Antithrombotic Activity of Brewers’ Spent Grain Peptides and their Effects on Blood Coagulation Pathways

Antithrombotic activity of brewers’ spent grain peptides before and after simulated gastrointestinal digestion and their effects on blood coagulation pathways were evaluated. Two hydrolysates were produced using sequential enzymatic systems: alkaline protease + Flavourzyme (AF) and neutral protease + Flavourzyme (PF). Simulation of gastrointestinal digestion of AF and PF hydrolysates was made using porcine pepsin and pancreatin enzymes, obtaining the corresponding digested samples: AFD and PFD, respectively. Peptides were fractionated by ultrafiltration using a 1 kDa cut-off membrane. Hydrolysates had peptides with medium and low molecular weights (2100 and 500 Da, respectively), and Glu, Asp, Leu, Ala, and Phe were the most abundant amino acids. Gastrointestinal digested hydrolysates presented high proportion of small peptides (~500 Da), and higher amount of Val, Tyr, and Phe than hydrolysates. Mass spectrum (HDMS Q-TOF) of AFD-ultrafiltered fraction <1 kDa exhibited peptides from 500 to 1000 Da, which are not present in AF. PFD showed the generation of new peptides from 430 to 1070 Da. All samples showed thrombin inhibitory activity. However, no effect was observed on prothrombin time. Peptides <1 kDa from hydrolysates and digested samples delayed thrombin and thromboplastin time respect to the control (~63%). Also the samples showed thrombin inhibitory activity at common pathway level. Thus, brewers’ spent grain peptides exerted their antithrombotic activity by inhibiting the intrinsic and common pathways of blood coagulation. This is the first report to demonstrate that brewers’ spent grain peptides are able to delay clotting time after simulated gastrointestinal digestion.     

Evaluating the performance of xanthophyll,chlorophyll and structure-sensitive spectral indices to detect water stress in five fruit tree species

This study assessed the capability of several xanthophyll, chlorophyll and structure-sensitive spectral indices to detect water stress in a commercial farm consisting of five fruit tree crop species with contrasting phenology and canopy architecture. Plots irrigated and non-irrigated for eight days of each species were used to promote a range of plant water status. Multi-spectral and thermal images were acquired from an unmanned aerial system while concomitant measurements of stomatal conductance (g_s), stem water potential (Ψ_s) and photosynthesis were taken. The Normalized Difference Vegetation Index (NDVI), red-edge ratio (R₇₀₀/R₆₇₀), Transformed Chlorophyll Absorption in Reflectance Index normalized by the Optimized Soil Adjusted Vegetation Index (TCARI/OSAVI), the Photochemical Reflectance Index using reflectance at 530 (PRI) and 515 nm [PRI_(570–515)] and the normalized PRI (PRI_norm) were obtained from the narrow-band multi-spectral images and the relationship with the in-field measurements explored. Results showed that within the Prunus species, Ψ_s yielded the best correlations with PRI and PRI_(570–515) (r² = 0.53) in almond trees, with TCARI/OSAVI (r² = 0.88) in apricot trees and with PRI_norm, R₇₀₀/R₆₇₀ and NDVI (r² from 0.72 to 0.88) in peach trees. Weak or no correlations were found for the Citrus species due to the low level of water stress reached by the trees. Results from the sensitivity analysis pointed out the canopy temperature (T_c) and PRI_(570–515) as the first and second most sensitive indicators to the imposed water conditions in all the crops with the exception of apricot trees, in which Ψ_s was the most sensitive indicator at midday. PRI_norm was the least sensitive index among all the water stress indicators studied. When all the crops were analyzed together, PRI_(570–515) and NDVI were the indices that better correlations yielded with Crop Water Stress Index, g_s and, particularly, Ψ_s (r² = 0.61 and 0.65, respectively). This work demonstrated the feasibility of using narrow-band multispectral-derived indices to retrieve water status for a variety of crop species with contrasting phenology and canopy architecture.     

