Digestive enzyme activity at different developmental stages of blackspot seabream, Pagellus bogaraveo (Brunnich 1768)

Blackspot seabream, Pagellus bogaraveo (Brunnich), has been identified as a potential species to diversify European aquaculture production. Although rearing aspects have been widely investigated, little information exists on the nutritional requirements for this species. The aim of this study was to build up information on the activity of digestive enzymes at certain developmental stages of blackspot seabream in order to understand the nutritional needs of larvae and post larvae. Fish larvae were reared from hatching to 55 days after hatching (dah), and the feeding plan consisted in rotifers (5–35 dah), Artemia naupli (30–35 dah) metanaupli (35–45) and Gemma microdiet (45–55 dah). At 7, 11, 21, 45 and 55 days after hatching (dah), pooled samples of fish larvae were collected for analysis of trypsin, amylase, lipase, alkaline phosphatase and leucine–alanine peptidase activity. Up to 21 dah, the whole larvae body was used for enzymatic analysis, whereas in older larvae only the dissected abdominal cavity was used. Blackspot seabream body dry weight growth was exponential, increasing from 60 μg at 5 dah to 30±9.7 mg at 55 dah. Amylase specific activity decreased significantly during development, exhibiting at 11 dah (0.6 U mg^?1 protein) an average value 2.7 times lower than at 7 dah, and remaining stable between 45 and 55 dah (0.7 U mg protein^?1). Trypsin specific activity remained constant until 21 dah (between 38 and 44 mU mg protein^?1), which could be related to the larvae feeding regime. At later stages of development, lipase‐specific activity exhibited a significant increase (P<0.05), being three times higher at 55 dah (8 U mg protein^?1) than at 45 dah. The total activity of the studied digestive enzymes increased significantly during larval development (until 21 dah), whereas afterwards only lipase and leucine–alanine peptidase increased significantly between 45 and 55 dah. The pattern of digestive enzymes activity was related to organogenesis and the type of food used at different developmental stages.     

Antioxidant and Antihypertensive Potential of Protein Fractions from Flour and Milk Substitutes from Canary Seeds (<Emphasis Type="Italic">Phalaris canariensis</Emphasis> L.)

Canary seed (Phalaris canariensis) is used to feed birds but it has been recently considered a promising cereal with nutraceutical potential for humans. The aim of this work was to analyze the protein fractions from canary seed flour and from milk substitutes (prepared by soaking the seeds in water 12 and 24 h), and to evaluate antioxidant and antihypertensive capacity of peptides obtained after in vitro digestion. Prolamins were the major protein fraction, followed by glutelins. After digestion, albumins and prolamins fractions from milks presented higher levels of peptides than flour, globulins showed more peptides in flour and glutelins were found in similar concentrations in all samples; 24 h milk prolamins had the highest concentration of peptides. Purification by high performance liquid chromatography (HPLC), sequencing of peptides, in vitro antioxidant ABTS (2,2′-azino-bis, 3-ethylbenzothiazoline-6-sulphonic acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) assays, and antihypertensive capacity (angiotensin converting enzyme (ACE) assay), indicated that peptides from canary seed prolamins were the most efficient compounds with antioxidant and antihypertensive activity. Canary seeds may be considered an accessible and cheap source to prepare milk substitutes with high contents of bioactive peptides with remarkable functional properties to promote better human health and healthy ageing.     

Distribution patterns and population structure of the blue shark (Prionace glauca) in the Atlantic and Indian Oceans

The blue shark (Prionace glauca) is the most frequently captured shark in pelagic oceanic fisheries, especially pelagic longlines targeting swordfish and/or tunas. As part of cooperative scientific efforts for fisheries and biological data collection, information from fishery observers, scientific projects and surveys, and from recreational fisheries from several nations in the Atlantic and Indian Oceans was compiled. Data sets included information on location, size and sex, in a total of 478,220 blue shark records collected between 1966 and 2014. Sizes ranged from 36 to 394 cm fork length. Considerable variability was observed in the size distribution by region and season in both oceans. Larger blue sharks tend to occur in equatorial and tropical regions, and smaller specimens in higher latitudes in temperate waters. Differences in sex ratios were also detected spatially and seasonally. Nursery areas in the Atlantic seem to occur in the temperate south‐east off South Africa and Namibia, in the south‐west off southern Brazil and Uruguay, and in the north‐east off the Iberian Peninsula and the Azores. Parturition may occur in the tropical north‐east off West Africa. In the Indian Ocean, nursery areas also seem to occur in temperate waters, especially in the south‐west Indian Ocean off South Africa, and in the south‐east off south‐western Australia. The distributional patterns presented in this study provide a better understanding of how blue sharks segregate by size and sex, spatially and temporally, and improve the scientific advice to help adopt more informed and efficient management and conservation measures for this cosmopolitan species.     

