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Investigation of the nutritional and functional roles of a combinational use of xylanase and β-glucanase on intestinal health and growth performance of nursery pigs

Hyunjun Choi*, Yesid Garavito Duarte*, Guilherme Pasquali†,

and Sung Woo Kim*

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*North Carolina State University, Raleigh, NC

†BASF SE, Ludwigshafen, Germany

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DEPARTMENT OF ANIMAL SCIENCE

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Non-starch polysaccharide (NSP) in plant feedstuffs for pigs

  • The use of plant feedstuffs, such as corn distiller’s dried grains with solubles (DDGS) and soybean meal, has been increased
    • High availability and economic advantages
  • A notable issue arises with the increased use of plant feedstuffs
    • Non-starch polysaccharides (NSP) especially soluble NSP
      • Negative impacts on intestinal health and growth of nursery pigs

(Stein and Shurson, 2009; Chen et al., 2020; Baker et al., 2024)

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Soluble NSP effects on intestinal health of nursery pigs

  • The NSP, once ingested, increase the viscosity of jejunal digesta
    • Mainly caused by the soluble fraction of NSP inhibiting endogenous enzymes from accessing feed particles in the small intestine
  • Increased digesta viscosity
    • Reduce nutrient utilization and growth (Hung et al., 2022)
    • Increase the risk of pathogenic bacterial overgrowth (Kim et al., 2012)
      • Viscous digesta slows down the passage rate and accelerates the proliferation of ammonia-producing bacteria in the luminal contents (Mcdonald et al., 2001) and mucosa (Mcdonald et al., 1999; Moita et al., 2022)
      • E. coli attachment in ileum was increased (Hopwood et al., 2004)

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Arabinoxylan and β-glucan in plant feedstuffs

  • Arabinoxylan (AX)
    • Included at 40 to 50% in corn and corn byproducts
    • Soluble AX is a high portion of soluble NSP in corn and corn byproducts
  • β-glucans
    • Constitute soluble NSP fraction, influencing digesta viscosity

(Knudsen et al., 1999 and 2014; Jaworski et al., 2015; Rodehutscord et al., 2016)

Item

Total

NSP, %

Total

AX, %

Total AX

to NSP

Soluble NSP, %

Soluble

AX, %

Soluble AX to sNSP

β-glucan,

%

Corn

9.7

4.6

47.0

1.1

0.6

56.0

0.1

DDGS

29.7

14.1

47.4

3.8

3.2

84.6

0.6

SBM

20.9

4.4

21.1

4.5

1.1

24.7

0.3

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Soluble arabinoxylan and β-glucan effects on small intestine

  • Soluble arabinoxylan
    • Increased viscosity of digesta (Chen et al., 2020; Lee et al., 2023)
    • Exhibited a strong correlation with viscosity in high-molecular weight fractions (Le Gall et al., 2010)
  • β-glucans
    • Cause viscous digesta, thereby increasing population of pathogenic bacteria such as E. coli (Li et al., 2006; Choi and Kim, 2023)
    • Primarily reduce the ileal digestibility of nutrients, decreased butyrate contents (Metzler-Zebeli and Zebeli, 2013)

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Functional role of NSPase on small intestine of pigs

  • Xylanase
    • Increase hydrolysis of arabinoxylans (Laerke et al., 2015), reducing digesta viscosity (Duarte et al., 2019; Moita et al., 2022)
    • Release xylo-oligosaccharides (XOS) in the small intestine (Laerke et al., 2015; Baker et al., 2021)
  • β-glucanase
    • Hydrolyze β-glucans by cleaving β-glycosidic linkages into gluco-oligosaccharides (GOS) in the small intestine (Yan et al., 2018)
    • Increased ileal digestibility of β-glucans, reducing digesta viscosity (Jensen et al., 1998)

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Importance of the jejunum in nursery pigs

  • Jejunum
    • Major portion of the small intestine to digest and absorb nutrients
    • Contains more immune cells than large intestine (Wiarda et al., 2020)
    • Nursery pigs with immature intestines encounter significant challenges from dietary compounds that activate intestinal immune responses and cause oxidative damages to jejunal mucosa (Kim and Duarte, 2021)
    • Microbiota produces bioactive compounds, altering jejunal immune system, barrier function, and cell proliferation (Duarte and Kim, 2022)

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Jejunal mucosa-associated microbiota

  • Has direct correlation with the intestinal health of host animals than the luminal microbiota in jejunum
  • Is distinctly different from luminal microbiota
  • Is the important parameter to investigate the intestinal health and growth of pigs

(Duarte and Kim, 2022; Adhikari et al., 2019)

