Data_Sheet_1_Dietary bile acids supplementation modulates immune response, antioxidant capacity, glucose, and lipid metabolism in normal and intrauterine growth retardation piglets.docx

Intrauterine growth retardation (IUGR) results in intestinal dysfunction contributing to metabolic syndrome and growth lag of piglets. Bile acid (BA) presents various bioactivities, including regulation roles in antioxidant, anti-inflammation, and glucose and lipid metabolism. Forty-eight weaned piglets were allocated to four groups in a 2 × 2 factorial arrangement with the effects of BA supplementation and IUGR challenge. Twenty-four IUGR piglets and 24 normal birth weight (NBW) piglets were allocated into two groups, respectively, including the control group fed with a basal diet, and the treatment group fed a basal diet supplemented with 400 mg/kg BA. The experiment lasted 28 days. The results indicated that BA improved liver and spleen indexes in IUGR piglets, whereas decreased blood RDW-CV and RDW-SD regardless of IUGR (P 2O2 and decreased liver T-AOC in weaned piglets (P < 0.05). In addition, IUGR downregulated liver Nrf1 and Nrf2 expression levels, while BA supplementation upregulated the Nrf2 expression of liver in weaned piglets (P < 0.05). Dietary BA decreased (P < 0.05) jejunal GSH concentration and ileal CAT activity regardless of IUGR. Furthermore, IUGR upregulated (P < 0.05) jejunal SOD and CAT expression levels; however, dietary BA upregulated ileal Nrf1 (P < 0.05) and Keap1 (P = 0.07) expression levels in piglets regardless of IUGR. Moreover, IUGR upregulated the liver lipid synthesis (FAS) and downregulated HSL and SCD1 expression levels, while dietary BA downregulated liver FAS and SCD1 expression levels (P < 0.05). However, BA supplementation could enhance liver gluconeogenesis by upregulating (P < 0.05) the liver G6PC and PCK1 expression levels in the NBW piglets but not in the IUGR piglets. Collectively, these findings suggest that BA could regulate the redox status of weaned piglets by regulating the Nrf2/Keap1 pathway and improving liver glucose and lipid metabolism of IUGR piglets. These findings will provide a reference for the application of BA in swine production; moreover, considering the physiological similarity between pigs and humans, these findings will provide a reference for IUGR research in humans.</p

Effects of dietary nutrient levels on microbial community composition and diversity in the ileal contents of pregnant Huanjiang mini-pigs

<div><p>The mammalian gut microbiota influences various metabolic and physiological processes. Substantial metabolic changes occur during a healthy pregnancy that may be related to microbiota composition dynamics. However, the effect of diet on intestinal microbiota composition and diversity during pregnancy remains unclear. We examined the ileal contents of Huanjiang mini-pigs at two pregnancy stages to determine the effects of dietary nutrient levels on such microbial communities. Animals received either a higher-nutrient (HN) diet formulated to meet US National Research Council requirements or a lower-nutrient (LN) diet that met the Chinese National Feeding Standard recommendations. On day 45 or 75 of pregnancy, sows were euthanized and their ileal contents sampled. Experimental diet and pregnancy stage did not affect ileal bacterial richness or diversity, as determined by Chao1 and ACE species richness measures and Shannon and Simpson indices, respectively. The phyla Firmicutes and Proteobacteria, accounting for 69.99–85.44% and 5.82–15.17% of the total reads, respectively, predominated regardless of diet. At the genus level, diet significantly affected the abundance of <i>Lactobacillus</i> species, which was greater in pigs given HN feed (<i>P <</i> 0.05), but had little impact on that of <i>Megasphaera</i> species (<i>P</i> = 0.096). Pregnancy stage had a minimal effect on Proteobacteria numbers (<i>P</i> = 0.053). The number of bacteria of the phylum Firmicutes and genus <i>Lactobacillus</i> decreased, while that of the phylum Proteobacteria, family Enterobacteriaceae, and genus <i>Bacteroides</i> increased between days 45 and 75 of pregnancy. Of the short-chain fatty acids (SCFAs) measured, only propionate levels changed significantly, with higher concentrations observed on day 45 than on day 75. Our findings indicate that Firmicutes and Proteobacteria dominate pregnant sow ileal bacterial profiles. Excepting a tendency for the number of Proteobacteria to increase as pregnancy progressed, pregnancy stage and diet had little effect on ileal microbiotic composition and diversity and luminal SCFA concentrations.</p></div

PCoA based on the UniFrac distance metric.

<p>To evaluate similarities between bacterial communities, graphs A, B, and C were generated using OTUs, based on the UniFrac distance metrics PC1 and PC2, PC3 and PC2, and PC1 and PC3 (on a two-dimensional array), respectively. Samples from each treatment group are represented as follows: ►, HN45; ■, HN75; ▲, LN45; and ●, LN75. HN45 and HN75 represent samples obtained from Huanjiang mini-pigs fed the higher-nutrient diet for 45 and 75 days, respectively. LN45 and LN75 indicate samples obtained from those given the lower-nutrient diet for 45 and 75 days, respectively.</p

Principal component analysis of bacterial OTUs in ileal contents and the 10 predominant OTUs.

<p>Principal component analysis of bacterial OTUs in ileal contents and the 10 predominant OTUs.</p

Effect of diet and pregnancy stage on ileal SCFA and BCFA concentrations (mg/g) in Huanjiang mini-pigs.

<p>Effect of diet and pregnancy stage on ileal SCFA and BCFA concentrations (mg/g) in Huanjiang mini-pigs.</p

Rarefaction curves of bacterial species abundance in the ileal luminal contents of pregnant Huanjiang mini-pigs.

<p>HN45 and HN75 represent samples obtained from Huanjiang mini-pigs fed a higher-nutrient diet for 45 and 75 days, respectively. LN45 and LN75 represent samples obtained from those fed a lower-nutrient diet for 45 and 75 days, respectively.</p

Composition of ileal bacterial communities at the genus level in Huanjiang mini-pigs at different stages of pregnancy (%).

<p>Composition of ileal bacterial communities at the genus level in Huanjiang mini-pigs at different stages of pregnancy (%).</p

Alpha diversity indices of ileal bacterial communities in Huanjiang mini-pigs at different stages of pregnancy^a.

<p>Alpha diversity indices of ileal bacterial communities in Huanjiang mini-pigs at different stages of pregnancy<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0172086#t002fn001" target="_blank">^a</a>.</p

Effect of diet and pregnancy stage on OTU levels (%) in the ileal bacterial communities of Huanjiang mini-pigs^*.

<p>Effect of diet and pregnancy stage on OTU levels (%) in the ileal bacterial communities of Huanjiang mini-pigs^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0172086#t005fn001" target="_blank">*</a>}.</p

Overlap in genes and compounds between Pharos and the BDRH.

a) more gene targets were found in the BDRH than via Pharos; b) Pharos had more compounds than BDRH.</p