Effect of farmers' seed selection on genetic variation of a landrace population of pepper (Capsicum annuum L.), grown in North-West Italy

Recognition by the European Community of élite landraces encourages farmers to grow these and earn more than is possible by growing modern varieties. However, farmers often exercise strong selection by collecting seed from a limited number of plants which best embody a few characters of prime interest. This, in the long run, may be responsible for considerable genetic erosion. In Southern Piedmont, North-West Italy, a local landrace of pepper (Capsicum annuum L.) known as Cuneo is grown; it deserves particular attention because of its hardiness, late production and fruit quality. We used random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers to assess the effect, on genetic composition of a population, of seed sampling carried out using the selection criteria adopted by the farmer. After two reproductive cycles using selection, it was already possible to detect loss in genetic variation and a change in allele frequencies, while no significant effect was found after two cycles of random sampling. Over this period, farmer selection pressure led to disappearance of eight low-frequency alleles, while only three alleles were lost in randomly sampled populations. Our results stress the need to adopt appropriate techniques for seed sampling in order to avoid genetic erosion of local landrace gene pools.     

Flumequine Uptake and the Aquatic Duckweed, Lemna minor L.

Phytotoxicity of Flumequine (F) on the aquatic duckweed, Lemna minor L., and plant drug uptake were evaluated by a simple ecotoxicological test. Flumequine, at all concentrations between 50 and 1000 μg L^-1 tested, affected the plant growth: leaves and roots were damaged, but duckweed continued to grow on a five weeks period. Furthermore, increasing drug concentrations decreased the chlorophyll b content in plants. These effects depend on F uptake by plants, which is quite high (from 0.72 to 13.93 μg g^-1 plant dry weight). Based on this activity, Lemna can be taken into consideration as a tool for in situ remediation of drug contaminated waters: the presence of Lemna significantly lower the F concentration in culture media on a five weeks period. Results strongly support its remediation capability.     

SSR-based identification key of cultivars of Olea europaea L. diffused in Southern-Italy

Olive (Olea europaea L.) is an important fruit species in Italy and Mediterranean basin constituted by a wide germplasm with a large number of cultivars present in all the Mediterranean area. SSRs are becoming the markers of choice for variability studies in olives as they are simple to perform, transferable, hypervariable, highly polymorphic and show a high information content.Olive genetic diversity was studied using 16 SSR markers on 30 cultivars diffused in Southern-Italy. Resolving Power (RP) and Power of Discrimination (PD) were calculated to evaluate the efficiency of the SSR markers investigated in studies of cultivars fingerprinting. Based on their high efficiency, two SSR markers, UDO43 and DCA16 were chosen to set up an identification key to distinguish the entire set of cultivars, confirming the high biodiversity of the Southern-Italian olive germplasm and the suitability of SSR markers in studies of cultivar diagnosis.     

A Metataxonomic Approach Reveals Diversified Bacterial Communities in Antarctic Sponges

Marine sponges commonly host a repertoire of bacterial-associated organisms, which significantly contribute to their health and survival by producing several anti-predatory molecules. Many of these compounds are produced by sponge-associated bacteria and represent an incredible source of novel bioactive metabolites with biotechnological relevance. Although most investigations are focused on tropical and temperate species, to date, few studies have described the composition of microbiota hosted by Antarctic sponges and the secondary metabolites that they produce. The investigation was conducted on four sponges collected from two different sites in the framework of the XXXIV Italian National Antarctic Research Program (PNRA) in November–December 2018. Collected species were characterized as Mycale (Oxymycale) acerata, Haliclona (Rhizoniera) dancoi, Hemigellius pilosus and Microxina sarai by morphological analysis of spicules and amplification of four molecular markers. Metataxonomic analysis of these four Antarctic sponges revealed a considerable abundance of Amplicon Sequence Variants (ASVs) belonging to the phyla Proteobacteria, Bacteroidetes, Actinobacteria and Verrucomicrobia. In particular, M. (Oxymycale) acerata, displayed several genera of great interest, such as Endozoicomonas, Rubritalea, Ulvibacter, Fulvivirga and Colwellia. On the other hand, the sponges H. pilosus and H. (Rhizoniera) dancoi hosted bacteria belonging to the genera Pseudhongella, Roseobacter and Bdellovibrio, whereas M. sarai was the sole species showing some strains affiliated to the genus Polaribacter. Considering that most of the bacteria identified in the present study are known to produce valuable secondary metabolites, the four Antarctic sponges could be proposed as potential tools for the discovery of novel pharmacologically active compounds.     

Expression of verocytotoxic Escherichia coli antigens in tobacco seeds and evaluation of gut immunity after oral administration in mouse model

Verocytotoxic Escherichia (E.) coli strains are responsible for swine oedema disease, which is an enterotoxaemia that causes economic losses in the pig industry. The production of a vaccine for oral administration in transgenic seeds could be an efficient system to stimulate local immunity. This study was conducted to transform tobacco plants for the seed-specific expression of antigenic proteins from a porcine verocytotoxic E. coli strain. Parameters related to an immunological response and possible adverse effects on the oral administration of obtained tobacco seeds were evaluated in a mouse model. Tobacco was transformed via Agrobacteium tumefaciens with chimeric constructs containing structural parts of the major subunit FedA of the F18 adhesive fimbriae and VT2e B-subunit genes under control of a seed specific GLOB promoter. We showed that the foreign Vt2e-B and F18 genes were stably accumulated in storage tissue by the immunostaining method. In addition, Balb-C mice receiving transgenic tobacco seeds via the oral route showed a significant increase in IgA-positive plasma cell presence in tunica propria when compared to the control group with no observed adverse effects. Our findings encourage future studies focusing on swine for evaluation of the protective effects of transformed tobacco seeds against E. coli infection.     

