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Proteome of liver, intestine and muscle of weaning piglets in response to dietary inclusion of Chlorella vulgaris or its combination with Enterococcus lactis probiotic

ABSTRACT: Maize and soybean meal are widely used in monogastric animal diets. However, their production raises sustainability concerns. Microalgae, like Chlorella vulgaris, are a potential alternative feedstuff due to their interesting nutritional profile, particularly a high crude protein content. As part of a study investigating the individual and combined effects of dietary C. vulgaris and probiotic Enterococcus lactis SF68 on weaning piglet growth performance, dietary utilization, digestibility, organ morphometrics and physiology by metabolomics, lipidomics and proteomics, the submitted dataset includes the proteomics approach. At weaning (28 days old), Piétrain x (Landrace x Large White) piglets were obtained from a commercial farm. Animals were housed in groups of three in 2m2 pens and, for five weeks, fed one of three diets ad libitum: a control diet based on cereals and soybean meal (Control); a diet with 5% C. vulgaris (CV5); and a diet with 5% C. vulgaris and E. lactis SF68 probiotic (CV5P). Liver, ileum, and muscle tissues were sampled and proteomic analysis performed by LC-MS/MS. Proteomics analysis identified 1966 proteins in the liver, 1592 in the intestine and 1020 in the muscle. In general, multivariate analyses were able to identify broad proteome differences in the liver, but not in the intestine nor muscle. Nonetheless, all organs exhibited notable changes in levels of individual proteins by univariate analyses. Analyses of differential abundance revealed that the probiotic was a major driver of the observed effects whereas the combination of C. vulgaris and probiotic resulted in fewer differentially abundant proteins than expected from an additive effect of individual factors. This suggests the existence of complex interactions between the two dietary components. Overall, results favor the combination of C. vulgaris with the probiotic. In the liver, the combined diet effect resulted in immune attenuation as well as maintained protein catabolism, attenuated mobilization of lipid reserves, and modulation of cellular activity. These effects could enhance the systemic availability of amino acids and lipids to support muscle development during the post-weaning growth stage. In the intestine, evidence of lower ketone oxidation suggests an improved nutritional state, along with evidence of immune priming against potential bacterial infections in piglets fed the CV5P diet. In the muscle, the combined diet effect drives changes in muscle fiber types by affecting connective tissue, mitochondrial respiration and regulation of lipid composition. These findings suggest a positive outcome in expected meat quality, potentially improving tenderness and nutritional value. The output of this dataset discloses the importance of considering the intricate interplay between dietary components and their combined effects on multi-organ systems.

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ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Intestine, Liver, Muscle

SUBMITTER: Hugo Osorio

LAB HEAD: André Almeida

PROVIDER: PXD062608 | Pride | 2026-02-10

REPOSITORIES: pride

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Project description:The aim of the present study was to generate an experimental model to characterize the nutrigenomic profile of a plant-derived nutritional stress (S30 = 300 g Kg-1 Soybean Meal). Our results provided: a) a snapshot of molecular signatures describing a chronic and advanced nutritional stress to which future nutrigenomic studies might refer to; and b) a platform for the identification of candidate genes for the molecular phenotyping of several physiological parameters in liver and distal intestine. Atlantic salmon was used as a model. The nutritional stress was induced by inclusions of dietary defatted soybean meal (SBM) up to a level of 300 g kg-1, being this ingredient extensively demonstrated to induce reduced performance and enteropathy in the distal intestine (Baeverfjord and Krogdahl, 1996;Urán et al., 2009;URÁN et al., 2008). A control treatment with no SBM (S0) as well as intermediate levels of inclusion (100 g kg-1 and 200 g kg-1 SBM) were included to span a range of optimal and sub-optimal conditions. Performance parameters were measured and impaired growth was taken as an indicator of pronounced and chronic nutritional stress. Molecular analyses were performed in two tissues, liver and distal intestine. Distal intestine was chosen for being the site most morphologically and physiologically affected during the development of intestinal pathologies associated with plant ingredients such as SBM (Baeverfjord and Krogdahl, 1996;Kortner et al., 2011), while liver for being arguably the most metabolic active tissue. These tissues have been the most investigated targets in nutritional studies on fish so far. To the best of our knowledge, this study is the most comprehensive of its kind to report on the transcriptomic profile of the distal intestine and the liver, hence highlighting the supporting role of this tissue, in fish undergoing SBM-induced nutritional stress. Skugor et al (, 2011) described gene expression profiles of liver and intestine in fish fed 200 g kg-1 SBM inclusion using a 11K trout array. By investigating a larger number of probes (44K) in a more severe nutritional stress (300 g kg-1), our work will add further nutrigenomic information to the current literature.

