The Effect of an Essential Oil Blend on Growth Performance, Intestinal Health, and Microbiota in Early-Weaned Piglets.

Essential oils (EO) are promising feed additives for their antibacterial, antioxidant, and immune-enhancing abilities with low toxicity. Carvacrol, thymol, and cinnamaldehyde are commonly used to synthesize EO. However, few studies focus on combining these three EO in early-weaned piglets. In the present study, 24 piglets weaned at 21 d of age were randomly divided into 2 groups (6 replicate pens per group, 2 piglets per pen). The piglets were fed a basal diet (the control group) and a basal diet supplemented with 400 mg/kg EO (a blend consisting of carvacrol, thymol, and cinnamaldehyde, the EO group) for 28 days. At the end of the experiment, one piglet per pen was randomly chosen to be sacrificed. Growth performance, hematology, plasma biochemical indices, antioxidant capacity, intestinal epithelial development and immunity, colonic volatile fatty acids (VFA), and microbiota were determined. The results indicated that the diet supplemented with EO significantly improved average daily feed intake (ADFI, p < 0.01) and average daily gain (ADG, p < 0.05) in the day 0 to 28 period. EO supplementation led to a significant decrease in plasma lysozyme (p < 0.05) and cortisol levels (p < 0.01). Additionally, EO significantly promoted jejunal goblet cells in the villus, jejunal mucosa ZO-1 mRNA expression, ileal villus height, and ileal villus height/crypt depth ratio in piglets (p < 0.05). The ileal mucosal TLR4 and NFκB p-p65/p65 protein expression were significantly inhibited in the EO group (p < 0.05). Colonic digesta microbiota analysis revealed that bacteria involving the Erysipelotrichaceae family, Holdemanella genus, Phascolarctobacterium genus, and Vibrio genus were enriched in the EO group. In conclusion, these findings indicate that the EO blend improves ADG and ADFI in the day 0 to 28 period, as well as intestinal epithelial development and intestinal immunity in early-weaned piglets, which provides a theoretical basis for the combined use of EO in weaned piglets.

Effects and interaction of dietary electrolyte balance and citric acid on growth performance, intestinal histomorphology, digestive enzyme activity and nutrient transporters expression of weaned piglets.

Fifty-six piglets were weaned at 21 days and randomly assigned to 1 of 8 dietary treatments with 7 replicate pens for a 14-day experimental period. The eight experimental diets were prepared via a 2 × 4 factorial arrangement with citric acid (CA; 0 and 0.3%) and dietary electrolyte balance (dEB, Na +K - Cl mEq/kg of the diet; -50, 100, 250, and 400 mEq/kg). Varying dEB values were obtained by altering calcium chloride and sodium bicarbonate contents. Dietary CA significantly increased (p < .05) villus height (VH) and villus height:crypt depth (VH:CD) in the jejunum. Piglets fed a 250 mEq/kg diet increased (p < .05) VH and VH:CD values in the duodenum. Jejunal VH and VH:CD increased (quadratic; p < .05), and ileal VH:CD (liner and quadratic; p < .05) decreased as dEB was increased in diets without CA, but no such effect was observed on the diets containing CA (dEB ×CA; p < .05). The CD in jejunum (quadratic; p < .05) increased as dEB was increased in diets containing CA, whereas it was decreased (linear; p < .05) in the diets without CA (dEB ×CA; p < .001). Dietary CA increased maltase activity and reduced the number of Ki67-positive cells (p < .05). Increasing dEB values in diets without CA increased sucrose and lactase activities (quadratic; p < .05), but no such effect was observed in the diets with CA (dEB ×CA; p < .05). An interaction effect between dEB and CA on the number of Ki67-positive cells was observed (p < .001). In conclusion, 250 mEq/kg dEB diet with CA improved piglet intestinal digestion and absorption function by improving intestinal morphology and increasing digestive enzyme activities. However, these improvements were also observed in piglets fed the 100 mEq/kg dEB diet without CA.

Effects and interaction of dietary electrolyte balance and citric acid on the intestinal function of weaned piglets.

