Combination of Mechanical/Physical Pretreatments with Trypsin or Pancreatin on Arthrospira platensis Protein Degradation

Research Areas: AgricultureABSTRACT - Arthrospira platensis contains high protein content and quality, which makes it a promising feed source for livestock animals. However, this microalga presents a recalcitrant peptidoglycan cell wall, and its main proteins form protein-pigment complexes attached to the algal thylakoid membrane. The purpose of the present study was to evaluate the effect of mechanical/physical pretreatments (bead milling, extrusion, freeze-drying, heating, microwave and sonication) combined with commercial enzymes (trypsin or pancreatin) on the degradation of A. platensis proteins. Protein degradation was assessed through the quantification of total protein and protein fractions (18–26 kDa, 40–48 kDa and others) on SDS-PAGE gels and the evaluation of the coefficient of protein degradation (CPD). The results showed that no significant differences were found among treatments for CPD values, except for an increase related to 18–26 kDa protein (phycocyanin subunits) with extrusion combined with pancreatin. In addition, extrusion and microwave caused a decrease of total protein in algal supernatant probably due to a denaturation/aggregation and reduction of solubility. Overall, extrusion is a promising pretreatment for A. platensis cell wall disruption and protein bioaccessibility. Further studies could elucidate how the effect of extrusion on protein solubility affects the activity of peptidases on protein degradationinfo:eu-repo/semantics/publishedVersio

Current knowledge and future perspectives of the use of seaweeds for livestock production and meat quality : a systematic review

Research Areas: Agriculture ; Veterinary SciencesABSTRACT - The effects of dietary macroalgae, or seaweeds, on growth performance and meat quality of livestock animal species are here reviewed. Macroalgae are classified into Phaeophyceae (brown algae), Rhodophyceae (red algae) and Chlorophyceae (green algae). The most common macroalga genera used as livestock feedstuffs are: Ascophyllum, Laminaria and Undaria for brown algae; Ulva, Codium and Cladophora for green algae; and Pyropia, Chondrus and Palmaria for red algae. Macroalgae are rich in many nu-trients, including bioactive compounds, such as soluble polysaccharides, with some species being good sources of n-3 and n-6 polyunsaturated fatty acids. To date, the in-corporation of macroalgae in livestock animal diets was shown to improve growth and meat quality, depending on the alga species, dietary level and animal growth stage. Generally, Ascophyllum nodosum can increase average daily gain (ADG) in ruminant and pig mostly due to its prebiotic activity in animal's gut. A. nodosum also enhances marbling score, colour uniformity and redness, and can decrease saturated fatty acids in ruminant meats. Laminaria sp., mainly Laminaria digitata, increases ADG and feed efficiency, and improves the antioxidant potential of pork. Ulva sp., and its mixture with Codium sp., was shown to improve poultry growth at up to 10% feed. Therefore, seaweeds are promising sustainable alternatives to corn and soybean as feed ingre-dients, thus attenuating the current competition among food-feed-biofuel industries. In addition, macroalgae can hinder eutrophication and participate in bioremediation. However, some challenges need to be overcome, such as the development of large- scale and cost-effective algae production methods and the improvement of algae digestibility by monogastric animals. The dietary inclusion of Carbohydrate-Active enZymes (CAZymes) could allow for the degradation of recalcitrant macroalga cell walls, with an increase of nutrients bioavailability. Overall, the use of macroalgae as feedstuffs is a promising strategy for the development of a more sustainable livestock production.info:eu-repo/semantics/publishedVersio

Impact of dietary Chlorella vulgaris and carbohydrate-active enzymes incorporation on plasma metabolites and liver lipid composition of broilers

