Young-Dal Jang, Ph.D.

University of Kentucky, Lexington, US

yd.jang@uky.edu

September 21, 2015

2015 Allen D. Leman Conference

• Cereal grains and oil seed meals:

– Undigestible portion: arabinoxylans and phytate

2

Arabinoxylans, % DM Phytate P, % as-fed

Corn 3.8 0.21

SBM 4.5* 0.38

Corn germ meal (CGM) 18.5 0.58¶

Wheat middlings 18.8 0.61

Corn DDGS (>10%) 12.3 0.26

(Arabinose + xylose)

Jaworski et al., 2015; *Bach Knudsen, 1997

NRC, 2012;¶Rojas et al., 2013

Usage of by-product the content of arabinoxylans and phytate

• Arabinoxylans: A major NSP in wheat

– The main component of wheat cell wall (aleurone layer and pericarp)

– Increase endogenous loss/reduce digestibility

– Increase viscosity and reduce passage rate of digesta

– Proliferate gut microbes

• Xylanase: a glycosidase enzyme

– Hydrolyzes arabinoxylan structure

– Improved fiber digestibility in corn-SBM (Passos et al., 2015),

wheat-based diet (Atakora et al., 2011) and in the diet

containing corn DDGS (Jang et al., 2013).

3

• Phytate (phytic acid): A storage form of P in cereal grains and oil seeds

– The main accumulation site is the aleurone layer

– Phytic acid bound to Ca, Mg (Chelate), fat, protein, and starch

– Reduce the availability of P and the other nutrients

• Phytase: Sequential removal of phosphate groups and release P from phytate.

4

5

• The effect of phytase is very clear in improving P digestibility.

• But…Super-dosing phytase??

– Using phytase at over 500 FTU/kg feed

• Completely degrade all phytate in the diet

• Remove anti-nutrient effects of phytate

– Effect…

• Improved growth and nutrient digestibility

• Additional mineral release

• Reduced energy costs of digestion

• Phytic acid and arabinoxylans are both found in the aleurone layer of wheat.

• Xylanase increases the permeability of the aleurone layer (the site of phytic acid storage).

• Then, the accessibility and action of phytase to the phyticacid in the cell wall can be improved.

• However, the results are inconsistent and research on the inclusion of xylanase and phytase to corn-based diets containing multiple by-products is limited.

6

• When pigs were fed high fiber diets containing corn DDGS,

wheat middlings, and corn germ meal (CGM),

• Objective 1.

– The effect of xylanase supplementation on ATTD and energy

release for growing pigs fed diets containing phytase.

• Objective 2.

– The effect of the graded level of phytase supplementation on

growth performance, carcass characteristics, and ATTD for growing

pigs fed diets with or without xylanase supplementation.

7

• 25 pigs (mean initial BW: 76.5 kg)

• 5 treatments in 5 replicates:

– PC: A corn-SBM basal diet with 15% each of corn DDGS, CGM, and wheat middlings

– NC: low ME diet by 103 kcal/kg from the PC diet

– NX1: NC + 8,000 BXU of xylanase/kg diet

– NX2: NC + 16,000 BXU of xylanase/kg diet, and

– NX3: NC + 24,000 BXU of xylanase/kg diet

• All diets contained 250 FTU/kg of phytase (Quantum Blue; AB vista Feed Ingredients, Marlborough, UK).

• Xylanase used: Econase XT; AB vista Feed Ingredients, Marlborough, UK

8

• 10-12 d of adaptation in the farm (ad libitum)

• Daily feed allowance: 2.7% of BW

• After a 7-d adaption period to the metabolic crates, feces and urine were collected for 5 consecutive days (total collection method).