Osteogenic potential of sorted equine mesenchymal stem cell subpopulations

The objectives of this study were to use non-equilibrium gravitational field-flow fractionation (GrFFF), an immunotag-less method of sorting mesenchymal stem cells (MSCs), to sort equine muscle tissue-derived mesenchymal stem cells (MMSCs) and bone marrow-derived mesenchymal stem cells (BMSC) into subpopulations and to carry out assays in order to compare their osteogenic capabilities. Cells from 1 young adult horse were isolated from left semitendinosus muscle tissue and from bone marrow aspirates of the fourth and fifth sternebrae. Aliquots of 800 × 10³ MSCs from each tissue source were sorted into 5 fractions using non-equilibrium GrFFF (GrFFF proprietary system). Pooled fractions were cultured and expanded for use in osteogenic assays, including flow cytometry, histochemistry, bone nodule assays, and real-time quantitative polymerase chain reaction (qPCR) for gene expression of osteocalcin (OCN), RUNX2, and osterix. Equine MMSCs and BMSCs were consistently sorted into 5 fractions that remained viable for use in further osteogenic assays. Statistical analysis confirmed strongly significant upregulation of OCN, RUNX2, and osterix for the BMSC fraction 4 with P < 0.00001. Flow cytometry revealed different cell size and granularity for BMSC fraction 4 and MMSC fraction 2 compared to unsorted controls and other fractions. Histochemisty and bone nodule assays revealed positive staining nodules without differences in average nodule area, perimeter, or stain intensity between tissues or fractions. As there are different subpopulations of MSCs with different osteogenic capacities within equine muscle- and bone marrow-derived sources, these differences must be taken into account when using equine stem cell therapy to induce bone healing in veterinary medicine.     

Preparation of antioxidant enzymatic hydrolysates from alpha-lactalbumin and beta-lactoglobulin. Identification of active peptides by HPLC-MS/MS

We have investigated the antioxidant activity of hydrolysates from whey proteins bovine alpha-lactalbumin (alpha-La) and beta-lactoglobulin A (beta-Lg A) by commercial proteases (pepsin, trypsin, chymotrypsin, thermolysin, and Corolase PP). Corolase PP was the most appropriate enzyme to obtain antioxidant hydrolysates from alpha-La and beta-Lg A (ORAC-FL values of 2.315 and 2.151 micromol of Trolox equivalent/mg of protein, respectively). A total of 42 peptide fragments were identified by HPLC-MS/MS in the beta-Lg A hydrolysate by Corolase PP. One of the sequences (Trp-Tyr-Ser-Leu-Ala-Met-Ala-Ala-Ser-Asp-Ile) possessed radical scavenging (ORAC-FL value of 2.621 micromol of Trolox equivalent/micromol of peptide) higher than that of butylated hydroxyanisole (BHA). Our results suggest that whey protein hydrolysates could be suitable as natural ingredients in enhancing antioxidant properties of functional foods and in preventing oxidation reaction in food processing.     

Assessment of the spectrophotometric method for determination of angiotensin-converting-enzyme activity: influence of the inhibition type

A set of in vitro assay conditions were selected for the determination of ACE-inhibitory activity, and the need was demonstrated to standardize this assay so that the results obtained by different authors may be comparable. The conditions selected were as follows: 10 mM HHL concentration in 0.2 M potassium phosphate buffer and 0.3 M NaCl and 26 mU of ACE/mL as reaction medium; incubation time, 80 min at 37 degrees C. The method was applied to the study of ACE-inhibitory activity of dairy product and wine samples. Of the samples assayed, it was infant formulae whey that produces the greatest ACE inhibition. Red wine also presents a high inhibition percentage. This latter sample has an important matrix effect that must be corrected in the calculation. ACE-inhibition type was also studied, using a yogurt whey and a Captropil solution as substrates. The whey produced noncompetitive inhibition and the Captropil competitive inhibition.     

Starch fine structure and physicochemical properties of specialty rice for canning

The long-grain, specialty rice cultivars, Bolivar, Cheniere, Dixiebelle, and L-205 are used for wet-pack canning. These cultivars have similar apparent amylose content but showed differences in canning, pasting, and gelatinization properties. Starch fine structures were analyzed to rationalize observed differences in functionality. Cheniere amylopectin had the lowest weight-average molar mass (M_w), shortest average chain length (CL), smallest z-average radius of gyration (R_z), lowest proportion of long chains (DP 37–65), and highest polydispersity; while its amylose had the largest M_w and R_z. These structural features were associated with more leached solids in the canning broth, lower volume expansion, lower peak and final viscosity, and lower gelatinization temperature and enthalpy. Bolivar amylopectin had the largest M_w, longest average CL, largest R_z, highest proportion of long chains (DP 25–65), and lowest proportion of short chains (DP 6–12); while its amylose had the smallest M_w and lowest polydispersity. These structures were associated with lower levels of leached solids, higher volume expansion, and higher peak and final viscosity. L-205 was similar to Bolivar in most structural and functional properties; those of Dixiebelle were either comparable to Bolivar or intermediate to Bolivar and Cheniere. These findings point to the importance of the molar mass of amylopectin and the proportion of long and short chains on the canning stability of rice.     

Effects of Two Energy-Restricted Diets Containing Different Fruit Amounts on Body Weight Loss and Macronutrient Oxidation

The consumption of specific foods in energy-restricted diets may affect the weight loss process. The purpose of this research was to evaluate whether obese women following two hypocaloric diets with distinct fruit content differ in weight loss and metabolic responses. Fifteen obese women were included, who were randomly assigned to follow a low or a high-fruit energy-restricted diet for 8 weeks. The main outcome variables were weight and fat losses. Metabolic measurements concerning macronutrient oxidation were also assessed by using ¹³C labelled fructose and indirect calorimetry. The induced weight loss was similar for both diets (6.9 ± 2% vs. 6.6 ± 2%, p = 0.785). Both experimental diets similarly improved the lipid plasma profile in the participants, but the cholesterol fall was higher in obese subjects receiving the diet containing more fruit. No statistical differences in lipids carbohydrates and ¹³C labelled fructose utilisation were observed, but protein oxidation was differently affected by the experimental diets. The compensatory effects of the associated fibre/fructose intake may explain the lack of a specific effect of the fruit amount on hypocaloric diets designed to weight loss, although the increased fibre content from enriched fruit diets may be involved in the favourable effects on cholesterol plasma levels.