Assessment of genetic variation in a working collection of Vigna vexillata (L.) A. Rich. by isozyme and RAPD analyses

Genetic variation based on isozyme and RAPD analyses was investigated in 47 and 34 accessions respectively of Vigna vexillata from different geographical origins and belonging to three botanical varieties. A total of 9 enzyme systems were studied, accounting for 14 putative loci, 8 of which were polymorphic. The analysis of genetic diversity revealed a low level of within accession variation (H_S=0.013), while between accession diversity (D_ST) was 0.120. Coefficient of gene differentiation (G_ST) was 0.905, indicating that most variation was among accessions. Nei's genetic distances were calculated on the basis of allelic frequencies and a UPGMA dendrogram was constructed. Twenty arbitrary 10-mer oligonucleotides were used in RAPD analysis. Amplification profiles disclosed a higher level of polymorphism than isozymes. Based on amplification patterns, the similarity index of Jaccard was calculated and a dendrogram constructed on the basis of the similarity matrix. The final clustering based on RAPD data was similar to the one obtained using isozyme allelic frequencies. The classification in botanical varieties did not reflect the allelic constitution of the different samples. On the other hand, referring to geographical origin, most accessions from Africa and from Latin America were distributed respectively in two distinct clusters in the dendrogram. This grouping might also reflect the differences observed in the germination behaviour of V. vexillata from the two continents.     

Interactions between temperature and size on the growth,size heterogeneity,mortality and cannibalism in cultured larvae and juveniles of the Asian catfish,Pangasianodon hypophthalmus (Sauvage)

Temperature effects vary throughout the ontogeny, but are proportionally more variable during the early life stages than in older fish. The larvae of a few species have been studied but contrasting trajectories were observed, thereby making it difficult to predict how temperature impacts on the growth, survival and size heterogeneity in a particular species. This study examined these interactions in young [0.9 mg–1 g wet mass (WM)] Pangasianodon hypophthalmus, one of the most extensively cultured tropical fish. Fish were raised at five temperatures from 23 to 33 °C in a recirculated water system, fed in excess with a high‐energy feed (Artemia then a formulated feed, >50% protein), and examined at 1‐day (first feeding days) or 4‐day intervals (older fish). The temperature that produced the fastest growth (T°_opt) was 31 °C at the start of exogenous feeding (0.9 mg); it increased to 32.7 °C at 8 mg then decreased by 0.7 °C for each 10‐fold increase of WM. Size heterogeneity was lower and survival was higher (70–85% from 0.9 mg to 1 g) when the temperature was close to T°_opt. Comparisons with other species suggest that the slope of the relationship between T°_opt and fish size has a strong latitudinal component, and may also reflect the fish sensitivity to oxygen.     

Ecological classification of land and conservation of biodiversity at the national level: The case of Italy

The aims of this study are to describe the ecological classification of land in Italy and to show how the resulting land units can act as reliable frameworks for coarse scale environmental analyses that can be used to implement national conservation strategies. We first collected, homogenised and drew physical thematic maps, which were then linked to biological and human features. We then performed a gap analysis of land heterogeneity compared with Natural Protected Areas and Natura2000 network on the basis of three categories: Total gaps, Partial gaps, and Protected. Moreover, we assessed the conservation status of the land units by summarising the environmental quality using the Index of Landscape Conservation. We identified and mapped 3 Land Regions, 24 Land Systems, and 149 Land Facets. Total gaps account for 28% of the country, Partial gaps for 38% and Protected for 34%. The Natura2000 network is more representative than the system of National Protected Areas of the overall land heterogeneity as regards both the types (18 out of 24) and extent (72%) of the Land Systems. Low conservation status prevails in the Land Facets of the Mediterranean Region located along the coasts and plains on sedimentary deposits, whereas high and very high conservation status is found along the higher belts of the Alpine and Apennine chains. These results highlight the potential use of ecological land classification for biodiversity monitoring and conservation purposes, e.g. when identifying land units that need to be recovered or targeted for enhanced biodiversity and ecosystem services protection.     

Occurrence of pipecolic acid and pipecolic acid betaine (homostachydrine) in Citrus genus plants

The presence of pipecolic acid and pipecolic acid betaine, also known as homostachydrine, is herein reported for the first time in Citrus genus plants. Homostachydrine was found in fruits, seeds, and leaves of orange, lemon, and bergamot (Citrus bergamia Risso et Poit). As homostachydrine was not commercially available, as a comparative source, extracts of alfalfa leaves ( Medicago sativa L.) were used, in which homostachydrine is present at high concentration. Then, the results where confirmed by comparison with an authentic standard synthesized and purified starting from pipecolic acid. The synthesized standard was characterized by a ESI-MS/MS study using a 3D ion-trap mass spectrometer. When subjected to MS/MS fragmentation in positive ion mode, homostachydrine, unlike its lower homologue proline betaine (also known as stachydrine), showed a pattern of numerous ionic fragments that allowed unambiguous identification of the compound. For the quantitation in the plant sources, high sensitivity and specificity were achieved by monitoring the transition (158 → 72), which is absent in the fragmentation patterns of other major osmolytes commonly used as markers for studies of abiotic stress. As for the metabolic origin of homostachydrine, the occurrence in citrus plants of pipecolic acid leads to the hypothesis that it could act as a homostachydrine precursor through direct methylation.     