9

Item Positive control Negative control

Ingredient

Corn 26.52 26.52

SBM (dehulled; 48% crude protein) 19.10 19.10

Corn germ meal 15.00 15.00

Corn DDGS 15.00 15.00

Wheat middlings 15.00 15.00

Corn starch 2.00 4.50

Grease 4.50 2.00

Others1 2.88 2.88

Total 100.00 100.00

Calculated composition, %

ME, kcal/kg 3,360 3,256

Crude protein 21.40 21.40

Lysine 0.982 0.982

Methionine 0.376 0.376

Calcium 0.66 0.66

STTD P 0.29 0.29

Phytate P, analyzed 0.34 0.34

Available P, analyzed2 0.28 0.281 Others included L-lysineHCl, L-threonine, limestone, salt, vitamin and trace mineral premix, Santoquin, and AB-20 (clay). 2Available P was calculated by subtracting phytate P from total P.

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11

Treatment: PC NC NX1 NX2 NX3

Items1 Xylanase, BXU/kg: 0 0 8,000 16,000 24,000

Xylanase, BXU/kg ND2 ND 10,800 22,300 31,800

Phytase, FTU/kg 659 565 514 542 609

1BXU = xylanase unit; FTU = phytase units.2ND = none detected. The limit of detection was 2,000 BXU/kg.

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• Apparent Total Tract Digestibility of

– Dry matter (DM), gross energy (GE),

– Ether extract (EE), crude protein (N),

– Neutral detergent fiber (NDF), acid detergent fiber (ADF), hemicellulose

– Ca, and P

• Retention

– GE, N, Ca, and P

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• ANOVA (Proc GLM of SAS)

• Least squares means (PDIFF option of SAS)

• Orthogonal polynomial contrasts (linear and quadratic) with increasing xylanase level (0, 8000, 16000, and 24000 BXU/kg)

• P < 0.05: significant, P < 0.10: tendency

Treatment: PC NC NX1 NX2 NX3

Items Xylanase, BXU/kg: 0 0 8,000 16,000 24,000 SEM

ATTD, %

DM 81.15 81.49 81.47 81.37 82.40 0.41

GE 81.76 81.26 81.18 81.16 82.08 0.38

N 84.68 83.88 83.62 84.10 84.66 0.53

EE 92.71a 86.38b 86.78b 88.06b 87.33b 1.08

Ca 43.10b 57.76a 60.28a 56.70a 61.59a 2.95

P 48.84 50.34 51.07 48.90 49.65 2.05

DE in diet, kcal/kg 3,567.3a 3,383.0b 3,379.8b 3,378.9b 3,417.2b 15.85

ME in diet, kcal/kg 3,437.7a 3,225.8b 3,230.8b 3,214.8b 3,256.8b 17.41

a,b P < 0.05.

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• PC treatment had the greatest ATTD of EE, DE and ME but the lowest ATTD of Ca among dietary treatments (P < 0.05).

• No xylanase effects in ATTD of DM, GE, N, EE, Ca and P.

15

• ATTD of hemicellulose was improved by increasing xylanase level (*linear, P < 0.05).

• ATTD of NDF in NX3 treatment has a numerical improvement (linear, P=0.15).

57.7

61.9

64.6

58.7

60.862.2

57.4

60.5

62.6

58.0

60.3

61.8

57.6

64.0

68.3

50.0

52.0

54.0

56.0

58.0

60.0

62.0

64.0

66.0

68.0

70.0

72.0

ADF NDF Hemicellulose

PC NC NX1 NX2 NX3Xylanase, BXU/kg: 0 0 8,000 16,000 24,000

%

*

Treatment: PC NC NX1 NX2 NX3

Items Xylanase, BXU/kg: 0 0 8,000 16,000 24,000 SEM

Retention, g/d

GE, kcal/d 7,158a 6,624b 6,651b 6,602b 6,738b 97.1

N 29.57 26.18 26.87 22.41 26.64 2.26

Ca 5.39b 7.65a 7.90a 7.37a 8.08a 0.49

P 5.69 5.60 5.52 5.30 5.39 0.33

Retention, % of intake

GE 78.79c 77.48d 77.60d 77.22d 78.23cd 0.41

N 40.41 36.80 37.36 32.10 36.84 2.72

Ca 38.34b 54.88a 56.94a 52.83a 57.85a 2.93

P 44.12 43.74 43.13 41.46 41.53 2.20a,b P < 0.05. c,d P < 0.10.

