Establishment of cryopreserved gene banks of European chestnut and cork oak

Cryopreservation of selected genotypes of European chestnut and cork oak was carried out in two laboratories in a project involving conservation of field collections. Plant material was selected on the basis of disease resistance (chestnut), growth habit, phytosanitary performance and cork quality (cork oak). The cryopreservation technique comprised of vitrification of shoot apices isolated from in vitro stock shoot cultures (chestnut) and somatic embryos (cork oak). Forty-three out of 46 chestnut genotypes assayed survived the freezing process, but only 63% recovered their capacity to produce new shoots. After completion of multiplication and rooting steps, the surviving shoots produced plants that were morphologically identical to those derived from non-supercooled material. All 51 cork oak genotypes withstood freezing and were able to produce new somatic embryos through a process of secondary embryogenesis. Multiplication and germination of the recovered embryos enabled production of plants that were morphologically identical to those derived from non-supercooled material. In light of the results obtained, long-term cryopreservation of these species is feasible, thereby ensuring conservation of valuable genotypes during field evaluation.     

Structure and species composition of novel forests dominated by an introduced species in northcentral Puerto Rico

The African tulip tree, Spathodea campanulata Beauv., is an introduced species forming novel forest types in Puerto Rico. These forests develop naturally after deforestation, agricultural use and land abandonment, and there are many questions as to their ecological characteristics. We sampled structure and species composition of large, small, and juvenile trees (≥10, ≥2.5 to <10, and <2.5 cm diameter at breast height, respectively) in nine secondary forests dominated by S. campanulata on alluvial, karst, and volcanic substrates in northcentral Puerto Rico. No differences were found in S. campanulata forest structure between substrates. Of a total of 79 species found, 17 were introduced. Forests on karst and alluvium had the highest and lowest global species richness, respectively. Species richness increased from large to small to juvenile trees in most sites, but more so on karst. The percentage of introduced species was inversely related to species richness of tree size classes on all substrate types. The dominance of S. campanulata in the large tree size class was highest and lowest in alluvial and volcanic sites, respectively, and decreased from large to small to juvenile trees on all substrate types. Species richness of S. campanulata forests is lower than that of native forests on equivalent substrates. Although land use history affects composition, the juvenile tree species established in S. campanulata forests seems to correspond to the geological substrate were sites are found. This study shows S. campanulata restores forest structure and native tree species on abandoned agriculture and grazing lands in Puerto Rico. Management of these novel forests should consider them as resources were natural processes have the potential of making them more diverse.     

Light quality affects in vitro adventitious rooting and ex vitro performance of cherry rootstock Colt

The effect of light quality on in vitro rooting of cherry rootstock “Colt” was studied to evaluate adventitious root development, ex vitro acclimatization and plant growth performance. Adventitious rooting was dependent on the light quality to which the microcutting were exposed. Highest number of roots per microcuttings was recorded under dichromatic light (blue + red). This response could be ascribed to the control of a strong synergistic interaction between phytochromes and blue light photoreceptors. Red light was more effective on root elongation than blue light. In this response a strong synergistic interaction between phytochromes and blue light photoreceptors was suggested. The effect of light quality on the number of root/explant affected the plant during acclimation, scoring the highest level of plant survival in the blue-, red- and blue + red-exposed plantlets. The light quality effect was also observed under greenhouse culture conditions, as shown by growth parameters at the end of six months in plants growth. No specific role was observed for the photoequilibrium of phytochrome. The results reported in this work show that plantlets exposed to different light quality, during the in vitro rooting phase, retain the light quality effects in the subsequent plant acclimatization and greenhouse plant growth phase.     

Epicatechin: endothelial function and blood pressure

Epidemiological studies indicate an inverse relationship between flavanol intake and the risk of cardiovascular disease. Potential mechanisms include their effects on endothelial function and hypertension. A number of studies have shown that flavanol-rich cocoa reduces blood pressure and endothelial dysfunction, whereas black tea may have opposite effects. These results highlight the importance of the different effects of the multitude of phytochemical constituents in these foods and the need for studying the individual flavanols. Epicatechin seems to be a major bioactive constituent of cocoa and other flavanol-rich foods and beverages. It has been shown to improve endothelial function in animals and humans. In salt-sensitive animal models of hypertension, epicatechin lowers blood pressure and the associated end-organ damage. Nitric oxide (NO) seems to play a key role in the protection of both hypertension and endothelial dysfunction.     

