Genetic stability at nuclear and plastid DNA level in regenerated plants of <Emphasis Type="Italic">Solanum</Emphasis> species and hybrids

In this work we detected the extent of variability at nuclear and cytoplasmic DNA level of regenerated plants belonging to Solanum genotypes with a different genetic background and somatic chromosome number. As for the nuclear characterization, a total of 66 (18.5%) polymorphic bands were scored using 13 ISSR primers on 45 randomly selected regenerants. Our results show that the regenerants obtained from clone cmm 1T and, at lower level, those from cph 1C are unstable under in vitro conditions or rather more prone to in vitro-induced stress leading to somaclonal variation than the other genotypes used. Two types of changes were observed: disappearance of parental ISSR fragments, termed “loss”; appearance of novel ISSR fragments, termed “gain”. The most frequent event occurring in the regenerants was the loss of fragments (41 bands). Regenerated plants were analyzed with seven plastid universal primers to determine the cytoplasmic composition at chloroplast level. All cpDNA primer pairs tested produced amplicons of the same size in all genotypes analyzed and no polymorphic fragments were observed with any universal primers used. Our results show that under in vitro culture conditions genotype affects the integrity of the genome. In addition, the absence of polymorphism at plastid level confirms the greater genetic stability of cytoplasmic DNA.     

Sowing time and prediction of flowering of different hemp (Cannabis sativa L.) genotypes in southern Europe

Fibre hemp can be grown under a wide range of agro-ecological conditions, but it requires special attention for several physiological features and crop management. A management strategy in order to reduce inputs and thus achieve acceptable yield could be achieved optimizing sowing time. With this respect, the effects of sowing date on hemp biology and yield was studied, using two monoecious and two dioecious genotypes. Field experiments were carried out in two subsequent years (2003-2004) in South of Italy, using drip irrigation system. Sowing time, in the two year period, ranged between March 10th and July 22nd. Optimal sowing time was observed between the end of April and the first three weeks of May; in that range, the dioecious Fibranova yielded the most in terms of aboveground biomass and stem dry yield, followed by Tiborszallasi, while the two monoecious showed the lowest yield. On the contrary, before and after that period, the shorter day length caused an early floral induction that strongly reduced stem and fibre elongation, and thus aboveground dry biomass and consequently stem yields. Based on this study a simulation model focusing on flowering prediction in Mediterranean environment was developed.     

Efficacy of Vaccination Against Streptococcus iniae During Artificial Spawning of the Red-Tail Black Shark (Epalzeorhynchos bicolor,fam. Cyprinidae)

Artificial spawning may precipitate high mortalities of red-tail black shark (Epalzeorhynchos bicolor) broodstock from Streptococcus iniae infection. Two vaccine formulations (aluminum or oil adjuvant) were evaluated for effectiveness. Fish (mean weight 21.2 ± 5.7 g) were harvested, acclimatized, and vaccinated by intracoelomic injection. After 21 days, fish were moved to a biosecure laboratory and, one day later, simultaneously spawned and challenged by bath or by intracoelomic injection with S. iniae. Both formulations increased relative% survival (RPS) in unspawned (RPS [formulation]: 80 [aluminum]; 54 [oil]) and spawned fish (87–95 [aluminum]; 73–75 [oil]), suggesting that vaccination increases survival of red-tail black shark broodstock after spawning.     

Proteomic profiling of sea bass muscle by two-dimensional gel electrophoresis and tandem mass spectrometry

In this study, the proteome profile of European sea bass (Dicentrarchus labrax) muscle was analyzed using two-dimensional electrophoresis (2-DE) and tandem mass spectrometry with the aim of providing a more detailed characterization of its specific protein expression profile. A highly populated and well-resolved 2-DE map of the sea bass muscle tissue was generated, and the corresponding protein identity was provided for a total of 49 abundant protein spots. Upon Ingenuity Pathway Analysis, the proteins mapped in the sea bass muscle profile were mostly related to glycolysis and to the muscle myofibril structure, together with other biological activities crucial to fish muscle metabolism and contraction, and therefore to fish locomotor performance. The data presented in this work provide important and novel information on the sea bass muscle tissue-specific protein expression, which can be useful for future studies aimed to improve seafood traceability, food safety/risk management and authentication analysis. This work is also important for understanding the proteome map of the sea bass toward establishing the animal as a potential model for muscular studies.     

Isolation and characterization of Avenin-like protein type-B from durum wheat

Proteomic techniques were applied for the investigation of “avenin-like protein type-B” in tetraploid wheat. Protein was extracted from endosperm, according to a classical chromatographic procedure and its identity confirmed by liquid chromatography-tandem mass spectrometry analysis. The characterization of avenin-like protein type-B was in good agreement with the gene-derived sequence, with the exception of glutamine as N-terminus. The presence in the glutenin extract and the high number of cysteine residues suggested that avenin-like protein type-B is integrated into gluten polymers via inter-chain disulphide bonds. This raises the question of whether the protein could play a role in determining the functional properties of gluten.     

