Integrated Omics analysis of pig muscle metabolism under the effects of dietary Chlorella vulgaris and exogenous enzymes

Monogastric feeding is dependent on costly conventional feedstuffs. Microalgae such as Chlorella vulgaris are a sustainable alternative; however, its recalcitrant cell wall hinders monogastric digestion. Carbohydrate Active Enzyme (CAZyme) supplementation is a possible solution. The objective of this work was to evaluate the effect of 5% dietary C. vulgaris (CV) and enzymatic supplementation (CV + R— Rovabio® Excel AP; CV + M—four CAZyme mix) on muscle transcriptome and proteome of finishing pigs, in an integrated approach. Control pigs increased the abundance of contractile apparatus (MYH1, MYH2, MYH4) and energy metabolism (CKMT1, NDUFS3) proteins, demonstrating increased nutrient availability. They had increased expression of SCD, characteristic of increased glucose availability, via the activation of SREBP-1c and ChREBP. CV and CV + R pigs upregulated proteolytic and apoptotic genes (BAX, DDA1), whilst increasing the abundance of glucose (UQCRFS1) and fatty acid catabolism (ACADS) proteins. CV + R pigs upregulated ACOT8 and SIRT3 genes as a response to reduced nutrient availability, maintaining energy homeostasis. The cell wall specific CAZyme mix, CV + M, was able to comparatively reduce Omics alterations in the muscle, thereby reducing endogenous nutrient catabolism compared to the CV + R and CVinfo:eu-repo/semantics/publishedVersio

Expression of genes controlling fat deposition in two genetically diverse beef cattle breeds fed high or low silage diets

Articles in International JournalsBackground: Both genetic background and finishing system can alter fat deposition, thus indicating their influence on adipogenic and lipogenic factors. However, the molecular mechanisms underlying fat deposition and fatty acid composition in beef cattle are not fully understood. This study aimed to assess the effect of breed and dietary silage level on the expression patterns of key genes controlling lipid metabolism in subcutaneous adipose tissue (SAT) and longissimus lumborum (LL) muscle of cattle. To that purpose, forty bulls from two genetically diverse Portuguese bovine breeds with distinct maturity rates, Alentejana and Barrosã, were selected and fed either low (30% maize silage/70% concentrate) or high silage (70% maize silage/30% concentrate) diets. Results: The results suggested that enhanced deposition of fatty acids in the SAT from Barrosã bulls, when compared to Alentejana, could be due to higher expression levels of lipogenesis (SCD and LPL) and β-oxidation (CRAT) related genes. Our results also indicated that SREBF1 expression in the SAT is increased by feeding the low silage diet. Together, these results point out to a higher lipid turnover in the SAT of Barrosã bulls when compared to Alentejana. In turn, lipid deposition in the LL muscle is related to the expression of adipogenic (PPARG and FABP4) and lipogenic (ACACA and SCD) genes. The positive correlation between ACACA expression levels and total lipids, as well trans fatty acids, points to ACACA as a major player in intramuscular deposition in ruminants. Moreover, results reinforce the role of FABP4 in intramuscular fat development and the SAT as the major site for lipid metabolism in ruminants. Conclusions: Overall, the results showed that SAT and LL muscle fatty acid composition are mostly dependent on the genetic background. In addition, dietary silage level impacted on muscle lipid metabolism to a greater extent than on that of SAT, as evaluated by gene expression levels of adipogenic and lipogenic factors. Moreover, the response to diet composition evaluated through mRNA levels and fatty acid composition showed interesting differences between Alentejana and Barrosã bulls. These findings provide evidence that the genetic background should be taken into account while devising diet-based strategies to manipulate fatty acid composition of beef cattle tissues

Quality Traits and Nutritional Value of Pork and Poultry Meat from Animals Fed with Seaweeds

ReviewSeaweeds have caught the attention of the scientific community in recent years. Their production can mitigate the negative impact of anthropogenic activity and their use in animal nutrition reduces the dependency on conventional crops such as maize and soybean meal. In the context of monogastric animals, novel approaches have made it possible to optimise their use in feed, namely polysaccharide extraction, biomass fermentation, enzymatic processing, and feed supplementation with carbohydrate-active enzymes (CAZymes). Their bioactive properties make them putative candidates as feed ingredients that enhance meat quality traits, such as lipid oxidation, shelf-life, and meat colour. Indeed, they are excellent sources of essential amino acids, polyunsaturated fatty acids, minerals, and pigments that can be transferred to the meat of monogastric animals. However, their nutritional composition is highly variable, depending on species, harvesting region, local pollution, and harvesting season, among other factors. In this review, we assess the current use and challenges of using seaweeds in pig and poultry diets, envisaging to improve meat quality and its nutritional valueinfo:eu-repo/semantics/publishedVersio