16

• GE retention (% of intake) in NX3 treatment slightly increased (P<0.10).

• Xylanase supplementation to the high fiber diet also containing phytase can improve some aspects of fiber digestibility and thereby, energy utilization.

17

• 45 individually-fed pigs (mean initial BW: 26.4 kg)

• 9 treatments in 5 replicates (1 + 2 x 4 factorial arrangement)

– High and low-energy control diets (PC and NC) same with Exp. 1

– Factor 1: Phytase level (0, 500, 1,000, and 2,000 FTU/kg)

– Factor 2: Xylanase supplementation (0 vs. 24,000 BXU/kg)

• This level was decided based on the result of Exp. 1

• Phytase and xylanase used: same products as used in Exp.1

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Treatment: PC NC NP500 NP1000 NP2000 NX NXP500 NXP1000 NXP2000

Phytase, FTU/kg: 0 0 500 1,000 2,000 0 500 1,000 2,000

Xylanase, BXU/kg: 0 0 0 0 0 24,000 24,000 24,000 24,000

• 4 phase feeding (ad libitum): – Phase 1: 25-50 kg (d 0-28), Phase 2: 50-75 kg (d 29-56),

Phase 3: 75-100 kg (d 57-81), and Phase 4: 100-125 kg

(d 82-105)

• 0.3% TiO2 was included in the diets of Phase 4 as an indicator for ATTD calculation– Feces was collected at the end of Phase 4 (avg. BW: 118 kg).

• Real-time ultrasound scan was performed when pigs reached at 120 kg BW.

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20

Phase 1 Phase 2 Phase 3 Phase 4

Item, %Positive

control

Negative

control

Positive

control

Negative

control

Positive

control

Negative

control

Positive

control

Negative

control

Ingredient

Corn 27.672 27.672 33.800 33.800 38.474 38.474 43.106 43.106

SBM (dehulled, 48% CP) 21.000 21.000 15.000 15.000 10.500 10.500 6.000 6.000

Corn germ meal 13.000 13.000 13.000 13.000 13.000 13.000 13.000 13.000

Corn DDGS 15.000 15.000 15.000 15.000 15.000 15.000 15.000 15.000

Wheat middlings 15.000 15.000 15.000 15.000 15.000 15.000 15.000 15.000

Corn starch 1.500 4.000 1.500 4.000 1.500 4.000 1.500 4.000

Grease 4.000 1.500 4.000 1.500 4.000 1.500 4.000 1.500

Others1 2.828 2.828 2.700 2.700 2.526 2.526 2.394 2.394

Total 100.000 100.000 100.000 100.000 100.000 100.000 100.000 100.000

Calculated composition, %

ME, kcal/kg 3,344 3,241 3,355 3,251 3,365 3,262 3,374 3,271

Crude protein 21.90 21.90 19.56 19.56 17.78 17.78 16.01 16.01

Lysine 0.980 0.980 0.850 0.850 0.730 0.730 0.610 0.610

Methtionine 0.373 0.373 0.346 0.346 0.326 0.326 0.307 0.307

Calcium 0.67 0.67 0.59 0.59 0.52 0.52 0.46 0.46

STTD P 0.29 0.29 0.28 0.28 0.27 0.27 0.26 0.26

Phytate P, analyzed 0.45 0.40 0.40 0.37 0.40 0.38 0.37 0.40

Available P, analyzed2 0.18 0.22 0.21 0.21 0.19 0.19 0.19 0.181 Others included L-lysineHCl, limestone, salt, vitamin and trace mineral premix, Santoquin, and AB-20 (clay). 2Available P was calculated by subtracting phytate P from total P.

