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Investigation of nutritional and functional roles of β-mannanase with low and high doses of phytase on intestinal health and growth of nursery pigs fed low-cost formulated feeds

Hyunjun Choi*, Zixiao Deng*, Adebayo Sokale†, Anna Fickler‡, Ahmed Amerah§, and Sung Woo Kim*

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*North Carolina State University, Raleigh, NC; †BASF Corp, Florham Park, NJ, USA; ‡BASF SE, Ludwigshafen, Germany; §Cargill Animal Nutrition, United Kingdom

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Nursery feeds in soybean meal (SBM)

  • Feeds for nursery pigs use expensive, highly digestible feedstuffs to help pigs adapt to solid feeds, whereas SBM is a cost-effective protein supplement.
  • However, SBM contains anti-nutritional compounds, including phytate and non-starch polysaccharides, which can negatively impact the intestinal health and growth of nursery pigs.

(Skinner et al., 2014; Hansen et al., 1993; Hong et al., 2004; Deng and Kim, 2024)

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Anti-nutritional compounds: phytate and β-mannan

  • Phytate
    • Binds to nutrients such as minerals and amino acids, reducing nutrient digestibility
    • ↓ Efficiency of endogenous digestive enzymes, leading to an increase in undigested protein in the small intestine
  • β-mannan
    • ↑ Digesta viscosity, negatively influencing mucosal microbiota
    • ↑ Immune responses due to its structural similarity to pathogenic microbes

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(Eeckhout and De Paepe, 1994; Baker et al., 2024; Moita and Kim, 2023)

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Feed enzymes targeting for anti-nutritional compounds

  • Phytase
    • ↓ Phytate, ↑ nutrient utilization, and ↑ growth performance
    • ↓ Endogenous losses of energy and amino acids
    • ↓ Myoinositol, increasing its absorption
  • β-mannanase
    • ↓ β-mannan in feeds, reducing digesta viscosity and releasing short manno-oligosaccharides, which exhibit prebiotic effects on intestinal health and growth of pigs
    • ↓ Activation of immune responses in the intestine of pigs

(Kies et al., 2006; Kiarie et al., 2022; Moita and Kim, 2023; Jang et al., 2024)

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Hypothesis and objective

  • This study aimed to test the hypothesis that low-cost formulated feeds (LCF) negatively influence the mucosa-associated microbiota, intestinal health, and growth performance of nursery pigs, and supplementing β-mannanase with a high dose of phytase in LCF could mitigate these negative impacts as well as decreasing feed cost compared to control feeds (CON).
  • The objective of this study was to investigate the impacts of LCF and the effects of β-mannanase with a high dose of phytase on the mucosa-associated microbiota, intestinal health, and growth performance of pigs fed LCF.

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Materials and methods: animals, design, and feeds

  • A total of 72 pigs (36 barrows and 36 gilts; 6.2 ± 0.2 kg BW)
    • Individually housed in a pen (n = 12 per treatment).
  • Randomized complete block design, with initial BW and sex as blocks
  • The LCF contained 14% more SBM than the CON.
  • Phytase dose: 750 vs. 2,000 FTU/kg (low vs. high); β-mannanase dose: 0 vs. 800 TMU/kg (without vs. with)
  • Treatment arrangements:
    • 2 (750 or 2,000 FTU/kg in CON) + 2 × 2 factors, with phytase (750 or 2,000 FTU/kg in LCF) and β-mannanase (0 or 800 TMU/kg in LCF).
  • Experimental feeds were fed in 3 phases for 42 d (10, 10, and 22 d, respectively).

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Materials and methods: composition of CON and LCF

 

Phase 1

 

Phase 2

 

Phase 3

Item

CON

LCF

 

CON

LCF

 

CON

LCF

Feedstuffs, %

​

​

​

​

​

​

​

​

Corn, yellow dent

44.24

39.21

​

56.33

51.24

​

70.13

62.59

Soybean meal (SBM)

16.00

30.00

​

17.00

31.00

​

20.50

34.50

Enzyme-treated SBM

6.00

-

​

6.00

-

​

6.00

-

Fish meal

4.00

3.00

​

2.00

2.00

​

-

-

Poultry meal

3.00

2.00

​

2.00

-

​

-

-

Blood plasma

4.00

3.00

​

1.50

-

​

-

-

Poultry fat

-

-

​

-

0.55

​

-

-

Whey permeate

20.00

20.00

​

12.00

12.00

​

-

-

Others

2.76

2.79

​

3.17

3.21

​

3.37

2.91

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Materials and methods: data and sample collection

  • BW and feed intake were measured on d 0, 10, 20, and 42.
  • Fecal score was measured during the experimental period (at 2-d interval).
  • Pigs were euthanized to collect samples on d 20 and 42.
    • Jejunal digesta for viscosity
    • Jejunal mucosa and jejunal tissues for intestinal health
      • Alpha and beta diversity and relative abundance (RA) of mucosa-associated microbiota
      • TLR2, TLR4, NOD1, NOD2, CD14, NF-κB, and IFN-γ
      • IgA, IgG, IL-6, IL-8, TNF-α, protein carbonyl, and malondialdehyde
      • Intestinal morphology and crypt cell proliferation
    • Ileal digesta for apparent ileal digestibility of nutrients
      • DM, GE, CP, EE, NDF, and ADF

