Diet supplementation with donkey milk upregulates liver mitochondrial uncoupling,reduces energy efficiency and improves antioxidant and antiinflammatory defences in rats

Dietary PUFA, mainly those of the n‐3 family, are known to play essential roles in the maintenance of energy balance and in the reduction of body fat deposition through the upregulation of mitochondrial uncoupling that is the main source of reactive oxygen species. We hypothesized that rat supplementation with raw donkey's milk (DM), characterized by low‐fat content and higher n3:n6 ratio, may affect energy balance, lipid metabolism, and prooxidant status as compared to animals treated with cow's milk. In the present study, the effects of drinking raw DM (for 4 weeks) on energy balance, lipid metabolism, antiinflammatory, and antioxidant/detoxifying defences was compared to that produced by rat intake of an iso‐energetic amount of raw cow's milk. The hypolipidemic effect produced by DM paralleled with the enhanced mitochondrial activity/proton leakage and with the increased activity or expression of mitochondrial markers namely, carnitine palmitoyl transferase and uncoupling protein 2. The association of decreased energy efficiency with reduced proinflammatory signs (TNF‐α and LPS levels) with the significant increase antioxidant (total thiols) and detoxifying enzyme activities (glutathione‐S‐transferase NADH quinone oxidoreductase) in DM‐treated animals, indicated that beneficial effects were attributable, at least in part, to the activation of nuclear factor 2 erythroid‐related factor 2 pathway.     

Digestive protection of probiotic Lacticaseibacillus rhamnosus in Ricotta cheese by monoglyceride structured emulsions

This research aimed at studying the potential use of monoglyceride (MG) structured emulsions (MSEs) as delivery and protective systems for probiotic bacteria in Ricotta cheese. To this purpose, a low-fat commercial Ricotta cheese was added with MSEs formulated with milk, as water phase, and sunflower oil (MSE-SO) or anhydrous milk fat (MSE-AMF), as lipid phase. A commercial whole milk Ricotta cheese (W-RC) was considered as reference. A probiotic Lacticaseibacillus rhamnosus strain was inoculated as free cells in W-RC or embedded into the MSEs and added to the low-fat Ricotta at the same reference fat content. After physico-chemical characterisation, L. rhamnosus viability and sample destructuring behaviour upon in vitro digestion were evaluated. At the end of in vitro digestion, both W-RC and sample containing MSE-SO were unable to protect cells. By contrast, sample with AMF ensured a sufficient probiotic viability, even after 14 days of storage at 4 °C. This result was attributed to system composition and structure. During the gastric phase, the presence of caseins and MG-AMF mixed structures induced the formation of clots, entrapping and protecting cells against the acidic pH of the stomach, as confirmed by confocal micrographs and particle size. During the intestinal phase, cell viability was guaranteed by the formation of mixed micelles promoted by MG. It was demonstrated that microbial cells located near MG structures where they found protection.     

Autochthonous white grape pomaces as bioactive source for functional jams

Seeds and skins from grape pomaces of Pecorello and Mantonico cv underwent extraction (ultrasound–assisted or maceration), in order to obtain added-value ingredients for the production of a functional pear jam. The antioxidant features of the extracts were tested by in vitro colorimetric assays. Among seeds, Mantonico by maceration (MSC) showed the best results, as well as Mantonico by ultrasound-assisted extraction (MBs) among skin extracts. The selected extracts were fully characterised by NMR and MS techniques, confirming the presence of many polyphenols, flavonoids and tannins among others. Pectin was then derivatised by the grafting procedure with the active molecules of MBs and MSC. The latter produced the best antioxidant polymer also without toxicity evaluated using Caco-2 cells and was used for the jam preparation. The functional pear jam showed improved antioxidant performances in comparison with its non-functional counterparts as well as its maintenance over time (15 days).     

