Changes in foliar carbon isotope composition and seasonal stomatal conductance reveal adaptive traits in Mediterranean coppices affected by drought

We estimated water-use efficiency and potential photosynthetic assimilation of Holm oak(Quercus ilex L.) on slopes of NW and SW aspects in a replicated field test examining the effects of intensifying drought in two Mediterranean coppice forests. We used standard techniques for quantifying gas exchange and carbon isotopes in leaves and analyzed total chlorophyll, carotenoids and nitrogen in leaves collected from Mediterranean forests managed under the coppice system. We postulated that responses to drought of coppiced trees would lead to differential responses in physiological traits and that these traits could be used by foresters to adapt to predicted warming and drying in the Mediterranean area. We observed physiological responses of the coppiced trees that suggested acclimation in photosynthetic potential and water-use efficiency:(1) a significant reduction in stomatal conductance(p0.01) wasrecorded as the drought increased at the SW site;(2) foliar δ13C increased as drought increased at the SW site(p0.01);(3) variations in levels of carotenoids and foliar nitrogen, and differences in foliar morphology were recorded, and were tentatively attributed to variation in photosynthetic assimilation between sites. These findings increase knowledge of the capacity for acclimation of managed forests in the Mediterranean region of Europe.     

Reversing agriculture from intensive to sustainable improves soil quality in a semiarid South Italian soil

Intensive agriculture (IA) is widespread in South Italy, although it requires frequent tillage, large amounts of fertilizers and irrigation water. We have assessed the efficacy of reversing IA to sustainable agriculture (SA) in recovering quality of a typical South Italy soil (Lithic Haploxeralf). This reversion, lasting from 2000 to 2007, replaced 75% of nutrients formerly supplied inorganically by farmyard manuring and reduced the tillage frequency. Several chemical and biochemical properties, functionally related to C and N mineralisation–immobilisation processes and to P and S nutrient cycles, were monitored annually from 2005 to 2007 in the spring. Reversing IA to SA decreased soil bulk density, almost doubled the soil organic matter (SOM) as favoured the immobilisation of C and N, increased most soil microbial indicators but decreased contents of nitrate, mineral N and K₂SO₄-extractable C. The K₂SO₄-extractable C/K₂SO₄-extractable organic N ratio suggested that substrate quality rather than the mass of readily available C and N affected biomass and activity of soil microflora. Also, the largely higher 10-day-evolved CO₂–C-to-inorganic N ratio under SA than IA indicated that higher C mineralisation, associated with higher microbial biomass N immobilisation, occurred under SA than IA. Decreases in most soil enzyme activities under IA, compared to SA, were much higher than concomitant decreases in SOM content. Soil salinity and sodicity were always higher in IA than SA soil, although not critically high, likely due to the intensive inorganic fertilisation as irrigation waters were qualitatively and quantitatively the same between the two soils. Thus, we suggest that the cumulative small but long-term saline (osmotic) and sodic (dispersing) effects in IA soil decreased the microbial variables more than total organic C and increased soil bulk density.     

Leishmania DNA load and cytokine expression levels in asymptomatic naturally infected dogs

The factors responsible for the clinical progress of visceral leishmaniasis (VL) in dogs have not been yet established. The starting hypothesis was the possibility of associating the changing level of a specific type of cytokines with the evolution of the infection towards infection-manifested disease or resistant behaviour. For this purpose the authors have established a connection between Leishmania load, cytokine mRNA accumulation, and the progression of the disease in naturally infected asymptomatic dogs. We made use of real-time (RT) PCR system to detect the expression of cytokine mRNA levels during all the phases of the infection. In particular, we measured the amount of parasites in samples such as blood, lymph nodes and skin, and the expression levels of IFN-gamma, IL-2, IL-4, IL-10, IL-12 and IL-18 cytokines in the blood. We employed different targeted real-time PCR assay on 40 naturally infected dogs, initially asymptomatic; 20 of these progressed to overt disease, and the 20 remaining dogs remained asymptomatic throughout the period of study (2 years). Two other groups included: 20 naturally infected dogs with clinical signs of VL, and 20 healthy dogs living in a non-endemic area. All these animals were employed as positive and negative controls, respectively. The overall results obtained demonstrate that the simultaneous evaluation of parasites and cytokine levels represents a reliable tool for predicting disease development, and thus for choosing the best treatment for the asymptomatic form of the disease.     

