulphate in mine waters at Vigonzano displays lower δ ³⁴S values in the range 0.6 to 1.5‰. The δ ³⁴S signature of massive ore specimens is within the range reported for most volcanic-hosted massive sulphide deposits, including Cyprus-type deposits.