Existence of two O-serotypes in the fish pathogen Pseudomonas anguilliseptica

The serological characteristics of a group of 32 Pseudomonas anguilliseptica strains isolated in Spain from seabream (Sparus aurata) and turbot (Scophthalmus maximus) were compared with a total of 18 collection strains of this bacterial species with different geographical and host origin. The employment of different techniques, including slide agglutination, microagglutination and dot blot, allowed us to establish two serological groups, one comprising practically all the eel isolates, and the other including the majority of isolates from other fish species. The study of the lipopolysaccharides (LPS) and outer membrane proteins (OMP) corroborated these results, indicating that the serological differences among strains are due to the somatic antigen and not to antigenic determinants of protein nature. Therefore, a serological scheme of two "O" serotypes is proposed for P. anguilliseptica. The results obtained will be of importance for epidemiological studies as well as for the design of adequate vaccine formulations.     

Gastrointestinal parasites in greater rheas (Rhea americana) and lesser rheas (Rhea pennata) from Argentina

Few data exist on the parasites of ratites, especially from regions within their natural range. It is only recently that extensive studies on the parasites of ostriches (Struthio camelus) have been published, mainly from European countries where commercial farming has expanded. Two species of ratites are native in South America: the lesser rhea also known as Darwin's rhea (Rhea pennata) and the greater rhea (Rhea americana). Both species are considered near threatened by the IUCN and are included in the CITES’ Appendices I and II, respectively. Parasitological studies have conservation implications, as they allow us to assess the risk of transmission of pathogens from farmed ratites to wild populations. In this study 92 faecal samples from greater rheas and 55 faecal samples from lesser rheas from different localities in Argentine were analyzed to determine their gastrointestinal parasites. In greater rheas the protozoa (Balantidium coli-like and Entamoeba spp.) and helminths (Fasciola hepatica and Deletrocephalus spp.). The protozoa had not previously been cited as parasites of greater rheas in South America. Cysts and/or trophozoites of B. coli-like were found in 16.3% of the samples, while the prevalence of the remaining parasites was below 10%. Lesser rheas harbored the protozoa B. coli-like, Entamoeba spp. and Chilomastix spp. as well as F. hepatica and nematode eggs and larvae. B. coli-like cysts were found in 20.0% of the samples, while the prevalence of the other parasites remained below 5%. Some of them had not been cited as infecting lesser rheas yet.     

Deletion of the GI-2 integrase and the wbkA flanking transposase improves the stability of Brucella melitensis Rev 1 vaccine

Brucella melitensis Rev 1 is the best vaccine available for the prophylaxis of small ruminant brucellosis and, indirectly, for reducing human brucellosis. However, Rev 1 shows anomalously high rates of spontaneous dissociation from smooth (S) to rough (R) bacteria, the latter being inefficacious as vaccines. This S-R instability results from the loss of the O-polysaccharide. To overcome this problem, we investigated whether some recently described mechanisms promoting mutations in O-polysaccharide genes were involved in Rev 1 S-R dissociation. We found that a proportion of Rev 1 R mutants result from genome rearrangements affecting the wbo O-polysaccharide loci of genomic island GI-2 and the wbkA O-polysaccharide glycosyltransferase gene of the wbk region. Accordingly, we mutated the GI-2 int gene and the wbk IS transposase involved in those arrangements, and found that these Rev 1 mutants maintained the S phenotype and showed lower dissociation levels. Combining these two mutations resulted in a strain (Rev 2) displaying a 95% decrease in dissociation with respect to parental Rev 1 under conditions promoting dissociation. Rev 2 did not differ from Rev 1 in the characteristics used in Rev 1 typing (growth rate, colonial size, reactivity with O-polysaccharide antibodies, phage, dye and antibiotic susceptibility). Moreover, Rev 2 and Rev 1 showed similar attenuation and afforded similar protection in the mouse model of brucellosis vaccines. We conclude that mutations targeting genes and DNA sequences involved in spontaneous O-polysaccharide loss enhance the stability of a critical vaccine phenotype and complement the empirical stabilization precautions taken during S Brucella vaccine production.     

