Development In Neonatal Piglets Research Articles

An insight into the temporal dynamics in the gut microbiome, metabolite signaling, immune response, and barrier function in suckling and weaned piglets under production conditions.

Little information is available on age- and creep-feeding-related microbial and immune development in neonatal piglets. Therefore, we explored age- and gut-site-specific alterations in the microbiome, metabolites, histo-morphology, and expression of genes for microbial signaling, as well as immune and barrier function in suckling and newly weaned piglets that were receiving sow milk only or were additionally offered creep feed from day of life (DoL) 10. The experiment was conducted in two replicate batches. Creep feed intake was estimated at the litter level. Piglets were weaned on day 28 of life. Gastric and cecal digesta and jejunal and cecal tissue were collected on DoL 7, 14, 21, 28, 31, and 35 for microbial and metabolite composition, histomorphology, and gene expression. In total, results for 10 piglets (n = 5/sex) per dietary group (sow milk only versus additional creep feed) were obtained for each DoL. The creep feed intake was low at the beginning and only increased in the fourth week of life. Piglets that were fed creep feed had less lactate and acetate in gastric digesta on DoL 28 compared to piglets fed sow milk only (p < 0.05). Age mainly influenced the gastric and cecal bacteriome and cecal mycobiome composition during the suckling phase, whereas the effect of creep feeding was small. Weaning largely altered the microbial communities. For instance, it reduced gastric Lactobacillaceae and cecal Bacteroidaceae abundances and lowered lactate and short-chain fatty acid concentrations on DoL 31 (p < 0.05). Jejunal and cecal expression of genes related to microbial and metabolite signaling, and innate immunity showed age-related patterns that were highest on DoL 7 and declined until DoL 35 (p < 0.05). Weaning impaired barrier function and enhanced antimicrobial secretion by lowering the expression of tight junction proteins and stimulating goblet cell recruitment in the jejunum and cecum (p < 0.05). Results indicated that age-dependent alterations, programmed genetically and by the continuously changing gut microbiome, had a strong impact on the expression of genes for gut barrier function, integrity, innate immunity, and SCFA signaling, whereas creep feeding had little influence on the microbial and host response dynamics at the investigated gut sites.

Influence of high- and low-fermentable dietary fibres in sows' diet on the colostrum potential against Clostridioides difficile toxin-induced effects in IPEC-J2 cells.

Sow colostrum has been reported to protect the IPEC-J2 cells and piglet colon tissues from detrimental effect of Clostridioides difficile toxins. Since dietary fibre can influence the colostrum composition in sows, we hypothesised that it can also differentially affect the colostrum potential against C. difficile toxin-induced effects in IPEC-J2. IPEC-J2 were incubated with colostrum from sows fed either high-fermentable sugar beet pulp (SBP) or low-fermentable lignocellulose (LNC) fibres and in combination with the toxins and analysed by trans-epithelial electrical resistance (TEER) and cell viability using propidium iodide in flow cytometry. Toxins drastically decreased the integrity of IPEC-J2. Colostrum from the sows fed either SBP or LNC exerted protective effect against toxins on IPEC-J2 integrity and this effect was numerically superior in the SBP group. Differences in the percentages of TEER between different treatments were noted after 2 h (p = 0.043), 3 h (p = 0.017) and 4 h (p = 0.017) of incubation and a tendency for differences was noted after 5 h of incubation (p = 0.071). Colostrum from either SBP- or LNC-fed sows did not protect the IPEC-J2 from toxin-induced death. Colostrum of the sows fed either high-fermentable or low-fermentable fibres has a potential to protect IPEC-J2 from the loss of integrity, which may be important in protection from C. difficile-infection development in neonatal piglets.

Immunocyte Populations Observed from Birth to Weaning in Blood, Spleen and Mesenteric Lymph Nodes of Piglets.