Anthocyanidins decrease endothelin-1 production and increase endothelial nitric oxide synthase in human endothelial cells

Epidemiological and intervention studies correlate anthocyanin-rich beverages and a low incidence of coronary heart diseases. Since endothelin-1 (ET-1) and nitric oxide (NO) produced by endothelial NO synthase (eNOS) are vascular tension regulators secreted by endothelial cells, we studied the influence of two anthocyanidins, namely cyanidin (CY) and delphinidin (DP), on the regulation of ET-1 and eNOS in cultured human umbilical vein endothelial cells (HUVECs). Aglycon anthocyanidin forms, such as CY and DP, may be present in vivo after the first deglycosylation step occurring in the jejunum and in the liver. DP showed a major action compared to CY inducing a significant dose-dependent inhibitory effect on both protein and mRNA levels of ET-1. CY and DP both increased the protein level of eNOS, but DP showed the major effect raising eNOS protein in a dose-dependent manner. To correlate the vasoprotective effect of CY and DP with their antioxidant activity, we analysed also the antioxidant effect of anthocyanidins both in vitro and in HUVECs. In particular, we examined the effect of anthocyanidins on endothelial heme oxygenase-1 (HO-1), an inducible stress protein. In all tests, DP showed a higher antioxidant activity than CY. Finally, the antiproliferative effect induced by DP was detected in HUVECs. DP and CY differ in the number and position of hydroxyl groups in their structure; therefore, the greater biological activity by DP, compared with CY, seems to be due to the presence of the three hydroxyl groups on the B ring in the molecular structure of DP.     

Nonstarter lactobacilli isolated from soft and semihard Argentinean cheeses: genetic characterization and resistance to biological barriers

Nonstarter lactic acid bacteria isolated from Argentinean cheeses were identified and characterized by focusing on their resistance to biological barriers, along with other physiological features of potential interest, in the search for future probiotic organisms. Lactobacilli were enumerated and isolated from semihard and soft cheeses made with multistrain Streptococcus thermophilus starters. Lactobacilli counts in 1-week-old cheeses were between 10(5) and 10(7) CFU/g and then reached 10(7) CFU/ g in all 1-month samples, while streptococci were always above 10(9) CFU/g. A total number of 22 lactobacilli isolates were retained, identified, and characterized by in vitro tests. Species identity was determined by carbohydrate metabolism and species-specific PCR assays. Genetic diversity was explored by random amplified polymorphic DNA (RAPD) PCR analysis. The Lactobacillus strains were assigned to the species L. casei, L. plantarum, L. rhamnosus, L. curvatus, L. fermentum, and L. perolens. All the strains studied tolerated 25 ppm of lysozyme, and most of them showed resistance to 0.3% bile. After incubation in gastric solution (pH 2.0), counts decreased by several log units, ranging from 3.2 to 7.0. The strains were able to grow in the presence of bile salts, but only three isolates were capable of deconjugation. The nonstarter lactobacilli that were assayed fermented the prebiotic substrates (especially lactulose and inulin). Some strains showed high cell hydrophobicity and beta-galactosidase activity, as well as inhibitory activity against pathogenic bacteria. It was concluded that most of the lactobacilli isolated in this study demonstrated resistance to biological barriers and physiological characteristics compatible with probiotic properties, which make them suitable for further research in in vivo studies aimed at identifying new probiotic organisms.     

Visual evaluation of cattle cleanliness and correlation to carcass microbial contamination during slaughtering

The aim of the study was to establish whether the visual cleanliness of cattle slaughtered was correlated to hide and carcass contamination as indicated by aerobic colony count (ACC), Enterobacteriaceae count (EC) and Escherichia coli count (ECC). Cattle in a slaughterhouse were visually inspected and assigned to a category from 1 (very clean) to 5 (very dirty) based on cleanliness. Fifteen animals for each category were randomly selected, hide and carcass sampled and analyzed for ACC, EC and ECC. Results showed that increasing dirt on cattle was associated with higher ACC, EC and ECC on hide and carcasses. Carcass ACC and ECC belonging to animals classified in cleanliness categories 3, 4 or 5 have a higher probability of exceeding the limits set by the Reg. EU 2073/2005. The study supports the conclusion that the pre-slaughter visual evaluation of animal cleanliness and application of corrective actions can be an effective aid to reduce carcass contamination.     

