Water distribution and microstructure in enhanced pork

Water characteristics and meat microstructure of NaHCO3-enhanced pork were compared with NaCl- and Na4O7P2-enhanced pork using low-field proton NMR relaxometry, advanced microscopy techniques, and traditional meat quality measurements. Porcine samples were enhanced at 4 degrees C for 48 h with sodium salts individually and in the following combinations: (i) 5% NaCl, (ii) 5% Na4O7P2, (iii) 3% NaHCO3, (iv) 5% NaCl and 5% Na4O7P2, (v) 5% NaCl and 3% NaHCO3, (vi) 5% Na4O7P2 and 3% NaHCO3, and (vii) 5% NaCl, 5% Na4O7P2, and 3% NaHCO3. Independently of the marinade used, the water-binding capacity was improved, cooking loss was reduced, and the yield was enhanced compared with nonmarinated pork samples. This was also reflected in the water mobility within the samples measured by proton NMR relaxometry. Visualization of samples by confocal laser scanning microscopy (CLSM) revealed salt-dependent microstructural changes in the green pork samples treated with NaHCO3, giving rise to nearly complete disintegration of overall structures. High-resolution visualization by atomic force microscopy (AFM) further suggested that a higher cooking loss in sodium chloride-enhanced samples could be ascribed to less solubilization and higher heat-induced protein denaturation compared with phosphate- and bicarbonate-enhanced samples.     

Geography limits island small‐scale fishery production

Interacting social and ecological processes shape productivity and sustainability of island small‐scale fisheries (SSF). Understanding limits to productivity through historical catches help frame future expectations and management strategies, but SSF are dispersed and unaccounted, so long‐term standardized data are largely absent for such analyses. We analysed 40 years of trade statistics of a SSF product that enter international markets (sea cucumber) from 14 Pacific Island Countries and Territories (PICT) against response variables to test predictors of fishery production: (i) scale, (ii) productivity and (iii) socio‐economics. Combined production in PICT peaked over 20 years ago, driven by exploitation trends in Melanesia that accounted for 90% of all production since 1971. The size of island fisheries (as measured by total exports), and the duration and magnitude of fishery booms were most influenced by ungovernable environmental variables, in particular land area. The large and high islands of Melanesia sustained larger booms over longer periods than atoll nations. We hypothesize that land area is a proxy for land‐based nutrient availability and habitat diversity, and therefore the productivity of the shallow water areas where SSF are operating. PICT need to tailor management based on the intrinsic productivity of shallow inshore habitats: harvests from atoll nations will need to be smaller per unit area than at the high islands. Particularly countries with low productivity fisheries must consider the crucial economic “safety nets” that export SSF make up for dispersed island populations and incorporate them into broader development and island resilience strategies.     

Functional and ultrastructural evaluation of neutrophils from foals and lactating and nonlactating mares

Neutrophils from 4 pony foals, 3 lactating pony mares, and 3 nonlactating mares were evaluated ultrastructurally and by in vitro function tests. Neutrophils from foals had significantly (P = 0.05) less random migration than neutrophils from mares; values in tests for iodination and Staphylococcus aureus ingestion were also lower with foal neutrophils. Neutrophils from lactating mares had lower responses to iodination, antibody-dependent cell-mediated cytotoxicity, and random migration tests than did neutrophils from nonlactating mares. Ultrastructurally, granule concentration did not differ significantly among groups. A slight decrease in primary granules and a corresponding increase in granules with a flocculent matrix indicates partial spontaneous neutrophil degranulation in foals and lactating mares.     

Characterisation of duck thromhocytes

Duck thrombocytes were initially identified in peripheral blood mononuclear cells (PBMCS) purified from whole blood on Ficoll-Paque density gradients by examining stained smears of these cells. These thrombocytes could be readily purified from lymphocytes on the FACStar cell sorter by their increased side-scatter. They were similar to chicken thrombocytes in both appearance and function; they had a diameter of 4.5–6 μm and contained large vacuoles and were able to phagocytose carbon and Staphylococcus aureus. A monoclonal antibody (mAb) BA3 was generated which binds specifically to duck thrombocytes and has facilitated the characterisation of these cells which comprise up to 50–60 per cent of cells in Ficoll-Paque purified duck PBMCS.     