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Treatment: PC NC NP500 NP1000 NP2000 NX NXP500 NXP1000 NXP2000

Phytase1, FTU/kg: 0 0 500 1,000 2,000 0 500 1,000 2,000

Xylanase1, BXU/kg: 0 0 0 0 0 24,000 24,000 24,000 24,000

Xylanase, BXU/kg

Phase 1 <2000 <2000 <2000 <2000 <2000 39,300 39,000 36,000 31,600

Phase 2 <2000 <2000 <2000 <2000 <2000 36,000 31,700 34,500 32,900

Phase 3 <2000 <2000 <2000 <2000 <2000 33,400 38,500 36,600 35,000

Phase 4 <2000 <2000 <2000 <2000 <2000 38,100 34,000 35,200 33,300

Phytase, FTU/kg

Phase 1 <50 <50 436 1,170 2,110 <50 492 1,120 1,620

Phase 2 <50 <50 484 1,290 1,940 <50 706 1,220 2,490

Phase 3 <50 <50 618 1,140 1,870 <50 551 1,070 1,950

Phase 4 <50 <50 538 830 1,910 <50 408 955 2,2601FTU = phytase units; BXU = xylanase unit.

36,475

33,775

35,875

35,150

Average

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• Growth performance

– ADG, ADFI, and G:F ratio

• Carcass characteristics

– Backfat thickness, Longissimus muscle (LM) area,

carcass lean percent, lean daily gain, lean gain to feed ratio

• Apparent Total Tract Digestibility of

– DM, GE, EE, N, NDF, ADF, hemicellulose, Ca, and P

23

• ANOVA (Proc GLM of SAS): Main effects of Xyl and Phyand interaction

• Least squares means (PDIFF option of SAS)

• Single degree of freedom contrast (PC vs. NC)

• Orthogonal polynomial contrasts (linear and quadratic) with increasing phytase level (0, 500, 1,000, and 2,000 FTU/kg)

• For carcass measurement data, scan weight was considered as a covariate

• P < 0.05: significant, P < 0.10: tendency

0.76

0.80

0.84

0.88

0.92

PC NC NP500 NP1000 NP2000 NX NXP500 NXP1000 NXP2000

ADG, kg/d

0.30

0.32

0.34

0.36

0.38

PC NC NP500 NP1000 NP2000 NX NXP500 NXP1000 NXP2000

G:F ratio

24

P<0.05

P<0.05

25

• No sig. xylanase effect.• Only numerical

improvement in ADG and G:F ration.

0.880

0.890

0.900

0.910

0.920

Overall

ADG, kg/d

PC 0 24,000 BXU/kg

0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

Overall

ADFI, kg/d

PC 0 24,000 BXU/kg

0.33

0.34

0.35

0.36

0.37

0.38

0.39

Overall

G:F ratio

PC 0 24,000 BXU/kg

+1.0%

+2.3%

-1.4%

0.780

0.820

0.860

0.900

0.940

Overall

ADG, kg/d

PC 0 500 FTU/kg 1,000 FTU/kg 2,000 FTU/kg

2.200

2.300

2.400

2.500

2.600

2.700

Overall

ADFI, kg/d

PC 0 500 FTU/kg 1,000 FTU/kg 2,000 FTU/kg

0.310

0.320

0.330

0.340

0.350

0.360

0.370

0.380

0.390

Overall

G:F ratio

PC 0 500 FTU/kg 1,000 FTU/kg 2,000 FTU/kg26

* Linear (P < 0.05)¶ Quadratic (P < 0.05)

¶

*

+8.8% +5.5%+7.5%

+5.0% +5.9%+2.9%

+3.7%

-0.9%

+4.4%

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0.32

0.33

0.34

0.35

0.36

0.37

0 500 1,000 2,000

Phytase, FTU/kg

G:F ratio within phytase treatment

0 24,000 BXU/kg

+4.6%-0.3%+5.0%

• No statistical interaction observed.• Up to 1,000 FTU phytase/kg, xylanase numerically increased G:F ratio.

