An accurate high-resolution melting method to genotype bovine β-casein

In the present study, we used high-resolution melting analysis to simultaneously identify the two mutations that taken together give us the genotype of the most common bovine β-casein variants. Allelic variants are detected in real time, and no post-PCR manipulations are required, therefore limiting costs and possible carryover contamination. Data can be copied to a Microsoft Excel spreadsheet for semiautomatic determination of the genotype. The method was validated using different DNA sources, in 122 samples from 12 different bovine populations, obtaining the first report on bovine β-casein genotypes for some of them.     

A disaster-severity assessment DSS comparative analysis

This paper aims to provide a comparative analysis of fuzzy rule-based systems and some standard statistical and other machine learning techniques in the context of the development of a decision support system (DSS) for the assessment of the severity of natural disasters. This DSS, which will be referred to as SEDD, has been proposed by the authors to help decision makers inside those Non-Governmental Organizations (NGOs) concerned with the design and implementation of international operations of humanitarian response to disasters. SEDD enables a relatively highly accurate and interpretable assessment on the consequences of almost every potential disaster scenario to be obtained through a set of easily accessible information about that disaster scenario and historical data about similar ones. Thus, although SEDD’s methodology is rather sophisticated, its data requirements are small, which, therefore, enables its use in the context of NGOs and countries requiring humanitarian aid. In this sense, SEDD opposes to some current tools which focuses on one phenomena-one place disaster scenarios (earthquakes in California, hurricanes in Florida, etc.) and/or have extensive and/or technologically sophisticated data requirements (real-time remote sensing information, exhaustive building census, etc.). Moreover, although focused on disaster response, SEDD can also be useful in other phases of disaster management, as disaster mitigation or preparedness. Particularly, the predictive accuracy and interpretability of SEDD fuzzy methodology is compared here in a disaster severity assessment context with those of multiple linear regression, linear discriminant analysis, classification trees and support vector machines. After an extensive validation over the EM-DAT disaster database, it is concluded that SEDD outperforms the methods above in the task of simultaneously providing an accurate and interpretable inference tool for the evaluation of the consequences of disasters.     

Potential of wine-associated lactic acid bacteria to degrade biogenic amines

Some lactic acid bacteria (LAB) isolated from fermented foods have been proven to degrade biogenic amines through the production of amine oxidase enzymes. Since little is known about this in relation to wine micro-organisms, this work examined the ability of LAB strains (n = 85) isolated from wines and other related enological sources, as well as commercial malolactic starter cultures (n = 3) and type strains (n = 2), to degrade histamine, tyramine and putrescine. The biogenic amine-degrading ability of the strains was evaluated by RP-HPLC in culture media and wine malolactic fermentation laboratory experiments. Although at different extent, 25% of the LAB isolates were able to degrade histamine, 18% tyramine and 18% putrescine, whereas none of the commercial malolactic starter cultures or type strains were able to degrade any of the tested amines. The greatest biogenic amine-degrading ability was exhibited by 9 strains belonging to the Lactobacillus and Pediococcus groups, and most of them were able to simultaneously degrade at least two of the three studied biogenic amines. Further experiments with one of the strains that showed high biogenic amine-degrading ability (L. casei IFI-CA 52) revealed that cell-free extracts maintained this ability in comparison to the cell suspensions at pH 4.6, indicating that amine-degrading enzymes were effectively extracted from the cells and their action was optimal in the degradation of biogenic amines. In addition, it was confirmed that wine components such as ethanol (12%) and polyphenols (75 mg/L), and wine additives such as SO₂ (30 mg/L), reduced the histamine-degrading ability of L. casei IFI-CA 52 at pH 4.6 by 80%, 85% and 11%, respectively, in cell suspensions, whereas the reduction was 91%, 67% and 50%, respectively, in cell-free extracts. In spite of this adverse influence of the wine matrix, our results proved the potential of wine-associated LAB as a promising strategy to reduce biogenic amines in wine.     

Study of the shelf life of a dry fermented sausage "salchichon" made from raw material enriched in monounsaturated and polyunsaturated fatty acids and stored under modified atmospheres

The microbiological, physico-chemical and sensory properties of salchichon with high unsaturated fat content, packed under vacuum and 20/80% CO(2)/N(2) modified atmosphere, were evaluated to determine its quality changes during storage under refrigeration. These sausages were manufactured with pork meat and pork backfat obtained from pigs fed with three different diets (control diet-CO, high oleic diet-HO and high linoleic diet-HL). In general, few significant differences were found in counts of different groups of microorganism between the three types of sausages and no difference between the packaging methods. A reduction in pH values was observed during storage and no great differences were determined by storage period on water activity (a(w)). Both parameters (pH and a(w)) presented similar results to those found in different Spanish sausages and other European dry fermented products. The sensory results denoted that sensory quality gradually decreased during storage under both packaging conditions (vacuum and 20/80% CO(2)/N(2)), so it is not advisable to store longer than 150days. On the other hand, fermented sausages with high content of unsaturated fatty acids had similar sensory properties to those of conventional sausages, and even a comparable sensory stability. In conclusion, the results showed healthier salchichons (HO and HL) similar to the traditional (CO) one could be manufactured and stored under refrigeration after slicing for a reasonable period, but the advantage of the gas mixture packaging (20/80% CO(2)/N(2)) versus vacuum packaging was not clear.     