Susceptibility to flavescence dorée of different Vitis vinifera genotypes from north-western Italy

The aim of this work was to evaluate the susceptibility to flavescence dorée (FD) of 12 Vitis vinifera cultivars grown in Piedmont, and representative of the wine-making tradition of this area. The experiments were conducted under controlled conditions to ensure constant infection pressure. Test plants were ex vitro potted vines, singly inoculated with four Scaphoideus titanus infected by FD-C phytoplasma (FDp), under greenhouse conditions. Vines were tested for FDp at 5 and 8 weeks postinoculation (wpi) and the phytoplasma load was measured in leaves and roots at 8 wpi. Within the 14 V. vinifera accessions (belonging to 12 cultivars), three susceptibility clusters were identified. Cultivars within the low susceptibility group showed low phytoplasma loads and low percentages of infected plants, suggesting a tolerant behaviour. To confirm these results, four Vitis cultivars, representing extremes of FD susceptibility from low to high, were grafted onto Kober 5BB rootstocks and inoculated with laboratory-infected S. titanus, under semi-field conditions. The transmission experiments onto grafted cuttings confirmed that susceptibility to the disease depends on the scion genotype. The data indicated that none of the tested V. vinifera genotypes are resistant to FD, although some cultivars with low susceptibility are available, and can be explored for identifying genetic traits involved in disease tolerance/resistance. Moreover, ranking Vitis genotypes for their susceptibility to FD is in itself a valuable tool to support vine growers in their decision management, by helping them to choose the most appropriate varieties according to their specific FD epidemiological contexts.     

Post-traumatic peripheral arteriovenous fistula manifesting as digital haemorrhages in a cat: diagnosis with contrast-enhanced 3D CT imaging

Arteriovenous fistulae (AVF) are defined as congenital or acquired abnormal direct communications between an artery and a vein leading to abnormal blood circulation. This report describes an unusual manifestation of acquired peripheral AVF in a cat for which the diagnosis was confirmed by computed tomographic (CT) imaging and three-dimensional (3D) reconstruction. A 10-year-old female spayed domestic shorthaired cat was presented with a 2-month history of nonhealing, crusting, erosive and ulcerative skin lesions on the dorsal right forepaw. Severe chewing and biting, but not lameness, had been reported. Systemic abnormalities were not noted. Histopathology revealed increased numbers of thin-walled and slightly grouped vascular profiles in the superficial and mid-dermis, which were often markedly dilated and partially obscured by prominent hyaline deposits. There were a few pyknotic nuclear fragments and haemorrhages in vascular walls as well as multifocal luminal thrombosis with or without recanalization. Differential diagnoses included progressive angiomatosis with trauma or AVF with secondary regional venous hypertension. Computed tomographic images were acquired using a 16-slice Siemens Somotom Sensation CT scanner, and 3D images were created using the Voxar 3D software. Image reconstruction revealed tortuous aberrant vasculature on the medial aspect of the radius and around the carpus compared to normal vascularization on the contralateral limb. These changes were suggestive of the diagnosis of acquired peripheral AVF. The differential diagnosis for localized, nonhealing, haemorrhagic, crusted, erosive or ulcerative distal extremity skin lesions in cats should include acquired AVF, and diagnosis may be confirmed with contrast-enhanced CT imaging.     

Intrinsic innervation of the horse ileum

This paper describes the morphology and distribution of the enteric nervous system (ENS) cells and fibres immunoreactive for choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), substance P (SP), calcitonin gene-related peptide (CGRP), NF200 kDa (NF200), and S100 protein. The percentages of subclasses of enteric neurons in the total neuronal population were investigated by the use of anti-PGP 9.5 or anti-NSE antibodies.ChAT-IR myenteric plexus (MP) and submucosal plexus (SMP) neurons were 66 ± 7% and 74 ± 15%, respectively, whereas those cells expressing nNOS-IR were 38 ± 7% and 5 ± 1%, respectively. MP and SMP neurons expressing both phenotypes were also present. SP-IR was expressed by 14 ± 13% of MP and 66 ± 8% of SMP neurons whereas CGRP-IR was observed only in the SMP (43 ± 6%). NF200-IR was expressed by 61 ± 15% and 91 ± 10% of the MP and SMP neurons, respectively. The majority of the CGRP-IR SMP neurons expressed also SP-IR. Almost all SP-IR neurons in both the plexuses were cholinergic. The present study quantifies the main neuronal subpopulations of the ENS of the horse ileum; these data might be utilized to understand the neuronal modifications which occur in several gastrointestinal tract disorders.     

Differential scanning calorimetry evidence of the enhancement of beta-sitosterol absorption across biological membranes mediated by beta-cyclodextrins

beta-Sitosterol is a plant sterol that has received much attention because of its effectiveness in reducing the absorption of dietary cholesterol, as well as in offering protection from cardiovascular diseases and cancer development. Thus, the knowledge of the interaction of beta-sitosterol with biological membranes can help in understanding its mechanism of action. In the present paper, the differential scanning calorimetry technique has been used to study the interaction of beta-sitosterol with a biomembrane model constituted by dimyristoylphosphatidylcholine multilamellar vesicles. Furthermore, kinetic experiments have been carried out to follow the uptake of beta-sitosterol by biomembranes and the effect of beta-cyclodextrins on such a process. Our results indicate that opportune concentrations of beta-cyclodextrins improve the uptake of beta-sitosterol by phospholipid membranes.