Multiplex real-time PCR SYBR Green for detection and typing of group III Clostridium botulinum

Clostridium botulinum type C and type D belonging to the group III organisms, are mainly responsible for animal botulism outbreaks. Clinical signs alone are often insufficient to make a diagnosis of botulism and a laboratory confirmation is required. Laboratory confirmation can be performed by demonstrating the presence of botulinum neurotoxins in serum, gastrointestinal contents, liver, wound of sick or dead animals, or by demonstrating the presence of C. botulinum in gastrointestinal contents, liver, and wound. Demonstration of spores in gastrointestinal contents or tissue of animals with clinical signs indicative of botulism reinforces the clinical diagnosis. With the aim of detecting and typing C. botulinum group III organisms, a multiplex real-time PCR SYBR Green was developed and in-house validated. Selectivity, limit of detection, relative accuracy, relative specificity, relative sensitivity, and repeatability of the method were investigated. The multiplex real-time PCR SYBR green used showed a 100% selectivity, 100% relative accuracy, 100% relative specificity, 100% relative sensitivity and a limit of detection of 277 and 580 DNA copies for C. botulinum type C and C. botulinum type D, respectively. The method reported here represents a suitable tool for laboratory diagnosis of type C and D botulism and for testing a large number of samples collected during the animal botulism surveillance and prevention activities.     

Cystic liver disease related to high Platynosomum fastosum infection in a domestic cat

Platynosomum fastosum is a small fluke found in the biliary ducts and gallbladder of cats. Its lifecycle includes the snail Sublima octona as intermediate host, and lizards, toads and geckos as paratenic hosts. Affected cats are usually adult and acquire the parasite by feeding on infected lizards. This parasite occurs across the world but is more frequent in tropical areas. The clinical signs range from none to obstruction of the biliary tract, with hepatic failure and death, reinforcing the necessity of including the liver fluke Platynosomum fastosum in the differential diagnosis of hepatic diseases in cats. This report describes an unusual case of a cat with a polycystic hepatic disease and a severe infestation by Platynosomum fastosum and presents a review of the literature.     

Multinucleated giant cells with an osteoclast phenotype derived from caprine peripheral blood mononuclear cells

Formation of multinucleated giant cells (MGCs) by macrophage fusion is a typical cytopathic effect of lentiviral replication in caprine monocytes and MGC formation from cultured caprine peripheral blood mononuclear cells (PBMCs) has been considered to be diagnostic for small ruminant lentivirus (SRLV) infection. In this study, formation of MGCs was observed after 7–14 days when PBMCs were cultured from healthy goats free from SRLV infection. These MGCs expressed tartrate-resistant acid phosphatase, calcitonin receptor, integrin αVβ3, cathepsin K and matrix metalloproteinase 9 and were able to resorb bone in vitro in the absence of RANKL and macrophage colony stimulating factor, consistent with an osteoclast phenotype.     

A dynamic risk assessment model (FUMAgrain) of fumonisin synthesis by Fusarium verticillioides in maize grain in Italy

Fumonisin contamination of maize grain starts in the field. Forecasting Fusarium infection and fumonisin synthesis could allow operators in the field to control contamination during the growing season and to make the best agronomic decisions for high quality yields while respecting the limits imposed by the European Union.A research project to develop a decision support system for the control of field-phase fumonisin contamination began in Italy in 2003. This paper presents a preliminary version of the engine of the decision support tool: FUMAgrain, a dynamic risk assessment model developed with data from the north of Italy. The structure of FUMAgrain is based on the pathosystem formed by maize, F. verticillioides and Ostrinia nubilalis (European Corn Borer). The elements of the pathosystem are simulated by three sub-models: (i) maize development, (ii) F. verticillioides infection and fumonisin synthesis, (iii) European Corn Borer wounding activity on maize grain. Inputs to the model are (i) planting date, (ii) hourly meteorological data including temperature, relative humidity, wind speed and rain intensity, (iii) information on the phenological development of the hybrid planted (flowering and dry-down), and (iv) information about the chemical treatment against European Corn Borer. FUMAgrain gives an initial risk alert at the end of flowering based on the meteorological conditions during this phase. A second alert follows maturation when an assessment is made from (i) maize grain moisture, (ii) European Corn Borer damage to the ear, and (iii) fumonisin synthesis risk. Following calibration and validation with data FUMAgrain demonstrated its good capability to simulate fumonisin synthesis in maize grain in Italy (calibration: R² = 0.70; validation: R² = 0.71) and its usefulness for determining the optimal harvest date while respecting grain safety levels required by the international market and limiting moisture content, hence drying costs.     

Novel Prunus rootstock somaclonal variants with divergent ability to tolerate waterlogging

Plants require access to free water for nutrient uptake, but excess water surrounding the roots can be injurious or even lethal because it blocks the transfer of free oxygen between the soil and the atmosphere. Genetic improvement efforts in this study were focused on the increased tolerance in roots to waterlogging. Among a pool of clones generated in vitro from leaf explants of rootstock Mr.S.2/5 of Prunus cerasifera L., the S.4 clone was flood tolerant whereas the S.1 clone was sensitive. The S.4 clone formed adventitious roots on exposure to flooding. Moreover, the chlorophyll content and mitochondrial activity in the leaf and root, soluble sugar content, alcohol dehydrogenase activity and ethylene content were different between the clones. The sorbitol transporter gene (SOT1) was up-regulated during hypoxia, the alcohol dehydrogenase genes (ADH1 and ADH3) were up-regulated in the leaves and down-regulated in the roots of the S.4 clone during hypoxia, and the 1-aminocyclopropane-1-oxidase gene (ACO1) was up-regulated in the leaves and roots of the S.4 clone during hypoxia and down-regulated in the wild-type roots. In addition, in the S.4 root, hypoxia induced significant down-regulation of a glycosyltransferase-like gene (GTL), which has a yet-undefined role. Although the relevant variation in the S.4 genome has yet to be determined, genetic alteration clearly conferred a flooding-tolerant phenotype. The isolation of novel somaclonals with the same genomic background but with divergent tolerance to flooding may offer new insights in the elucidation of the genetic machinery of resistance to flooding and aid in the selection of new Prunus rootstocks to be used in various adverse environments.     