Effect of muscular hypertrophy on physico-chemical, biochemical and texture traits of meat from yearling bulls

The effect of the presence of the double-muscling gene either homozygous (mh/mh) or heterozygous (mh/+) on the physico-chemical, biochemical and texture traits of Longissimus thoracis muscle of yearling bulls of the Asturiana de los Valles (AV) breed was studied. Meat of mh/mh bulls had lower amount of intramuscular fat (p<0.001) and also lower total (p<0.01) and insoluble collagen (p<0.05), although the double-muscling genotype did not affect collagen solubility. Homozygous animals had lower pigment content (p<0.05) and a lighter meat, showing lower water holding capacity, estimated as expressible juice under pressure (p<0.001). Genotype affected significantly the metabolic traits of muscle, with mh/mh animals having higher muscle glycolytic metabolism, assessed by a higher (p<0.001) lactate dehydrogenase (LDH) activity and lower (p<0.001) oxidative activity of the isocitrate dehydrogenase (ICDH). The percentage of myosin heavy chain I in muscle was lower, although not significantly, for mh/mh bulls compared to mh/+ bulls. Texture measurements performed on raw material showed that meat of mh/mh bulls had lower resistance to total and 80% compression, which means lower background or collagen toughness. However, there were no differences between genotypes on shear force of cooked meat. Therefore, the physico-chemical, biochemical and texture traits of meat from mh/mh and mh/+ bull are in general significantly different, which could affect the sensorial quality of meat and hence the consumer acceptance.     

Study of Spray System Applications of Edible Coating Suspensions Based on Hydrocolloids Containing Cellulose Nanofibers on Grape Surface (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.)

An edible coating is a useful technology to preserve fruit quality by covering them with a protective layer which improves the appearance and provides a semipermeable barrier for gases and water vapor transfer, allowing extension of their shelf life. In this study, edible coatings based on hydroxypropyl methyl cellulose, k-carrageenan, glycerol, and cellulose nanofibers were formulated. The operational conditions of the spray system were studied to obtain a coating with optimal adhesion on grape (Vitis vinifera L.) surface. Furthermore, the physico-chemical properties of grapes covered with edible coatings were evaluated during refrigerated storage. A full factorial 2³ experimental design was applied, where liquid suspension flow (1–5 L h^?1), air pressure (50–200 kPa), and height of impact (0.3–0.5 m) were evaluated as independent variables, whereas the percentage of coating and thickness of coating was the response variables. Both mechanical and physico-chemical properties were evaluated during 41 days in both coated and uncoated grapes as shelf life criteria. Throughout the storage time, noticeable changes in pH and total soluble solids were not found in grapes covered with edible coatings and they showed the highest stability for the evaluated mechanical properties. Moreover, coated grapes showed final weight loss and water vapor permeability values of approximately 30 and 34% lower, respectively, than uncoated grapes, suggesting a shelf life extension.     

Stimulation of microsomal cholesterol ester hydrolase by glucagon,cyclic AMP analogues,and vasopressin in isolated rat hepatocytes

Short-term activation of microsomal cholesterol ester hydrolase by glucagon, cAMP analogues, and vasopressin in isolated rat hepatocytes is described. Glucagon led to a dose-and time-dependent activation of cholesteryl oleate hydrolysis, but values returned to basal levels within 120 min. Exposure of isolated hepatocytes to 0.5 mM concentrations of dibutyryl-cAMP or 8-[4-chlorophenylthio]-cAMP, or 25 μM forskolin caused persistent activation of cholesterol ester hydrolase activity after a lag period of 30 min. The three agents resulted in early marked intracellular accumulation of cAMP that declined progressively, and moderate and sustained reductions in the diacylglycerol content. The actions of glucagon on hepatocytes were inhibited by pretreatment of cells with 10 nM [8-arginine] vasopressin. Vasopressin elicited a consistent and sustained increase in cholesterol ester hydrolase activity and diacylglycerol without affecting cAMP while reducing the effect of glucagon on cAMP. Furthermore, the effects of glucagon and vasopressin on the activation of cholesterol ester hydrolase were not additive despite the similarity of their stimulation of diacylglycerol formation. Blockade of vasopressin-mediated activation of cholesterol ester hydrolase and diacylglycerol content were induced by excess prazosin. These data suggest that stimulation of microsomal cholesterol ester hydrolase in isolated liver cells may involve at least two signal transduction systems.     

Biodegradation of 2-fluorobenzoate and dichloromethane under simultaneous and sequential alternating pollutant feeding

Two up-flow fixed-bed reactors (UFBRs), inoculated with activated sludge and operated for 162 days, were fed 1mmolL(-1)d(-1) with two model halogenated compounds, 2-fluorobenzoate (2-FB) and dichloromethane (DCM). Expanded clay (EC) and granular activated carbon (GAC) were used as biofilm carrier. EC did not have any adsorption capacity for both model compounds tested, whereas GAC could adsorb 1.3mmolg(-1) GAC for 2-FB and 4.5mmolg(-1) GAC for DCM. Both pollutants were degraded in both reactors under simultaneous feeding. However, biodegradation in the EC reactor was more pronounced, and re-inoculation of the GAC reactor was required to initiate 2-FB degradation. Imposing sequential alternating pollutant (SAP) feeding caused starvation periods in the EC reactor, requiring time-consuming recovery of 2-FB biodegradation after resuming its feeding, whereas DCM degradation recovered significantly faster. The SAP feeding did not affect performance in the GAC reactor as biodegradation of both pollutants was continuously observed during SAP feeding, indicating the absence of true starvation.