Effect of some phenolic compounds and beverages on pepsin activity during simulated gastric digestion

The effect of some polyphenols (resveratrol, catechin, epigallocatechin-3-gallate, and quercetin) and beverages (red wine and green tea) on the enzymatic activity of pepsin during the digestion of three different substrates (pork meat, insoluble azocasein, and denatured hemoglobin) has been investigated. The tested polyphenols and beverages increase the initial velocity of the reaction, and the activating effect is concentration dependent. The order of effectiveness of polyphenols in increasing the initial velocity of the reaction is resveratrol > or = quercetin > epigallocatechin-3-gallate > catechin. The kinetic data obtained with soluble denatured hemoglobin show that the K(m) for the substrate is not changed, whereas the V(max) of the reaction is increased. Pepsin activity follows a simple Michaelis-Menten kinetic suggesting that k(3) is increased by polyphenols. To the authors' knowledge, the present study is the first demonstration that some polyphenols and related beverages are able to enhance the enzymatic activity of pepsin.     

Quantitative aspects of solid-state C-NMR spectra of humic substances from soils of volcanic systems

Cross-Polarisation Magic Angle Spinning Carbon-13 Nuclear Magnetic Resonance spectroscopy (CPMAS ¹³C-NMR) represents one of the most powerful tools to investigate soil organic matter (SOM) mainly because of its inherent capacity to provide a semi-quantitative evaluation of carbon distribution. A critical parameter during acquisition of CPMAS ¹³C-NMR spectra is the contact time required to obtain the cross-polarisation between proton and carbon nuclei. The procedure to evaluate the best contact time for the acquisition of a quantitative CPMAS ¹³C-NMR spectrum is to perform Variable Contact Time (VCT) experiments. In this work the structural features of a number of purified humic substances from Italian and Costarican volcanic soils were investigated by CPMAS ¹³C-NMR spectroscopy after having performed preliminary VCT experiments. The VCT experiments showed that the average contact times vary according to the origin and chemical structure of the humic material. The optimal contact times (OCT) for nine humic samples were between 250 and 800 μs These values were different from the time of 1000 μs that is commonly applied as the best average contact time for humic materials. Moreover, by comparing the NMR data to those obtained by elemental analysis (C/H ratio), it appeared that the efficiency of the cross-polarisation between protons and carbons, and hence the contact time, is affected not only by the number of protons, but also by their distribution over the molecules. The evaluation of errors in quantitative estimation of the different carbons revealed that the use of OCT generally reduced by half the loss of signals occurring when the average contact time of 1000 μs is used in CPMAS ¹³C-NMR spectra of humic substances.     

Effect of split marketing on the welfare, performance, and carcass traits of finishing pigs

The aim of this study was to compare a split marketing (SM) strategy, in which the heaviest pigs in a group are removed and slaughtered earlier than the others, with an all-out (AO) marketing strategy, in which all pigs are removed from the pen simultaneously and slaughtered on the same day, in terms of welfare, performance, and carcass traits of noncastrated (i.e., intact) male and female pigs. The experimental treatments were arranged in a 2 × 2 factorial array with 1) marketing strategy (SM vs. AO) and 2) sex (males vs. females), which yielded 4 treatment groups of 14 pigs (73.1 ± 4.8 kg): male SM, male AO, female SM, and female AO (7 replicates/group). Pigs in AO groups were all slaughtered after 6 wk on trial, whereas in SM groups the 3 heaviest pigs were removed and slaughtered 2 wk before the remainder of the group, which were slaughtered at the same time as the AO pigs. Pigs were fed a liquid diet from a long trough 3 times daily. Behavioral observations were conducted before and after SM, the day of SM, and 1 and 2 wk later. Behavior was recorded both during and between feed events, and skin lesions were scored on all, except the 3 pigs removed from SM groups before and 2 wk after SM. Growth performance, feed efficiency, and carcass traits were recorded. The number of aggressive interactions during feed events decreased after the 3 pigs were removed from SM groups. This reduction in aggressive interactions was observed on the day of SM in male groups (before SM: 24.3 vs. the day of SM: 14.7, SED = 3.31, P < 0.05 for interaction) and in subsequent observations in female groups (before SM: 21.4 vs. days after SM: 13.4, SED = 3.31, P < 0.05 for interaction). However, SM had no effect on behaviors recorded between feed events or on the number and severity of skin lesions (P > 0.10). There were no differences between the 11 remaining pigs in SM groups and the 14 pigs in AO groups in terms of growth performance, feed efficiency, and carcass traits of female or intact male pigs (P > 0.10). However, reduced within-pen CV in carcass weight was detected in pigs from SM groups compared with pigs from AO groups (8.6 vs. 10.9, SEM = 0.72, P < 0.05). Therefore, in restrictively fed pigs, a SM strategy improved the welfare of both female and intact male pigs by reducing aggressive interactions during feeding but had no effect on performance or carcass traits.     