Simple SummaryUnderstanding the immunocyte development in neonatal piglets is crucial because susceptibility to pathogen infections and efficacy of vaccination highly depend on the immune status of the piglet. However, the immunocyte information on the period between birth and weaning remains scarce, especially that on immune tissues other than blood, such as mesenteric lymph nodes (MLN) and spleens. Our results indicated that the percentage of innate immunity cells such as myeloid cells, most of them possibly being monocytes, declined from the first day after birth in blood, MLN and spleens. By contrast, other innate immunocytes such as natural killer cells and adaptive immunocytes such as T and B cells were produced while the piglets still suckled in the three organs. Weaning did not seemingly affect the dynamics of the studied immunocytes in a drastic manner, at least during the 14 days post weaning. We foresee that the information from the present study warrants the design of better programs for the administration of vaccines and/or innate immunity stimulators to suckling and weaned piglets.Susceptibility to pathogen infections and efficacy of vaccination highly depend on the immune status of the piglet. Here, we measured immunocytes in piglets from birth to weaning to elucidate how immunocyte populations change during development and are affected by weaning. Crossbred piglets were used. Suckling piglets were euthanized at 1, 7, 14, 21, 28 or 35 days old (3~4 piglets at each time point). In addition, seven piglets were weaned at 21 days old, with four being euthanized at 28 days old and the remaining at 35 days old. Piglet carcasses were dissected, and blood, mesenteric lymph nodes (MLN) and spleen were sampled. In total, seven antibodies were used to stain the immunocyte population. Dynamics of myeloid (CD3–SWC3+CD16+), natural killer (NK; CD3–SWC3–CD16+), killer T (CD3+CD8+), helper T (CD3+CD4+) and B (CD3–CD21+) cells were analyzed. Percentage of innate immunity cells such as myeloid cells declined (p < 0.05) from the first day after birth. In contrast, percentage of NK cells increased in piglets while they were still suckling. Killer T, helper T, and B cell populations increased around 2~3 weeks after birth. No significant differences in the populations of the evaluated cell types were observed between suckling and weaned piglets at least for 14 days post weaning.

The effect of maternal antibiotic use in sows on intestinal development in offspring.

The objective of this study is to investigate the effect of a maternal antibiotic administration during the last week of gestation on the early life intestinal development in neonatal piglets. Colonization of the gut with bacteria starts during birth and plays a major role in the intestinal and immunological development of the intestine. We demonstrate that maternal interventions induced changes in the sows (n = 6 to 8 per treatment) fecal microbiota diversity around birth (P < 0.001, day 1). Whole-genome microarray analysis in small intestinal samples of 1-d old piglets (n = 6 to 8 per treatment) showed significantly expressed genes (Padj < 0.05) which were involved in processes of tight junction formation and immunoglobulin production. Furthermore, when performing morphometry analysis, the number of goblet cells in jejunum was significantly (P < 0.001) lower in piglets from amoxicillin administered sows compared with the respective control piglets. Both significantly expressed genes (Padj < 0.05) and significant morphometry data (jejunum P < 0.05 and ileum P < 0.01) indicate that the crypts of piglets from amoxicillin administered sows deepen around weaning (day 26) as an effect of the amoxicillin administration in sows. The latter might imply that the intestinal development of piglets was delayed by maternal antibiotic administration. Taken together, these results show that maternally oral antibiotic administration changes in early life can affect intestinal development of the offspring piglets for a period of at least 5 wk after the maternal antibiotic administration was finished. These results show that modulation of the neonatal intestine is possible by maternal interventions.

Early Supplementation of Phospholipids and Gangliosides Affects Brain and Cognitive Development in Neonatal Piglets

As human breast milk is a rich source of phospholipids (PHOS) and gangliosides (GLS), and breastfed infants have improved learning compared to formula fed, the importance of dietary PHOS and GLS for brain development is of interest. Herein, we sought to determine the effects of these lipids on brain and cognitive development. Male and female piglets were artificially reared and fed formula containing 0% (control), 0.8%, or 2.5% LACPRODAN® (PL‐20, Arla Foods Ingredients, DK), a PHOS/GLS supplement, from postnatal day (PD) 2 to PD28. Beginning PD14, performance in a spatial T‐maze task was assessed. At PD28, brain MRI data were acquired and piglet hippocampal tissue was obtained for metabolic profiling. Diet affected maze performance with piglets receiving 0.8% and 2.5% PL‐20 making fewer errors than 0% (80% vs 75% correct on average) (P&lt;0.05) and taking less time to make a choice (3 seconds vs 5 seconds per trial) (P&lt;0.01). Mean brain weight was 5% higher for piglets receiving 0.8% and 2.5% PL‐20 (P&lt;0.05) compared to 0%; and voxel‐based morphometry revealed multiple brain areas with greater volumes and more gray and white matter in piglets receiving 0.8% and 2.5% PL‐20 compared to 0%. Metabolic profiling of hippocampal tissue revealed that multiple phosphatidylcholine metabolites were altered by diet. In summary, dietary PHOS and GLS improved spatial learning and affected brain growth and composition. (Supported by Arla Foods Ingredients and R01 HD069899).

