Betaine hydrochloride addition in Bama mini-pig's diets during gestation and lactation enhances immunity and alters intestine microbiota of suckling piglets.

BACKGROUND: Maternal nutrition during gestation and lactation is essential for offspring's health. The present study aimed to investigate the effects of betaine hydrochloride addition to sow diets during gestation and lactation on suckling piglet's immunity and intestine microbiota composition. Forty Bama mini-pigs were randomly allocated into two groups and fed a basal diet (control group) and a basal diet supplemented with 3.50 kg ton-1 betaine hydrochloride (betaine group) from day 3 after mating to day 21 of lactation. After 21 days of the delivery, 12 suckling piglets from each group with similar body weight were selected for sample collection. RESULTS: The results showed that maternal betaine hydrochloride addition decreased (P < 0.05) the plasma levels of interleukin (IL)-1ß, IL-2, IL-6, and tumor necrosis factor-α in suckling piglets. Furthermore, dietary betaine hydrochloride addition in sow diets increased (P < 0.05) the villus height (VH) and VH to crypt depth ratio in the jejunum and ileum of suckling piglets. In the piglets' intestinal microbiota community, the relative abundances of Roseburia (P < 0.05) and Clostridium (P = 0.059) were lower in the betaine group compared to those in the control group. Moreover, betaine hydrochloride addition in sow diets decreased the colonic tyramine (P = 0.091) and skatole (P = 0.070) concentrations in suckling piglets. CONCLUSION: Betaine hydrochloride addition in sow diets enhanced the intestinal morphology, improved immunity, and altered intestinal microbiota of suckling piglets. These findings indicated that betaine hydrochloride addition in sow diets during gestation and lactation will impact suckling piglets' health. © 2021 Society of Chemical Industry.

Minipigs as a neonatal animal model for tuberculosis vaccine efficacy testing.

Many vaccines against childhood diseases are administered early after birth, but vaccine development studies frequently test efficacy in adult rather than in neonatal animal models. In countries with endemic tuberculosis (TB), Bacillus Calmette-Guerin (BCG) is administered as part of the neonatal vaccine regimen because it prevents against the disseminated form of TB in children, although it has variable efficacy against pulmonary TB. Several promising new vaccines against TB are currently being tested in adult animal models. Here we evaluated neonatal piglets as an animal model to test vaccine efficacy. For this purpose, minipigs were vaccinated or not with BCG 48 h after birth and their immune response followed longitudinally until adolescence. We characterized the memory and activation phenotype of T cells, cytokine profile, and monocyte activation in response to BCG stimulation from 4 weeks of age into adolescence- age of 24 weeks. Immunological responses in vaccinated and non-vaccinated animals were further monitored upon infection with a low dose exposure to Mycobacterium tuberculosis strain HN878 via the aerosol route. Comparing the immunological response elicited by BCG vaccination in minipigs vs similar studies in infants, suggest that minipigs have the potential to serve as an effective neonatal animal model for vaccine development.

Nonclinical evaluation of immunological safety in Göttingen Minipigs: The CONFIRM initiative.

There is a growing need to consider non-rodent species for the immunological safety evaluation of drug candidates. The EU Framework-6 RETHINK Project demonstrated that the Göttingen Minipig is a relevant animal model for regulatory toxicology studies. Extensive knowledge on the immune system of domestic pigs is available and fewer differences from humans have been identified as compared to other species, such as mice or non-human primates. Minipig data are too scarce to allow for claiming full immunological comparability with domestic pigs. Another gap limiting minipig use for immunological safety evaluation is the lack of a qualified and validated database. However, available data lend support to the use of minipigs. The need for a COllaborative Network For Immunological safety Research in Minipigs (the CONFIRM Initiative) was obvious. It is intended to trigger immunological safety research in Göttingen Minipigs, to assist and synergize fundamental, translational and regulatory investigative efforts relevant to the immunological safety evaluation of pharmaceuticals and biologics, and to spread current knowledge and new findings to the scientific and regulatory toxicology community.

Reduced expression of SLA Class 1 antigens by intestinal epithelium of newborn piglets.

Expression of major histocompatibility complex Class 1 by the small intestine mucosa of piglets was compared by indirect immunofluorescence during the 4 days following birth with both maternal and artificial feeding. The duodenal epithelium did not express Class I antigen during these 4 days. The jejunal epithelium did not express Class I antigen at Day 0 after birth but expression developed from Day 1. However ileal epithelium expressed Class 1 antigen throughout the study period. In contrast, cells from the lamina propria of all samples expressed Class 1 antigen. There was no difference between piglets receiving maternal colostrum and artificially reared piglets. This lack of Class 1 expression occurs at a time when the intestinal epithelium constitutes an interface between piglet and colostral maternal cells. This can be of biological relevance for mother-newborn interactions.

Intestinal absorption of immunologically intact macromolecules in germfree colostrum-deprived piglets maintained on total parenteral nutrition.

