Project description:Microbiome analysis has relied largely on metagenomics to characterize microbial populations and predict their functions. Here, we used a TMT LC-MSMS metaproteomic analysis of the fecal microbiome in piglets before and after weaning to compare protein abundances as they pertain to microbial populations specific to either a milk- or plant-based diet. Fecal samples were collected from six piglets on the day of weaning and four weeks after transitioning to a standard nursery diet. Using the 12,554 protein groups identified in samples, we confirmed the shift in protein composition that takes place in response to the microbial succession following weaning and demonstrated the redundancy in metabolic processes between taxa. We identified taxa with roles as primary degraders based on corresponding proteins synthesized, thereby providing evidence for cross-feeding. Proteins associated with the breakdown of milk-specific carbohydrates were common among pre-weaned pigs, whereas the proteome of post-weaned piglets contained a greater abundance of proteins involved in the breaking down plant-specific carbohydrates. Furthermore, output revealed that production of propionate takes place via the propionaldehyde pathway in pre-weaned piglets, but changes to production via the succinate pathway in post-weaned piglets. Finally, a disproportionate quantity of carbohydrate-active enzymes (CAZymes) (~8%) were produced by fungi, which typically only represent ~0.1% of the microbiome taxa. Information gathered through this characterization of the metaproteome before and after weaning revealed important differences regarding the role of members in the microbial community, thereby providing information for the optimization of diets and products for both piglet and microbiome health.
Project description:Early-weaning-induced stress causes diarrhea, thereby reduces growth performance of piglets. Gut bacterial dysbiosis emerges as a leading cause of post-weaning diarrhea. The present study was aimed to investigate the effect of capsulized fecal microbiota transportation (FMT) on gut bacterial community, immune response and gut barrier function of weaned piglets. Thirty-two were randomly divided into two groups fed with basal diet for 21 days. Recipient group was inoculated orally with capsulized fecal microbiota of health Tibetan pig daily morning during whole period of trial, while control group was given orally empty capsule. The results showed that the F/G ratio, diarrhea ratio, diarrhea index, and histological damage score of recipient piglets were significantly decreased. FMT treatment also significantly increased the colon length of piglets. Furthermore, the relative abundances of Firmicutes, Euryarchaeota, Tenericutes, Lactobacillus, Methanobrevibacter and Sarcina in colon of recipient piglets were increased, and the relative abundances of Campylobacter, Proteobacteria, and Melainabacteria were significantly decreased compared with control group.
Project description:Weaning is a very critical period for piglets, typically accompanied by lower feed intake, weight loss after weaning and increased mortality. At weaning, piglets are exposed to many stressors, such as loss of mothering, mixing with other litters, end of lactational immunity, and a change in their environment and gut microbiota. After weaning, morphological and histological changes occur in the small intestine of piglets producing a rapid change of feeding regime which is critical for the immature digestive system. Sixteen female piglets were weaned to assess the effect of sorbic acid supplementation on the small intestine tissue transcriptome. At weaning day (T0), 4 piglets were sacrified and tissue samples collected. The remaining 12 piglets were weighted and randomly assigned to different post weaning (T5) diets. Diet A (n=6) contained 5 g/kg of sorbic acid. Diet B (n=6) is the same as Standard diet. Total RNA was isolated from ileum samples to be analyzed using the a CombiMatrix CustomArrayTM 90K platform . Even though diet had no detectable effect during the first 5 days after weaning, outcomes from this study highlighted some of the response mechanisms to the stress of weaning occurring in the piglet gut. A total of 205 differentially expressed genes were used for functional analysis using bioinformatics through BLAST2GO, Ingenuity Pathway Analysis 8.0, and the Dynamic Impact Aproach (DIA). Bioinformatics analysis revealed that Apoptosis, RIG-I-like and NOD-like receptor signaling were altered as a result of weaning. Results suggest that immune and inflammatory responses were activated and likely are a cause of small intestine atrophy as revealed by a decrease in villus height and villus/crypt ratio. Keywords: weaning, gut, gene expression, sorbic acid, microarray analysis
Project description: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). They were fed ad libitum for 4 weeks, after which 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.
Project description:The brown seaweed Laminaria digitata is a novel feedstuff for weaned piglets. It can help prevent dysbiosis in addition to improve overall health and performance. However, it has a recalcitrant cell wall that is not easily digested by the piglet digestive system. Alginate lyase has promising effects for the supplementation of in vivo diets in order to address this issue. The objective of this study is to evaluate the effect of 10% dietary Laminaria digitata inclusion and alginate lyase supplementation on the hepatic proteome and metabolome of weaned piglets in a physiological study. The diets caused incipient differences on the metabolome of piglets, with the proteome having the most significant changes. Feeding seaweed provided a source of n-3 PUFA that accumulated in the liver, signalling for increased fatty acid oxidation (FABP, ACADSB, ALDH1B1). This contributed at least in part to reduce oxidative stability of the tissue, demanding the higher abundance of GST to maintain it. Reactive oxygen species possibly damaged proteins, which caused hepatocytes to increase proteasome activity (LAPTM4B, PSMD4), recycling their amino acids. Providing alginate lyase to the diet increased the number of differentially abundant proteins, including GBE1 and LDHC that contributed to the maintenance of circulating glucose through mobilization of glycogen stores and branched chain amino acids. Enzymatic supplementation enhanced the baseline effects of feeding seaweed alone.
