Lymphocyte activation as cytokine gene expression and secretion is related to the porcine reproductive and respiratory syndrome virus (PRRSV) isolate after in vitro homologous and heterologous recall of peripheral blood mononuclear cells (PBMC) from pigs vaccinated and exposed to natural infection

The present study evaluated the lymphocyte activation in PRRSV-vaccinated pigs subsequently exposed to natural infection by in vitro stimulation of peripheral blood mononuclear cells (PBMC) with homologous vaccine and two heterologous PRRSV isolates. The responsiveness was assessed by determining IFN-γ secreting cells by ELISpot assay, lymphocyte CD8 phenotype by intracellular staining/flow cytometry, cytokine gene expression by real-time quantitative PCR and cytokine secretion by ELISA. Conventional pigs were weaned at 28 days of age and inoculated intramuscularly (IM) or needle-less intradermally (ID) with a modified-live PRRSV vaccine suspended in adjuvant, while control pigs were injected with adjuvant alone (ADJ). Blood samples were collected at vaccination, 35 days post-vaccination and after 35 days post-exposure to natural infection by a heterologous field strain. Thirty-five days post-vaccination, PRRSV vaccine induced a low but significant virus-specific IFN-γ secreting cell response upon stimulation with both the vaccine strain and the two isolates in vaccinated pigs. Conversely, after 35 days post-exposure, only the vaccine strain and the BS/114/S isolate triggered this response. Intracellular staining showed that PRRSV-specific immune cells reacting upon vaccine strain and BS/114/S stimulation were mostly CD8⁺ IFN-γ producing cells whereas the stimulation with BS/55 isolate induced an IFN-γ production associated to the CD8^?IFN-γ⁺ phenotype. At 35 days post-vaccination, PBMC from vaccinated pigs showed lower IL-10 expression and release, and higher TNF-α gene expression upon stimulation with both the vaccine and viral isolates. After infection, both cytokines were not differently modulated in different groups. Immune parameters give evidence that IFN-γ secreting cells in the peripheral blood can be elicited upon PRRSV infection although vaccination itself does not stimulate high levels of these reactive cells. Moreover, the cross-reactivity against divergent PRRS viruses can show a different intensity and be differently associated with cytotoxic CD8⁺IFN-γ⁺ as well as CD8^?IFN-γ⁺ cells. Overall, the obtained data confirmed that the immune activation against PRRSV is not dependent on the genetic divergence of the virus. Especially after infection, a different immune reactivity was evident upon stimulation with the different isolates in terms of frequency and CD8 phenotype of PRRSV-specific IFN-γ producing cells. The modulation of cytokines in vaccinated pigs appeared to be more dependent on vaccination or infection conditions than on stimulation by different isolates, and the changes of IL-10 more relevant than those of TNF-α at gene and protein levels. Moreover, under the conditions of this study, the PRRSV vaccine administered via the intradermal route by a needle-less device was confirmed to induce an immune response comparable or in some cases higher than the intramuscular route.     

Large-scale in vitro propagation of giant reed (Arundo donax L.), a promising biomass species

Summary

Giant reed (Arundo donax L.), a promising energy crop, is vegetatively-propagated from fragments of stems and rhizomes. This may limit large-scale cultivation, since it is time-consuming and involves considerable cost and effort. Tissue culture is an alternative to conventional methods of vegetative propagation and may represent a useful tool for large-scale propagation of plants for biomass production programmes. This report describes a protocol for the large-scale in vitro propagation of giant reed by adventitious bud formation. Stem nodes with dormant buds proved to be the most effective to initiate in vitro cultures giving the highest percentage of differentiated shoots (77%) compared to the other plant fractions tested. A sterilisation procedure using 5 g l^–1 HgCl₂ enabled the production of sterile explants. Moreover, early results indicated that late Autumn excision dates not only gave a higher percentage of well-developed shoots ( > 80%), but also a lower level of bacterial contamination (15 – 20%). 6-Benzylaminopurine (BAP) at 3.0 mg l^–1 was most effective in promoting shoot multiplication when added to a basal medium containing Murashige and Skoog (MS) macro- and micro-nutrients, Morel’s vitamins, 30 g l^–1 sucrose, and 7.0 g l^–1 bacteriological agar supplemented with 1.0 mg l^–1 indole-3-acetic acid (IAA) and 0.05 mg l^–1 gibberellic acid (GA₃). Rooting was successfully induced on the same basal medium used for proliferation, modified by halving the MS macro-nutrients and replacing BAP and the other growth regulators with 2.0 mg l^–1 1-naphthaleneacetic acid (NAA). Successful acclimatisation (> 95% survival) of plantlets was carried out, even in late Winter, in a cold greenhouse or under simpler facilities such as shade nets.     

Population dynamics and spread of Unaspis yanonensis in Calabria, Italy

The occurrence of Unaspis yanonensis (Kuwana) (Hemiptera: Diaspididae) in Calabria (southern Italy) poses a serious threat to citrus cultivation. A regional survey was performed to record information about its distribution in local citrus orchards. Field population dynamics were also studied. The arrowhead scale, U. yanonensis, was recorded in the two main citrus cultivation areas of Calabria. In the investigated environments U. yanonensis completed two generations per year; a typical bi-modal oviposition was recorded. The arrowhead scale overwintered as second-instar males and pre-ovipositing females.     

