Isolation,growth and differentiation of equine mesenchymal stem cells: effect of donor,source, amount of tissue and supplementation with basic fibroblast growth factor

Mesenchymal stem cells (MSC) are increasingly used as therapeutical aid for the orthopaedic injuries in the horse. MSC populate different tissues but the most commonly used for clinical purposes are isolated from bone marrow or adipose tissue. The first objective of this study was to investigate if the donor animal, the tissue of origin and the technique of isolation could influence the number of MSC available for transplantation after a short-term expansion. The second aim was to devise a culture system capable of increasing MSC lifespan and we tested the effect of basic fibroblast growth factor (bFGF). Results indicate that MSC can be efficiently isolated from both sources and supplementation of bFGF enhances proliferation rate maintaining differentiation potential. In addition, this study shows that collection, expansion and storage of frozen MSC can be performed for later therapeutic use.     

Application of high‐throughput single nucleotide polymorphism genotyping for assessing the origin of Engraulis encrasicolus eggs

1. Single nucleotide polymorphism (SNP) markers in anchovy (Engraulis encrasicolus) egg samples were analysed to detect their origin on a small spatial scale (200 km) by assigning genotypes to adult anchovy stocks. The novelty of this work is the application of a rapid high‐throughput method for genotyping each single anchovy egg, in a single execution, using a set of 96 genome‐wide SNPs in a dynamic array system with microfluidic technology (Fluidigm 96.96).
2. The existence of two ecotypes in E. encrasicolus had already been identified based on SNP polymorphism in the Atlantic Ocean and in the Mediterranean Sea, showing that habitat type (offshore versus coastal/estuarine) is the most important component of genetic differentiation among populations of anchovy.
3. In this work, anchovy egg genotypes from areas of the Western Mediterranean were assigned to adult populations. Only two localities in which adult anchovies were sampled represented donor populations for the coastal/estuarine egg genotypes. Although some degree of mixing among the hauls could exist, the assignment of egg groups to adult populations led to distinguishing the contributions of distinct ecotypes to new wild generations. We can conclude that the high rate of egg dispersion caused by marine currents and the different degrees of local retention could explain the genetic heterogeneity observed in the adult populations, where eggs from neighbouring spawning sites tend to mix.
4. The results highlight that this technique represents a new and useful tool for addressing evolutionary questions, breed recognition, assignment, and connectivity assessment of individual eggs, and anchovy population dynamics, for the management of stocks.

     

Extranodal gammadelta-T-cell lymphoma in a dog with leishmaniasis

An 8-year-old intact male mongrel dog with alopecia and weight loss was referred to the Veterinary Faculty of Naples. The dog had pale mucous membranes, enlarged prescapular lymph nodes, and splenomegaly. Laboratory abnormalities included anemia, thrombocytopenia, and hyperglobulinemia. Bone marrow aspirate smears contained numerous Leishmania amastigotes and an immunofluorescent antibody titer was strongly positive (1:1280) for leishmaniasis. The dog was treated with a combination of meglumine antimoniate and allopurinol for 60 days and showed clinical improvement. Two months after the end of treatment the dog was again referred because of relapse of leishmaniasis and the presence of a firm subcutaneous mass on the medial right thigh. Based on cytologic examination of fine needle aspirates of the mass, a diagnosis of large-cell lymphoma was made. Flow cytometry of tumor cells revealed gammadelta-T-cell lymphoma with a CD5+, CD3+, TCRgammadelta+, CD4-, CD8-, CD45RA+ immunophenotype. Using nested PCR, amastigotes were not detected in the neoplastic tissue. An association between leishmaniasis and hematopoietic tumors has been described rarely. gammadelta-T cells may be involved in the host response to this parasite, and prolonged antigenic stimulation and chronic immunosuppression (typical of leishmaniasis) play a crucial role in the etiopathogenesis of T-cell lymphoma.     

