Effect of dietary protein and

protease supplementation

on performance and gut

health of broiler chicks

P. Buttin, F. Yan, J. Dibner, C. D. Knight,

M. Vazquez-Anon, N. Odetallah, S. Carter

2

Novus international in few words

• Global company specialized in feed additive

– North American, Japanese capital Mitsui/ Nippon Soda

– 1.3 billion turnover , 850 employees of witch 150 R&D

• Born in 1991 out of the scientific heritage of Monsanto

– With 2 major products : methionine analogue / ethoxiquine

• Since developed into a multiproduct company

– Methionine sources, antioxidants, enzymes, gut health,

chelated trace elements, pigments,

3

20 years “stability “

150 to 330 $ / ton

480 to 550

$ /ton

bonddad.blogspot.com

Soybean price last 25 years

Continuous increase in feed protein cost

4

Feed 67%

Grower 14%

Chick 12 %

Other7%

Feed 51%

Grower 20 %

Chick 17 %

Other 12 %

Feed now represents two thirds

of broiler production costs

2000 2011

Broiler cost

of production

Agristat

5

0

50

100

150

200

250

300

350

400

450

2011 2012 2013 2014 2015 2016

Phytase

Mannanase

NSP/Xylanase

Protease

Increasing interest in feed enzymes strong growth expected in protease use

Mill

ion

$

6

Cibenza DP100 a protease for feed application

• Recently introduced in animal feed

• FDA and AAFCO approved

• Applying for registration in EU

• Heat stable from natural thermophillic

Bacillus licheniformis PWD-1

• Broad spectrum activity: casein, collagen,

keratin, elastin

7

3 potential contributions of protease use

in animal production

1- Performance

protease

Increased amino acid availability

2- Gut health 3- Environment

Less protein undigested

Less protein

in feed

Higher amino

acid availability

Maintain weight gain

& feed efficiency

Less protein

in feed

Same protein

retention

Less protein

excretion

Less undigested

protein

Less proteolytic

fermentation

Less biogenic amines

& bacterial toxins

8

3 treatments x 8 pens x 45 broilers

protease DP100 at 0.5 kg/ton

-7.5% CP

-7.5% CP

+ DP100

Control

a b

a

1

1.2

1.4

1.6

1.8

2

2.2

2.4

2.6

35d 42d

Live weight (kg)

P<0.05

b

ab

a

a

c

b

1

1.1

1.2

1.3

1.4

1.5

1.6

1.7

1.8

1.9

35d 42d

Feed conversion ratio

P<0.05

P= 0.065

Gutierrez - Southern Poultry, Atlanta 2011

Reduction in feed protein reduced feed cost

9

ATA for “growth promotion”

Objective to reduce antibiotics used as GP

• Work with antibacterial

– New additives mimicking in some way antibiotics

Generalized crossed resistance to expect ?

• Work on gut flora subtract (flora nutrition)

– What ingredients? (like corn vs wheat)

– What formulation? like low protein vs high protein

– Digestibility aids like protease

10

Objective of study 1

To evaluate the effect of inclusion of

protease at two dietary crude protein

levels on broiler performance and gut

health under stress conditions

11

Materials and methods

• Floor pen study, 288 male broilers, Ross 708

• 4 treatments : 9 replicate of 8 birds

– 2x Protein : normal (22.1% CP ) &high (30% CP by adding poultry meal)

– 2x Protease (DP100) : with and without

• Stress challenge

– Diet based on wheat and rye, no xylanase

– Coccidia challenge on day 7 (3x non attenuated vaccine)

• Measurements

– Performance at day 7, 14, 21, 28

– Ileal Clostridium perfringens count at day 15

– Gut histology at day 14

– Serum α-1-acid glycoprotein level at day 22

12

Experimental diets

Ingredient Normal

protein

High

protein

% %

Soybean meal (48%) 32.36 31.87

Wheat 25.00 25.00

Rye 20.00 20.00

Poultry meal 0.00 14.36

Corn 13.32 4.57

Soybean oil 4.32 3.29

Dicalcium phosphate 1.89 0.00

Limestone 1.33 0.00

Others 1.78 0.91

Total 100.00 100.00

Calc. nutrients Normal

protein

High

protein

% %

ME (kcal/kg) 3000 3000

Crude protein (%) 22.12 30

d Lys,% 1.21 1.39

d TSAA,% 0.89 0.89

d Thr, % 0.79 0.86

13

Effect of dietary CP and protease inclusion

on performance index

150

170

190

210

230

250

270

Low CP High CP

Performance index at day 28

Control

Protease

Source of Variation P-value

CP 0.260

Protease 0.171

CP x Protease 0.607

•Performance Index = ((Livability*((Body

Weight*1000) /Day of study)*10) / (Feed to

Gain corrected for dead bird weight)).

