Prenatal exposure to innately preferred D-limonene and trans-anethole does not overcome innate aversion to eucalyptol, affecting growth performance of weanling piglets

Funding: Animal Nutrition and Welfare Service (SNIBA) of the Autonomous University of Barcelona.In the present research, two studies were performed to determine the effects of specific botanical compounds (BCs) on the innate feed preference and feed intake of piglets, as follows: Exp. 1 studied the innate feed preferences of post-weaning piglets using a double-choice feeding test. A total of 828 weaned piglets were distributed into 36 pens (23 pigs/pen) and assigned to three dietary pair choice feeding options (n = 12): unsupplemented prestarter diets (reference) versus reference plus D-limonene, trans-anethole, or eucalyptol. Piglets showed a preference for diets with D-limonene (53.8%) and trans-anethole (54.5%), and an aversion to eucalyptol (41.6%) (p < 0.05). Exp. 2 studied whether the prenatal and perinatal exposure to D-limonene, trans-anethole, and eucalyptol influences the feed intake and growth of newly-weaned piglets. Twenty-eight gestating and lactating sows were distributed into two dietary treatments (n = 14): unsupplemented Control diets or Control plus a blend of BCs (BBC; containing D-limonene, trans-anethole, and eucalyptol). D-limonene, trans-anethole, and eucalyptol were transferred into the placental fluid, and D-limonene and trans-anethole into the milk (p < 0.05). Furthermore, weanling piglets (n = 200; Control) and (n = 203; BBC) received the same treatment as their mothers in prestarter diets. The early response after weaning showed that piglets' post-weaning BW gain was higher in the Control (p < 0.05) group than in those exposed to BBC. In conclusion, prenatal exposure to preferred D-limonene and trans-anethole, or familiarity to eucalyptol did not help to overcome the innate aversion to eucalyptol and its negative effect on weanling piglets' BW

Phytogenic Compounds Supplemented to Gestating Hyperprolific Sows Affects the Gut Health-Related Gene Expression and Histological Responses in Neonate Piglets

This research aims to determine whether a specific blend of phytogenic compounds (BPC) supplemented in gestating hyperprolific sow diets can promote prenatal maternal effects in terms of piglet gut function and morphology. Twenty-eight (Landrace × Yorkshire) gilts and sows (parity 0 to 7) were randomly distributed by parity number and body weight into two dietary treatments: unsupplemented Control (CON) (n = 14) or CON diet supplemented with 1 g/kg feed of BPC during gestation (n = 14). The BPC supplementation during gestation of sows downregulated the neonate piglets' jejunal genes involved in oxidation (SOD2) and nutrient transport (SLC16A1/MCT1, SLC11A2/DMT1, and SLC39A/ZIP4), while IFNG and CLDN4 related to immune response and barrier function, respectively, were upregulated (q < 0.10). In addition, the jejunal villus height and the ratio of the villus height to crypt depth tended to increase (p < 0.10), while goblet cell volume density was higher (p < 0.05) in BPC compared to CON. In conclusion, dietary supplementation of BPC in gestating diets for hyperprolific sows influences neonatal histomorphology and expression of genes related to the intestinal function and health

Evidence from Studies with Heat-Stressed Caco-2 Cells, C. elegans and Growing Broilers

Climatic changes and heat stress have become a great challenge in the livestock industry, negatively affecting, in particular, poultry feed intake and intestinal barrier malfunction. Recently, phytogenic feed additives were applied to reduce heat stress effects on animal farming. Here, we investigated the effects of ginseng extract using various in vitro and in vivo experiments. Quantitative real-time PCR, transepithelial electrical resistance measurements and survival assays under heat stress conditions were carried out in various model systems, including Caco-2 cells, Caenorhabditis elegans and jejunum samples of broilers. Under heat stress conditions, ginseng treatment lowered the expression of HSPA1A (Caco-2) and the heat shock protein genes hsp-1 and hsp-16.2 (both in C. elegans), while all three of the tested genes encoding tight junction proteins, CLDN3, OCLN and CLDN1 (Caco-2), were upregulated. In addition, we observed prolonged survival under heat stress in Caenorhabditis elegans, and a better performance of growing ginseng-fed broilers by the increased gene expression of selected heat shock and tight junction proteins. The presence of ginseng extract resulted in a reduced decrease in transepithelial resistance under heat shock conditions. Finally, LC-MS analysis was performed to quantitate the most prominent ginsenosides in the extract used for this study, being Re, Rg1, Rc, Rb2 and Rd. In conclusion, ginseng extract was found to be a suitable feed additive in animal nutrition to reduce the negative physiological effects caused by heat stress. View Full-Tex

