Effects of ionic strength and sulfate upon thermal aggregation of grape chitinases and thaumatin-like proteins in a model system

Consumers expect white wines to be clear. During the storage of wines, grape proteins can aggregate to form haze. These proteins, particularly chitinases and thaumatin-like proteins (TL-proteins), need to be removed, and this is done through adsorption by bentonite, an effective but inefficient wine-processing step. Alternative processes are sought, but, for them to be successful, an in-depth understanding of the causes of protein hazing is required. This study investigated the role played by ionic strength (I) and sulfate toward the aggregation of TL-proteins and chitinases upon heating. Purified proteins were dissolved in model wine and analyzed by dynamic light scattering (DLS). The effect of I on protein aggregation was investigated within the range from 2 to 500 mM/L. For chitinases, aggregation occurred during heating with I values of 100 and 500 mM/L, depending on the isoform. This aggregation immediately led to the formation of large particles (3 μm, visible haze after cooling). TL-protein aggregation was observed only with I of 500 mM/L; it mainly developed during cooling and led to the formation of finite aggregates (400 nm) that remained invisible. With sulfate in the medium chitinases formed visible haze immediately when heat was applied, whereas TL-proteins aggregated during cooling but not into particles large enough to be visible to the naked eye. The data show that the aggregation mechanisms of TL-proteins and chitinases are different and are influenced by the ionic strength and ionic content of the model wine. Under the conditions used in this study, chitinases were more prone to precipitate and form haze than TL-proteins.     

Effects of a selection of additives on in vitro ruminal methanogenesis and in situ and in vivo NDF digestibility

The effects of 18 essential oils, yeast, Quebracho tannin and Quillaja saponin on ruminal methane and gas production (GP) were studied in vitro. A lactating cow diet was incubated with rumen inoculum. Doses of the additives (mg/L) were: essential oils = 500; tannin = 67; yeast = 8.35 and 16.7; and saponin = 300. Lemongrass, estragole, eugenol, geraniol, limonen, thyme oil and thymol produced less gas (overall mean 33.8 mL/200 mg dry matter (DM)) than control (43.6 mL/200 mg DM; P < 0.001). Methane produced (mL/200 mg DM) by guaiacol (10.7), lemongrass (9.6), limonene (11.4), thyme oil (10.9) and thymol (2.1) was lower than control (12.5) (P < 0.001). Methane percentage on total GP was lower (P < 0.001) for cinnamic alcohol (25.4), guaiacol (24.5), thymol (19.7) and vanillin (26.3) than control (28.8). In a second experiment, thymol, guaiacol and yeast were added to the diet of dry fistulated cows to determine in situ neutral detergent fiber digestibility (NDFD) of six forages and in vivo dietary NDFD. Thymol and yeast decreased in situ NDFD after 24 h (33.9% and 33.5% vs. 38.1%; P = 0.008). Thymol decreased in vivo total tract NDFD (40.8% vs. 51.4%; P = 0.02). Differences in GP and methane levels were registered within classes of additives. A careful selection of additives may allow for the manipulation of ruminal fermentation.     

Customizing Properties of β-Chitin in Squid Pen (Gladius) by Chemical Treatments

The squid pen (gladius) from the Loligo vulgaris was used for preparation of β-chitin materials characterized by different chemical, micro- and nano-structural properties that preserved, almost completely the macrostructural and the mechanical ones. The β-chitin materials obtained by alkaline treatment showed porosity, wettability and swelling that are a function of the duration of the treatment. Microscopic, spectroscopic and synchrotron X-ray diffraction techniques showed that the chemical environment of the N-acetyl groups of the β-chitin chains changes after the thermal alkaline treatment. As a consequence, the crystalline packing of the β-chitin is modified, due to the intercalation of water molecules between β-chitin sheets. Potential applications of these β-chitin materials range from the nanotechnology to the regenerative medicine. The use of gladii, which are waste products of the fishing industry, has also important environmental implications.     

Environmental conditions in a lagoon and their possible effects on shellfish contamination by Giardia and Cryptosporidium

Over a 1-year period, we carried out an analysis for the presence of Giardia and Cryptosporidium (oo)cysts in 934 specimens taken from four species of shellfish farmed in the Varano Lagoon. We also studied the chemical, physical and meteo-climatic parameters in the same environment, to evaluate the possible effects of environmental conditions on protozoan contamination in farmed shellfish. Using both IF and molecular tests, all 38 pools of shellfish gave negative values for Giardia and Cryptosporidium oo/cysts, thus confirming previous investigations. The environmental factors considered having a negative influence on the survival of Giardia and Cryptosporidium are high water temperatures (8–29°C) and high salinity levels (23–32?ppt). However, the small amount of freshwater discharged into the lagoon (up to 0.0091?m³ s^?1), the long-lasting solar irradiation (mean of 896?W?m^?2) and the shallowness of the lagoon (mean 2.5?m) may also have a synergic effect in limiting the presence of Giardia and Cryptosporidium in the lagoon.     