Early Supplementation of Phospholipids and Gangliosides Affects Brain and Cognitive Development in Neonatal Piglets

Background: Because human breast milk is a rich source of phospholipids and gangliosides and breastfed infants have improved learning compared with formula-fed infants, the importance of dietary phospholipids and gangliosides for brain development is of interest.Objective: We sought to determine the effects of phospholipids and gangliosides on brain and cognitive development.Methods: Male and female piglets from multiple litters were artificially reared and fed formula containing 0% (control), 0.8%, or 2.5% Lacprodan PL-20 (PL-20; Arla Foods Ingredients), a phospholipid/ganglioside supplement, from postnatal day (PD) 2 to PD28. Beginning on PD14, performance in a spatial T-maze task was assessed. At PD28, brain MRI data were acquired and piglets were killed to obtain hippocampal tissue for metabolic profiling.Results: Diet affected maze performance, with piglets that were fed 0.8% and 2.5% PL-20 making fewer errors than control piglets (80% vs. 75% correct on average; P < 0.05) and taking less time to make a choice (3 vs. 5 s/trial; P < 0.01). Mean brain weight was 5% higher for piglets fed 0.8% and 2.5% PL-20 (P < 0.05) than control piglets, and voxel-based morphometry revealed multiple brain areas with greater volumes and more gray and white matter in piglets fed 0.8% and 2.5% PL-20 than in control piglets. Metabolic profiling of hippocampal tissue revealed that multiple phosphatidylcholine-related metabolites were altered by diet.Conclusion: In summary, dietary phospholipids and gangliosides improved spatial learning and affected brain growth and composition in neonatal piglets.

Oral Administration of Lactobacillus fermentum I5007 Favors Intestinal Development and Alters the Intestinal Microbiota in Formula-Fed Piglets

The present study was conducted to evaluate the effects of early administration of Lactobacillus fermentum I5007 on intestinal development and microbial composition in the gastrointestinal tract using a neonatal piglet model. Full-term 4 day old piglets, fed with milk replacer, were divided into a control group (given placebo of 0.1% peptone water) and a L. fermentum I5007 group (dosed daily with 6 × 10(9) CFU/mL L. fermentum I5007). The experiment lasted 14 days. On day 14, a significant increase in the jejunum villous height (583 ± 33 vs 526 ± 18) and increases in the concentrations of butyrate (7.55 ± 0.55 vs 5.33 ± 0.39) and branched chain fatty acids in the colonic digesta were observed in piglets in the L. fermentum I5007 treatment (P < 0.05). mRNA expression of IL-1β (1.29 ± 0.29 vs. 0.62 ± 0.07) in the ileum were lower after 14 days of treatment with L. fermentum I5007. Denaturing gradient gel electrophoresis (DGGE) revealed that L. fermentum I5007 affected the colonic microbial communities on day 14 and, in particular, reduced numbers of Clostridium sp. L. fermentum I5007 play a positive role in gut development in neonatal piglets by modulating microbial composition, intestinal development, and immune status. L. fermentum I5007 may be useful as a probiotic for application in neonatal piglets.

Effects of Dietary Supplementation with Combined Arginine and Glutamine on Growth Performance and Small Intestinal Development in Neonatal Piglets

Effects of Dietary Supplementation with Combined Arginine and Glutamine on Growth Performance and Small Intestinal Development in Neonatal Piglets

Effect of dietary yeast β‐glucan on immune development in neonatal piglets

Purified yeast (1,3/1,6)‐β‐D‐glucan (βG) is a biological response modifier meaning that it does not have direct cytotoxic activities, but is able to boost host defense mechanisms. Herein, the hypothesis that dietary βG would modulate the intestinal and systemic immune development was tested. Piglets (n=5–6/group) were fed formula containing 0 (control), 1.8, 18 or 90 mg βG/kg body weight. Piglets were euthanized at 7 and 21 days of age. Ileal Peyer's Patches (IPP), spleen, mesenteric lymph nodes (MLN) and ascending colon (ASC) were collected for analysis. Piglet weight, formula intake and small intestinal length and weight were unaffected by dietary βG. In addition, dectin (a receptor for βG), IL‐10, IL‐12 and TNF‐α mRNA expression in IPP, spleen, MLN and ASC were unaffected by age or dietary βG. T‐cell phenotypes (CD45+CD3+CD4+ and CD45+CD3+CD8+) were examined in IPP, spleen, MLN, and peripheral blood mononuclear cells (PBMC) by flow cytometry. No differences in IPP or MLN T‐cells were observed. Spleen and PBMC T‐cells showed an interaction between age and dietary βG. Thus, dietary βG had little effect on intestinal mucosal immune development in neonatal piglets. (Supported by Abbott Nutrition)