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Materials and methods: statistical analysis

  • Randomized complete block design
    • Fixed effects: the LCF, high dose of phytase in LCF, β-mannanase in LCF, and the interaction between high dose of phytase and β-mannanase in LCF
    • Random effects: initial BW and sex
    • Experimental unit: pen
  • Significance: P < 0.05; tendency: 0.05 ≤ P < 0.10

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Viscosity of jejunal digesta

LCF: 0.060;

Phy: 0.364; Man: <0.05;

Phy×Man: 0.480

LCF: 0.685;

Phy: 0.706; Man: 0.780;

Phy×Man: 0.109

Phase 2 (d 20)

Phase 3 (d 42)

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Alpha diversity of jejunal mucosal microbiota (d 20)

LCF: 0.338;

Phy: 0.565; Man: 0.949; Phy×Man: 0.904

LCF: 0.379;

Phy: 0.637; Man: 0.234; Phy×Man: 0.711

LCF: 0.379;

Phy: 0.637; Man: 0.234; Phy×Man: 0.711

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Alpha diversity of jejunal mucosal microbiota (d 42)

LCF: 0.682;

Phy: 0.168; Man: 0.629; Phy×Man: 0.990

LCF: 0.713;

Phy: 0.274; Man: 0.751; Phy×Man: 0.193

LCF: 0.693;

Phy: 0.361; Man: 0.803; Phy×Man: 0.072

AB

AB

A

B

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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RA of jejunal mucosal microbiota (d 20)

LCF: <0.05;

Phy: 0.346; Man: <0.05;

Phy×Man: 0.235

LCF: <0.05;

Phy: 0.443; Man: <0.05;

Phy×Man: 0.575

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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RA of jejunal mucosal microbiota (d 42)

LCF: 0.602;

Phy: 0.470; Man: <0.05;

Phy×Man: 0.625

LCF: 0.633;

Phy: 0.664; Man: 0.130;

Phy×Man: 0.434

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Relative expression of jejunal pattern recognition receptor (d 20)

LCF: 0.804;

Phy: 0.059; Man: 0.204; Phy×Man: 0.370

LCF: 0.834;

Phy: 0.354; Man: 0.977; Phy×Man: 0.070

LCF: 0.520;

Phy: 0.305; Man: 0.231; Phy×Man: 0.233

AB

AB

A

B

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Relative expression of jejunal pattern recognition receptor (d 20)

LCF: 0.520;

Phy: 0.305; Man: 0.231; Phy×Man: 0.233

LCF: 0.557;

Phy: 0.738; Man: 0.890; Phy×Man: 0.388

LCF: 0.067;

Phy: 0.782; Man: 0.249; Phy×Man: 0.211

Diet:

Phy:

Man:

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Relative expression of jejunal pattern recognition receptor (d 42)

LCF: <0.05;

Phy: 0.848; Man: 0.963; Phy×Man: 0.831

LCF: 0.704;

Phy: 0.440; Man: 0.851; Phy×Man: <0.05

LCF: <0.05;

Phy: 0.945; Man: 0.468; Phy×Man: 0.089

AB

AB

B

A

AB

A

B

AB

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Relative expression of jejunal pattern recognition receptor (d 42)

LCF: <0.05;

Phy: 0.981; Man: 0.534; Phy×Man: 0.904

LCF: <0.05;

Phy: 0.820; Man: 0.650; Phy×Man: 0.666

LCF: 0.154;

Phy: 0.422; Man: 0.460; Phy×Man: 0.359

Diet:

Phy:

Man:

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Immune responses in the jejunal mucosa (d 20)

LCF: 0.584;

Phy: 0.675; Man: 0.823; Phy×Man: 0.760

LCF: 0.808;

Phy: 0.915; Man: <0.05; Phy×Man: 0.125

LCF: 0.630;

Phy: 0.311; Man: 0.977; Phy×Man: 0.516

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Immune responses in the jejunal mucosa (d 20)

LCF: 0.495;

Phy: 0.185; Man: 0.991;

Phy×Man: 0.273

LCF: 0.085;

Phy: 0.498; Man: 0.204;

Phy×Man: 0.175

Diet:

Phy:

Man:

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Immune responses in the jejunal mucosa (d 42)

LCF: 0.146;

Phy: 0.257; Man: 0.988; Phy×Man: 0.328

LCF: 0.249;

Phy: 0.684; Man: 0.508; Phy×Man: 0.347

LCF: 0.129;

Phy: 0.588; Man: 0.588; Phy×Man: 0.065

A

B

A

AB

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Immune responses in the jejunal mucosa (d 42)