14

b

a

b b

0

1

2

3

4

5

Normal CP High CP

Ileal Clostridium perfringens (log cfu) at day 15

Control

Protease

Effect of dietary CP and protease inclusion

on ileal Clostridium perfringens

Source of Variation P-value

CP 0.128

Protease 0.107

CP x Protease 0.043

15

600

650

700

750

800

850

900

950

1000

1050

Low CP High CP

Serum α-1-acid glycoprotein level day 22

Control

Protease

Effect of dietary CP and protease inclusion on a

liver acute phase protein production

Source of variation P-value

CP 0.717

Protease 0.022

CP x Protease 0.738

16

Effect of dietary CP and protease inclusion

on gut morphology (villus/crypt ratio)

Source of variation P-value

CP 0.821

Protease 0.054

CP x Protease 0.295

Duodenum villus /crypt ratio

day 14

0

2

4

6

8

10

12

Low CP High CP

Control

Protease

17

Conclusions study 1

Excess dietary protein together with a

coccidial challenge resulted in

– Intestinal wall erosion

– Ileal dysbacteriosis

– Increased serum acute phase proteins

Inclusion of DP 100 restored performance

to the levels shown by the low protein group

18

Objectives of study 2

To evaluate the effect of inclusion of protease

at two dietary crude protein levels on broiler

performance under stress conditions based

on bird density

19

Materials and methods

• Floor pen study, 1104 male broilers Ross 708

• 6 treatments with 8 replicates of 21 or 25 birds /pen – Normal protein, low protein , low protein + protease (DP100)

– No stress, stress

• Stress challenge – Normal conditions: 21 bird/pen (14.3/m²) , no feed outage

– Stress conditions: 25 birds/pen (17.2/m²), 8 hr feed outage at d0 and d14

• Measurements – 14 and 27

– BW, FI, FCR, mortality

20

Composition of starter diet

Ingredients normal protein

low protein

Corn 54.04 58.19

Soybean meal, 48% 34.96 31.29

DDGS 5.00 5.00

Soybean oil 2.32 1.75

L-LYSINE HCL 78% 0.18 0.20

MHA - 84% 0.24 0.22

Threonine 0.08 0.07

Phytase 0.05 0.05

Protease 0.00 0.05

Total 100 100

Nutrient normal protein

low protein

ME, kcal/kg 3040 3040

Crude protein, % 22.50 21.10

Dig Lys, % 1.19 1.11

Dig TSAA, % 0.89 0.83

Dig Thr, % 0.78 0.72

Crumble - 0 to14 days

21

Composition of grower diet

Ingredients normal protein

low protein

Corn 59.11 63.23

Soybean meal, 48% 29.94 26.27

DDGS 5.00 5.00

Soybean oil 2.58 2.01

L-LYSINE HCL 78% 0.18 0.21

MHA - 84% 0.21 0.19

Threonine 0.06 0.06

Phytase 0.05 0.05

Protease/corn starch 0.00 0.05

Total 100 100

Nutrient normal protein

low protein

ME, kcal/kg 3120 3120

Crude protein, % 20.50 19.10

Dig Lys, % 1.07 0.99

Dig TSAA, % 0.82 0.76

Dig Thr, % 0.70 0.64

Pellet - 14 to 27 days

22

no stress stress

LW kg

Effect of protein level at 27 days

a ab

c

b

1.28

1.32

1.36

1.40

1.44

1.48

1.52

NP

LP

LP

+D

P1

00

NP

LP

LP

+D

P1

00

P<0.05

no stress stress

FCR

c

a

bc

ab

1.40

1.44

1.48

1.52

NP

LP

LP

+D

P1

00

NP

LP

LP

+D

P1

00

23

no stress stress

LW

… and effect of protease

a ab

a

c

b b

1.28

1.32

1.36

1.40

1.44

1.48

1.52

NP

LP

LP

+D

P1

00

NP

LP

LP

+D

P1

00

P<0.05

no stress stress

FCR

c

a

ab

bc

ab

ab

1.40

1.44

1.48

1.52

NP

LP

LP

+D

P1

00

NP

LP

LP

+D

P1

00

24

Effect of the protease in the low protein diets

on feed conversion ratio

Source of Variation P value SEM

Stress 0.1455 0.006

Protease 0.0813 0.006

Str x Pr 0.6694 0.008

1.40

1.42

1.44

1.46

1.48

1.50

1.52

no stress Stress

LP LP+DP100

FCR at 27 days

25

Conclusions Study 2

• Stressed birds had a better growth on the low

protein diets

• Low protein diet worsen feed efficiency in non

stressed birds

• Inclusion of the protease resulted in improvement

in feed efficiency in birds fed low protein diets in

both the stressed and non-stressed groups

(p<0.1)

26

Overall conclusions

• A range of stress factors were shown to have an effect on

bird performance

– High protein diets together with a coccidial challenge resulted in

dysbacteriosis and changes in gut structure

– Increased bird density reduced performance although this effect

was less apparent in birds on a low protein diet

• Inclusion of the protease Cibenza DP100

– reduced the dysbacteriosis and improved gut morphology in birds

fed high protein diets

– In low protein diets, improved feed efficiency in both control and

stressed birds

27