Insights into the virulence‐related genes of Edwardsiella tarda isolated from turbot in Europe: genetic homogeneity and evidence for vibrioferrin production

Edwardsiella tarda has long been known as a pathogen that causes severe economic losses in aquaculture industry. Insights gained on E. tarda pathogenesis may prove useful in the development of new methods for the treatment of infections as well as preventive measures against future outbreaks. In this report, we have established the correlation between the presence of virulence genes, related with three aspects typically involved in bacterial pathogenesis (chondroitinase activity, quorum sensing and siderophore‐mediated ferric uptake systems), in the genome of E. tarda strains isolated from turbot in Europe and their phenotypic traits. A total of 8 genes were tested by PCR for their presence in 73 E. tarda isolates. High homogeneity was observed in the presence/absence pattern of all the strains. Positive results in the amplification of virulence‐related genes were correlated with the detection of chondroitinase activity in agar plates, in vivo AHL production during fish infection and determination of type of siderophore produced by E. tarda. To the best of our knowledge, this is the first study carried out with European strains on potential virulence factors. Furthermore, we demonstrated for the first time that E. tarda produces the siderophore vibrioferrin.     

New insights into radiata pine seedling root infection by Fusarium circinatum

Pine root infection by Fusarium circinatum has been reported in the literature, but the underlying pathogenic interaction is poorly understood. A green fluorescent protein (GFP)‐tagged F. circinatum isolate, together with confocal microscopy, was used in order to monitor the events associated with root infection of Pinus radiata seedlings. It was found that in order to reach and successfully infect pine roots, F. circinatum employed features that are similar to those previously described for other root‐infecting pathogens, such as mycelial strands, single runner hyphae and simple hyphopodia as well as other features that are reminiscent of those that are known to be involved in biotrophic invasion, such as bulbous invasive hyphae and filamentous invasive hyphae. Abundant sporulation was observed at the root surface as well as inside tracheids both in roots and in the root collar region. The fungus can spread from the roots to the aerial parts of the plant, and once there, colonization appears to be similar to the process that occurs when the pathogen is inoculated in the stem. Wilting symptoms and plant demise may be the result of a reduction in water uptake by roots and of the blockage of the vascular system by fungal hyphae and resin.     

The effect of Saccharomyces cerevisiae on digestion and mortality in the volcano rabbit(Romerolagus diazi)

An experiment was conducted to evaluate whether supplementation with a probiotic could enhance digestion and reduce mortality in the volcano rabbit in captivity. Two enclosures at Chapultepec Zoo, Mexico(114 individuals) were used in a crossover design(two periods of 60 days) with the following treatments: control group and supplementation with Saccharomyces cerevisiae(2×108 CFU/exhibit/day). Supplementation with the probiotic negatively affected(P0.01) the digestibility of dry matter, organic matter, neutral detergent fiber(NDF) and energy. Mortality increased(P0.04) following supplementation with the probiotic(4.26% vs. 8.89%), primarily in the juvenile rabbits. The results indicate that yeast supplementation in the volcano rabbit negatively affects digestion and mortality in captivity.     

Direct-fed microbes: A tool for improving the utilization of low quality roughages in ruminants

For many years, ruminant nutritionists and microbiologists have been interested in manipulating the microbial ecosystem of the rumen to improve production efficiency of different ruminant species. Removal and restriction of antibiotics subtherapeutic uses from ruminant diets has amplified interest in improving nutrient utilization and animal performance and search for more safe alternatives. Some bacterial and fungal microorganisms as a direct-fed microbial(DFM) can be the most suitable solutions. Microorganisms that are commonly used in DFM for ruminants may be classified mainly as lactic acid producing bacteria(LAB), lactic acid utilizing bacteria(LUB), or other microorganism's species like Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Bacillus, Propionibacterium, Megasphaera elsdenii and Prevotellabryantii, in addition to some fungal species of yeast such as Saccharomyces and Aspergillus. A definitive mode of action for bacterial or fungal DFM has not been established; although a variety of mechanisms have been suggested. Bacterial DFM potentially moderate rumen conditions, and improve weight gain and feed efficiency. Fungal DFM may reduce harmful oxygen from the rumen, prevent excess lactate production, increase feed digestibility, and alter rumen fermentation patterns. DFM may also compete with and inhibit the growth of pathogens, immune system modulation, and modulate microbial balance in the gastrointestinal tract. Improved dry matter intake, milk yield, fat corrected milk yield and milk fat content were obtained with DFM administration. However, the response to DFM is not constant; depending on dosages, feeding times and frequencies, and strains of DFM. Nonetheless, recent studies have supported the positive effects of DFM on ruminant performance.