We have compared the neonatal absorption of anti-bovine gamma-globulin (BGG) antibody supplied in colostrum or saline in three groups of piglets born and maintained under different environmental conditions to determine the effect of these conditions on the cessation of intestinal absorption of macromolecules (anti-BGG antibody), termed "closure." An enzyme-linked immunosorbent assay was used to estimate the concentration of anti-BGG antibody in sera from each group of piglets. Three stages of macromolecular absorption through the piglet's intestine could be detected. The first stage is a nonselective massive absorption of macromolecules (in milligram levels) that lasts up to 3 days in germfree (GF) colostrum-deprived or conventional colostrum-fed piglets but up to 5 days in GF piglets maintained on total parenteral nutrition. In this stage, absorption was significantly (r = .05) higher in piglets fed anti-BGG serum with colostrum than in piglets fed anti-BGG serum without colostrum on GF day 0 (31.28% vs 15.59%) and GF-total parenteral nutrition day 3 (3.08% vs 0.11%). Thus, whenever there was the ability to absorb a massive amount of macromolecules, the sow colostrum had an enhancing affect. Although there was a minor effect of environmental or orally received stimuli in delaying closure, absorption of macromolecules decreased in all piglets maintained either parenterally or enterally after day 3. Thus, intestinal closure to massive absorption of macromolecules in piglets is primarily time (age)-dependent. The second stage is a selective absorption of immunoglobulins in much smaller quantities (microgram levels), inasmuch as absorption of 0.02% to 0.1% was determined in all 5-day-old piglets.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunomodulating effects of intestinal absorbed maternal colostral leukocytes by neonatal pigs.

Intestinal absorption of fluorescein isothiocyanate-labelled maternal colostral leukocytes (FITC-CL) was studied in 49 neonatal colostrum-deprived (CD) pigs from nine Minnesota miniature sows. Within 2 h postfeeding (pf), maternal FITC-CL were absorbed from the sibling's digestive tract and migrated into blood. The peak appearance of FITC-CL in blood occurred in samples at 5 and 7 h pf. By 24 h pf, cells were detected in liver, lung, lymph nodes, spleen and gastrointestinal tissues. To confirm intercellular migration of FITC-CL, gastrointestinal explant cultures from neonatal CD pigs were used. Maternal FITC-CL were observed to intercellularly migrate in 24 to 48 h pf between duodenal- and jejunal-epithelial cells to lamina propria cells and submucosal spaces. Fluorescein isothiocyanate-labelled maternal colostral leukocytes were not absorbed via ileal explant cultures. Unlike FITC-CL, maternal FITC-peripheral blood mononuclear leukocytes (FITC-PBL) were not absorbed either in vivo or in vitro by gastrointestinal tissues. When maternal FITC-PBL were intravenously administered to siblings they were distributed in blood and organs similar to FITC-CL. Following exposure to FITC-labelled cells, treated- and mock (untreated)-pigs were compared on the basis of PBL proliferative responses to phytomitogens. Sibling CD-pigs fed maternal FITC-CL showed higher PBL T-cell responses to phytohemagglutinin (PHA) and concanavalin A (ConA), and a significant stimulation (p < or = 0.01) of B-cell responses to pokeweed mitogen (PWM). Pigs fed FITC-PBL showed little PBL responses to PHA, ConA and PWM over PBL from mock pigs. Similarly, the influence of noncellular constituents of colostrum were also assessed by proliferative studies.(ABSTRACT TRUNCATED AT 250 WORDS)

[Cellular immunity and enzyme activity of xenobiotic metabolism in the lymphocytes of normal minipigs and in alcohol and anabol exposures]. / Kletochnyi immunitet i aktivnost' fermentov metabolizma ksenobiotikov v limfotsitakh u mini-svinei v norme i pri deistvii alkogolia i anabola.

Immunological indices and activity of xenobiotic metabolism enzymes in lymphocytes were studied on minipigs under normal conditions, under conditions of chronic alcoholic intoxication and after administration of anabol (an immunomodulator) to normal healthy animals and to animals with alcohol intoxication. Age-related differences with respect to the number of T-lymphocytes and activity of lymphocyte glutathione S-transferase were observed in the normal animals, the other indices such as activity of natural killer cells, K-cells, blast cell transformation with concanavalin A and activity of cytochrome c-reductase being independent of the age. Administration of anabol to healthy animals did not alter their immunoenzymatic status. Chronic alcohol intoxication was accompanied by development of secondary immune deficiency characterized by lower immunological indices and lower activity of xenobiotic metabolism enzymes in lymphocytes. Daily exposure to 0.8 g of anabol for 12 days at this background resulted in normalization of the above indices.

Delayed-type skin hypersensitivity and in vitro lymphocyte immunostimulation responses of swine following inoculation with Mycobacterium avium cell walls and a mycobacterial immunopotentiating glycolipid.

Miniature swine (n = 5 per group) were inoculated intradermally with mineral oil-in-water emulsions containing either 150 micrograms of mycobacterial immunopotentiating glycolipid P3 (EP3), 150 micrograms of lyophilized Mycobacterium avium (serotype 8) cell walls (E-MaCW), or 150 micrograms P3 and 150 micrograms M. avium cell walls (EP3-MaCW). Swine vaccinated with E-MaCW and EP3-MaCW developed antigen-sensitive lymphocytes detectable with delayed-type hypersensitivity (DTH) skin tests and lymphocyte transformation assays. Swine injected with EP3 were not sensitized. In general EP3-MaCW evoked a more pronounced in vivo DTH tuberculin skin test and in vitro lymphocyte transformation responses than E-MaCW. Time-course studies indicated a more persistent response in swine injected with EP3-MaCW than in those given E-MaCW. Commercial type Yorkshire swine (n = 5) inoculated intradermally with EP3-MaCW developed cell-mediated immune (CMI) responses to avian tuberculin detectable in vivo with delayed-type skin hypersensitivity and in vitro with lymphocyte immunostimulation responses.