Development of a polymerase chain reaction and restriction typing assay for the diagnosis of bovine herpesvirus 1, bovine herpesvirus 2, and bovine herpesvirus 4 infections.

A multiplex polymerase chain reaction (PCR) method coupled with a restriction analysis of PCR products (PCR with restriction fragment length polymorphism) was developed for the simultaneous detection of bovine herpesvirus 1, bovine herpesvirus 2, and bovine herpesvirus 4 infections. The specificity, sensitivity, and practical diagnostic applicability of this method were evaluated. This assay may be also adapted to the diagnosis of suid herpesvirus 1 and equine herpesviruses 1 and 3 and could become a powerful diagnostic tool.     

Nematological problems in the production of conifers

Several conifer nurseries were visited in Piedmont (Italy) and soil and root-samples were taken and examined for plant-parasitic nematodes. In these investigations damage to the mycorrhizae caused by phytohelminths, most frequently through their trophic activity on the tissue of the host plant, has been observed.     

Separation of flavanone-7-O-glycoside diastereomers and analysis in citrus juices by multidimensional liquid chromatography coupled with mass spectrometry

The major flavanone-7-O-glycoside constituents in citrus fruit juices (naringin, hesperidin, neohesperidin, narirutin, and eriocitrin) were separated as diastereomers by multidimensional liquid chromatography. The method consisted of coupling two HPLC columns: a reversed-phase (RP(18)) column was used for the separation of flavanone glycosides, which were, then, individually switched into a carboxymethylated beta-cyclodextrin (beta-CD)-based column and resolved as the corresponding stereoisomers. The method was used for the full analysis of flavanone glycosides in fresh hand-squeezed and commercial fruit juices by combining the quantitative estimation with the diastereomeric analysis. Quantitative data were in general consistent with previously reported data in this field. CC-LC isomer analysis was carried out by coupling the beta-CD column with a mass spectrometer operated with negative ion electrospray ionization (ESI-MS). The results showed that hesperidin was present in orange juices almost exclusively as the 2S isomer, whereas narirutin had mainly the 2R configuration. In grapefruit juices (2S)-naringin prevailed with the respect to the 2R isomer, whereas the opposite was true for narirutin. Lemon juices contained eriocitrin stereoisomers in equal amount (50% each), but hesperidin was almost exclusively found as the 2S isomer. Significant differences of the diastereomeric ratios were observed between freshly squeezed juices and juices from commercial sources.     

Vineyards in Terraced Landscapes: New Opportunities from Lidar Data

Vineyard landscapes are a relevant part of the European culture, and several authors concluded that they are the agricultural practice that causes the highest soil loss. Grape quality depends on the availability of water, and soil erosion is an important parameter dictating the vineyard sustainability; therefore, soil and water conservation measures are often implemented. Among them, the construction of terraces is the most widely used system. However, while favouring agricultural activities, terraces if not properly maintained can lead to local instabilities creating hazards for settlements and cultivations, and for the related economy. Terraced fields are also served by agricultural roads that can have deep effects on water flows triggering surface erosion. The goal of this research is to use lidar elevation data for a hydro‐geomorphological analysis of terraced vineyards. The work is divided in two parts. At first, the Relative Path Impact Index is tested in two vineyards to identify terrace‐induced and road‐induced erosions. Statistical thresholds of the Relative Path Impact Index are then defined to label the most critical areas. On the second step, using the index and the defined thresholds, we simulate different scenarios of soil conservation measures, establishing the optimal solution to reduce erosion. The results highlight the effectiveness of high‐resolution topography in the analysis of surface erosion in terraced vineyards, when the surface water flow is the main factor triggering the instabilities. The proposed analysis can help in scheduling a suitable planning to mitigate the consequences of the anthropogenic alterations induced by the terraces and agricultural roads. Copyright © 2014 John Wiley & Sons, Ltd.     

DYSPLASIA EPIPHYSEALIS HEMIMELICA IN A BOXER PUPPY

A Boxer puppy had an unusual dysplastic lesion of the distal epiphysis of the left femur. Biopsy and CT examination were performed. A diagnosis of dysplasia epiphysealis hemimelica (DEH) was made. To the investigators knowledge, this condition has not been described before in animals. DEH is a growth disorder involving preferentially the medial compartment of the lower limbs, and it is associated with epiphyseal hypertrophy and delayed mineralization.     

Monitoring the effects of storage in caviar from farmed Acipenser transmontanus using chemical, SEM, and NMR methods

The effects of storage at 4 degrees C on the quantity and quality of chemical components in the caviar from farmed Acipenser transmontanus have been analyzed by SEM, chemical methods, and NMR and MRI techniques. Particular attention has been focused on the lipid components, the distribution and mobility of which were strongly affected by the storage time. MRI and relaxation data indicated that lipids are endowed with two different mobility regimes, one slow (short T1) and one fast (long T1), both lengthening with the storage time. Chemical analysis assessed a total fat content that remained practically unchanged and a constant fatty acid composition during the total storage time. The combination of the two methods allowed one (a) to suppose that a mechanism of lipid hydrolysis (faster in unsalted than in salted eggs) is still occurring during storage of caviar at 4 degrees C for up to approximately 4 months and (b) to exclude that an intensive oxidative process is active in the same storage period.     