Chemical conversion of phenylethylamine into phenylacetaldehyde by carbonyl-amine reactions in model systems

The chemical conversion of phenylethylamine into phenylacetaldehyde in the presence of lipid oxidation products (LOPs) was studied to investigate the possibility that biogenic amines can be converted into Strecker aldehydes upon processing. Model systems of phenylethylamine and methyl 13-hydroperoxyoctadeca-9,11-dienoate (HP), 2,4-decadienal (DD), 4,5-epoxy-2-heptenal (EH), 4,5-epoxy-2-decenal (ED), 4-oxo-2-hexenal (OH), 4-oxo-2-nonenal (ON), or 4-hydroxy-2-nonenal (HN) were heated for 1 h at 180 °C and pH 3. Although HN and EH did not produce more phenylacetaldehyde than when phenylethylamine was heated alone, all other lipid oxidation products assayed increased the amount of phenylacetaldehyde produced by 300-900%, with ON being the most reactive compound for this reaction. The reaction was mainly produced at acidic pH values (<6) and was dependent upon the concentration of the LOPs involved, and the phenylacetaldehyde produced increased linearly as a function of the time and temperature. The E(a) values for the reactions between phenylethylamine and DD and ON were 54.8 and 53.8 kJ/mol, respectively. The reaction is proposed to take place by the formation of an imine between the phenylethylamine and the LOPs, which is later converted into another imine by an electronic rearrangement. This new imine is the origin of phenylacetaldehyde by hydrolysis. These results show a new pathway for Strecker aldehyde formation. This route provides a potential way to reduce biogenic amine content in foods when they can be thermally processed before consumption.     

Food-derived peptides stimulate mucin secretion and gene expression in intestinal cells

In this study, the hypothesis that food-derived opioid peptides besides β-casomorphin 7 might modulate the production of mucin via a direct action on epithelial goblet cells was investigated in HT29-MTX cells used as a model of human colonic epithelium. Seven milk whey or casein peptides, a human milk peptide, and a wheat gluten-derived peptide with proved or probable ability to bind μ- or δ-opioid receptors were tested on the cell culture. Significantly increased secretion of mucins was found after exposure to six of the assayed peptides, besides the previously described β-casomorphin 7, as measured by an enzyme-linked lectin assay (ELLA). Human β-casomorphin 5 and α-lactorphin were selected to study the expression of mucin 5AC gene (MUC5AC), the HT29-MTX major secreted mucin gene. α-Lactorphin showed increased expression of MUC5AC from 4 to 24 h (up to 1.6-fold over basal level expression), although differences were statistically different only after 24 h of exposure. However, this increased expression of MUC5AC did not reach significance after cell treatment with human β-casomorphin 5. In conclusion, six food-derived peptides have been identifed with described or probable opioid activity that induce mucin secretion in HT29-MTX cells. Concretely, α-lactorphin is able to up-regulate the expression of the major secreted mucin gene encoded by these cells.     

Effects of adrenocorticotropic hormone challenge and age on hair cortisol concentrations in dairy cattle

Dairy cattle suffer stress from management and production; contemporary farming tries to improve animal welfare and reduce stress. Therefore, the assessment of long-term hypothalamic-pituitary-adrenal function using non-invasive techniques is useful. The aims in this study were: to measure cortisol concentration in cow and calves hair by radioimmunoassay (RIA), to test cortisol accumulation in bovine hair after adrenocorticotropic hormone (ACTH) challenges, and determine the influence of hair color on cortisol concentrations. Fifteen Holstein heifers were allotted to 3 groups (n = 5 each): in control group (C), just the hair was sampled; in the saline solution group (SS), IV saline solution was administered on days 0, 7, and 14; and the ACTH group was challenged 3 times with ACTH (0.15 UI per kg of body weight) on days 0, 7, and 14. Serum samples from the SS and ACTH groups were obtained 0, 60 and 90 min post-injection. Serum cortisol concentration was greater 60 and 90 min after injection with ACTH. Hair was clipped on days 0, 14, 28, and 44. Hair cortisol was methanol extracted and measured by RIA. Hair cortisol was preserved for 11 mo. Hair cortisol concentrations in the ACTH group were greater than in the saline and control groups on days 14 and 28, but not on day 44. Concentrations were greater in calves than in cows and greater in white hair than in black hair. Cortisol accumulated in bovine hair after ACTH challenges, but the concentration was affected by both age and hair color. If hair color effects are taken into account, assessing cortisol concentration in hair is a potentially useful non-invasive method for assessing stress in cattle.     