Glucoraphasatin and glucoraphenin, a redox pair of glucosinolates of brassicaceae, differently affect metabolizing enzymes in rats

Brassica vegetables are an important dietary source of glucosinolates (GLs), whose breakdown products exhibit anticancer activity. The protective properties of Brassicaceae are believed to be due to the inhibition of Phase-I or induction of Phase-II xenobiotic metabolizing enzymes (XMEs), thus enhancing carcinogen clearance. To study whether GLs affect XMEs and the role of their chemical structure, we focused on two alkylthio GLs differing in the oxidation degree of the side chain sulfur. Male Sprague-Dawley rats were supplemented (per oral somministration by gavage) with either glucoraphasatin (4-methylthio-3-butenyl GL; GRH) or glucoraphenin (4-methylsulfinyl-3-butenyl GL; GRE), at 24 or 120 mg/kg body weight in a single or repeated fashion (daily for four consecutive days), and hepatic microsomes were prepared for XME analyses. Both GLs were able to induce XMEs, showing different induction profiles. While the inductive effect was stronger after multiple administration of the higher GRH dosage, the single lower GRE dose was the most effective in boosting cytochrome P-450 (CYP)-associated monooxygenases and the postoxidative metabolism. CYP3A1/2 were the most affected isoforms by GRH treatment, whereas GRE induced mainly CYP1A2 supported oxidase. Glutathione S-transferase increased up to approximately 3.2-fold after a single (lower) GRE dose and UDP-glucuronosyl transferase up to approximately 2-fold after four consecutive (higher) GRH doses. In conclusion, the induction profile of these GLs we found is not in line with the chemopreventive hypothesis. Furthermore, the oxidation degree of the side chain sulfur of GLs seems to exert a crucial role on XME modulation.     

Influence of water quench cooling on degassing and aroma stability of roasted coffee

Coffee roasting experiments with air cooling versus water quench cooling were carried out on laboratory scale with a fluidized-bed hot air roasting system (200 g batch size) and on production scale with a rotating bowl roaster (320 kg batch size). Two series of coffees with different water contents resulted, which were stored at 25 degrees C under normal atmospheric conditions. Carbon dioxide desorption was followed and stability of selected aroma compounds was tested with headspace solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) and stable isotope labeled compounds as internal standards. Degassing is faster in water-quenched coffees with higher moisture content, but pore size distribution in the different coffee samples did not correlate with degassing behavior. Bean firmness, which increases with increasing moisture content, might have an influence on degassing. Air- and water-quenched coffees exhibit similar stability of most aroma compounds despite different degassing behavior. However, evolution of dimethyl trisulfide was different in coffees with increased water content. This suggests higher thiol oxidation rates, a factor that is cited to be related to a faster loss of freshness attributes.     

Genetic variability of the prion protein gene (PRNP) in wild ruminants from Italy and Scotland

The genetics of the prion protein gene (PRNP) play a crucial role in determining the relative susceptibility to transmissible spongiform encephalopathies (TSEs) in several mammalian species. To determine the PRNP gene variability in European red deer (Cervus elaphus), roe deer (Capreolus capreolus) and chamois (Rupicapra rupicapra), the PRNP open reading frame from 715 samples was analysed to reveal a total of ten single nucleotide polymorphisms (SNPs). In red deer, SNPs were found in codons 15, 21, 59, 78, 79, 98, 136, 168 and 226. These polymorphisms give rise to 12 haplotypes, and one of which is identical to the PRNP of American wapiti (Rocky Mountain elk, Cervus elaphus nelsoni). One silent mutation at codon 119 was detected in chamois and no SNPs were found in roe deer. This analysis confirmed that European wild ruminants have a PRNP genetic background that is compatible with TSE susceptibility, including chronic wasting disease.     