Congenital malformations of the lumbosacral vertebral column are common in neurologically normal French Bulldogs,English Bulldogs,and Pugs,with breed‐specific differences

Congenital vertebral malformations are common findings on diagnostic imaging of the vertebral column in “screw‐tailed” brachycephalic dogs. The aims of this study were to evaluate the prevalence and anatomical characteristics of lumbosacral congenital vertebral malformations in French Bulldogs, English Bulldogs, and Pugs presenting for problems unrelated to spinal disease, as well as possible associations with the degree of tail malformation, lumbosacral intervertebral disc herniation, or spondylosis deformans. In this retrospective cross‐sectional study, CT scans of vertebrae L6 to S3 and of the coccygeal vertebrae were reviewed for type of congenital vertebral malformations (hemivertebrae, block vertebrae, lumbosacral transitional vertebrae, and spina bifida), lumbosacral intervertebral disc herniation, lumbosacral spondylosis deformans, and degree of tail malformation. In 76 (51.0%) of the 149 included dogs (53 French Bulldogs, 37 English Bulldogs, and 59 Pugs) at least one type of congenital vertebral malformations was found, with lumbosacral transitional vertebrae being the most common (34.2%). There was a significantly higher prevalence of lumbosacral transitional vertebrae (54.2%) and lower prevalence of hemivertebrae (1.7%) in Pugs compared to English (13.5% and 24.3%, respectively) and French Bulldogs (26.4% and 32.0%, respectively). Tail malformation was significantly more severe in dogs with evidence of hemivertebrae. Congenital vertebral malformations are a common finding in the lumbosacral vertebral column of French Bulldogs, English Bulldogs, and Pugs. These anatomical variances need to be considered when interpreting diagnostic studies and when planning for neurosurgical and neurodiagnostic procedures. Furthermore, this study suggests a possible association between the degree of tail malformation and lumbosacral hemivertebrae.     

Natural variability in bovine milk oligosaccharides from Danish Jersey and Holstein-Friesian breeds

Free oligosaccharides are key components of human milk and play multiple roles in the health of the neonate, by stimulating growth of selected beneficial bacteria in the gut, participating in development of the brain, and exerting antipathogenic activity. However, the concentration of oligosaccharides is low in mature bovine milk, normally used for infant formula, compared with both human colostrum and mature human milk. Characterization of bovine milk oligosaccharides in different breeds is crucial for the identification of viable sources for oligosaccharide purification. An improved source of oligosaccharides can lead to infant formula with improved oligosaccharide functionality. In the present study we have analyzed milk oligosaccharides by high-performance liquid chromatography chip quadrupole time-of-flight mass spectrometry and performed a detailed data analysis using both univariate and multivariate methods. Both statistical tools revealed several differences in oligosaccharide profiles between milk samples from the two Danish breeds, Jersey and Holstein-Friesians. Jersey milk contained higher relative amounts of both sialylated and the more complex neutral fucosylated oligosaccharides, while the Holstein-Friesian milk had higher abundance of smaller and simpler neutral oligosaccharides. The statistical analyses revealed that Jersey milk contains levels of fucosylated oligosaccharides significantly higher than that of Holstein-Friesian milk. Jersey milk also possesses oligosaccharides with a higher degree of complexity and functional residues (fucose and sialic acid), suggesting it may therefore offer advantages in term of a wider array of bioactivities.     

Impact of high-fat and high-carbohydrate diets on liver metabolism studied in a rat model with a systems biology approach

The aim of the present study was to investigate the use of an integrated metabolomics and proteomics approach in the elucidation of diet-induced effects on hepatic metabolism in a rat model. Nuclear magnetic resonance (NMR)-based metabolomics of liver extracts revealed a pronounced effect of a high-fat diet on the hepatic betaine content, whereas a carbohydrate-rich diet induced increases in hepatic glucose. In addition, the metabolomic investigations revealed that the high-fat diet was associated with increased hepatic lipid levels, which was not evident with the carbohydrate-rich diet. The proteomic investigations revealed strong high-fat diet effects on the expression of 186 proteins in the liver including malate dehydrogenase. Comparison of malate dehydrogenase expression determined by proteomics and NMR metabolite profiles revealed correlations between malate dehydrogenase and lactate, glucose, and glutamine/glutamate signals, thereby demonstrating a diet-induced regulation that was evident at both proteomic and metabolomic levels.     