Control of postharvest diseases on citrus fruit by preharvest applications of biocontrol agent Pantoea agglomerans CPA-2: Part II. Effectiveness of different cell formulations

Infection of citrus fruit by postharvest pathogens often occurs in the field prior to harvest; therefore, it could be advantageous to apply biocontrol agents before harvest, which would reduce initial infection and then remain active and control pathogens in storage and under commercial conditions. The objective of the present work was to evaluate the effectiveness of different formulations of Pantoea agglomerans applied preharvest for controlling postharvest diseases on citrus. Results confirmed the protective effect of the additive Fungicover (FC) on populations of P. agglomerans exposed to non-conducive field conditions. In general, when osmotic-adapted and lyophilised P. agglomerans cells were used in bacterial treatments, these treatments showed greater survival rates than treatments with non-osmotic-adapted or fresh cells under field conditions. However, this superiority was only found when Fungicover was also added to suspensions of bacterial treatments. Therefore, bacterial treatments with Fungicover had population levels of P. agglomerans cells 1.2 and 2.8 log CFU cm⁻² higher than bacterial treatments without Fungicover during field experiments. These results allowed us to conclude that it is possible to improve environmental stress tolerance and ecological competence of P. agglomerans cells by integrating certain formulation strategies. Consequently, the improved formulation of P. agglomerans provided an effective control for orange fruit against natural postharvest pathogen infections and artificial infections of Penicillium digitatum with values of decay reduction higher than 50%. These latter results also demonstrated that it is possible to control postharvest pathogens using bacterial preharvest treatments.     

Influence of different anaesthetic protocols over the sperm quality on the fresh,chilled (4°C) and frozen‐thawed epididymal sperm samples in domestic dogs

This study assessed the influence of three different anaesthetic protocols on semen quality obtained from the epididymis. Sixty male dogs undergoing to routine sterilization were assigned to three anaesthetic protocols: thiopental group (TG, n = 20), propofol group (PG, n = 20) and ketamine–dexmedetomidine group (KDG, n = 20). Immediately after orchidectomy, the cauda epididymides and vas deferent ducts were isolated and then a retrograde flushing was performed to collect spermatozoa. In experiment 1, after the initial evaluation of the semen (sperm concentration, sperm motility and the percentages of live spermatozoa, abnormal spermatozoa and acrosome membrane integrity), semen samples were diluted in Tris‐glucose‐egg yolk extender and chilled for 48 hr, and the sperm motility was assessed at 6, 24 and 48 hr. In experiment 2, semen samples were diluted in Tris‐glucose‐egg yolk extender and chilled for 24 hr, and then samples were frozen in two extenders with different glycerol concentrations, to reach a final concentration of 50–100 × 10⁶ spermatozoa ml^?1, 20% egg yolk, 0.5% Equex and 4% and 5% glycerol, respectively. Mean values of total sperm concentration, sperm viability and the percentages of intact acrosome and abnormal spermatozoa were not significantly different between experimental groups, and therefore, the anaesthetic protocols assessed did not affect sperm parameters mentioned above. However, our study confirmed a detrimental effect of the use of thiopental (TG) over the total sperm motility (p < 0.05) and progressive sperm motility (p < 0.05) of the fresh and chilled epididymal sperm samples. The anaesthetic protocols including the application of propofol or ketamine–dexmedetomidine can be used to recover sperm in domestic canids without significant changes in sperm quality compared when semen is collected routinely and these techniques could be applicable to endangered wild canids.