Identification of Insulin-Mimetic Plant Extracts: From an In Vitro High-Content Screen to Blood Glucose Reduction in Live Animals

Type 2 diabetes mellitus (T2DM) is linked to insulin resistance and a loss of insulin sensitivity, leading to millions of deaths worldwide each year. T2DM is caused by reduced uptake of glucose facilitated by glucose transporter 4 (GLUT4) in muscle and adipose tissue due to decreased intracellular translocation of GLUT4-containing vesicles to the plasma membrane. To treat T2DM, novel medications are required. Through a fluorescence microscopy-based high-content screen, we tested more than 600 plant extracts for their potential to induce GLUT4 translocation in the absence of insulin. The primary screen in CHO-K1 cells resulted in 30 positive hits, which were further investigated in HeLa and 3T3-L1 cells. In addition, full plasma membrane insertion was examined by immunostaining of the first extracellular loop of GLUT4. The application of appropriate inhibitors identified PI3 kinase as the most important signal transduction target relevant for GLUT4 translocation. Finally, from the most effective hits in vitro, four extracts effectively reduced blood glucose levels in chicken embryos (in ovo), indicating their applicability as antidiabetic pharmaceuticals or nutraceuticals

Impact of a High-Fat or High-Fiber Diet on Intestinal Microbiota and Metabolic Markers in a Pig Model

To further elaborate interactions between nutrition, gut microbiota and host health, an animal model to simulate changes in microbial composition and activity due to dietary changes similar to those in humans is needed. Therefore, the impact of two different diets on cecal and colonic microbial gene copies and metabolic activity, organ development and biochemical parameters in blood serum was investigated using a pig model. Four pigs were either fed a low-fat/high-fiber (LF), or a high-fat/low-fiber (HF) diet for seven weeks, with both diets being isocaloric. A hypotrophic effect of the HF diet on digestive organs could be observed compared to the LF diet (p &lt; 0.05). Higher gene copy numbers of Bacteroides (p &lt; 0.05) and Enterobacteriaceae (p &lt; 0.001) were present in intestinal contents of HF pigs, bifidobacteria were more abundant in LF pigs (p &lt; 0.05). Concentrations of acetate and butyrate were higher in LF pigs (p &lt; 0.05). Glucose was higher in HF pigs, while glutamic pyruvic transaminase (GPT) showed higher concentrations upon feeding the LF diet (p &lt; 0.001). However, C-reactive protein (CRP) decreased with time in LF pigs (p &lt; 0.05). In part, these findings correspond to those in humans, and are in support of the concept of using the pig as human model

Ginseng Extract Ameliorates the Negative Physiological Effects of Heat Stress by Supporting Heat Shock Response and Improving Intestinal Barrier Integrity: Evidence from Studies with Heat-Stressed Caco-2 Cells, C. elegans and Growing Broilers

Climatic changes and heat stress have become a great challenge in the livestock industry, negatively affecting, in particular, poultry feed intake and intestinal barrier malfunction. Recently, phytogenic feed additives were applied to reduce heat stress effects on animal farming. Here, we investigated the effects of ginseng extract using various in vitro and in vivo experiments. Quantitative real-time PCR, transepithelial electrical resistance measurements and survival assays under heat stress conditions were carried out in various model systems, including Caco-2 cells, Caenorhabditis elegans and jejunum samples of broilers. Under heat stress conditions, ginseng treatment lowered the expression of HSPA1A (Caco-2) and the heat shock protein genes hsp-1 and hsp-16.2 (both in C. elegans), while all three of the tested genes encoding tight junction proteins, CLDN3, OCLN and CLDN1 (Caco-2), were upregulated. In addition, we observed prolonged survival under heat stress in Caenorhabditis elegans, and a better performance of growing ginseng-fed broilers by the increased gene expression of selected heat shock and tight junction proteins. The presence of ginseng extract resulted in a reduced decrease in transepithelial resistance under heat shock conditions. Finally, LC-MS analysis was performed to quantitate the most prominent ginsenosides in the extract used for this study, being Re, Rg1, Rc, Rb2 and Rd. In conclusion, ginseng extract was found to be a suitable feed additive in animal nutrition to reduce the negative physiological effects caused by heat stress

Phytogenic actives supplemented in hyperprolific sows: effects on maternal transfer of phytogenic compounds, colostrum and milk features, performance and antioxidant status of sows and their offspring, and piglet intestinal gene expression