Rearing environment affects development of the immune system in neonates

Early-life exposure to appropriate microbial flora drives expansion and development of an efficient immune system. Aberrant development results in increased likelihood of allergic disease or increased susceptibility to infection. Thus, factors affecting microbial colonization may also affect the direction of immune responses in later life. There is a need for a manipulable animal model of environmental influences on the development of microbiota and the immune system during early life. We assessed the effects of rearing under low- (farm, sow) and high-hygiene (isolator, milk formula) conditions on intestinal microbiota and immune development in neonatal piglets, because they can be removed from the mother in the first 24 h for rearing under controlled conditions and, due to placental structure, neither antibody nor antigen is transferred in utero. Microbiota in both groups was similar between 2 and 5 days. However, by 12-28 days, piglets reared on the mother had more diverse flora than siblings reared in isolators. Dendritic cells accumulated in the intestinal mucosa in both groups, but more rapidly in isolator piglets. Importantly, the minority of 2-5-day-old farm piglets whose microbiota resembled that of an older (12-28-day-old) pig also accumulated dendritic cells earlier than the other farm-reared piglets. Consistent with dendritic cell control of T cell function, the effects on T cells occurred at later time-points, and mucosal T cells from high-hygiene, isolator pigs made less interleukin (IL)-4 while systemic T cells made more IL-2. Neonatal piglets may be a valuable model for studies of the effects of interaction between microbiota and immune development on allergy.

Temporal Proteomic Analysis Reveals Continuous Impairment of Intestinal Development in Neonatal Piglets with Intrauterine Growth Restriction

Efficiency of nutrient utilization is reduced in neonates with intrauterine growth restriction (IUGR) compared with those with a normal birth weight (NBW). However, the underlying mechanisms are largely unknown. In this study, we applied temporal proteomic approach, coupled with histological and biochemical analyses, to study dynamic changes of the proteome in the small intestinal mucosa of IUGR piglets during the nursing period (Days 1, 7 and 21). We identified 56 differentially expressed protein spots between IUGR and NBW piglets. These proteins participate in key biological processes, including (1) absorption, digestion and transport of nutrients; (2) cell structure and motility; (3) glucose and energy metabolism; (4) lipid metabolism; (5) amino acid metabolism; (6) mineral and vitamin metabolism; (7) cellular redox homeostasis; (8) stress response; and (9) apoptosis. The results of our temporal proteomics analysis reveal continuous impairment of intestinal development in neonatal piglets with IUGR. The findings have important implications for understanding metabolic defects in the small intestine of IUGR neonates and are expected to provide new strategies to improve their survival and growth.

Soy‐based Formula Does Not Adversely Affect Immune System Development in Neonatal Piglets

Breast‐fed infants are less susceptible to infections and allergic reactions than those fed formula, suggesting that diet may influence neonatal immune system development. The goal of the present study was two fold: 1. to determine the effect of formula feeding relative to breast‐feeding on neonatal immunity; and 2) to determine whether soy‐based formula has any adverse effects on neonatal immune system development. Piglets were breast‐fed by the sow or weaned at 2 days to a milk‐ or soy‐based formula. At PND 21, splenocytes from the sow‐fed group showed 3 to 4 fold higher T lymphocyte proliferation compared to the formula‐fed groups. The increase in T‐lymphocyte proliferation was also reflected in circulating levels of IgG and IgM in sow‐reared piglets. Notably, both formula diets showed greater NK cell cytotoxicity than the sow‐fed piglets. Our findings suggest that the neonatal innate immune system in sow‐reared piglets is functionally better developed than that in formula‐fed piglets. Comparison between soy‐ and milk‐based formulas did not show any differences in NK cell cytotoxicity, total circulating antibody levels or lymphocyte proliferation, suggesting that soy formula does not have adverse effects on neonatal immune system development. Supported by ARS CRIS#6251‐51000‐005‐03S and in part by INBRE program funded by NIH.