Culture as an Engine of Local Development Processes: System‐Wide Cultural Districts I: Theory

Building on the early works of Alfred Marshall, analyses of local economies have emphasized the spatial function of clusters and industrial districts in terms of external economies of localization and agglomeration. Recent literature has emphasized the importance of culture and the complementarities between culture and local tangible and intangible assets. This paper aims to provide an analytical foundation for these processes with a view to developing tools for policy design, analysis and evaluation. By “system‐wide cultural districts,” we refer to a new approach to local development where cultural production and participation present significant strategic complementarities with other production chains. In this view, culture drives the accumulation of intangible assets such as human, social, and cultural/symbolic capital, thereby fostering economic and social growth and environmental sustainability.     

Acquisition capability of the grapevine Flavescence dorée by the leafhopper vector Scaphoideus titanus Ball correlates with phytoplasma titre in the source plant

Flavescence dorée (FD) is one of the most economically important grapevine diseases in Southern Europe, and it is associated with phytoplasmas, phloem-limited wall-less bacteria. Recovery from disease naturally occurs in infected grapevines during the following seasons after infection. The capability of the leafhopper vector Scaphoideus titanus to acquire FD phytoplasma (FDP) from recovered and infected grapevines of Barbera and Nebbiolo varieties was investigated in North-western Italy vineyards monitored from 2007 to 2011. Pathogen concentration was quantified by real-time PCR in FDP-infected grapevines and broad beans, also used as source plants under controlled conditions, to correlate acquisition capabilities and phytoplasma titre in source plants. S. titanus acquired FDP from infected, but not from recovered, grapevines. FDP titre was higher in Barbera than in Nebbiolo and higher in summer than in spring, and acquisition efficiency and pathogen titre in source plants were positively correlated, both in field and laboratory conditions. Recovered plants do not represent a source of inoculum for the vector and therefore do not contribute to FDP spread. The inability of recovered plants to serve as FDP acquisition sources for the vector as well as the effect of the season and of the two grapevine varieties on the FDP acquisition efficiency are relevant results to re-design disease management practices, especially since insecticide treatments against the vector are not fully effective, and newly designed successful control strategies are required.     

Assessing seed desiccation responses of native trees in the Caribbean

New Forests - Native trees from the Caribbean were tested for seed desiccation responses, by adapting the “100-seed test” protocol. Ninety-seven seed lots of 91 species were collected...     

Reclamation of Polluted Soil: Phytoremediation Potential of Crop-Related BRASSICA Species

Soils polluted by heavy metals can be reclaimed using a number of expensive tactics that either remove the contaminants or stabilize them within the soil. The value of metal accumulating plants for environmental remediation has recently been appreciated and promising results have been obtained. This paper reports a study on the behavior of Brassica napus, Brassica juncea, Raphanus sativus and Brassica carinata grown on a substrate contaminated by several heavy metals caused by the use of contaminated irrigation water. Data on carbon dioxide assimilation, biomass growth and the bioconcentration and translocation factor of each metal in each species were measured. The polluted substrate caused only a small variation in photosynthesis, however transpiration was more affected by the experimental substrate and in all three species of the genus Brassica the presence of metals in the substrate resulted in higher transpiration levels.Two bioconcentration factors were calculated respectively for the roots (BCF) and the shoots (BCF); the BCF was >1 for all the species for Cd, Cu, Ni and Zn without significant differences among species. All the values of BCF were lower than 0.5; among the metals, all Brassica species demonstrated a similar performance for Cd and Zn, whereas for other elements the bioconcentration factor was very low.     

Existence of different physiological forms within genetically diverse strains of <Emphasis Type="Italic">Ampelomyces quisqualis</Emphasis>

Powdery mildew fungi are parasitized by strains of the genetically distinct Ampelomyces quisqualis. To investigate whether differences in the phylogeny and other cultural, morphological and physiological characteristics of these different strains are related to differences in their geographic origins or the host species from which they were isolated, several A. quisqualis strains isolated from different species of Erysiphaceae collected in different countries and possessing different ITS rDNA sequences were selected and characterized. The results revealed some significant variation among the selected strains, which provides evidence for the existence of different physiological forms within the A. quisqualis species. Two groups that display differential growth on artificial media were identified. These groups also differ in the morphology of their mycelium, but not in the morphology of their pycnidia and conidia. Temperature greatly affected the in vitro growth of the A. quisqualis strains and growth rate was closely correlated to colony color. Differences in the conidial germination of distinct strains were observed during the recognition phase of the parasitic relationship. The germination of each of the investigated strains was greatly stimulated by all of the examined powdery mildew species and not only by the conidia of their original hosts. An Italian strain isolated from grapevine in the Trentino Alto-Adige region was identified as the strain that germinates the most quickly in the presence of powdery mildew conidia. Phylogenetic analysis revealed that these A. quisqualis strains can be classified into five different genetic groups, which generally correlate with the fungal host of origin and morphological and growth characteristics.