Project description:Scope: The âPredictive Adaptive Responseâ hypothesis suggests the in utero environment when mismatched with the post-natal environment can influence later life health. Underlying mechanisms are poorly understood, but may involve gene transcription changes, regulated via epigenetic mechanisms. Methods and Results: In a 2x2 factorial design, female C57Bl/6 mice were randomised to low or normal folate diets (0.4mg/ or 2mg folic acid/kg diet) prior to and during pregnancy and lactation with. At weaning, offspring were randomised to high or low fat diets at weaning. Genome-wide gene expression and promoter DNA methylation were measured using microarrays in adult male livers. Maternal folate depletion and high fat intake post-weaning influenced gene expression (1959 and 1612 genes respectively) and promoter DNA methylation (208 and 344 loci respectively) but changes in expression and methylation were poorly matched for both dietary interventions. Expression of 667 genes was altered in response to both maternal folate depletion and post-weaning high fat feeding. In addition, there was evidence that the combined dietary insult (i.e. maternal folate depletion followed by high fat post-weaning) exerted the largest expression change for most of these genes. Conclusion: Our observations align with, and provide evidence in support of a potential underlying mechanism for, the âPredictive Adaptive Responseâ hypothesis. Elucidation of these mechanisms may identify targets for interventions to mitigate effects of adverse nutrition exposures during early development on disease risk in later life.
Project description:Abstract Scope: The âPredictive Adaptive Responseâ hypothesis suggests the in utero environment when mismatched with the post-natal environment can influence later life health. Underlying mechanisms are poorly understood, but may involve gene transcription changes, regulated via epigenetic mechanisms. Methods and Results: In a 2x2 factorial design, female C57Bl/6 mice were randomised to low or normal folate diets (0.4mg/2mg folic acid/kg diet) prior to and during pregnancy and lactation with, offspring randomised to high or low fat diets at weaning. Genome-wide gene expression and promoter DNA methylation were measured using microarrays in adult male livers. Maternal folate depletion and high fat intake post-weaning influenced gene expression (1959 and 1612 genes respectively) and promoter DNA methylation (208 and 344 loci respectively) but changes in expression and methylation were poorly matched for both dietary interventions. Expression of 667 genes was altered in response to both maternal folate depletion and post-weaning high fat feeding. In addition, there was evidence that the combined dietary insult (i.e. maternal folate depletion followed by high fat post-weaning) exerted the largest expression change for most of these genes. Conclusion: Our observations align with, and provide evidence in support of a potential underlying mechanism for, the âPredictive Adaptive Responseâ hypothesis. Elucidation of these mechanisms may identify targets for interventions to mitigate effects of adverse nutrition exposures during early development on disease risk in later life.
Project description:probiotics and prebiotics to maternal diets is related to decreased incidence of diarrhea and greater weight gain during lactation. Our objective was to determine the impact of adding whole ground oat as a prebiotic alone or in combination with postbiotic yeast culture (YC) (Saccharomyces cerevisiae) to sow gestation and lactation rations on milk composition, piglet growth, and incidence of post weaning diarrhea (PWD). Diets: control (CON), CON + yeast culture (YC) [5g/kg], CON + oat (15% inclusion rate) (Oat) or CON+ YC [5g/kg] + Oat (15%) were fed during the last 30 days of gestation and throughout lactation (18-21 days). Shotgun proteome analysis of day 4 and 7 postpartum milk samples found 36 differentially abundant proteins (P-adj <0.1) in both Oat and YC supplemented sows relative to CON. Notable was increased expression of antimicrobial proteins, lactoferrin and chitinase. IgG in milk of Oat supplemented sows was lower than YC supplemented sows (p<0.05) but had greater E. coli-antigen reactivity. Piglet weights at birth were similar. At weaning YC + Oat piglets weighed less and gained less weight (p<0.05) postweaning than CON. The incidence of PWD was lowest in the YC and Oat groups compared to CON and YC+ Oat groups. These data suggest that Oat or YC culture supplementation alters milk immune and antimicrobial associated proteins that can impact piglets but may have negative effects on piglet growth when given in combination.
Project description:This study examines whether maternal low ω6:ω3 ratio diet and offspring seaweed (SW) supplementation can improve offspring immunity and performance by elucidating the effects on piglet serum proteome. A total of 16 sows were given either a standard (CR, 13:1) or low ω6:ω3 ratio diet (LR, 4:1) during pregnancy and lactation and their male weaned piglets were supplemented with SW powder (4 g/kg, SW) or not (CT) in a 21-day post-weaning (PW) diet. Four PW piglet groups were then identified based on dam and piglet treatment, namely CRCT, CRSW, LRCT, and LRSW (n = 10 each). Piglet serum collected at weaning and d21 PW were analyzed (n = 5 each) using TMT-based quantitative proteomics and validated by appropriate assays.