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

ResearchBackground: 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 supplementationinfo:eu-repo/semantics/publishedVersio

Effect of dietary inclusion of Spirulina on production performance, nutrient digestibility and meat quality traits in post-weaning piglets

The effect of Spirulina (Arthrospira platensis), individually or in combination with two commercial carbohydrases, in piglet diets was assessed on growth performance, nutrient digestibility and meat quality traits. Forty post-weaned male piglets from Large White × Landrace sows crossed with Pietrain boars with an initial live weight of 12.0 ± 0.89 kg were used. Piglets were assigned to one of four dietary treatments (n = 10): cereal and soya bean meal base diet (control), base diet with 10% Spirulina (SP), SP diet supplemented with 0.005% Rovabio® Excel AP (SP + R) and SP diet supplemented with 0.01% lysozyme (SP + L). Animals were slaughtered after a 4-week experimental period. Growth performance was negatively affected by the incorporation of Spirulina in the diets, with an average decrease of 9.1% on final weight, in comparison with control animals. Total tract apparent digestibility (TTAD) of crude protein was higher (p < .05) in the control group than in other groups. In addition, lysozyme increased TTAD of crude fat and acid detergent fibre, relative to the SP and control groups, respectively. In addition, the incorporation of Spirulina, individually and supplemented with enzymes, did not impair meat quality traits. Surprisingly, no protective effect against lipid oxidation was observed with the inclusion of Spirulina in pork after 7 days of storage. This study indicates that growth performance of post-weaning piglets was impaired by the incorporation of 10% Spirulina in the diets, which is mediated by an increase in digesta viscosity and a lower protein digestibility, as a consequence of the resistance of microalga proteins to the action of endogenous peptidases. In addition, it also indicates that lysozyme, in contrast to Rovabio® Excel AP, is efficient in the degradation of Spirulina cell wall in piglet's intestine. However, the digestion of proteins liberated by Spirulina cell wall disruption is still a challengeinfo:eu-repo/semantics/publishedVersio

Integrated transcriptomics and proteomics analysis reveals muscle metabolism effects of dietary Ulva lactuca and ulvan lyase supplementation in weaned piglets

Abstract Seaweeds, including the green Ulva lactuca, can potentially reduce competition between feed, food, and fuel. They can also contribute to the improved development of weaned piglets. However, their indigestible polysaccharides of the cell wall pose a challenge. This can be addressed through carbohydrase supplementation, such as the recombinant ulvan lyase. The objective of our study was to assess the muscle metabolism of weaned piglets fed with 7% U. lactuca and 0.01% ulvan lyase supplementation, using an integrated transcriptomics (RNA-seq) and proteomics (LC–MS) approach. Feeding piglets with seaweed and enzyme supplementation resulted in reduced macronutrient availability, leading to protein degradation through the proteasome (PSMD2), with resulting amino acids being utilized as an energy source (GOT2, IDH3B). Moreover, mineral element accumulation may have contributed to increased oxidative stress, evident from elevated levels of antioxidant proteins like catalase, as a response to maintaining tissue homeostasis. The upregulation of the gene AQP7, associated with the osmotic stress response, further supports these findings. Consequently, an increase in chaperone activity, including HSP90, was required to repair damaged proteins. Our results suggest that enzymatic supplementation may exacerbate the effects observed from feeding U. lactuca alone, potentially due to side effects of cell wall degradation during digestion

Dietary Arthrospira platensis improves systemic antioxidant potential and changes plasma lipids without affecting related hepatic metabolic pathways in post-weaned piglets