Screening and selection of eggplant and wild related species for resistance to <Emphasis Type="Italic">Leveillula taurica</Emphasis>

Screening for resistance to powdery mildew of Solanum melongena and wild related species was made in the field under natural infection conditions. A total of 172 accessions originating from several geographical parts of the world were tested. Single plant selection for resistance was carried out and open-pollination was used. Most S. melongena accessions were susceptible or highly susceptible to the disease. By S₀ to S₃ selection, an increase in the overall powdery mildew resistance level of S. melongena population was obtained and four S. melongena lines possessing a high level of resistance were obtained. Among the wild Solanum species, S. laciniatum and S. nigrum showed to be non-host plants of L. taurica. S. quinquangolare showed no symptoms of powdery mildew, S. linnaeanum, S. aculeatissimum, S. aviculare, S. pseudocapsicum were highly resistant, S. spinosissimum was resistant, S. gilo, S. capsicoides were susceptible or highly susceptible, and plants of S. sisymbriifolium showed a widely variable disease reaction. Four S. melongena resistant lines were obtained: PAVEG 10187 S₃, PAVEG 10196 S₃, P.I. 230279 S₃ and P.I. 419198 S₃. These S. melongena lines together with the resistant wild species could be used for genetic studies, classical breeding programs and biotechnological applications.     

Effect of maturity stage at harvest on the ensilability of maize hybrids in the early and late FAO classes,grown in areas differing in yield potential

The aim of this study was to assess the effects of earliness (according to FAO class), maturity stage at harvest and environmental conditions on the ensilability of maize hybrids described by their fermentation products and by a fermentation quality index (FQI). Maize hybrids belonging to early (n = 14) and late (n = 15) FAO classes were grown in low, medium and high potential yield areas and harvested at an early (EH), medium (MH) and late maturity stage (LH), that is, at 1/3, 2/3 and 5 d after the 2/3 milk line stage, respectively, according to a split‐plot design. Upon harvest, each sample (n = 522) was analysed for dry matter (DM) and water‐soluble carbohydrates (WSC) before being ensiled in vacuum‐packed bags (n = 1,044). After 60 days of conservation, samples were analysed for DM and fermentation products. In the pre‐ensiling phase, DM was higher in early hybrids (p = .001), low yield areas (p < .001) and at LH maturity (p < .001), whereas WSC contents were higher in early hybrids (p < .001), medium yield areas (p < .001) and at EH maturity (p < .001). With regard to silages, early hybrids had a higher FQI (p < .001), which was highest in areas with a high yield potential (p < .01) and at EH maturity (p < .01). Late hybrids proved to be better suited for low yield areas compared with early hybrids (p < .01) and had a higher FQI at EH and MH than at LH maturity (p < .01).     

Identification of herbaceous hosts of the <Emphasis Type="Italic">Grapevine Pinot gris virus</Emphasis> (GPGV)

Grapevine Pinot gris Virus (GPGV) is a single stranded RNA of the genus Trichovirus infecting grapevine (Vitis vinifera) and associated with stunting, chlorotic mottling and leaf deformation symptoms. During a monitoring of GPGV infection in vineyards of the Trentino region in Italy, we have detected the virus in the herbaceous plants Silene latifolia subsp. Alba (Mill.) (bladder campion) and Chenopodium album L. (white goosefoot), which showed symptoms of viral infection. The full-length GPGV RNA genome, amplified from these infected hosts, was sequenced and a phylogenetic analysis revealed that its closest relative is the strain SK13, recently isolated in Slovakia. Our results indicate that herbaceous plants can be considered as a reservoir for the GPGV virus. This finding is important for studying the epidemiological aspects of GPGV disease and to formulate appropriate control measures.     

Modelling self-pruning and branch attributes for young Quercus robur L. and Fagus sylvatica L. trees

The aim of this study was to develop statistical models for first order branchiness in young planted forest stands of pedunculate oak (Quercus robur L.) and European beech (Fagus sylvatica L.), and to give an ecological and silvicultural interpretation to these models. The reported models focus on the lower most-valuable stem part (i.e. until 6 m height), and cover different tree development classes to capture the development of branchiness over time. For each species 30 study plots were selected spread over two nearby forests in Flanders (northern Belgium), minimising site and genetic variability. Branches were counted on a total of 399 oak and 376 beech trees. On a subsample of 30 trees per species (one tree per plot), detailed non-destructive branch measurements were performed, yielding data for 555 oak and 438 beech branches. For both species, models for tree self-pruning (i.e. total branch number and dead branch portion), branch mortality and branch architecture (i.e. branch diameter and branch insertion angle) were built. A generalised linear mixed modelling approach was adopted. The models for total branch number and dead branch portion may be interpreted in terms of four processes contributing to self-pruning: (1) stand and tree development, (2) tree competitive status, (3) stand density and (4) site humidity. The reported models reveal similar self-pruning rates in oak and beech, but with different driving factors: early branch dying and slow shedding for oak and the other way around for beech. Mortality of individual branches is further determined by branch position and branch dimension. Branch diameter and branch insertion angle of both species are mainly related to branch cord length and relative branch position. All modelled effects are consistent with known ecological and ecophysiological processes. Silvicultural implications for stand establishment and early tree selection are discussed. The reported models can be used to fine-tune operational silvicultural choices for quality timber production. This is a first step towards the integration of branchiness models for oak and beech into forest growth simulators.     

Combining isotopic signatures of n(Sr)/n(Sr) and light stable elements (C,N, O,S) with multi-elemental profiling for the authentication of provenance of European cereal samples

The aim of this work (from the FP6 project TRACE) was to develop methods based on the use of geochemical markers for the authentication of the geographical origin of cereal samples in Europe (cf. EC regulations 2081/92 and 1898/06). For the first time, the potential usefulness of combining n(⁸⁷Sr)/n(⁸⁶Sr) and δ¹³C, δ¹⁵N, δ¹⁸O and δ³⁴S isotopic signatures, alone or with key element concentrations ([Na], [K], [Ca], [Cu] and [Rb], progressively identified out of 31 sets of results), was investigated through multiple step multivariate statistics for more than 500 cereal samples collected over 2 years from 17 sampling sites across Europe representing an extensive range of geographical and environmental characteristics.