Diet:

Phy:

Man:

LCF: 0.381;

Phy: 0.586; Man: 0.071;

Phy×Man: 0.785

LCF: 0.836;

Phy: 0.666; Man: 0.577;

Phy×Man: 0.488

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Intestinal morphology in the jejunum (d 20)

Diet:

Phy:

Man:

LCF: 0.977;

Phy: 0.519; Man: 0.297; Phy×Man: 0.174

LCF: <0.05;

Phy: 0.752; Man: 0.100; Phy×Man: 0.724

LCF: 0.099;

Phy: 0.800; Man: 0.680; Phy×Man: 0.117

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Intestinal morphology in the jejunum (d 42)

Diet:

Phy:

Man:

LCF: 0.646;

Phy: 0.397; Man: 0.738; Phy×Man: 0.685

LCF: 0.276;

Phy: <0.05; Man: 0.969; Phy×Man: 0.062

LCF: 0.182;

Phy: 0.507; Man: 0.816; Phy×Man: 0.076

AB

A

B

AB

AB

AB

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Jejunal crypt cell proliferation (d 20 and 42)

Diet:

Phy:

Man:

Phase 2 (d 20)

Phase 3 (d 42)

LCF: 0.727;

Phy: <0.05; Man: 0.146;

Phy×Man: 0.503

LCF: 0.896;

Phy: 0.354; Man: 0.590;

Phy×Man: 0.959

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Apparent ileal digestibility of nutrients (d 42)

Diet:

Phy:

Man:

LCF: 0.726;

Phy: 0.680; Man: 0.864; Phy×Man: 0.796

LCF: 0.951;

Phy: 0.483; Man: 0.975; Phy×Man: 0.626

LCF: 0.590;

Phy: 0.791; Man: 0.369; Phy×Man: 0.244

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Apparent ileal digestibility of nutrients (d 42)

Diet:

Phy:

Man:

LCF: <0.05;

Phy: 0.162; Man: <0.05; Phy×Man: 0.695

LCF: 0.135;

Phy: 0.701; Man: 0.089; Phy×Man: 0.666

LCF: 0.286;

Phy: 0.589; Man: 0.169; Phy×Man: 0.881

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Growth performance in phase 1 (d 0 to 10)

Diet:

Phy:

Man:

LCF: <0.05;

Phy: 0.414; Man: 0.264; Phy×Man: 0.152

LCF: <0.05;

Phy: 0.959; Man: 0.121; Phy×Man: 0.058

LCF: <0.05;

Phy: 0.050; Man: 0.761; Phy×Man: 0.454

AB

B

AB

A

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Growth performance in phase 2 (d 10 to 20)

Diet:

Phy:

Man:

LCF: 0.708;

Phy: 0.194; Man: 0.431; Phy×Man: 0.266

LCF: 0.131;

Phy: 0.948; Man: 0.114; Phy×Man: 0.442

LCF: <0.05;

Phy: <0.05; Man: 0.221; Phy×Man: 0.740

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Growth performance in overall period (d 0 to 42)

Diet:

Phy:

Man:

LCF: 0.882;

Phy: 0.286; Man: 0.659; Phy×Man: 0.256

LCF: 0.354;

Phy: 0.453; Man: 0.466; Phy×Man: 0.196

LCF: 0.166;

Phy: 0.679; Man: 0.587; Phy×Man: 0.889

Phy: high dose of phytase (n =12)

Man: β-mannanase (n =12)

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Summary (1)

  • The Low-cost formulated feeds:
    • ↑ Jejunal digesta viscosity, with ↑ RA of fiber-degrading bacteria in the small intestine
    • ↑ Expression of jejunal interferon-γ, ↑ jejunal TNF-α
    • ↓ Intestinal morphology
    • ↓ Growth performance of pigs during phases 1 and 2
  • High dose of phytase in LCF
    • ↑ Jejunal crypt cell proliferation
    • ↑ G:F during phases 1 and 2

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Summary (2)

  • Phases 1 and 2: supplementing β-mannanase in low-cost formulated feeds mitigated the negative impacts of LCF on the intestinal health of pigs:
    • ↓ Jejunal digesta viscosity
    • ↓ fiber-degrading bacteria
    • ↓ Expression of jejunal NOD1
    • ↑ Jejunal IgG
    • Decreased jejunal crypt depth
  • Phase 3: supplementing β-mannanase in low-cost formulated feeds
    • ↑ Fiber-degrading bacteria
    • ↓ Jejunal IL-6
    • ↑ Fat and fiber utilization

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Conclusion

  • The LCF containing high SBM negatively affected jejunal digesta viscosity, intestinal health, and growth performance of nursery pigs, whereas supplementing β-mannanase with a high dose of phytase in LCF mitigated the negative impacts of LCF on intestinal health by improving jejunal digesta viscosity, mucosal microbiota, immune response, and fat digestion.

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Thank you!� Thoughts? Questions?

Kim Lab 2025

Acknowledgments:

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