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Central hypothesis

  • NSP, especially soluble NSP, increasing jejunal digesta viscosity and risk of pathogenic bacterial overgrowth would:
    • Alter jejunal mucosal microbiota
    • Increase microbial sensing by enterocytes
    • Increase inflammatory cytokine response in the jejunum
    • Increase humoral immune response in the jejunum
    • Increase oxidative damage products in the jejunum
    • Increase jejunal mucosa damage and jejunal tissue repair
    • Reduced nutrient utilization and growth performance of nursery pigs

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(Kim and Duarte, 2021; Duarte and Kim, 2022)

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Central hypothesis

In the small intestine

Microbiota

Microbial sensing

Immune response

Oxidative damages

Tissue damages

Tissue repairing

Digestion & absorption

Growth

NSP challenge

Dietary intervention (NSPase)

NSP to XOS & GOS

Reduced viscosity

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Xylanase effects on intestinal health (jejunum)

  • Xylanase supplementation
    • Positively modulated jejunal mucosa-associated microbiota
      • Chao 1 index was increased (Baker et al., 2024)
      • Prevotella was increased, whereas Helicobacter and Lactobacillus were decreased (Baker et al., 2024)
      • Succinivibrio and Pseudomonas were linearly increased (Moita et al., 2022)
    • Reduced oxidative damage products in the jejunum (Duarte et al., 2019)
    • Improved intestinal morphology and tissue repair in the jejunum
    • Improved nutrient utilization and growth of nursery pigs

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(Tiwari et al., 2018; Duarte et al., 2019; Boontiam et al., 2022; Moita et al., 2022)

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β-glucanase effects on intestinal health (jejunum)

  • β-glucanase supplementation
    • Positively modulated jejunal mucosa-associated microbiota (Duarte et al., 2021)
      • Lachnospiraceae and Ruminococcaeceae were linearly increased
      • Helicobacter_rappini and Streptococcus_alactolyticus were linearly decreased
    • Decreased enterotoxigenic E. coli in the ileum (Hopwood et al., 2004)
    • Improved intestinal morphology in the jejunum (Duarte et al., 2021)
    • Improved growth of nursery pigs (Toress-Pitarch et al., 2017; Duarte et al., 2021)

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Synergistic effects of feed enzymes on hydrolysis of NSP

  • Considering the complex nature of NSP fractions and structures, the combinational use of feed enzymes targeting NSP has shown synergistic effects on
    • Hydrolysis of NSP related to digesta viscosity
    • Release of nutrients entrapped by NSP
  • These feed enzymes’ effects on the reduction of digesta viscosity and increase in oligosaccharides would positively impact jejunal mucosa-associated microbiota, intestinal health, and growth of pigs

(Kiarie et al., 2013; Tsai et al., 2017; Baker et al., 2021; Duarte and Kim, 2022)

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Hypothesis

  • It is hypothesized that a combinational use of xylanase and β-glucanase in pig feeds would have positive impacts on the composition of mucosa-associated microbiota and intestinal health in the jejunum, with reduced digesta viscosity, improved ileal digestibility of nutrients, and consequently improved growth of nursery pigs.

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Objective

  • To evaluate the intestinal health and growth performance of nursery pigs affected by dietary supplementation of increasing levels of combinational use of xylanase and β-glucanase
  • To estimate the optimal dose of combinational use of xylanase and β-glucanase in feeds for nursery pigs

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Materials and methods: animals, design, and diets

  • A total of 40 pigs (20 barrows and 20 gilts; 6.5 ± 0.4 kg BW)
    • Individually housed in a pen (n = 8/trt)
  • RCBD (initial BW and sex as blocks)
  • Basal diet: 750 FTU/kg of phytase
    • Ca: 0.17% and STTD P: 0.10% unit below nutrient requirement by NRC (2012)
  • Dietary treatments: 5 levels of xylanase and glucanase combination (XG)
    • 0, 280/125, 560/250, 840/375, and 1,120/500 TXU/TGU/kg
  • Experimental diets fed in 3 phases for 35 d (10, 10, and 15 d).

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Materials and methods: composition of basal diets

1 Others included poultry fat, L-Lys HCI, L-Met, L-Trp, L-Val, L-Iie, dicalcium phosphate, limestone, sodium chloride, vitamin premix, and mineral premix.