Cooking effects on water distribution in potatoes using nuclear magnetic resonance relaxation

Continuous low-field (LF) (1)H NMR relaxometry was used to monitor the structural changes during cooking of potatoes with two different dry matter (DM) contents. A principal component analysis of the relaxation decay curves revealed major events related to water mobility during cooking, which occur at 53 and 60 degrees C for potatoes with medium and low DM contents, respectively. Exponential analysis of the relaxation decays reveals two major water populations in the potato: a slow-relaxing (assigned to water in cytoplasm and extracellular cavities) water component, T(22) ( approximately 350-550 ms), and a fast-relaxing component (primarily assigned to water associated with starch and cell walls), T(21) ( approximately 45-65 ms). Significant DM dependent shifts in both the T(21) and T(22) relaxation time constants were observed during cooking, indicating that starch gelatinizes between 53 and 70 degrees C with water exchanging with the hydroxyls of starch (transition in T(21)) and cells start to disrupt with an increase in diffusion volumes at approximately 60 degrees C (transition in T(22)). The study reveals that continuous LF NMR measurement is an excellent and highly sensitive method to study changes in water mobility and water populations during the cooking of potatoes.     

Dynamic high-resolution 1H and 31P NMR spectroscopy and 1H T2 measurements in postmortem rabbit muscles using slow magic angle spinning

Postmortem changes in rabbit muscle tissue with different glycogen status (normal vs low) were followed continuously from 13 min postmortem until 8 h postmortem and again 20 h postmortem using simultaneous magic angle spinning (1)H and (31)P NMR spectroscopy together with measurement of the transverse relaxation time, T(2), of the muscle water. The (1)H metabolite spectra were measured using the phase-altered spinning sidebands (PASS) technique at a spinning rate of 40 Hz. pH values calculated from the (31)P NMR spectra using the chemical shifts of the C-6 line of histidine in the (1)H spectra and the chemical shifts of inorganic phosphate in the (31)P spectra confirmed the different muscle glycogen status in the tissues. High-resolution (1)H spectra obtained from the PASS technique revealed the presence of a new resonance line at approximately 6.8 ppm during the postmortem period, which were absent in muscles with low muscle glycogen content. This new resonance line may originate from the aminoprotons in creatine, and its appearance may be a result of a pH effect on the exchange rate between the amino and the water protons and thereby the NMR visibility. Alternatively, the new resonance line may originate from the aromatic protons in tyrosine, and its appearance may be a result of a pH-induced protein unfolding exposing hydrophobic amino acid residues to the aqueous environment. Further studies are needed to evaluate these hypotheses. Finally, distributed analysis of the water T(2) relaxation data revealed three relaxation populations and an increase in the population believed to reflect extramyofibrillar water through the postmortem period. This increase was significantly reduced (p < 0.0001) in samples from animals with low muscle glycogen content, indicating that the pH is controlling the extent of postmortem expulsion of water from myofibrillar structures. The significance of the postmortem increase in the amount extramyofibrillar water on the water-holding capacity was verified by centrifugation, which showed a reduced centrifugation loss in muscles with low preslaughter glycogen status (0.9 vs 1.9%, p = 0.07).     

Combined high-field 13C CP MAS NMR and low-field NMR relaxation measurements on post mortem porcine muscles

Changes in postmortem muscle characteristics are investigated in muscles from eight pigs exposed to different combinations of preslaughter stress (exercise on treadmill) and stunning method (CO(2) vs electrical stunning). Solid-state (13)C cross-polarization (CP) magic-angle spinning (MAS) NMR experiments are carried out on a total of 16 rapidly frozen M. longissimus muscle biopsies taken in vivo the day before slaughter and at 45 min postmortem. Simultaneously, low-field NMR T(2) relaxation time measurements are carried out on samples from M. longissimus. Glycogen and lactate are estimated from the (13)C CP MAS spectra, and correlations of r = 0.89 and r = 0.70, respectively, to subsequent biochemical determinations using partial least squares regression (PLSR) are established. Moreover, PLSR reveals that, besides the 72 ppm signal (carbons in glycogen), a signal around 38 ppm, which increases concomitantly with lactate, is also significantly correlated to changes in glycogen/lactate. With the assumption that the 38 ppm signal reflects CH(2) in phosphocreatine/creatine, altered mobility of creatine as a result of dephosphorylation is indicated. Finally, PLSR on the 45 min (13)C CP MAS spectra also reveals correlation (r = 0.54) to the slowest relaxing T(2) population (50 min postmortem), known to reflect extra-myofibrillar water. Subsequently, evaluation of the loading plot in the PLSR analysis reveals that the correlation exclusively is associated to the 52 ppm resonance intensity. With the assumption that this resonance reflects methyl groups in choline/phosphatidyl choline, the intensity changes in the 52 ppm resonance imply alterations in membrane properties. Accordingly, the data indicate a relationship between membrane properties and the amount of water being expelled from muscle cells postmortem, which supports the hypothesis that disruption of membranes is implicated in the postmortem mobilization of muscle water.