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The main goal of this study was to test a rational combination of pre‐selected carbohydrate‐active enzymes (CAZymes) and sulphatases, individually or in combination, in order to evaluate its capacity to disrupt Arthrospira platensis cell wall, allowing the release of its valuable nutritional bioactive compounds. By the end, a two‐enzyme constituted mixture (Mix), composed by a lysozyme and a α‐amylase, was incubated with A. platensis suspension. The microalga cell wall disruption was evaluated through the amount of reducing sugars released from the cell wall complemented with the oligosaccharide profile by HPLC. An increase of the amount of reducing sugars up to 2.42 g/L in microalgae treated with the Mix relative to no treatment (p <.05), as well as a 7‐fold increase of oligosaccharides amount (p <.001), were obtained. With resort of fluorescence microscopy, a 36% reduction of fluorescence intensity (p <.001) was observed using Calcofluor White staining. In the supernatant, the Mix caused a 1.34‐fold increase in protein content (p =.018) relative to the control. Similarly, n‐6 polyunsaturated fatty acids (PUFA) (p =.007), in particular 18:2n‐6 (p =.016), monounsaturated fatty acids (MUFA) (p =.049) and chlorophyll a (p =.025) contents were higher in the supernatant of microalgae treated with the enzyme mixture in relation to the control. Taken together, these results point towards the disclosure of a novel two‐enzyme mixture able to partial degrade A. platensis cell wall, improving its nutrients bioavailability for monogastric diets with the cost‐effective advantage use of microalgae in animal feed industry. [ABSTRACT FROM AUTHOR]

Seasonal weight loss (SWL) is a major constraint in extensive animal production systems in the tropics and Mediterranean. The objective of this study was to characterize the amino acid profile of muscle and hepatic tissues of Australian Merino, Damara and Dorper lambs under restricted feeding to evaluate the impact of SWL at the metabolic and physiological levels. SWL induced generalized muscle protein breakdown among restricted groups of all breeds, with varying intensity. Dorper breed mobilized less muscle amino acids when under these conditions, with the Damara having frequent significant differences, namely by having lower amino acid concentrations in the muscle of restricted lambs. Damara lambs showed greater ability to catabolize branched‐chain amino acids in the muscle tissue, which indicates yet another mechanism that provides the Damara with the necessary tools to endure harsh conditions. Overall, the Damara breed mobilized more muscle amino acids than the other breeds, with a better capacity to catabolize branched‐chain amino acids in the muscle, while maintaining muscle structural integrity. [ABSTRACT FROM AUTHOR]

EPA (20 : 5 n- 3) and DHA (22 : 6 n- 3) fatty acids have weight-reducing properties with physiological activity depending on their molecular structure – that is, as TAG or ethyl esters (EE). Aquaporins (AQP) are membrane protein channels recognised as important players in fat metabolism, but their differential expression in white adipose tissue (WAT) and brown adipose tissue (BAT), as well as their modulation by dietary n- 3 long-chain PUFA (LCPUFA) such as EPA and DHA, has never been investigated. In this study, the transcriptional profiles of AQP3, AQP5, AQP7 and selected lipid markers of WAT (subcutaneous and visceral) and BAT (interscapular) from hamsters fed diets containing n- 3 LCPUFA in different lipid structures such as fish oil (FO, rich in EPA and DHA in the TAG form) and FO-EE (rich in EPA and DHA in the EE form) were used and compared with linseed oil (LSO) as the reference group. A clear effect of fat depot was observed for AQP3 and leptin (LEP), with the lowest values of mRNA found in BAT relative to WAT. The opposite occurred for PPARα. AQP7 was affected by diet, with FO-fed hamsters having higher mRNA levels compared with LSO-fed hamsters. The relative gene expression of AQP5 , adiponectin (ADIPO), GLUT4 and PPARγ was influenced by both fat tissue and diet. Taken together, our results revealed a differential expression profile of AQP and some markers of lipid metabolism in both WAT and BAT in response to feeding n- 3 LCPUFA in two different structural formats: TAG v. EE. [ABSTRACT FROM AUTHOR]