Sex ratio variation and sex determination in the mallee dragon Ctenophorus fordi

Recent evidence suggests that many Australian agamids show temperature‐dependent sex determination (TSD) with variation in sex determining mechanisms among closely related taxa. However, as shown in other vertebrates, sex ratios can also be influenced by genetic or phenotypic differences among females in their propensity to produce sons or daughters, and these influences might confound any thermal effects of incubation per se. To address these issues, we investigated the determinants of sex ratios in the mallee dragon Ctenophorus fordi, together with a detailed analysis of karyotypes. There was no detectable variation in sex ratios arising from variation among females, clutches or incubation temperatures, which might indicate genetic sex determination for this species. However, there was no evidence of cytologically distinct sex chromosomes using standard banding techniques. The sex ratio pattern in C. fordi strongly contrasts with the results for the congener Ctenophorus pictus, where sex ratios show variation among females. Thus, Australian agamids offer promising opportunities to address fundamental issues in sex ratio biology.     

Alternative pathways in the life history of Gracilaria gracilis (Gracilariales, Rhodophyta) from north-eastern Sicily (Italy)

The reproductive phenology of Gracilaria gracilis has been followed closely in a brackish water lake in Sicily. The life history of this red alga deviated from the typical Polysiphonia-type cycle and was characterized by the predominance of the tetrasporophytic phase over gametophytes, in situ development of germinated gametophytes and the presence of sterile pleustophytic thalli. Release of spherical agglomerates of cells produced by young tetrasporophytes was observed as well. Their ability to germinate forming new thalli appears to represent an alternative reproductive strategy of this organism. The population structure appears to reflect a predominance of the diploid phase except during the warmer summer season when only vegetative juveniles were found in the lake. Release and early stages of development of carpospores and tetraspores were critically observed in culture. A Dumontia type pattern of germination was observed for both types of spores, showing an erect thallus arising from a multicellular basal disk. Compared to tetraspores, carpospores are less readily released resulting in a subsequent delay in the development of new gametophytes.     

Resistance screening essay of wine lactic acid bacteria on lysozyme: efficacy of lysozyme in unclarified grape musts

In wine making, the bacteriolytic activity of lysozyme has primarily been used to control the malolactic fermentation in wines. The use of lysozyme in musts before settling and the beginning of the alcoholic fermentation to inhibit the growth of lactic acid bacteria could be very beneficial. In a resistance test carried out in MT/b broth, lysozyme had greater antimicrobial activity toward Oenococcus oeni than Lactobacillus species. Several strains of wine bacteria belonging to Oenococcus proved sensitive to the bacteriolytic activity of lysozyme at low concentrations in both synthetic medium (MT/b) (50 mg/L), white must, or red must made with or without the skins (100 mg/L). Lactobacillus and Pediococcus strains survived at lysozyme concentrations of 200-500 and 500 mg/L, respectively, in MT/b and musts. Suspended solids in unclarified musts may strongly bind to lysozyme thereby causing its removal by filtration or centrifugation. One hour after lysozyme was added to musts, it was quantified by HPLC and found after centrifugation to be 40-50% and only 10% in musts made with or without the skins, respectively. Although appreciable amounts of lysozyme were bound to wine components, this did not appear to be a serious hindrance to lysozyme activity.     

On sweet acorn (<Emphasis Type="Italic">Quercus</Emphasis> spp.) cake tradition in Italian cultural and ethnic islands

The CNR-IGV, over the last 15 years, has organized and performed expeditions aimed at collecting germplasm and local knowledge on plants used by local populations in isolated areas, such as geographical islands, or local communities that for ethnic or cultural reasons have maintained to a certain degree their isolation from surrounding populations (cultural islands). In two such cultural islands (Bovesia, Calabria region, south-west mainland Italy, and Ogliastra, Sardinia island, Italy) a tradition regarding the use of acorns for preparing a cake was recorded. In Ogliastra, an elder lady still retains the knowledge regarding the collections, maintenance and utilization of acorns for preparing a cake, a knowledge once common and now quite completely disappeared. The present paper reports on the culture associated to acorn used as a food for humans.