Molecular and physiological characterisation of psychrotrophic hydrocarbon-degrading bacteria isolated from Terra Nova Bay (Antarctica)

A set of 21 Antarctic marine bacteria isolated from the Ross Sea and able to utilise diesel fuel as the sole carbon and energy source was characterised. Isolates were analysed by amplified 16S rDNA restriction analysis using the enzyme AluI, resulting in two different groups corresponding to different bacterial species. These species were assigned to the genera Rhodococcus and Alcaligenes, on the basis of 16S rDNA sequencing. This low degree of inter-specific biodiversity was parallel to a low intra-specific biodiversity, as shown by Random Amplified Polymorphic DNA analysis. Then, a 550-bp DNA fragment coding for the inner region of alkane mono-oxygenase was PCR-amplified from the genome of each strain. The phylogenetic analysis of the sequence of the putative AlkB protein coded for by the amplified DNA fragment revealed that these alkB genes were very likely inherited by horizontal gene transfer. Lastly, the analysis of the biodegradation ability of four strains revealed two different strategies of hydrocarbon uptake, mediated either by bio-surfactants and peculiar of Rhodococcus isolates, or by membrane modifications and shown by Alcaligenes isolates. In order to understand the interrelationships between hydrocarbon-degrading isolates, the dynamics of two strains, belonging to Rhodococcus and Alcaligenes, grown together in a co-culture was also followed over a seventeen days period.     

Effect of ochratoxin A-producing Aspergilli on stilbenic phytoalexin synthesis in grapes

Berries of Vitis vinifera L. cv. Barbera were infected, at veraison and during ripening, by a conidial suspension of A. japonicus, A. ochraceus, A. fumigatus and two isolates of A. carbonarius to control ochratoxin A production and stilbene induced synthesis. The experimental design provided also for intact and punctured berries and incubation temperature of 25 degrees C and 30 degrees C. All the tested fungi, except A. fumigatus, significantly increased trans-resveratrol synthesis over the control, while trans-piceid was not affected; only A. ochraceus significantly elicited the berries to synthesize piceatannol. The two isolates of A. carbonarius produced higher amounts of ochratoxin A than did the other fungi. A positive correlation between ochratoxin A and trans-resveratrol synthesis occurred. trans-Resveratrol and piceatannol showed fungicidal activity against A. carbonarius, being able to completely inhibit fungal growth at a concentration of 300 microg/g and 20 microg/g, respectively.     

Extraction of azadirachtin A from neem seed kernels by supercritical fluid and its evaluation by HPLC and LC/MS

A new supercritical extraction methodology was applied to extract azadirachtin A (AZA-A) from neem seed kernels. Supercritical and liquid carbon dioxide (CO(2)) were used as extractive agents in a three-separation-stage supercritical pilot plant. Subcritical conditions were tested too. Comparisons were carried out by calculating the efficiency of the pilot plant with respect to the milligrams per kilogram of seeds (ms/mo) of AZA-A extracted. The most convenient extraction was gained using an ms/mo ratio of 119 rather than 64. For supercritical extraction, a separation of cuticular waxes from oil was set up in the pilot plant. HPLC and electrospray mass spectroscopy were used to monitor the yield of AZA-A extraction.     

Identification of Vigna spp. through specific seed storage polypeptides

Summary Seed globulins of ten Vigna species were separated by means of SDS electrophoresis. Both inter and intraspecific variation were observed.Groups of specific bands which do not vary in the species being present in all the accessions of the species were identified. These species-specific bands, allow the identification of the 10 Vigna spp. analyzed.     

Variation within improved cultivars and landraces of lucerne in Central Italy

Summary A variety must be distinguishable, uniform, stable and of sufficient productivity to be officially registered. In Italy landraces of lucerne are still widely used because they are characterized by a good persistence and productivity in their area of origin and adaptation.The landrace from Casalina (20 km south of Perugia, Central Italy) has higher dry matter production than many other commercial cultivars. The objective of this paper was to measure the variation of the landrace Casalina and compare it with that of the commonest cultivars available on the market and with some lines selected from the landrace Casalina.The experiment consisted of 11 entries: Casalina, two selections from Casalina (one for high seed yield and one for frequent cuttings), the registered landrace Italia Centrale and 7 registered cultivars. Each entry was represented by 80 genotypes transplanted in the field 60×40 cm apart in a randomized block design with 4 replications. In 1991 the following characters were recorded on a single plant basis: time of first flowering, height, number of shoots and dry matter yield at the first harvest, time of second flowering, leafiness, height, number of shoots and dry matter yield at the second harvest, height, number of shoots and dry matter yield at the third and fourth harvest. Height, number of shoots and dry matter yield at the first harvest, dry matter at the second and third harvest were recorded during the 1992 season.Casalina was as variable as the other entries in 77% of the comparisons; it was more variable in 8% of the comparisons and less variable in 15% of the comparisons. In conclusion, the variation of landrace Casalina is equal to or less than that of registered varieties so that it could be directly registered at the National Registry of Varieties.     