A new platform for Real-Time PCR detection of Salmonella spp., Listeria monocytogenes and Escherichia coli O157 in milk

Intoxications and infections caused by food-borne pathogens represent an increasing public health problem, and diagnostic tests in multiplex format are needed for the rapid identification of food contaminations caused by more than one microbial species. We have developed a multiple PCR-based platform for the simultaneous detection of the widespread milk-associated pathogens Salmonella spp., Listeria monocytogenes and Escherichia coli O157. The assay combines an enrichment step in a medium properly formulated for the simultaneous growth of target pathogens, a DNA isolation method, and a multiplex Real-Time PCR detection system based either on dual-labelled probes (mRT-PCR), or on melting curve analysis (mHRM). The second, producing a distinct peak for each amplification product, allows the qualitative assessment of pathogen presence. Moreover, the internal amplification control (IAC) included in the reaction, ensuring the reliability of results, complies with quality management programmes. Inclusivity and exclusivity were 100% each, with a detection limit of 1 CFU for each pathogen in a total of five 25 ml-aliquots of raw milk, and a duration of two working days.The assay represents an alternative approach for the qualitative detection of the cited bacterial species, suitable for a relatively inexpensive screening of several milk samples, reducing the turnaround time and the workload.     

Effect of water/fuel emulsions and a cerium-based combustion improver additive on HD and LD diesel exhaust emissions

One of the major technological challenges for the transport sector is to cut emissions of particulate matter (PM) and nitrogen oxides (NOx) simultaneously from diesel vehicles to meet future emission standards and to reduce their contribution to the pollution of ambient air. Installation of particle filters in all existing diesel vehicles (for new vehicles, the feasibility is proven) is an efficient but expensive and complicated solution; thus other short-term alternatives have been proposed. It is well known that water/diesel (W/ D) emulsions with up to 20% water can reduce PM and NOx emissions in heavy-duty (HD) engines. The amount of water that can be used in emulsions for the technically more susceptible light-duty (LD) vehicles is much lower, due to risks of impairing engine performance and durability. The present study investigates the potential emission reductions of an experimental 6% W/D emulsion with EURO-3 LD diesel vehicles in comparison to a commercial 12% W/D emulsion with a EURO-3 HD engine and to a Cerium-based combustion improver additive. For PM, the emulsions reduced the emissions with -32% for LD vehicles (mass/km) and -59% for the HD engine (mass/ kWh). However, NOx emissions remained unchanged, and emissions of other pollutants were actually increased forthe LD vehicles with +26% for hydrocarbons (HC), +18% for CO, and +25% for PM-associated benzo[a]pyrene toxicity equivalents (TEQ). In contrast, CO (-32%), TEQ (-14%), and NOx (-6%) were reduced by the emulsion for the HD engine, and only hydrocarbons were slightly increased (+16%). Whereas the Cerium-based additive was inefficient in the HD engine for all emissions except for TEQ (-39%), it markedly reduced all emissions for the LD vehicles (PM -13%, CO -18%, HC -26%, TEQ -25%) except for NOx, which remained unchanged. The presented data indicate a strong potential for reductions in PM emissions from current diesel engines by optimizing the fuel composition.     

A model study for release of plasticizers from polymer films through vapor phase

A model has been applied to release of plasticizers from polymer films through vapor phase, for which the overall rate of release may be generally determined by evaporation of the plasticizer from the surface of the film, and/or by migration to the surface at below saturation in the polymer. The experimental system employed made use of n-butyl formate, diethyl-phthalate, di-n-butylphthalate, and N,n-butylbenzenesulfonamide as model molecules in polycaprolactam films, both with the aims of studying undesirable loss of plasticizers, and of simulating a model able to envisage releasing, at a constant rate, of any desired additive, with relatively high solubility and vapor pressure, to the content of packaging films. Experimental measurements were made (at 298–333 K) by investigating in a suitable cell the permeation of plasticizers both at concentrations below and above the saturation limit. Rate of evaporation of plasticizers from the membrane surface above the saturation limit, measured in this cell, were found to coincide satisfactorily with those calculated by application of the mass transfer theory to evaporation from a stationary liquid into a stirred gas, at a known velocity of gas flowing past the surface. From these latter rates and solubilities, the mass transfer coefficients H for evaporation could be obtained, as well as from experimental time lags the diffusivities D_p through the polymer membrane. In the light of the theoretical model a correlation was found between activation energies of H and evaporation enthalpies of the model molecules.