Research Areas: Veterinary SciencesBackground: Chlorella vulgaris has been proposed as a sustainable green feedstock in poultry nutrition due to its ease of cultivation, minimal environmental impact and balanced nutritional composition. However, the majority of studies documents the use of C. vulgaris as a dietary supplement in broilers instead of a feed ingredient. To the best of our knowledge, no report has shown the effect of a high-level incorporation (>2 % in the diet) of C. vulgaris on plasma metabolites and hepatic lipid composition of broilers. One hundred and twenty Ross 308 male birds were housed in 40 wired-floor cages and randomly distributed by the following experimental diets at 22 days of age (n = 10) during 15 days: (1) a corn-soybean meal based diet (control); (2) based diet with 10% of C. vulgaris; (3) diet 2 supplemented with 0.005% Rovabio® Excel AP; and (4) diet 2 supplemented with 0.01% of a pre-selected four-CAZyme mixture. Results: The inclusion of C. vulgaris at 10% in the diet, regardless of the presence of exogenous CAZymes, changed plasma metabolites but did not compromise broilers growth. Plasma total lipids increased in broilers fed C. vulgaris combined with the two feed CAZymes (p < 0.001) compared with the control diet. Moreover, the supplementation with Rovabio® increased total cholesterol and LDL-cholesterol, while the addition of the four-CAZyme mixture increased triacylglycerols, VLDL-cholesterol and ALP activity. In opposition, HDL-cholesterol levels decreased in broilers fed microalga alone (p = 0.002). Regarding hepatic composition, the inclusion of C. vulgaris in broiler diets, individually or combined with exogenous CAZymes, had a minor effect on fatty acids but improved the n-6/n-3 ratio and total carotenoids. Conclusions: In summary, the inclusion of a high level (10%) of C. vulgaris in broiler´s diet, regardless of the presence of exogenous CAZymes, improved hepatic antioxidant composition and did not impair broiler’s performance. In addition, the feed supplementation with CAZymes increased broilers lipemia. Therefore, dietary C. vulgaris at this incorporation level seems to be safe for animal health and do not compromise performance traits, with no need of CAZymes supplementation.info:eu-repo/semantics/publishedVersio

Fluorinated surfactants in solution: Diffusion coefficients of fluorinated alcohols in water

Intra-diffusion coefficients of three fluorinated alcohols, 2,2,3,3,3-pentafluoropropan-1-ol (PFP), 2,2,3,3,4,4,4-heptafluorobutan-1-ol (HFB) and 2,2,3,3,4,4,5,5,5-nonafluoropentan-1-ol (NFP) in water have been measured by the PFG–NMR spin-echo technique as a function of temperature and composition, focusing on the alcohol dilute region. For comparison, intra-diffusion coefficients of 2,2,2- trifluoroethanol (TFE) and HFB have also been measured in heavy water using the same method and conditions. As far as we know, these are the first experimental measurements of this property for these binary systems. Intra-diffusion coefficients for NFP in water and for TFE and HFB in heavy water have also been obtained by molecular dynamics simulation, complementing those for TFE, PFP and HFB reported in a previous work. The agreement between experimental and simulated results for PFP, HFB and NFP in water is reasonable, although presenting higher deviations than for the TFE/water system. From the dependence of the intra-diffusion coefficients on temperature, diffusion activation energies were estimated for all the solutes in water and heavy water

An individual alginate lyase is effective in the disruption of Laminaria digitata recalcitrant cell wall

Research Areas: Science & Technology - Other TopicsIn the present study, 199 pre-selected Carbohydrate-Active enZymes (CAZymes) and sulfatases were assessed, either alone or in combination, to evaluate their capacity to disrupt Laminaria digitata cell wall, with the consequent release of interesting nutritional compounds. A previously characterized individual alginate lyase, belonging to the family 7 of polysaccharide lyases (PL7) and produced by Saccharophagus degradans, was shown to be the most efcient in the in vitro degradation of L. digitata cell wall. The alginate lyase treatment, compared to the control, released up to 7.11 g/L of reducing sugars (p< 0.001) and 8.59 mmol/100 g dried alga of monosaccharides (p< 0.001), and reduced cell wall fuorescence intensity by 39.1% after staining with Calcofuor White (p= 0.001). The hydrolysis of gel-forming polymer alginate by the alginate lyase treatment could prevent the trapping of fatty acids and release benefcial monounsaturated fatty acids, particularly 18:1c9 (p < 0.001), to the extracellular medium. However, no liberation of proteins (p > 0.170) or pigments (p > 0.070) was observed. Overall, these results show the ability of an individual alginate lyase, from PL7 family, to partially degrade L. digitata cell wall under physiological conditions. Therefore, this CAZyme can potentially improve the bioavailability of L. digitata bioactive compounds for monogastric diets, with further application in feed industry.info:eu-repo/semantics/publishedVersio