Research ArticleBackground: The ability of a high level of dietary Arthrospira platensis, individually or in combination with two exogenous carbohydrate-degrading enzymes (lysozyme and Rovabio®), to improve systemic antioxidant potential and hepatic lipid metabolism was tested in piglets. Forty male post-weaned piglets, sons of Large White × Landrace sows crossed with Pietrain boars, were allocated into 4 groups (n = 10) and fed during 28 days one of the following diets: 1) a control basal diet (cereal and soybean meal); 2) a basal diet with 10% of A. platensis (AP); 3) the AP diet supplemented with 0.005% of Rovabio® (AP + R); 4) the AP diet supplemented with 0.01% of lysozyme (AP + L). Results: Arthrospira platensis decreased BW gain of piglets, regardless the addition of feed enzymes. The majority of plasma metabolites were affected by diets. A. platensis increased total lipids, total cholesterol and LDL-cholesterol, without changing hepatic fatty acid content or modulating, in an expressive manner, the transcriptional profile of lipid sensitive mediators. The antioxidant potential in general, and total carotenoids in particular, were improved by the microalga, regardless lysozyme or Rovabio®. Conclusions: Summing up, A. platensis, individually and combined with feed enzymes, impacts negatively on piglets’ growth but improves the systemic antioxidant potential and changes plasma lipids with a minor modulation on related hepatic metabolic pathwaysinfo:eu-repo/semantics/publishedVersio

Mineral Content of Liver of Buffaloes (<i>Bubalus bubalis</i>) Reared in Different Ecosystems in the Eastern Amazon

This study aimed to evaluate the influence of different production ecosystems, three in native and cultivated pastures (extensive), at two seasons of the year (dry and rainy), and one in confinement (intensive) in the Eastern Amazon, on the mineral content of buffalo liver raised on these ecosystems. Twelve male buffalo (n = 12), aged between 24 and 36 months, slaughtered in commercial slaughterhouses, were used in each of the ecosystems considered: Marajó; Lower Amazon; Cultivated Pasture, and in confinement system, Pará, Brazil. Approximately 5 g of liver was collected, stored and frozen until lyophilization. Samples were analyzed for mineral content based on inductively coupled plasma optical emission spectrometry (ICPOES) readings. The relationship between extensive ecosystems and an intensive production system (p p < 0.05) the mineral values found in the liver of buffaloes raised in the ecosystems, for sodium (Na), K, calcium (Ca), magnesium (Mg), sulfur (S), copper (Cu), iron (Fe), manganese (Mn) and barium (Ba). The period of the year interacted with the values of Na, K, S and Cu; however, an interaction of local vs. period of the yer was observed for the values of K, Mg, P, S and Cu. It can be concluded that the buffalo liver is an excellent source of minerals and can be included in the human diet and that the ecosystem the animals are raised influences its content

Arginine supplementation modulates pig plasma lipids, but not hepatic fatty acids, depending on dietary protein level with or without leucine

Abstract Background In the present study, the effect of arginine and leucine supplementation, and dietary protein level, were investigated in commercial crossbred pigs to clarify their individual or combined impact on plasma metabolites, hepatic fatty acid composition and mRNA levels of lipid sensitive factors. The experiment was conducted on fifty-four entire male pigs (Duroc × Pietrain × Large White × Landrace crossbred) from 59 to 92 kg of live weight. Each pig was randomly assigned to one of six experimental treatments (n = 9). The treatments followed a 2 × 3 factorial arrangement, providing two levels of arginine supplementation (0 vs. 1%) and three levels of basal diet (normal protein diet, NPD; reduced protein diet, RPD; reduced protein diet with 2% of leucine, RPDL). Results Significant interactions between arginine supplementation and protein level were observed across plasma lipids. While dietary arginine increased total lipids, total cholesterol, HDL-cholesterol, LDL-cholesterol, VLDL-cholesterol and triacylglycerols in NPD, the inverse effect was observed in RPD. Overall, dietary treatments had a minor impact on hepatic fatty acid composition. RPD increased 18:1c9 fatty acid while the combination of leucine and RPD reduced 18:0 fatty acid. Arginine supplementation increased the gene expression of FABP1, which contributes for triacylglycerols synthesis without affecting hepatic fatty acids content. RPD, with or without leucine addition, upregulated the lipogenic gene CEBPA but downregulated the fat oxidation gene LPIN1. Conclusions Arginine supplementation was responsible for a modulated effect on plasma lipids, which is dependent on dietary protein level. It consistently increased lipaemia in NPD, while reducing the correspondent metabolites in RPD. In contrast, arginine had no major impact, neither on hepatic fatty acids content nor on fatty acid composition. Likewise, leucine supplementation of RPD, regardless the presence of arginine, promoted no changes on total fatty acids in the liver. Ultimately, arginine, leucine and dietary protein reduction seem to be unrelated with fatty liver development