Midazolam oxidation in cattle liver microsomes: The role of cytochrome P450 3A

In humans, the cytochrome P450 3A (CYP3A) subfamily is involved in midazolam (MDZ) biotransformation into 1′- and 4-hydroxy metabolites, and the former serves as a probe for CYP3A catalytic activity. In veterinary species is still crucial to identify enzyme- and species-specific CYP substrates; thus, the aim of this study was to characterize MDZ oxidation in cattle liver. A HPLC-UV method was used to measure 1′- and 4-hydroxy MDZ (1′- and 4-OHMDZ, respectively) formation in cattle liver microsomes and assess the role of CYP3A by an immunoinhibition study. Moreover, MDZ hydroxylation was evaluated in 300 cattle liver samples and results were correlated with testosterone hydroxylation. Formation of both metabolites conformed to a single-enzyme Michaelis–Menten kinetics. Values of V_max and K_m were 0.67 nmol/min/mg protein and 6.16 μM for 4-OHMDZ, and 0.06 nmol/min/mg protein and 10.08 μM for 1′-OHMDZ. An anti-rat CYP3A1 polyclonal antibody inhibited up to 50% and 94% 1′- and 4-OHMDZ formation, respectively. MDZ oxidation in liver microsomes was poorly correlated with testosterone hydroxylation. In conclusion, cattle metabolized MDZ to 1′-OHMDZ and 4-OHMDZ. The immunoinhibition results indicated a major contribution of CYP3As to 4-OHMDZ formation and the involvement of other CYPs in 1′-OHMDZ production, paving the way for further investigations.     

Wood properties of young Douglas-fir in Southern Italy: results over a 12-year post-thinning period

This paper describes the study of a 31-year-old Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) stand located in Southern Italy, which was thinned 19 years after planting. The aim of the study is to evaluate the influence of three thinning treatments (unthinned, selective, and geometrical) performed twelve years later on wood density (WD), moisture content, shrinkage, ring width (RW), latewood proportion (LW%), modulus of elasticity, compression (CS), and bending strength (BS). The WD was higher in the unthinned stand. LW% and BS were lowest in the selective thinning and in the geometrical thinning, respectively. No significant differences were found in other variables. In addition, the thinning processes mostly affected the medium tree class more than the dominant and suppressed ones. Regression analysis established a correlation between mechanical characteristics and WD, RW and LW%. Mechanical strength is strongly correlated with WD than other variables. The stepwise model showed that WD and RW are most closely related to the behavior of CS, whereas only WD explained variation in BS. Stand density reduction may improve the development of stands without greatly affecting wood quality.     

Effect of moisture on physical parameters of timber from Turkey oak (<Emphasis Type="Italic">Quercus cerris</Emphasis> L.) coppice in Central Italy

This study aims to investigate wood density at different levels of moisture, basic density and shrinkage of timber from Turkey oak (Quercus cerris L.) coppice forests growing in Central Italy. We also studied the variability in density in the trees within and among sites. Density shows no significant statistical differences in the tested population. A higher variability in the shrinkage than in the density was found. Wood moisture is referred to as dry mass and fresh mass, which is related to many performance characteristics of wood, i.e., energy production. Trends in moisture and water content were studied because these physical parameters play an important role in the specific area of firewood which requires an accurate estimation of mass, volume and energy content. This work is a contribution to improve xylo-energy estimates of small and medium forestry issues.     

DNA fingerprinting and quality traits of Corbarino cherry-like tomato landraces

Twenty-five landraces of the cherry-like tomato named Corbarino, a typical niche product grown in the Sarno valley of the Campania region, have been characterized. The landraces used have been compared to eight cultivars widely spread in the same area of cultivation. The genetic diversity within and between landraces was evaluated through the comparison of DNA fingerprints obtained with (GATA)(4) probe hybridized to TaqI digested genomic DNA. Twenty-two of the Corbarino landraces were homogeneous and were unequivocally characterized by their DNA fingerprints. The others are probably "population varieties" in that within each of them polymorphic DNA fragments were identified. According to the characterization of fruit shapes, four groups were identified. One landrace from each group, harvested at the same ripening degree, was further characterized for yield, fruit quality components, antioxidant activities, and carotenoid contents and compared to Faino F(1) and Tomito F(1) hybrids. The Corbarino landraces were all characterized by high yield, a high level of carbohydrates, and high levels of soluble and total solids. Interestingly, the biotype named ISCI 05 was the most suitable for canning. Landraces ISCI 07 and ISCI 05 gave the highest value of antioxidant activities and carotenoid content. These data, together with previously published results, suggested that ISCI 05 should be proposed for the institution of a Label of Origin.     

Hydrolysis of wine aroma precursors during malolactic fermentation with four commercial starter cultures of Oenococcus oeni

The ability of four commercial preparations of Oenococcus oeni lactic acid bacteria (EQ 54, Lalvin OSU, Uvaferm Alpha, and Lalvin 31) to hydrolyze wine aroma precursors was evaluated by measuring the concentration of free and bound aroma compounds at the end of malolactic fermentation carried out in model wines containing a mixture of glycosides extracted from Muscat wine. At pH 3.4 there was a decrease in glycosylated compounds matched by a concomitant increase in free forms in all starter cultures tested. When malolactic fermentation was carried out at pH 3.2, a significant decrease in the ability to hydrolyze aroma precursors was observed for two of the cultures tested (Uvaferm Alpha and Lalvin 31). Large differences in the extent of hydrolysis and in the specificity of this activity toward specific aroma precursors were observed and appeared to be related to the chemical structure of the aglycon as well as to individual characteristics of each starter culture. The amounts of glycosylated aroma compounds released during malolactic fermentation suggest that O. oeni can alter the sensory characteristics of wine through the hydrolysis of aroma precursors.