Effect of Dietary Laminaria digitata with Carbohydrases on Broiler Production Performance and Meat Quality, Lipid Profile, and Mineral Composition

Research Areas: Agriculture; Veterinary Sciences; ZoologyABSTRACT - We hypothesized that dietary inclusion of 15% Laminaria digitata, supplemented or not with carbohydrases, could improve the nutritional value of poultry meat without impairing animal growth performance. A total of 120 22-day old broilers were fed the following dietary treatments (n = 10) for 14 days: cereal-based diet (control); control diet with 15% L. digitata (LA); LA diet with 0.005% Rovabio® Excel AP (LAR); LA diet with 0.01% alginate lyase (LAE). Final body weight was lower and feed conversion ratio higher with LA diet than with the control. The ileal viscosity increased with LA and LAR diets relative to control but without differences between LAE and control. The pH of thigh meat was higher, and the redness value of breast was lower with LA diet than with control. Meat overall acceptability was positively scored for all treatments. The γ-tocopherol decreased, whereas total chlorophylls and carotenoids increased in meat with alga diets relative to control. The percentage of n-3 polyunsaturated fatty acids (PUFA) and accumulation of bromine and iodine in meat increased with alga diets compared with control. Feeding 15% of L. digitata to broilers impaired growth performance but enhanced meat quality by increasing antioxidant pigments, with beneficial effects on n-3 PUFA and iodineinfo:eu-repo/semantics/publishedVersio

Influence of dietary Spirulin inclusion and lysozyme supplementation on the longissimus lumborum muscle proteome of newly weaned piglets

Research Areas: Biochemistry & Molecular BiologyArthrospira platensis (Spirulina) is a microalga with a high content of crude protein. It has a recalcitrant cell wall that limits the accessibility of the animal endogenous enzymes to its intracellular nutrients. Enzymatic supplementation aiming to degrade cell walls could benefit microalgae digestibility. The objective of this study was to evaluate the impact of dietary Spirulina and lysozyme supplementation over the muscle proteome of piglets during the post-weaning stage. Thirty piglets were randomly distributed among three diets: control (no microalga), SP (10% Spirulina) and SP + L (10% Spirulina +0.01% lysozyme). After 4 weeks, they were sacrificed and samples of the longissimus lumborum muscle were taken. The muscle proteome was analysed using a Tandem Mass Tag (TMT)-based quantitative approach. A total of 832 proteins were identified. Three comparisons were computed: SP vs Ctrl, SP + L vs Ctrl and SP + L vs SP. They had ten, four and twelve differentially abundant proteins. Glycogen metabolism and nutrient reserves utilization are increased in the SP piglets. Structural muscle protein synthesis increased, causing higher energy requirements in SP + L piglets. Our results demonstrate the usefulness of proteomics to disclose the effect of dietary microalgae, whilst unveiling putative mechanisms derived from lysozyme supplementation. Data available via ProteomeXchange with identifier PXD024083. Significance: Spirulina, a microalga, is an alternative to conventional crops which could enhance the environmental sustainability of animal production. Due to its recalcitrant cell wall, its use requires additional measures to prevent anti-nutritional effects on the feeding of piglets in the post-weaning period, during which they endure post-weaning stress. One of such measures could be CAZyme supplementation to help degrade the cell wall during digestion. Muscle proteomics provides insightful data on the effect of dietary microalgae and enzyme activity on piglet metabolism.info:eu-repo/semantics/publishedVersio

Effects of Chlorella vulgaris as a feed ingredient on the quality and nutritional value of weaned piglets' meat