High Antioxidant Activity Mixture of Extruded Whole Quality Protein Maize and Common Bean Flours for Production of a Nutraceutical Beverage Elaborated with a Traditional Mexican Formulation

The objective of this study was to determine the best combination of extrusion process variables for the production of whole quality protein maize (EQPMF) and common bean (ECBF) flours to prepare a high antioxidant activity mixture (EQPMF + ECBF) suitable to produce a nutraceutical beverage with high acceptability elaborated with a traditional Mexican formulation. Processing conditions were obtained from a factorial combination of barrel temperature (BT?=?120–170 °C) and screw speed (SS?=?120–200 rpm). Response surface methodology was applied to obtain maximum values for antioxidant activity (A _ox A) of the flour mixture (EQPMF + ECBF) and acceptability (A) of the nutraceutical beverage. The best combinations of extrusion process variables for EQPMF and ECBF to prepare an optimized mixture (60%EQPMF?+?40%ECBF) were BT?=?98 °C/SS?=?218 rpm and BT?=?105 °C/SS?=?83 rpm, respectively. The optimized mixture had A _ox A?=?14,320 μmol Trolox equivalent (TE)/100 g sample dry weight (dw) and a calculated protein efficiency ratio (C-PER) of 2.17. A 200 ml portion of a beverage prepared with 25 g of the optimized flour mixture had A _ox A?=?3,222 μmol TE, and A?=?89 (level of satisfaction “I like it extremely”). This nutraceutical beverage could be used as an alternative to beverages with low nutritional/nutraceutical value, such as those prepared with water, simple sugars, artificial flavoring and colorants, which are widely offered in the market.     

Fatty Acid Profile,Total Carotenoids,and Free Radical-Scavenging from the Lipophilic Fractions of 12 Native Mexican Avocado Accessions

Plant Foods for Human Nutrition - Mexico is the world’s largest producer, exporter, and consumer of avocados. “Hass” avocado is the most commercialized cultivar, while the native...     

Antioxidant Activity Evaluation of New Dosage Forms as Vehicles for Dehydrated Vegetables

A dehydrated vegetables mixture loaded in four pharmaceutical dosage forms as powder, effervescent granulate, sugar granulate and gumdrops were investigated for their antioxidant capacity using 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging capacity assay, oxygen radical absorbance capacity assay and ferric reducing antioxidant potential assay. Total phenolic content of dehydrated vegetables powder mixture was also measured by the Folin-Ciocalteu method, so as to evaluate its contribution to their total antioxidant function. The effect of different temperatures on stability of these systems after 90 days storage was also evaluated. These formulations presented strong antioxidant properties and high phenolic content (279 mg gallic acid equivalent/g of sample) and thus could be potential rich sources of natural antioxidants. Antioxidant properties differed significantly among selected formulations (p?<?0.05). Generally, the losses were lower in samples stored under refrigeration. To interpret the antioxidant properties a kinetic approach was performed. Degradation kinetics for the phenolic content and antioxidant capacity followed a zero-order function. Effervescent granulate was the formulation which underwent faster degradation. Contrary, sugar granulate and gumdrops were much more slowly. Time required to halve the initial amount of phenolic compounds was 589?±?45 days for samples stored at 4?º C, and 312?±?16 days for samples stored at room temperature. These developed dosage forms are new and innovative approach for vegetable intakes in population with special requirements providing an improvement in the administration of vegetables and fruits.     