We hypothesised that the incorporation of docosahexaenoic acid (DHA) across adipose tissues will be higher when it is ingested as triacylglycerols (TAG) structured at the sn-2 position. Ten-week old male hamsters were allocated to 4 dietary treatments (n = 10): linseed oil (LSO-control group), fish oil (FO), fish oil ethyl esters (FO-EE) and structured DHA at the sn-2 position of TAG (DHA-SL) during 12 weeks. In opposition to the large variations found for fatty acid composition in retroperitoneal white adipose tissue (WAT), brown adipose tissue (BAT) was less responsive to diets. DHA was not found in subcutaneous and retroperitoneal WAT depots but it was successfully incorporated in BAT reaching the highest percentage in DHA-SL. The PCA on plasma hormones (insulin, leptin, adiponectin) and fatty acids discriminated BAT from WATs pointing towards an individual signature on fatty acid deposition, but did not allow for full discrimination of dietary treatments within each adipose tissue. [ABSTRACT FROM AUTHOR]

BACKGROUND The individual and combined effects of betaine and arginine supplemented to reduced protein diets were investigated on plasma metabolites, hepatic fatty acid composition and mRNA levels of lipid-sensitive factors in commercial pigs. Betaine has previously been shown to reduce carcass fat deposition and arginine improves meat quality of finishing pigs. Forty male crossbred pigs were randomly assigned to one of five diets ( n = 8): 160 g kg−1 of crude protein (NPD), 130 g kg−1 of crude protein (RPD), RPD with 3.3 g kg−1 of betaine, RPD with 15 g kg−1 of arginine, and RPD with 3.3 g kg−1 of betaine and 15 g kg−1 of arginine. RESULTS The restriction of dietary protein increased total lipids ( P < 0.001), total cholesterol ( P < 0.001), high-density lipoprotein-cholesterol ( P < 0.001) and low-density lipoprotein cholesterol ( P < 0.001). Betaine and arginine, individually or combined, reduced the majority of plasma lipids ( P < 0.05) without affecting total fatty acids in the liver and the overall gene expression pattern. CONCLUSION These findings suggest a positive effect of betaine and arginine, singly or combined, by reversing plasma lipids increase promoted by dietary protein restriction. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Microalgae, small-sized algae, have been studied as a natural marine resource for a number of economically relevant applications, including animal feed. In this review, we unveil the dietary microalgae effects currently known on production and meat quality of livestock species (ruminants, pigs, poultry and rabbits). Microalgae are classified into diatoms ( Bacillariophyceae ), green algae ( Chlorophyceae ), golden algae ( Chrysophyceae ) and blue-green algae cyanobacteria ( Cyanophyceae ). The most important phototrophic species belong to Arthrospira , Chlorella , Dunaliella and Haematocussus genus. In addition, heterotrophic marine organisms, such as Crypthecodinium , Schizochytrium and Ulkenia , have been successfully cultivated for n -3 long-chain polyunsaturated fatty acids ( n -3 LCPUFA) production. Microalgae are mainly composed by proteins, carbohydrates, lipids, vitamins, minerals and bioactive compounds, such as carotenoids. This variable nutrient composition depends on species, strain and algae growing conditions. Research evidence so far has shown that the inclusion of microalgae in animal diets could improve growth and meat quality in ruminants, pigs, poultry and rabbits. These findings are highly dependent on microalgae own composition and their amount in the diet. In a general overview, the inclusion of Arthrospira platensis in pig and poultry diets increases average daily gain but negatively affects feed conversion ratio. Regarding Schizochytrium sp., this microalga improves fatty acid composition in pork and poultry meat, essentially due to its high content in docosahexaenoic acid (DHA). Chlorella , at very low percentages in feed, benefits growth performance parameters of poultry. The use of microalgae as feed ingredients is very promising as an alternative to corn and soybean, thus mitigating the current competition among food-feed-biofuel industries. In addition, microalgae contribute for the protection of environment and natural resources, namely land degradation and water deprivation. Microalgae also provide a sustainable alternative for n -3 LCPUFA availability, thus protecting worldwide fatty fish stocks. However, the cost-effective production and use of microalgae is a major challenge in the near future. In fact, the current microalgae cultivation technology should be improved to reduce their production costs. In addition, we foresee that the efficiency of microalgae incorporation in monogastric diets could be largely improved by the use of Carbohydrate-Active enZymes (CAZymes). CAZymes will allow the increase of nutrients bioavailability, as a consequence of recalcitrant microalgae cell walls degradation. Overall, the inclusion of microalgae in feed represents a very promising strategy for the maintenance and development of livestock sector, as an environmental friendly alternative to balance food-feed-biofuel industries. [ABSTRACT FROM AUTHOR]