Fifty-six piglets (6.26 ± 0.64 kg BW) were weaned at 21 d and randomly assigned to one of the eight dietary treatments with seven replicate pens for a 14-d experimental period. The eight experimental diets were prepared via a 2 × 4 factorial arrangement with citric acid (CA; 0% and 0.3%) and dietary electrolyte balance (dEB, Na + K - Cl mEq/kg of the diet; -50, 100, 250, and 400 mEq/kg). Varying dEB values were obtained by altering the contents of calcium chloride and sodium bicarbonate. An interaction (P < 0.05) between dEB and CA in diarrhea score and the number of goblet cell in jejunum were observed. Ileum pH significantly decreased in weaned piglets fed 250 mEq/kg dEB diet compared with those fed -50 and 400 mEq/kg dEB diets (P < 0.05). Supplementation of 0.3% CA decreased the number of goblet cell in the ileal crypt (P < 0.05) and the relative mRNA expression of cystic fibrosis transmembrane conductance regulator, tumor necrosis factor-α, interferon-γ (IFN-γ), interleukin-1ß (IL-1ß), interleukin-10 (IL-10), zona occludens-1, and Claudin-1 (P < 0.05). Increasing dEB values increased the number of goblet cells in the jejunal crypt (P < 0.05). A 250-mEq/kg dEB diet decreased the relative mRNA expression of IFN-γ, IL-1ß, and IL-10 (P < 0.05) than 100-mEq/kg dEB diet. The interaction between dEB and CA on the relative abundances of Cyanobacteria and Saccharibacteria was observed (P < 0.05). Supplementation of 0.3% CA increased relative abundances of and Streptococcus hyointestinalis. Piglets fed 250-mEq/kg diet increased relative abundances of Firmicutes and Lactobacillus rennini, and decreased the relative abundance of Proteobacteria, Veillonella, Actinobacillus minor, and Escherichia-Shigella.In conclusion, supplementation of 0.3% CA resulted in differential expression of inflammatory cytokines, ion transporters, and tight junction proteins, and changes in the microbial community composition. A 250-mEq/kg dEB diet reduced gastrointestinal pH and promoted the enrichment of beneficial microbes in the gut microbiota, thereby suppressing inflammation and harmful bacteria. However, the addition of CA to diets with different dEB values did not promote intestinal function in weaned piglets.

Responses of Intestinal Microbiota and Immunity to Increasing Dietary Levels of Iron Using a Piglet Model.

Iron is an essential metal for both animals and microbiota. In general, neonates and infants of humans and animals are at the risk of iron insufficiency. However, excess dietary iron usually causes negative impacts on the host and microbiota. This study aimed to investigate overloaded dietary iron supplementation on growth performance, the distribution pattern of iron in the gut lumen and the host, intestinal microbiota, and intestine transcript profile of piglets. Sixty healthy weaning piglets were randomly assigned to six groups: fed on diets supplemented with ferrous sulfate monohydrate at the dose of 50 ppm (Fe50 group), 100 ppm (Fe100 group), 200 ppm (Fe200 group), 500 ppm (Fe500 group), and 800 ppm (Fe800), separately, for 3 weeks. The results indicated that increasing iron had no significant effects on growth performance, but increased diarrheal risk and iron deposition in intestinal digesta, tissues of intestine and liver, and serum. High iron also reduced serum iron-binding capacity, apolipoprotein, and immunoglobin A. The RNA-sequencing analysis revealed that iron changed colonic transcript profile, such as interferon gamma-signal transducer and activator of transcription two-based anti-infection gene network. Increasing iron also shifted colonic and cecal microbiota, such as reducing alpha diversity and the relative abundance of Clostridiales and Lactobacillus reuteri and increasing the relative abundance of Lactobacillus and Lactobacillus amylovorus. Collectively, this study demonstrated that high dietary iron increased diarrheal incidence, changed intestinal immune response-associated gene expression, and shifted gut microbiota. The results would enhance our knowledge of iron effects on the gut and microbiome in piglets and further contribute to understanding these aspects in humans.

Dietary Lysozyme Alters Sow's Gut Microbiota, Serum Immunity and Milk Metabolite Profile.

The aim of current study was to determine variations in sow's gut microbiota, serum immunity, and milk metabolite profile mediated by lysozyme supplementation. Twenty-four pregnant sows were assigned to a control group without supplementation and two treatments with 0.5 kg/t and 1.0 kg/t lysozyme provided in formula feed for 21 days (n = 8 per treatment). Microbiota analysis and metagenomic predictions were based on 16s RNA high-throughput sequencing. Milk metabolome was assessed by untargeted liquid chromatography tandem mass spectrometry. Serum biochemical indicators and immunoglobulins were also determined. Gut microbial diversity of sows receiving 1.0 kg/t lysozyme treatment was significantly reduced after the trial. Spirochaetes, Euryarchaeota, and Actinobacteria significantly increased while Firmicutes showed a remarkable reduction in 1.0 kg/t group compared with control. Lysozyme addition rebuilt sow's gut microbiota to beneficial composition identified by reduced richness of Escherichia coli and increased abundance of Lactobacillus amylovorus. Accordingly, microbial metabolic functions including pyrimidine metabolism, purine metabolism, and amino acid related enzymes were significantly up-regulated in 1.0 kg/t group. Microbial metabolic phenotypes like the richness of Gram-positive bacteria and oxidative stress tolerance were also significantly reduced by lysozyme treatment. Serum alanine transaminase (ALT) activity and IgA levels were significantly down-regulated in the 1.0 kg/t group compared with control, but IgM levels showed a significantly increase in 1.0 kg/t group. Milk metabolites such as L-glutamine, creatine, and L-arginine showed significantly dose-dependent changes after treatment. Overall, lysozyme supplementation could effectively improve the composition, metabolic functions, and phenotypes of sow's gut microbiota and it also benefit sows with better serum immunity and milk composition. This research could provide theoretical support for further application of lysozyme in promoting animal gut health and prevent pathogenic infections in livestock production.