Extraction of grape seed oil using compressed carbon dioxide and propane: extraction yields and characterization of free glycerol compounds

The main objective of this work was to compare the extraction of grape seed oil with compressed carbon dioxide and propane on the extraction yields and chemical characteristics of free glycerol compounds. The experiments were performed in a laboratory scale unit in the temperature range of 30 to 60 degrees C and pressures from 60 to 254 bar. The results showed that propane is a more suitable solvent for grape seed oil extraction than carbon dioxide, as higher extractions yields and a very fast kinetic of extraction were achieved with this solvent. In relation to compressed carbon dioxide extractions, both temperature and density presented a very pronounced and positive effect on the extraction yield. The oils extracted were analyzed qualitatively and quantitatively with regard to the free glycerol compounds, mainly fatty acids, ethyl, and methyl esters. The results showed that these compounds are present in low concentration in vegetable oil (<3%) and that, in general, samples extracted with propane present a smaller amount of peaks of free glycerol compounds in the oil than samples extracted with carbon dioxide.     

Computed tomographic diagnosis of incomplete palmar cortical (fatigue) fracture of the third metacarpal bone in two young adult endurance horses

Two young adult endurance horses were presented for investigation of sudden-onset forelimb lameness during competition. Clinical examination revealed a severe forelimb lameness and pain on palpation of the proximal palmar metacarpal area. Initial radiographic survey of the affected forelimb was unremarkable in both cases. A week of box rest resulted in only a mild improvement in the lameness. A second radiographic examination did not reveal any significant abnormalities. In both cases, scintigraphic examination was suggested, but was declined by the owner, and the lameness had significantly improved after local infiltration of 3 mL of anaesthetic solution in the proximal suspensory region. Radiographic and ultrasonographic examinations of the carpus and proximal suspensory region were unremarkable in Case 2. Case 1 had increased thickness of the medial lobe of the proximal aspect of the suspensory ligament. A computed tomographic examination was performed and an incomplete longitudinal (fatigue) fracture of the proximal third metacarpal bone was detected. After 8 months of box rest and rehabilitation, re-examination revealed resolution of the lameness in both cases. Advanced diagnostic imaging should be considered in young endurance horses with severe lameness during competition or training, in which radiographic and ultrasonographic examinations are unremarkable or do not explain the severity of the lameness. Computed tomography imaging is considered a useful diagnostic imaging technique to detect incomplete longitudinal palmar metacarpal cortical (fatigue) fractures due to its ability to provide excellent bone detail.     

Antioxidant and other biological activities of olive mill waste waters.

During olive oil production, large volumes of water are generated and subsequently discarded. Olives contain a variety of bioactive components, and some of them, according to their partition coefficients, end up in the water phase. The current investigation aimed at comparing different methods for the extraction of biologically active components of the olive mill waste waters (OMWW) and evaluating the in vitro antioxidant and anti-inflammatory activities of the resulting extracts. The results indicate that OMWW extracts are able to inhibit human LDL oxidation (a process involved in the pathogenesis of atherosclerosis) and to scavenge superoxide anions and hypochlorous acid at concentrations as low as 20 ppm. Finally, two of the three extracts also inhibited the production of leukotrienes by human neutrophils. The potency of the extracts depended on their degree of refinement: extracts containing only low molecular weight phenols were the most effective.     

Protective effect of the AT137RQ and ARQK176 PrP alleles against classical scrapie in Sarda breed sheep

The susceptibility of sheep to scrapie is under the control of the host’s prion protein (PrP) gene and is also influenced by the strain of the agent. PrP polymorphisms at codons 136 (A/V), 154 (R/H) and 171 (Q/R/H) are the main determinants of susceptibility/resistance of sheep to classical scrapie. They are combined in four main variants of the wild-type ARQ allele: VRQ, AHQ, ARH and ARR. Breeding programmes have been undertaken on this basis in the European Union and the USA to increase the frequency of the resistant ARR allele in sheep populations. Herein, we report the results of a multi-flock study showing the protective effect of polymorphisms other than those at codons 136, 154 and 171 in Sarda breed sheep. All ARQ/ARQ affected sheep (n = 154) and 378 negative ARQ/ARQ controls from four scrapie outbreaks were submitted to sequencing of the PrP gene. The distribution of variations other than those at the standard three codons, between scrapie cases and negative controls, was statistically different in all flocks. In particular, the AT₁₃₇RQ and ARQK₁₇₆ alleles showed a clear protective effect. This is the first study demonstrating a protective influence of alleles other than ARR under field conditions. If further investigations in other sheep breeds and with other scrapie sources confirm these findings, the availability of various protective alleles in breeding programmes of sheep for scrapie resistance could be useful in breeds with a low frequency of the ARR allele and would allow maintaining a wider variability of the PrP gene.