Impact of aquatic acoustic noise on oxidative status and some immune parameters in gilthead sea bream Sparus aurata (Linnaeus, 1758) juveniles

This study evaluated the ambient noise effect of onshore and offshore aquaculture systems on oxidative status and on some immune parameters in gilthead sea bream (Sparus aurata) juveniles. The experiment was performed for 40 days, projecting two different playlists of acoustic stimuli: offshore aquaculture noise condition that created the typical acoustic field in proximity of an offshore sea cage for fish farming, using a random sequence of quite sea background and boat noises; and onshore aquaculture noise condition that represented the acoustic field inside an open concrete tank for fish farming. A third group without any acoustic projection was used as a control. After 40 days from the beginning of acoustic noise, fish exposed to the offshore aquaculture condition showed significantly higher levels of total oxidant status, lysozyme activity, antiprotease activity and white blood cells together with a lower albumin/globulin ratio in comparison to fish from the control group; whereas fish exposed to the onshore condition showed analytical data placed between the other groups, though never significantly different. These results demonstrate that the offshore aquaculture noise, and in particular the sea soundscape, adversely influences the oxidative status and the immune function of gilthead sea bream determining a mild stress condition that could affect the sea bream welfare.     

Could hypoxia influence basic biological properties and ultrastructural features of adult canine mesenchymal stem /stromal cells?

The aim of the present study was to compare canine adipose tissue mesenchymal stem cells cultured under normoxic (20% O₂) and not severe hypoxic (7% O₂) conditions in terms of marker expression, proliferation rate, differentiation potential and cell morphology. Intra-abdominal fat tissue samples were recovered from 4 dogs and cells isolated from each sample were cultured under hypoxic and normoxic conditions. Proliferation rate and adhesion ability were determined, differentiation towards chondrogenic, osteogenic and adipogenic lineages was induced; the expression of CD44, CD34, DLA-DQA1, DLA-DRA1 was determined by PCR, while flow cytometry analysis for CD90, CD105, CD45 and CD14 was carried out. The morphological study was performed by transmission electron microscopy. Canine AT-MSCs, cultured under different oxygen tensions, maintained their basic biological features. However, under hypoxia, cells were not able to form spheroid aggregates revealing a reduction of their adhesivness. In both conditions, MSCs mainly displayed the same ultrastructural morphology and retained the ability to produce membrane vesicles. Noteworthy, MSCs cultivated under hypoxya revealed a huge shedding of large complex vesicles, containing smaller round-shaped vesicles. In our study, hypoxia partially influences the basic biological properties and the ultrastructural features of canine mesenchymal stem /stromal cells. Further studies are needed to clarify how hypoxia affects EVs production in term of amount and content in order to understand its contribution in tissue regenerative mechanisms and the possible employment in clinical applications. The findings of the present work could be noteworthy for canine as well as for other mammalian species.     

Ascorbate dynamics in embryos and larvae of sea bass and sea bream, originating from broodstocks fed supplements of ascorbic acid

The transfer of L-ascorbic acid (AA) from broodstock to the fertilised eggs and it's dynamics in embryos and fasting larvae, has been studied in sea bass (Dicentrarchus labrax L.) and sea bream (Sparus aurata L.). Two types of diets were fed to the broodstocks: one containing sufficient ascorbate for normal growth and the other with an extra addition of a very high dose (2,000 mg/kg feed) of ethyl-cellulose-coated L-ascorbic acid.The concentration of AA and total ascorbate (L-ascorbic acid plus dehydroascorbic acid) was detected through several embryo and larval development stages, including larvae just before feeding. In sea bass and sea bream, the mean (SD) concentration of total ascorbate in fertilised eggs originating from broodstock fed AA supplemented diet, was 218.5 (17.7) and 122.4 (5.1) g/g wet weight respectively. This was significantly higher (P < 0.01) than the unsupplemented groups, which contained only 155.9 (6.9) and 103.9 (3.5) g/g wet weight respectively. A diet with a vitamin C content adequate for normal growth, may not be sufficient for broodstock when the goal is transfer of TAA to embryos. © Rapid Science Ltd. 1998     

Bioactivity Screening of Antarctic Sponges Reveals Anticancer Activity and Potential Cell Death via Ferroptosis by Mycalols

Sponges are known to produce a series of compounds with bioactivities useful for human health. This study was conducted on four sponges collected in the framework of the XXXIV Italian National Antarctic Research Program (PNRA) in November-December 2018, i.e., Mycale (Oxymycale) acerata, Haliclona (Rhizoniera) dancoi, Hemimycale topsenti, and Hemigellius pilosus. Sponge extracts were fractioned and tested against hepatocellular carcinoma (HepG2), lung carcinoma (A549), and melanoma cells (A2058), in order to screen for antiproliferative or cytotoxic activity. Two different chemical classes of compounds, belonging to mycalols and suberitenones, were identified in the active fractions. Mycalols were the most active compounds, and their mechanism of action was also investigated at the gene and protein levels in HepG2 cells. Of the differentially expressed genes, ULK1 and GALNT5 were the most down-regulated genes, while MAPK8 was one of the most up-regulated genes. These genes were previously associated with ferroptosis, a programmed cell death triggered by iron-dependent lipid peroxidation, confirmed at the protein level by the down-regulation of GPX4, a key regulator of ferroptosis, and the up-regulation of NCOA4, involved in iron homeostasis. These data suggest, for the first time, that mycalols act by triggering ferroptosis in HepG2 cells.