Chlorella vulgaris (CH) is usually considered a feed supplement in pig nutrition, and its use as an ingredient is poorly studied. Among many interesting characteristics, this microalga has high protein levels and can be a putative alternative for soybean meal. Our aim was to study the effect of a 5% CH incorporation in the diet, individually or combined with two carbohydrases, on meat quality traits and nutritional value. Forty-four post-weaned male piglets individually housed, with an initial live weight of 11.2 0.46 kg, were randomly distributed into four experimental groups: control (n = 11, without CH) and three groups fed with 5% CH incorporation, plain (n = 10), with 0.005% Rovabio® Excel AP (n = 10), and with 0.01% of a pre-selected four-CAZyme mixture (n = 11). After two weeks of trial, piglets were slaughtered and longissimus lumborum collected. CH had no effect on piglets’ growth performance. In turn, incorporation of CH improved the nutritional value of meat by increasing total carotenoids and n-3 PUFA content, thus contributing to a more positive n-6/n-3 fatty acid ratio. The supplementation with Rovabio® benefited tenderness and increased overall acceptability of pork. Our results show beyond doubt the viability of the utilization of this microalga as a feed ingredient for swine productioninfo:eu-repo/semantics/publishedVersio

Plasma Metabolites and Liver Composition of Broilers in Response to Dietary Ulva lactuca with Ulvan Lyase or a Commercial Enzyme Mixture

The effect of a high incorporation level of Ulva lactuca, individually and supplemented with a Carbohydrate-Active enZyme (CAZyme) on broilers’ plasma parameters and liver composition is assessed here. Twenty one-day-old Ross 308 male broilers were randomly assigned to one of four treatments (n = 10): corn/soybean meal based-diet (Control); based-diet with 15% U. lactuca (UL); UL diet with 0.005% of commercial carbohydrase mixture; and UL diet with 0.01% of recombinant ulvan lyase. Supplementing U. lactuca with the recombinant CAZyme slightly compromised broilers’ growth by negatively affecting final body weight and average daily gain. The combination of U. lactuca with ulvan lyase also increased systemic lipemia through an increase in total lipids, triacylglycerols and VLDL-cholesterol (p < 0.001). Moreover, U. lactuca, regardless of the CAZyme supplementation, enhanced hepatic n-3 PUFA (mostly 20:5n-3) with positive decrease in n-6/n-3 ratio. However, broilers fed with U. lactuca with ulvan lyase reduced hepatic a- and -tocopherol concentrations relative to the control. Conversely, the high amount of pigments in macroalga diets led to an increase in hepatic beta-carotene, chlorophylls and total carotenoids. Furthermore, U. lactuca, alone or combined with CAZymes, enhanced hepatic total microminerals, including iron and manganese. Overall, plasma metabolites and liver composition changed favorably in broilers that were fed 15% of U. lactuca, regardless of enzyme supplementationinfo:eu-repo/semantics/publishedVersio

Effect on Broiler Production Performance and Meat Quality of Feeding Ulva lactuca Supplemented with Carbohydrases

Macroalgae have been increasingly exploited worldwide for feed, food and biofuel applications, due to their nutritive and bioactive compounds. Green seaweeds belonging to the genus Ulva have high growth rates, which makes them suitable for being cultured in sustainable algae production, such as an integrated multi-trophic aquaculture system. This is expected to increase the use of Ulva sp. as an alternative source to conventional feedstuffs (e.g., cereals and soybean meal) for poultry. The objective of the current study was to assess if the incorporation of 15% Ulva lactuca in broiler chickens’ diet, combined or not with carbohydrate-active enzymes, would enhance meat nutritional quality without compromising animal growth performance. Overall, U. lactuca led to an accumulation of antioxidant carotenoids, n-3 PUFA and macrominerals, including magnesium, potassium and phosphorus, in the breast muscle, with likely health benefits, without significantly impairing growth performance. The supplementation of macroalgae with a recombinant ulvan lyase reduced ileal viscosity with possible beneficial effects on broiler digestibility. Although dietary U. lactuca showed potential to increase meat quality, it reduced meat overall acceptability, which suggests the use of a lower algae inclusion level to prevent a negative meat sensory perception for consumersinfo:eu-repo/semantics/publishedVersio