Background: To overcome the current overexploitation offish rich in n-3 long chain polyunsaturated fatty acids (LCPUFA), microalgae have become a promising marine lipid source. The purpose of this study was to assess eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), isolated or combined from distinct marine origins, on the promotion of neuroprotective effects. Methods: The experiment lasted for 10 weeks and involved 32 Wistar rats, divided into 4 diets (n = 8): a diet rich in milk fat was taken as control (Milk Fat) and compared to n-3 LCPUFA enriched diets, either in EPA + DHA form through fish oil (Fish Oil), or EPA through Nannochloropsis oil (Nanno), or DHA through Schizochytrium oil (Schyzo), while maintaining Milk Fat incorporation. Results: Plasma lipid profile and dopamine levels were more beneficial in Fish Oil diet. In addition, n-3 LCPUFA incorporation was found increased in liver and erythrocytes from Fish Oil fed rats, suggesting that fish oil is a better dietary source for fatty acids deposition in the organism than microalgae. The Forced Swimming Test revealed a positive behavioural action of EPA + DHA, in opposition to Milk Fat and Nanno diets, which had higher immobile times. mRNA levels of serotonin receptors, HT1A and HT2A along with CREB, the transmission factor for learning and memory, were higher in the hippocampus of rats fed n-3 LCPUFA diets comparative to Milk Fat. Conclusion: Taken together, the combination of EPA and DHA from fish oil can counteract the undesirable health effects of saturated fat based diets and benefit, in the long run, neurological function. [ABSTRACT FROM AUTHOR]

In the present study, it was hypothesized that the incorporation of fatty acids is distinct among ruminant tissues and that it could be modulated by diet composition. To test this hypothesis, fatty acid composition, including conjugated linoleic acid isomers, of the most relevant beef by-products (brain, heart, kidney, liver, pancreas and tongue) from young bulls those fed distinct silage levels was assessed. Data indicated a large variation in fatty acid profile and conjugated linoleic acid composition among edible by-products. The most abundant fatty acids were C16:0 (kidney), C18:0 (heart and liver) and C18:1c9 (brain, pancreas and tongue) followed by C20:4n-6, except in brain (C22:6n-3 predominates). Brain, as shown by principal component analysis, presents a distinct fatty acid composition compared to the other beef by-products analysed. In addition, high silage diet relative to low silage diet promoted an increase of n-3 polyunsaturated fatty acid, t11,t13 and t11,c13 conjugated linoleic acid in heart, kidney, liver and pancreas. Overall, the data suggested that beef by-products had, in general, high contents of cholesterol, saturated fatty acid and trans fatty acid, as well as high levels of conjugated linoleic acid. Therefore, from a nutritional point of view they are recommended only in small amounts as part of a balanced diet. [ABSTRACT FROM AUTHOR]

The influence of genotype (lean v. fatty) and dietary protein level (normal v. reduced) on plasma metabolites, hepatic fatty acid composition and mRNA levels of lipid-sensitive factors is reported for the first time, using the pig as an experimental model. The experiment was conducted on forty entire male pigs (twenty lean pigs of Large White X Landrace X Pietrain cross-breed and twenty fatty pigs of Alentejana purebreed) from 60 to 93 kg of live weight. Each pig genotype was divided into two subgroups, which were fed the following diets: a normal protein diet (NPD) equilibrated for lysine (17⋅5% crude protein and 0⋅7% lysine) and a reduced protein diet (RPD) not equilibrated for lysine (13⋅1% crude protein and 0⋅4% lysine). The majority of plasma metabolites were affected by genotype, with lean pigs having higher contents of lipids, whereas fatty pigs presented higher insulin, leptin and urea levels. RPD increased plasma TAG, free fatty acids and VLDL-cholesterol compared with NPD. Hepatic total lipids were higher in fatty pigs than in the lean genotype. RPD affected hepatic fatty acid composition but had a slight influence on gene expression levels in the liver. Sterol regulatory element-binding factor 1 was down-regulated by RPD, and fatty acid desaturase 1 (FADS1) and fatty acid binding protein 4 (FABP4) were affected by the interaction between genotype and diet. In pigs fed RPD, FADS1 was up-regulated in the lean genotype, whereas FABP4 increased in the fatty genotype. Although there is a genotype-specific effect of dietary protein restriction on hepatic lipid metabolism, lipogenesis is not promoted in the liver of lean or fatty pigs. [ABSTRACT FROM AUTHOR]