Effect of Flavonoids and Saponins Extracted from Black Bean (Phaseolus vulgaris L.) Seed Coats as Cholesterol Micelle Disruptors

Strategies for reducing plasma cholesterol have been focused on reducing the absorption or synthesis of cholesterol. The aim of this study was to correlate the content of flavonoids and saponins in black bean (Phaseolus vulgaris L.) seed coats extracts with a potential effect in lowering cholesterol absorption by the inhibition of cholesterol micellar solubility. Extracts with different flavonoids and saponins concentration were obtained from a Box-Behnken design used to optimize extraction temperature, stirring time and solvent composition. Major flavonoids and saponins were quantified by HPLC-PDA-ELSD and confirmed through mass spectrometry. Contrary to the flavonoid content, saponins were correlated to the inhibitory effect of cholesterol micelle solubility as an approach to a potential reduction of cholesterol absorption. Extracts with the highest saponin content strongly inhibited cholesterol micellization with values of 55.4?±?1.9 %, while stigmasterol used as control, only reached 12?±?2.3 % at the same concentration (5 mg/ml). The optimal extracting conditions for saponins were 25 °C, during 3 h in 85 % aqueous-methanol. Correlations of inhibitory effect of cholesterol micellar solubility with the content of each identified saponin suggested that the reduction of cholesterol micellization depends on the C-22 substitution of saponins.     

pH-Dependent Solution Structure and Activity of a Reduced Form of the Host-Defense Peptide Myticin C (Myt C) from the Mussel Mytilus galloprovincialis

Myticin C (Myt C) is a highly variable host-defense peptide (HDP) associated to the immune response in the mediterranean mussel (Mytilus galloprovincialis), which has shown to be active across species due to its strong antiviral activity against a fish rhabdovirus found in fish cells overexpressing this HDP. However, the potential antimicrobial properties of any synthetic analogue of Myt C has not yet been analysed. Thus, in this work we have synthesised the sequence of the mature peptide of Myt C variant c and analysed the structure activity relationships of its reduced (non-oxidized) form (red-MytCc). In contrast to results previously reported for oxidized isoforms of mussel myticins, red-MytCc was not active against bacteria at physiological pH and showed a moderate antiviral activity against the viral haemorrhagic septicaemia (VHS) rhabdovirus. However, its chemotactic properties remained active. Structure/function studies in neutral and acid environments by means of infrared spectroscopy indicated that the structure of red-MytCc is pH dependent, with acid media increasing its alpha-helical content. Furthermore, red-MytCc was able to efficiently aggregate artificial phospholipid membranes at low pH, as well as to inhibit the Escherichia coli growth, suggesting that this activity is attributable to its more structured form in an acidic environment. All together, these results highlight the dynamic and environmentally sensitive behavior of red-Myt C in solution, and provide important insights into Myt C structure/activity relationships and the requirements to exert its antimicrobial/immunomodulatory activities. On the other hand, the pH-dependent direct antimicrobial activity of Myt C suggests that this HDP may be a suitable template for the development of antimicrobial agents that would function selectively in specific pH environments, which are sorely needed in this “antibiotic-resistance era”.     

Climate Variability and Oceanographic Settings Associated with Interannual Variability in the Initiation of Dinophysis acuminata Blooms

In 2012, there were exceptional blooms of D. acuminata in early spring in what appeared to be a mesoscale event affecting Western Iberia and the Bay of Biscay. The objective of this work was to identify common climatic patterns to explain the observed anomalies in two important aquaculture sites, the Galician Rías Baixas (NW Spain) and Arcachon Bay (SW France). Here, we examine climate variability through physical-biological couplings, Sea Surface Temperature (SST) anomalies and time of initiation of the upwelling season and its intensity over several decades. In 2012, the mesoscale features common to the two sites were positive anomalies in SST and unusual wind patterns. These led to an atypical predominance of upwelling in winter in the Galician Rías, and increased haline stratification associated with a southward advection of the Gironde plume in Arcachon Bay. Both scenarios promoted an early phytoplankton growth season and increased stability that enhanced D. acuminata growth. Therefore, a common climate anomaly caused exceptional blooms of D. acuminata in two distant regions through different triggering mechanisms. These results increase our capability to predict intense diarrhetic shellfish poisoning outbreaks in the early spring from observations in the preceding winter.