Project description:Background: Consumption of high fat diets has negative impacts on health and well-being, some of which may be epigenetically regulated. Selenium and folate are two compounds which influence epigenetic mechanisms. We investigated the hypothesis that post-weaning supplementation with adequate levels of selenium and folate in mouse offspring fed a high fat, low selenium and folate diet during gestation and lactation will lead to epigenetic changes of potential importance for long-term health. Female offspring of mothers fed the experimental diet were either maintained on this diet (HF-low-low), or weaned onto a high-fat diet with sufficient levels of selenium and folate (HF-low-suf), for 8 weeks. Gene and protein expression, DNA methylation, and histone modifications were measured in colon and liver of female offspring. Results: Adequate levels of selenium and folate post-weaning affected gene expression in colon and liver of offspring, including decreasing Slc2a4 gene expression. Protein expression was only altered in the liver. There was no effect of adequate levels of selenium and folate on global histone modifications in the liver. Global liver DNA methylation was decreased in mice switched to adequate levels of selenium and folate, but there was no effect on methylation of specific CpG sites within the Slc2a4 gene in liver. Conclusions: Post-weaning supplementation with adequate levels of selenium and folate in female offspring of mice fed high-fat diets during gestation and lactation can alter global DNA methylation in liver. This may be one mechanism by which the negative effects of a poor diet during early life can be ameliorated. Further research is required to establish what role epigenetic changes play in mediating observed changes in gene and protein expression, and the relevance of these changes to health. Female wild type C57BL/6 mice (Animal Resource Centre, Western Australia) were fed a High Fat diet containing low levels of selenium and folate (HF-Low) for 7 days prior to mating with male C57BL/6 mice (Ruakura Small Animal Facility, Hamilton, New Zealand). Mothers were maintained on the HF-Low diet throughout gestation and lactation. Offspring of these female mice were randomly assigned to one of two different dietary treatments: either the same diet as the mothers (HF-Low), or a High Fat diet containing adequate selenium and folate (HF-Suf). At 12 weeks of age, mice were euthanized and colon and liver samples taken for microarray, proteomics, and DNA methylation analyses. Genomic DNA, total RNA and protein from whole colon and liver tissue was extracted using an AllPrepM-BM-. DNA/RNA/Protein mini kit (Qiagen, Cat number 80004). Colon and liver RNA from six female offspring on the HF-Low diet was compared with colon and liver RNA from six female offspring on the HF-Suf diet. All individual RNA samples were hybridized against a common reference RNA on separate arrays. The reference RNA was prepared by pooling in equimolar proportions RNA extracted from the intestine and liver of twelve female C57BL/6 mice, these being all of the mice from which samples were derived for microarray analysis in the current study.

			Action	DRS
	I109_CV5P.raw	Raw
	I113_CV5.raw	Raw
	I125_CV5P.raw	Raw
	I12_Control.raw	Raw
	I15_Control.raw	Raw

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Proteome of liver, intestine and muscle of weaning piglets in response to dietary inclusion of Chlorella vulgaris or its combination with Enterococcus lactis probiotic

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