Dimensional analysis of milk fat globules in sow milk: effects of the lactation stage and fat content and comparison with vaccine milk

To assess the differences in the granulometry of milk fat globules between swine and bovine species, milk samples from 30 lactating sows were analyzed for fat globule dimensions and compared with cow milk samples. Results showed differences between the fat globules: sow milk presents reduced globule diameters compared with cow milk (volume-weighted diameter 2.62 vs. 3.27 µm, p < 0.001) and reduced interglobular distance. A positive relationship was observed between milk fat content and globule diameter, while a slight, insignificant inverse trend was detected between the day of lactation and fat globule diameter. These complex interactions between milk lipids, globule membrane proteins, and globule dimensions provide a better understanding of digestion/absorption phenomena in the design of milk replacers.

     

Effects of two fractions of inspired oxygen on lung aeration and gas exchange in cats under inhalant anaesthesia

ObjectiveTo compare the effects of two fractions of inspired oxygen (FiO₂) (0.4 and 1) on lung aeration and gas exchange during general anaesthesia in cats.Study designRandomized, blinded, controlled study.AnimalsThirty healthy, mixed breed, client owned female cats.Materials and methodsCats were premedicated intramuscularly with acepromazine (0.03 mg kg^?1) and medetomidine (0.015 mg kg^?1). Anaesthesia was induced with propofol (5 mg kg^?1) and, after orotracheal intubation, maintained with isoflurane carried by either 100% oxygen (G100, n = 15) or an oxygen-air mixture with 40% oxygen (G40, n = 15). All cats were placed in dorsal recumbency and breathed spontaneously throughout the entire procedure. Following surgery (ovariectomy), a spiral computed tomography (CT) of the thorax was performed, arterial oxygen (PaO₂) and carbon dioxide (PaCO₂) tensions were measured and alveolar-arterial gradient of oxygen [P(A-a)O₂] calculated. The CT images were analysed for lung aeration by the analysis of radiograph attenuations (Hounsfield units, HU), according to the following classification: hyperinflated area (-1000 to -900 HU), normally aerated area (-900 to -500 HU), poorly aerated area (-500 to -100 HU) and non-aerated area (-100 to +100 HU). The groups were compared using one-way anova.ResultsCompared to G100, the normally-aerated lung area was significantly greater and the poorly-aerated and non-aerated areas were significantly smaller in G40. PaCO₂ was similar in both groups. PaO₂ and P(A-a)O₂ were significantly higher in G100. In both groups, pulmonary atelectasis developed preferentially in the caudal lung fields.ConclusionIn cats anaesthetised with isoflurane, the administration of an FiO₂ of >0.9 significantly impaired lung aeration and gas exchange as compared to an FiO₂ of 0.4.Clinical relevanceAn FiO₂ of 0.4 may better preserve lung aeration and gas exchange in anaesthetised spontaneously breathing cats but monitoring is essential to ensure oxygenation is adequate.     

Assessment of synthetic chemicals for disruption of Rhynchophorus ferrugineus response to attractant-baited traps in an urban environment

The red palm weevil (RPW), Rhynchophorus ferrugineus Olivier (Coleoptera, Curculionidae), is one of the most severe pests of ornamental palm species in urban areas of Mediterranean countries. Aiming to discover inhibitory semiochemicals for RPW population management in urban environments, we conducted electroantennographic (EAG) screenings of 17 commercially available synthetic compounds, representing three groups of plant volatiles (isoprenoids, phenyl propanoid derivatives and fatty acid derivatives) known for their repellent effects toward insects. These tests were followed by trap-based screenings of EAG-active menthone, α-pinene and methyl salicylate, singly and in combination, under urban conditions. In EAG bioassays, RPW antennae of both sexes showed positive dose-dependent responses to 13 of the 17 synthetic chemicals with significant differences among them. In field trapping experiments, conducted in the city of Palermo, Italy, from weeks 31 to 38 in 2010 and 2011, α-pinene, tested singly or in combination with methyl salicylate (2010) or menthone (2011), reduced trap catches by about 30% to 40%. Methyl salicylate and menthone alone were inactive. In conclusion, RPW is significantly affected by α-pinene. This isoprenoid is a promising disruptant for semiochemical-based management of this pest. In addition, identification of a large number of EAG-active chemicals could serve as a database for future design of active repellents or attractants of RPW adults.     

Presence of Cocaine in the Tissues of the European Eel, Anguilla anguilla, Exposed to Environmental Cocaine Concentrations

The presence of illicit drugs and their metabolites in surface waters has to be considered a new type of hazard, still unknown, for the aquatic ecosystem, due to the potent pharmacological activities of all the illicit drugs. Our research was therefore aimed at evaluating the impact of illicit drugs on the aquatic fauna, till now still undervalued. To this aim, we verified the ability of the European eel (Anguilla anguilla), a well-known biomonitor of environmental contamination, to bioaccumulate cocaine, one of the most abundant illicit drugs found in surface waters. Silver eels were exposed to a nominal cocaine concentration of 20?ng/l for 1?month; at the same time, control, carrier, and post-exposure recovery groups were made. Brains, gills, liver, kidney, muscle, gonads, spleen, digestive tract, and sections of dorsal skin were assayed by high-pressure liquid chromatography. Cocaine was found in the tissues of the treated eels and, at low concentrations, in almost all tissues of post-exposure recovery eels. These results indicate that cocaine is able to accumulate into the eel tissues; its presence suggests potential risks for eels since cocaine could affect their physiology and contribute to their decline, and for humans consuming contaminated fish.