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Xylanase, BXU/kg P-values

Item 0 24,000 SEM Xyl

Scan weight, kg 120.84 120.84 - -

Backfat, mm 15.35 15.35 0.52 0.99

LM depth, mm 62.29 60.46 1.17 0.28

LM area, % 41.47 40.37 0.70 0.28

Carcass lean, % 56.60 56.39 0.36 0.68

Lean gain, g/d 358.48 354.22 3.19 0.35

Lean gain/feed 0.139 0.140 0.003 0.76

• PC treatment was slightly greater in lean gain to feed ratio than NC treatment (P=0.06).

• No xylanase effect in carcass measurements.

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12.00

13.00

14.00

15.00

16.00

17.00

PC 0 500FTU/kg

1,000FTU/kg

2,000FTU/kg

Backfat thickness, mm

54.00

55.00

56.00

57.00

58.00

PC 0 500FTU/kg

1,000FTU/kg

2,000FTU/kg

Carcass lean, %

0.125

0.135

0.145

0.155

PC 0 500FTU/kg

1,000FTU/kg

2,000FTU/kg

Lean gain/feed

340

345

350

355

360

365

PC 0 500FTU/kg

1,000FTU/kg

2,000FTU/kg

Lean gain, g/d

*Linear (P < 0.05)**Linear (P < 0.10)

+2.9% +3.2%

+6.0% -13.4%

+2.8% +4.0%

+8.2% -10.8%

+1.4% +1.7%

+2.2% -6.1%

**

*

**

Xylanase, BXU/kg P-values2

Item, % 0 24,000 SEM Xyl

DM 78.58 78.54 0.24 0.91

GE 78.01 77.87 0.28 0.73

N 76.24 76.21 0.46 0.96

EE 71.61 70.60 1.48 0.46

ADF 51.93 50.70 0.91 0.35

NDF 53.12 52.67 0.56 0.58

Hemicellulose 53.96 54.09 0.87 0.92

Ca 45.64 47.83 1.52 0.32

P 47.84 46.94 1.05 0.55

30

• No xylanase effect of in ATTD.

30

35

40

45

50

55

60

Neutral detergent fiber Hemicellulose Phosphorus

PC 0 500 FTU/kg 1,000 FTU/kg 2,000 FTU/kg

60

65

70

75

80

Dry matter Ether extract

PC 0 500 FTU/kg 1,000 FTU/kg 2,000 FTU/kg

31

* Linear (P < 0.05)¶ Quadratic (P < 0.05)

*, ¶

¶¶

¶

¶

• PC treatment had greater ATTD of EE but lower ATTD of P than NC treatments (P < 0.05).

• No phytase effect of in ATTD of GE, N, ADF, and Ca.%

%+1.5 ~ 2.0%

+8.0 ~ 15.8%

+5.8 ~ 8.4%

+6.9 ~ 11.3% +11.3 ~

24.0%

• Xylanase supplementation to the high fiber diet has no effect on growth, carcass characteristics, and ATTD.

• Increasing phytase level enhanced growth rate, feed efficiency, carcass leanness, and ATTD.

• There was no statistically significant interaction observed between xylanase and phytase but potentially in G:F ratio.

32

• Xylanase can improve fiber utilization in the diets containing multiple by-products.

• Super-dosing phytase can be an useful strategy to enhance nutrient utilization resulting in improved growth performance, and carcass leanness.

• 1,000 FTU phytase/kg diet

• Further investigation may be needed to clearly confirm the xylanase effect and the potential interaction between phytase and xylanase.

– with a large number of pigs and different physiological statuses

33

• National Pork Board

• AB Vista

• Dr. Lindemann

• Dr. Boyd

• Dr. Wilcock

• James H. Monegue

• Dr. Inocencio

• Dr. Monegue

• Dr. Hung

• Mandy Thomas

• Ning Lu

• Jina Lim

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35

Thank you.