The effects of 2’-fucosyllactose and Bifidobacterium longum subspecies infantis supplementation on growth, intestinal development and immune development and function in piglets

Daniels, Victoria Catherine

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https://hdl.handle.net/2142/120471 Copy

Description

Title

The effects of 2’-fucosyllactose and Bifidobacterium longum subspecies infantis supplementation on growth, intestinal development and immune development and function in piglets

Author(s)

Daniels, Victoria Catherine

Issue Date

2021-07-12

Director of Research (if dissertation) or Advisor (if thesis)

Donovan, Sharon M

Committee Member(s)

• Dilger, Ryan N
• Steelman, Andrew

Department of Study

Nutritional Sciences

Discipline

Nutritional Sciences

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

M.S.

Degree Level

Thesis

Keyword(s)

• 2’-FUCOSYLLACTOSE
• BIFIDOBACTERIUM LONGUM SUBSPECIES INFANTIS

Abstract

The newborn infant has a naïve immune system and immature gastrointestinal system. Breastfeeding is the optimal feeding mode to stimulate the development of these systems in the infant due to the bioactive nature of breastmilk. However, globally many infants rely on infant formula to meet their nutritional requirements. 2′-fucosyllactose (2′-FL) is a human milk oligosaccharide (HMO) abundant in human milk and has been shown to have effects on intestinal and immune development and function. Bifidobacterium longum subsp. Infantis (B. infantis) is a commensal bacterium found in the microbiota of the breast-fed infant gut that has been shown to impact the development of the microbiota and gut immunity. A commercially available strain of B infantis, Bi-26, can use 2′-FL as a carbon source; however, a potential synbiotic effect remains to be investigated. The goal of this thesis research was to examine the impact of 2′-FL and Bi-26 on intestinal and immune development and function in the piglet model. The overall hypothesis of this work was that 2′-FL and Bi-26 would demonstrate a symbiotic effect to stimulate intestinal development. In addition, it was hypothesized that 2′-FL and Bi-26 would promote healthy immune development and improved function, examined ex vivo. Specific aim 1 investigated the impact of 1 g/L 2′-FL and 10⁹ CFU Bi-26 supplementation in formula on growth, tolerance, crypt and villus development in the small intestine, and abundance of Bifidobacterium ssp. and Bi-26, in a piglet model. Intact (non-castrated) male piglets on postnatal day (PND) 2 were randomized to receive either the control formula (CON) or formula supplemented with 1.0 g/L 2′-FL (FL), then further randomized to receive CON with Bi-26 (BI) or with the 2′-FL formula (FLBI). Body weights and formula intake were monitored from PND 2 to 33/34 and animals were sacrificed on PND 34/35. Intestine, liver and brain weights were assessed, and intestinal samples were collected for morphological analyses and disaccharidase activity. Luminal samples from cecum, ascending colon (AC) and rectum (RC) were analyzed for dry matter. Volatile fatty acids (VFA) and abundance of Bifidobacterium ssp. and B infantis, analyzed by qPCR were measured in AC and RC contents. There was no experimental treatment effect on weights of the intestine, liver or brain, although brain weight tended (p=0.060) to be lower in CON piglets per kg/BW. Jejunal and ileal lactase and sucrase activities were similar between treatment groups. Histomorphology measures in jejunum, ileum and AC were similar across treatment groups, except for a trend for increased crypt width (p=0.071) and crypt volume (P=0.069) in the ileum of FL piglets. There was no effect of treatment of dry matter in the cecum, AC or RC, but dry matter was greater in RC than AC and cecum (p<0.0001). In the AC, acetate concentration was greater in FL than CON (p=0.046), and acetate tended to be greater in FL than CON in RC (p=0.0627). When taken as a percentage of total SCFA, propionate tended to be different in AC (p=0.080). In RC, propionate made up a larger proportion of SCFA in CON than BI (p=0.005) and FL (P=0.049). Absolute quantity of Bifidobacterium ssp. was similar across treatments in AC and RC. Occurrences of B. infantis were significantly higher in BI and FLBI piglets than CON and FL piglets in AC (p=0.0006) and RC (P<0.0001) contents. Specific aim 2 investigated the effect of and 1g/L 2′-FL and 10⁹ Bi-26 in formula on peripheral blood mononuclear cells (PBMC), mesenteric lymph node (MLN) immune cell population and cytokine production in response to ex vivo stimulation in addition to serum cytokines and immunoglobulins. At sacrifice, blood was taken for serum cytokine analysis and immune cells from blood and mesenteric lymph nodes were isolated for ex vivo stimulation and flow cytometry analysis. Luminal contents from the AC and RC were taken for IgA quantification. PBMC and MLN immune cell populations were unaffected by experimental treatment. sIgA levels tended (p=0.073) to be greater in the RC than AC; but there was no treatment effect. Serum IL-1, IL-1RA, IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18 were higher in FL compared to CON. Serum IgG and IgM were similar across treatments. When unstimulated versus LPS stimulation were compared between treatment groups in PBMC, IFN-γ was higher in FL than CON, and IL-1RA and IL-6 were higher in FLBI than CON. When unstimulated versus LPS stimulation were compared between treatment groups in MLN, IFN-γ was higher in FLBI than CON. The addition of 2′-FL and Bi-26 in formula were well tolerated in the piglet and had no negative impact on growth, intestinal or immune development. 2′-FL showed prebiotic potential in the production of short chain fatty acids in the colon. However, there was little impact on small intestinal development. 2′-FL also showed a potential prebiotic effect on serum cytokines and ex vivo stimulation response. This prebiotic effect was modulated by Bi-26 when PBMC and MLN were stimulated with LPS. This data demonstrates 2′-FL and Bi-26 may have a synbiotic effect on the immune system during immune challenge. Further investigation may focus on the effects of 2′-FL and Bi-26 on response to immune challenge in vivo.

Graduation Semester

2021-08

Type of Resource

Thesis

Copyright and License Information

© 2021 Victoria Catherine Daniels

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