Item

Phase 1

Phase 2

Phase 3

Feedstuff, %

 

 

 

Corn, yellow dent

41.44

46.48

51.12

Soybean meal, 48% CP

17.00

15.50

14.00

Corn DDGS

5.00

15.00

30.00

Whey permeate, 80% lactose

20.00

12.00

-

Processed soybean meal

6.00

3.00

-

Blood plasma

6.00

3.00

-

Titanium dioxide

-

-

0.40

Others1

4.06

4.52

3.98

Supplement (corn and feed enzyme)

0.50

0.50

0.50

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Materials and methods: calculated composition of basal diets

Item

Phase 1

Phase 2

Phase 3

Dry matter, %

 91.53

90.62

89.29

Metabolizable energy, kcal/kg

3,400

3,400

3,350

Crude protein, %

22.07

20.53

20.21

SID1 Lys, %

1.50

1.35

1.23

SID Met + Cys, %

0.82

0.74

0.68

SID Thr, %

0.88

0.79

0.73

SID Trp, %

0.26

0.22

0.20

Total Ca, %

0.68

0.63

0.53

STTD2 P, %

0.36

0.30

0.23

1 SID = standardized ileal digestible

2 STTD = standardized total tract digestible

(NRC, 2012)

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Materials and methods: data and sample collection

  • BW and feed intake were measured on d 0, 10, 20, and 35.
  • Fecal score was measured during all the experimental period (2-d interval).
  • All pigs were euthanized to collect samples on d 35.
    • Jejunal digesta for viscosity
    • Jejunal mucosa and jejunal tissues for intestinal health
      • Relative abundance (RA) and alpha diversity of mucosa-associated microbiota
      • IgA, IgG, IL-8, TNF-α, protein carbonyl, and malondialdehyde (MDA)
      • Intestinal morphology and crypt cell proliferation
    • Ileal digesta for apparent ileal digestibility (AID) of nutrients
      • Dry matter, gross energy, crude protein, and ether extract

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Materials and methods: statistical analysis

  • Randomized complete block design
    • Experimental unit: pen
    • Fixed effect: dietary treatment
    • Random effects: initial BW and sex
  • Polynomial contrasts in Proc MIXED procedure of SAS 9.4
    • Linear and quadratic effects of XG
  • XG intake (TXU/TGU d): overall ADFI (kg/d) × XG level (TXU/TGU kg)
    • Linear and quadratic effects of increasing XG intake
    • Optimal levels of XG
  • Significance: P < 0.05; tendency: 0.05 ≤ P < 0.10

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Results: viscosity, α-diversity of mucosal microbiota (jejunum)

Linear: 0.128

Quadratic: 0.070

Linear: 0.663

Quadratic: 0.360

Linear: 0.906

Quadratic: 0.098

Linear: 0.642

Quadratic: 0.263

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Results: RA of mucosal microbiota (jejunum)

Linear: 0.975

Quadratic: <0.05

Linear: 0.497

Quadratic: <0.05

Linear: 0.358

Quadratic: <0.05

Family level

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Results: optimal levels of XG on viscosity and RA

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Results: immune status (jejunal mucosa)

Linear: 0.566

Quadratic: 0.536

Linear: 0.073

Quadratic: 0.669

Linear: 0.981

Quadratic: 0.668

Linear: 0.804

Quadratic: 0.900

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Results: intestinal morphology and crypt cell proliferation

Linear: 0.572

Quadratic: 0.530

Linear: 0.692

Quadratic: 0.166

Linear: 0.827

Quadratic: 0.085

Linear: 0.195

Quadratic: 0.138

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Results: AID of energy and nutrients

Linear: 0.137

Quadratic: 0.361

Linear: 0.072

Quadratic: 0.246

Linear: 0.187

Quadratic: 0.315

Linear: 0.444

Quadratic: 0.208

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Results: enzyme intake on AID of nutrients

y = 0.014 × XG intake (TXU/d) + 57.13

P value: Overall model: 0.065; intercept: <0.01; slope: 0.065.

y = 0.009 × XG intake (TXU/d) + 52.25

P value: Overall model: 0.087; intercept: <0.01; slope: 0.087.

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Results: overall growth performance

Linear: 0.183

Quadratic: 0.424

Linear: 0.200

Quadratic: 0.546

Linear: 0.918

Quadratic: 0.736

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Results: enzyme intake on growth performance

y = 0.09 × XG intake (TXU/d) + 392

P value: Overall model: 0.021; intercept: <0.01; slope: 0.021.

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Conclusions

  • A combinational use of xylanase and β-glucanase would hydrolyze the non-starch polysaccharide fractions, positively modulating jejunal mucosa-associated microbiota.
  • Increased intake of these enzyme combination reduced digesta viscosity and humoral immune response in the jejunum resulting in improved intestinal structure, and ileal digestibility of nutrients, and finally improving growth of nursery pigs.
  • The beneficial effects were maximized when xylanase and β-glucanase were supplemented at a range of 550 to 800 TXU/kg and 250 to 360 TGU/kg, respectively.

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Thank you!� Thoughts? Questions?

Acknowledgment:

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