Effects of different management practices on arbuscular mycorrhizal fungal diversity in maize fields by a molecular approach

As obligate mutualistic symbionts, arbuscular mycorrhizal fungi (AMF) colonize the roots of many agricultural crops, and it is often claimed that agricultural practices are detrimental to AMF. As a result, agroecosystems impoverished in AMF may not get the fully expected range of benefits from these fungi. Using molecular markers on DNA extracted directly from soil and roots, we studied the effects of different management practices (tillage and N fertilization) on the AMF communities colonizing an experimental maize field in Central Italy. Our molecular analysis based on three different nuclear rRNA regions (18S, 28S and ITS) allowed us to assess AMF biodiversity. Glomeraceae members were the main colonizer, and they co-occurred with Gigasporaceae and Paraglomus regardless of the management practices applied. Diversisporaceae and Entrophosporaceae members were instead detected in the N-fertilized soils and in the untreated soil, respectively. The results obtained indicated that the general AMF assemblages structure and composition in the maize field plots appear to be primarily influenced by N fertilization and, to a lesser extent, by tillage. This study also validates the usefulness of multiple molecular markers to consolidate and refine the assessment of the environmental AMF diversity.     

Spectroscopic properties of humic acids isolated from the rhizosphere and bulk soil compartments and fractionated by size-exclusion chromatography

The humic acids (HAs) isolated by conventional procedure from rhizosphere (r) and bulk (c) soils were analyzed by means of chemical and physico-chemical techniques. Two different crops were selected, tomato (T) and artichoke (A), and each HA was fractionated by size-exclusion chromatography (SEC) into three fractions with increasing molecular size, respectively, Fraction I (FrI) < Fraction II (FrII) < Fraction III (FrIII). Elemental analysis data indicated greater N and S contents in the rhizosphere T-HAs, with respect to rhizosphere A-HA, which suggests the occurrence in the former ones of a large amount of organic nitrogen- and sulfur-containing compounds that are released by the rhizodeposition processes. Further, the three HA fractions from the bulk soils of the two series showed a gradual increase of C, H, and N contents, and a decrease of O and S contents and C/N and C/H ratios. These results suggested that the lowest molecular size fractions are richer in oxygenated functional groups, whereas the higher molecular size fractions are richer in N-containing groups and structural C- and H-containing units. The three HA fractions from the rhizosphere soils of the two series showed a gradual decrease in C content, and an increase of H, N, and O contents, which suggests the possible incorporation into soil HAs of a multitude of C-containing compounds of low molecular size released by plant roots. The FT-IR data, in general, suggested that the contents of carboxylic, phenolic and N-containing groups and polysaccharide-like components in HAs from rhizosphere soils are larger than those of HAs from the corresponding bulk soils. Further, the FrI fraction consisted mainly of simple structural units, likely quinonic and phenolic units with a prevalent aromatic character, whereas the FrII and, especially, FrIII fractions featured a mixed aliphatic/aromatic nature and a greater molecular complexity. The extent of these differences appeared to depend on the plant species and age, and is mainly due to the partial incorporation into rhizosphere HAs of typical root exudate components, such as amino acids, amides, aliphatic and aromatic acids of low molecular size, polysaccharides and sugars, fatty acids and sterols, and enzymes.     

Microwave-induced cooling of a superconducting qubit

We demonstrated microwave-induced cooling in a superconducting flux qubit. The thermal population in the first-excited state of the qubit is driven to a higher-excited state by way of a sideband transition. Subsequent relaxation into the ground state results in cooling. Effective temperatures as low as approximately 3 millikelvin are achieved for bath temperatures of 30 to 400 millikelvin, a cooling factor between 10 and 100. This demonstration provides an analog to optical cooling of trapped ions and atoms and is generalizable to other solid-state quantum systems. Active cooling of qubits, applied to quantum information science, provides a means for qubit-state preparation with improved fidelity and for suppressing decoherence in multi-qubit systems.     

Properties of dissolved organic matter in forest soils: Influence of different plant covering

The nature of different forest plant litters and the effect of their degradation processes on the properties of soil dissolved organic matter is a very important environmental issue in protected areas such as the “Natural Oriented Reserve Bosco delle Pianelle” in Southern Italy, Chemical and spectroscopic techniques were used to evaluate the influence of four different tree covers on the properties of dissolved organic matter (DOM) isolated from litter and surface soil layers. Fourier transform infrared spectroscopy analysis showed that all litter samples feature a prevalent aromatic character. Ligninolitic components of litters produced by Pinus halepensis L. and a mixture of Quercus trojana Webb. and Q. ilex L. were more degraded than those produced by either Q. ilex or a mixture of Carpinus betulus L. and C. orientalis Miller. The larger C/N ratios of the former litters might account for the greater degradation. Further, a lesser lignin decomposition was shown also for the latter samples, likely due to adverse topographical factors, such as less intense sunlight and greater moisture content. Spectroscopic analysis of litter DOM showed that easily degraded components, such as water-soluble compounds and carbohydrates, were preferentially degraded resulting in DOM that was enriched in lignin-derived compounds. Fluorescence analysis data of all litter DOM samples showed the occurrence of fluorescent units qualitatively similar regardless of plant covering, which suggests that in all cases simple structural components of low-molecular weight and small degree of aromatic polycondensation and content of conjugated chromophores are present. Soil DOM featured several numerous fluorescent units that differ as a function of the parent litter and/or its decomposition processes. Along the soil profiles water-soluble aliphatic compounds of low-molecular weight are found down to the deepest layer, whereas aromatic–ligninolitic compounds are decreasing with depth, probably because of their adsorption by mineral soil components. A greater amount of aromatic units, likely lignin-derived compounds, was found in the DOM from P. halepensis L., which suggested a more extended lignin decomposition. Further, with increasing soil depth, DOM was characterized by a decrease of low-molecular weight organic molecules and lower degree of humification.     

Antioxidant Properties of Fermented Soy during Shelf Life

Plant Foods for Human Nutrition - Glycine max (soybean) is a fundamental food in human nutrition, largely utilized by the consumers, and in particular, fermented soy is mainly used. However, health...