The isolated or combined effects of betaine and arginine supplementation of reduced protein diets (RPD) on fat content, fatty acid composition and mRNA levels of genes controlling lipid metabolism in pig m. longissimus lumborum and subcutaneous adipose tissue (SAT) were assessed. The experiment was performed on forty intact male pigs (Duroc×Large White×Landrace cross-breed) with initial and final live weights of 60 and 93 kg, respectively. Pigs were randomly assigned to one of the following five diets (n 8): 16·0 % of crude protein (control), 13·0 % of crude protein (RPD), RPD supplemented with 0·33 % of betaine, RPD supplemented with 1·5 % of arginine and RPD supplemented with 0·33 % of betaine and 1·5 % of arginine. Data confirmed that RPD increase intramuscular fat (IMF) content and total fat content in SAT. The increased total fat content in SAT was accompanied by higher GLUT type 4, lipoprotein lipase and stearoyl-CoA desaturase mRNA expression levels. In addition, the supplementation of RPD with betaine and/or arginine did not affect either IMF or total fat in SAT. However, dietary betaine supplementation slightly affected fatty acid composition in both muscle and SAT. This effect was associated with an increase of carnitine O-acetyltransferase mRNA levels in SAT but not in muscle, which suggests that betaine might be involved in the differential regulation of some key genes of lipid metabolism in pig muscle and SAT. Although the arginine-supplemented diet decreased the mRNA expression level of PPARG in muscle and SAT, it did not influence fat content or fatty acid composition in any of these pig tissues. [ABSTRACT FROM PUBLISHER]

The cumulative effects of dietary arginine, leucine and protein levels on fat content, fatty acid composition and mRNA levels of genes controlling lipid metabolism in pig longissimus lumborum muscle and subcutaneous adipose tissue (SAT) were investigated. The experiment was performed on fifty-four intact male pigs (Duroc × Pietrain × Large White × Landrace crossbred), with a live weight ranging from 59 to 92 kg. The pigs were randomly assigned to one of six experimental treatments (n 9). The treatments followed a 2 × 3 factorial arrangement, with two levels of arginine supplementation (0 v. 1 %) and three levels of a basal diet (normal protein diet, NPD; reduced protein diet, RPD; reduced protein diet to achieve 2 % of leucine, RPDL). The results showed that dietary arginine supplementation did not affect the intramuscular fat (IMF) content and back fat thickness, but increased the total fat in SAT. This effect was associated with an increase in fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD) mRNA levels in SAT, which suggests that arginine might be involved in the differential regulation of some key lipogenic genes in pig muscle and SAT. The increase in IMF content under the RPD, with or without leucine supplementation, was accompanied by increased FASN and SCD mRNA levels. Arginine supplementation did not influence the percentage of main fatty acids, while the RPD had a significant effect on fatty acid composition in both tissues. Leucine supplementation of RPD did not change IMF, total fat of SAT and back fat thickness, but increased 16 : 0 and 18 : 1cis-9 and decreased 18 : 2n-6 in muscle. [ABSTRACT FROM PUBLISHER]

Abstract: Obesity consists in fat accumulation leading to increase in adipose cells number and size. Adipocyte membrane biophysical properties are critical to maintain cellular viability in metabolically healthy obesity. This study investigated the effect of the genetic background and dietary protein restriction on fat tissue lipid composition, adipocyte membrane fluidity and water permeability using the pig as experimental model. Twenty-four male pigs from distinct genotypes, lean and obese, were fed on normal and reduced protein diets within a 2×2 factorial arrangement (two genotypes and two diets). Backfat thickness was twofold higher in obese than in lean pigs but unrelated to dietary protein level. In contrast, total fatty acids in the subcutaneous adipose tissue were dependent on both breed and diet, with increased lipid content promoted by the fatty genotype and by the restriction of dietary protein. Adipose membranes isolated from obese pig’s subcutaneous fat tissue showed higher permeability to water, in line with an increased fluidity. Moreover, the reduced content of dietary protein influenced positively the fluidity of adipose membranes. Neither genotype nor diet affected total cholesterol concentration in the adipose membranes. Membrane-saturated fatty acids’ content was influenced by genotype, while membrane-polyunsaturated fatty acids, particularly from the n-6 family, was influenced by diet. The ratio of oleic (18:1c9)/linoleic (18:2n-6) acids was positively correlated with membrane fluidity. All together, these findings reinforce the genetic background as a determinant player on adipose membrane biophysical properties and point to the dietary protein level as an important factor for subcutaneous lipid deposition as well as for regulation of membrane function, factors that may have impact on human obesity and metabolic syndrome. [Copyright &y& Elsevier]