Differences in the fragrances of pollen,leaves, and floral parts of garland (Chrysanthemum coronarium) and composition of the essential oils from flowerheads and leaves

Headspace analyses of pollen, whole flowerheads, ligulate and tubular florets, flower buds, involucral bracts, and leaves have been performed on the food plant Chrysanthemum coronarium L. (Asteraceae). The analyses permitted differences in the pattern of volatiles emitted by the different floral parts to be observed and the site and phenological stage of emission of these chemicals to be verified. Camphor and cis-chrysanthenyl acetate were emitted mainly by ligulate and tubular florets; the production of myrcene and (Z)-ocimene was higher in the flower buds, whereas beta-caryophyllene, (E,E)-alpha-farnesene, and (E)-beta-farnesene seemed attributable mainly to the involucral bracts. The leaves showed a quite different volatile profile, with (Z)-ocimene as the main constituent. Pollen showed a completely different composition of its volatiles, with perilla aldehyde, cis-chrysanthenyl acetate, and camphor among the principal compounds; many carbonylic compounds and linear hydrocarbons have been detected exclusively in pollen. Furthermore, the essential oils obtained from flowerheads and leaves have been studied. These samples showed mainly quantitative differences. Camphor (22.1%) and cis-chrysanthenyl acetate (19.9%) were the main constituents of the oil from flowers, whereas the oil from the leaves contained mainly (Z)-ocimene (45.4%) and myrcene (28.2%).     

Response of Pinus leucodermis to climate and anthropogenic activity in the National Park of Pollino (Basilicata, Southern Italy)

Pinus leucodermis (=P. heldreichii var. leucodermis) is widespread in the Balkan Peninsula and is present as a post-glacial relict in Southern Italy. The oldest Italian populations of this species are located at high elevation in the National Park of Pollino, where grazing and logging had endangered their survival, especially during the 20th century. In 1993 the National Park was founded and anthropogenic activities were restricted. To understand the response of P. leucodermis at the upper tree-line to climatic and anthropogenic activity variations, we developed a tree-ring width chronology (1464-2003). For the period 1804-2003 separate chronologies for the earlywood and latewood were built, and resin duct density was assessed on total ring, earlywood and latewood. Age structure of saplings was also determined. After 1950 a steep decline in tree-ring width was followed by a recovery since 1981. During the 20th century radial growth response to climate was not strong and not stable. In the period 1953-2000 P. leucodermis radial growth seemed to take advantage of high temperatures and low precipitation. Resin duct density chronologies were not a reliable dendroecological variable, but they documented a metabolic trade-off between growth and differentiation processes. The gap in the age structure between long-lived trees (over 200 years old) and saplings around 40 years old, which mainly grow in protected microsites between rocks, could indicate a negative influence of grazing and related human activities. We suggest that the protection strategies introduced by the National Park could play a positive role in the recruitment of new saplings and in the ring growth recovery of old trees of P. leucodermis.     

Determination of free phenolic acids in wort and beer by coulometric array detection

The simultaneous determination of 19 phenolic compounds was performed directly in wort and beer by a combination of reverse-phase high-performance liquid chromatography coupled with coulometric array detection. Chromatographic separation was achieved with an appropriate gradient of flow and a binary solvent based on phosphate buffer, methanol, and acetonitrile in a 45-min run. Eight serial coulometric detectors were used for on-line generation of voltammetric data to resolve coeluting compounds. The method was reliable and sensitive, the regression coefficient of standard calibration curves is 0.972 < or = r < or = 1.000, and the standard deviation value ranges from 0.010 to 0.129 mg/L for wort and from 0.002 to 0.332 mg/L for beer. The mean concentrations of phenolic acids were 22.1 and 33.8 mg/L, respectively, in worts and beers produced in Italy. These amounts represent 5 and 10% of the non-tannic, non-flavonoid phenols in wort and beer, respectively.