Phytogenic actives (PA) are plant-derived natural bioactive compounds that may promote livestock health and well-being, as well as improve growth performance and production efficiency. The current study aims to evaluate their effects on sows and their offspring. Eighty-one hyperprolific sows (up to parity 7) were assigned to 3 experimental treatments. Control sows were offered a nonsupplemented diet during gestation and lactation, and treated sows were fed the control diet supplemented with 1 g/kg of a blend of PA (BPA) in lactation (L) or during gestation and lactation (GL). An evaluation was made of placental and milk maternal transfer of these BPA and colostrum–milk features, sows and piglets antioxidant status, reproductive performance (litter size), body weight (BW) changes, weaning-estrus interval, and litter performance. Finally, piglet´s jejunum gene expression was measured. The BPA supplementation during gestation (GL) increased the number of piglets born alive (P = 0.020) and reduced (P  0.05). Dietary phytogenic volatile compounds reached GL placental fluid, and milk of L and GL sows (P < 0.05). Moreover, colostrum protein in GL and milk fat content in L and GL were increased (P < 0.05). Milk of GL showed inhibitory activity against Bacillus subtilis and Staphylococcus aureus (P < 0.05). Antioxidant status of GL sows showed an enhanced (P < 0.05) of catalase (CAT) and total antioxidant capacity levels at early gestation (day 35), whereas higher levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) enzymes at late gestation (day 110). Likewise, GL newborn piglets showed higher CAT levels, whereas both CAT and SOD levels in suckling piglets, as well as CAT, SOD, and GSH-Px in weaned piglets, were increased in L and GL (P < 0.05). Jejunum messenger ribonucleic acid abundance of suckling piglets in L and GL groups showed overexpression of barrier function MUC2, digestive enzyme IDO, and immune response PPARGC-α, TNF-α, TGF-β1, and IL-10 genes (P < 0.05). In conclusion, dietary BPA supplementation in hyperprolific sows increased the litter size (born alive) and improved the composition and bioactivity of colostrum and milk, besides, modified the antioxidant status of sows and their offspring, as well as the suckling piglets gut health gene expression. Several BPA volatile compounds were prenatal and postnatal maternally transferred (placental fluid and milk).Peer reviewe

Intestinal Microbiota and Microbial Metabolites Are Changed in a Pig Model Fed a High-Fat/Low-Fiber or a Low-Fat/High-Fiber Diet

<div><p>The intestinal microbiota and its metabolites appear to be an important factor for gastrointestinal function and health. However, research is still needed to further elaborate potential relationships between nutrition, gut microbiota and host’s health by means of a suitable animal model. The present study examined the effect of two different diets on microbial composition and activity by using the pig as a model for humans. Eight pigs were equally allotted to two treatments, either fed a low-fat/high-fiber (LF), or a high-fat/low-fiber (HF) diet for 7 weeks. Feces were sampled at day 7 of every experimental week. Diet effects on fecal microbiota were assessed using quantitative real-time PCR, DNA fingerprinting and metaproteomics. Furthermore, fecal short-chain fatty acid (SCFA) profiles and ammonia concentrations were determined. Gene copy numbers of lactobacilli, bifidobacteria (<i>P</i><0.001) and <i>Faecalibacterium prausnitzii</i> (<i>P</i><0.05) were higher in the LF pigs, while <i>Enterobacteriaceae</i> were more abundant in the HF pigs (<i>P</i><0.001). Higher numbers of proteins affiliated to <i>Enterobacteriaceae</i> were also present in the HF samples. Proteins for polysaccharide breakdown did almost exclusively originate from <i>Prevotellaceae</i>. Total and individual fecal SCFA concentrations were higher for pigs of the LF treatment (<i>P</i><0.05), whereas fecal ammonia concentrations did not differ between treatments (<i>P</i>>0.05). Results provide evidence that beginning from the start of the experiment, the LF diet stimulated beneficial bacteria and SCFA production, especially butyrate (<i>P</i><0.05), while the HF diet fostered those bacterial groups which have been associated with a negative impact on health conditions. These findings correspond to results in humans and might strengthen the hypothesis that the response of the porcine gut microbiota to a specific dietary modulation is in support of using the pig as suitable animal model for humans to assess diet-gut-microbiota interactions.</p><p>Data are available via ProteomeXchange with identifier PXD003447.</p></div

Development of short-chain fatty acids (SCFA) and ammonia contents in feces of pigs (n = 4 per treatment) over time.

<p>Development of short-chain fatty acids (SCFA) and ammonia contents in feces of pigs (n = 4 per treatment) over time.</p

Mean values of bacterial numbers in feces of pigs (n = 4 per treatment) at the base status compared to week 1 of the experiment, HF and LF.

<p>HF, high-fat/low-fiber; LF, low- fat/high-fiber; FM, fresh matter. Values represent least squares means ± SEM. <i>P</i><0.05*, <i>P</i><0.01**, <i>P</i><0.001***.</p