Evaluation of genetic diversity and root traits of sea beet accessions of the Adriatic Sea coast

Thirty-nine sea beet [Beta vulgaris L. ssp. maritima (L.) Arcang.] accessions of the Adriatic coast were screened genetically and for their adaptive morpho-functional root traits in order to identify new sources of abiotic resistances for sugar beet breeding programs. Genetic diversity was evaluated with 21 microsatellites markers that identified 44 polymorphic alleles. Sea beets grouped into two main clusters: the West and the East Adriatic coast groups, with the latter showing higher genetic diversity. Among sea beet accessions with desirable root traits, four accessions have proved to be interesting for sugar beet [B. vulgaris (L.) ssp. vulgaris] breeding aimed to improve tolerance to nutritional stresses. Lastovo (ID 29) and Zut (ID 34) accessions were characterized by the highest values of RER, TRL, FRL and RSA still maintaining a high value of RTD, while Grado (ID 21) an Portic (ID 23) accessions were characterized by the highest RTD, but with low values of RER, TRL, FRL and RSA parameters.     

Classification of Italian honeys by 2D HR-NMR

The importance of honey has been recently increased because of its nutrient and therapeutic effects, but the adulteration of honey in terms of botanical origin has increased, too. The floral origin of honeys is usually determined using melisso-palynological analysis and organoleptic characteristics, but the application of these techniques requires some expertise. A number of papers have confirmed the possibility of characterizing honey samples by selected chemical parameters. In this study high-resolution nuclear magnetic resonance (HR-NMR) and multivariate statistical analysis methods were used to identify and classify honeys of five different floral sources. The 71 honey samples (robinia, chestnut, citrus, eucalyptus, polyfloral) were analyzed by HR-NMR using both 1H NMR and heteronuclear multiple bond correlation spectroscopy (HMBC). Spectral data were analyzed by application of unsupervised and supervised pattern recognition and multivariate statistical techniques such as principal component analysis (PCA) and general discriminant analysis (GDA). The use of 1H-(13)C HMBC coupled with appropriate statistical analysis seems to be an efficient technique for the classification of honeys.     

Immunochemical assays for direct sulfonamide antibiotic detection in milk and hair samples using antibody derivatized magnetic nanoparticles

Two direct enzyme-linked immunosorbent assays (ELISAs) have been developed for detection of sulfonamide antibiotic residues in milk samples. One of them is using magnetic nanoparticles (MNP) for target capture/enrichment (Ab-MNP-ELISA), and the second is performed using microtiter plates. Selective polyclonal antibodies, raised against 5-[6-(4-amino-benzenesulfonylamino)-pyridin-3-yl]-2-methyl-pentanoic acid (SA1), used in combination with an enzyme tracer prepared with the same hapten, has allowed us to reach a limit of detection (LOD) lower than 0.5 microg L(-1) for both ELISA formats. Sulfapyridine, sulfamethoxypyridazine, sulfathiazole, and sulfachloropyridazine are detected below the maximum residue limits established by the European Union for these antibiotics in milk (100 microg L(-1)). Matrix effects and accuracy studies performed with full-cream milk and hair extracts indicated a lack of interference from these sample matrices and very good recovery values, especially when using the Ab-MNP format. Milk samples and hair extracts can be measured without any previous treatment. The results demonstrate the high potential of these methods as screening tools for food safety and inspection controls.     

<Emphasis Type="Italic">Vicia faba</Emphasis> plants respond to oviposition by invasive <Emphasis Type="Italic">Halyomorpha halys</Emphasis> activating direct defences against offspring

The invasive stink bug Halyomorpha halys is established in many European and American agro-ecosystems, where it causes severe crop losses. Potential control measures might include enhancement of plant defences. When attacked by herbivorous insects that oviposit on it, the plant may respond by priming direct defences, which might affect the development of future brood. Halyomorpha halys attacks numerous plant species in the invaded areas. Here, we investigated whether Vicia faba plants challenged by H. halys females can impair the development of its offspring through the activation of induced direct defences. We measured the weight and dimension of nymphs that developed on oviposition-experienced plants after 7 and 17 days from hatching. Nymphs that developed on oviposition-experienced plants weighed less compared to those that developed either on control plants or on plants solely subjected to H. halys feeding, and third instars showed shorter dimensions (tibia length). In addition, when oviposition-experienced plants were attacked by nymphs, higher and more rapid expression of two jasmonic acid-dependent genes (cysteine proteinase inhibitor gene and NAI1) was detected, possibly due to a mechanism of priming. Increased expression of the salicylic acid-responsive PR1 gene was also detected in egg-experienced plants, although the response was delayed compared to JA-dependent genes. Our results suggest that V. faba plants recognize H. halys oviposition as a warning signal and pre-activate defences against future nymphal herbivory.