The research shows the results of using a protease (Ronozyme ProAct) in improving protein and amino acid digestibility of a conventional commercial 45% protein Meat and Bone Meal. Why not join the Feed Enzyme discussion on LinkedIn? http://www.linkedin.com/groups?home=&gid=4738175&trk=anet_ug_hm&goback=%25 Follow us: @DSMFeedTweet Or speak the expert: @Jobsorbara

1. Use of a protease to enhance the
utilization of meat and bone meal
amino acids by broilers.
J.C.C. Carvalho1, A.G. Bertechini1, F.R. Mesquita1, R.L. Rios1,
E.M.C. Lima1, J.O.B. Sorbara*2
1
Universidade Federal de Lavras, Lavras, MG, Brazil,
2
DSM Nutritional Products, São Paulo, SP, Brazil

2. Abstract

3. Introduction
• Meat and Bone Meal
(MBM)
– Main source Ca & P
– But also source of CP and AA
• CP and AA feed
contribution from MBM
–
–
–
–
–
–
–
8.0% CP
7.5% Dig Arg
5.5% Dig Lys
2.6% Dig TSAA
5.0% Dig Thr
4.8% Dig Ile
6.0% Dig Val

4. Digestibility of MBM
• MBM with 35% CP
• MBM with 55% CP
• MBM
– 73% CP
– 78% Arg
– 73% Lys
– 62% TSAA
– 66% Thr
– 63% Trp
– 68% Ile
– 71% Leu
– 67% Val
– 65% His
Source: Rostagno et al.,
2005
– 82% CP
– 84% Arg
– 81% Lys
– 73% TSAA
– 79% Thr
– 75% Trp
– 83% Ile
– 84% Leu
– 81% Val
– 82% His
Source: Rostagno et al.,
2005
– 65% CP
– 77% Arg 5% CV
– 69% Lys 8% CV
– 62% TSAA 10% CV
– 62% Thr 13% CV
– 55% Trp 16% CV
– 69% Ile
– 71% Leu
– 70% Val
– 71% His
Source: Lemme et al.,
2004
•Range of 5 to 16% difference in AA digestibility

5. Digestibility of MBM
Conclusions:
•MBM has very high variability in its CP&AA content and
digestibility.
•Also, it is well established that the amino acid digestibility of
animal by-products are dependent on processing conditions such as
temperature or ash content (Skurray, 1974).

6. Objective
• This study tested the ability of a commercially
available protease to improve the protein and amino
acid digestibility of a conventional commercial 45%
protein Meat and Bone Meal

7. Material and Methods
• Metabolism Trial
• 160 Male Cobb 500
• Experimental Design
• Experimental Period
– 14 to 18 d old
adaptation period
– 19 to 21 d old total
excrete collection
– 4 treatments
– 8 replicates of 5 birds each
• Analyses
Protease
(PROT/kg)
Tr.
Ingredients
1
Reference Diet
0
2
Reference Diet
15000
3
70% Reference Diet + 30% MBM
0
4
70% Reference Diet + 30% MBM
15000
– Samples were freezedried
– Amino Acids analyzed by
HPLC (White et al., 1986;
Hagen et al., 1989).

8. Material and Methods
• Calculate the Total Apparent Amino Acid Digestibility:
– TAAA (TD or RD) = ingested AA– excreted AA
ingested AA
– TAAA (Ingredient)= AARD + (AATD – AARD)
g/g substitution
Where: TD – Test Diet; RD – Reference Diet;
AARD – Amino Acid in Reference Diet;
AATD – Amino Acid in Test Diet.
• Data analyzed according to SAS procedures (2001).
– Student Test (P<0.05)

9. Material and Methods
• Product Specifications
– RONOZYME ProAct is a mono component protease with 75000
PROT/g originated from Nocardiopsis prasina and produced by
the bacteria Bacillus licheniformis
– Recommend Dose 15000 PROT/kg = 200 ppm
– One ProAct activity unit (PROT) is the amount of enzyme that
releases 1 µmol of p-nitroaniline from 1 mM substrate (Suc-AlaAla-Pro-Phe-pNA) per minute at pH 9.0 and 37 °C.

10. Material and Methods
• Reference Diet Composition
Ingredients
Corn
SBM
Fat
DCP 24/18
Limestone
Salt
Mineral Premix1
Vitamins Premix1
DL-Methionine 99%
L-Lysine 78%
L-Threonine 98%
Other Additives
Choline Chloride 70%
Total
%
53,749
38,747
3,283
2,151
0,951
0,457
0,100
0,100
0,247
0,063
0,007
0,088
0,057
100,000
Calculate Content
AME, Kcal
2980
CP, %
22,00
Ca, %
1,00
Av. P, %
0,50
Na, %
0,20
Dig Lys, %
1,15
Dig TSAA, %
0,85
Dig Thr, %
0,76

11. Material and Methods
• Meat and Bone Meal
Composition
Nutrient
Fat (%)
Peroxide Index (meq/kg)
Moisture (%)
Crude Protein (%)
Ash (%)
Ca (%)
P (%)
Ca:P Ratio
8.47
0.00
4.69
43.53
39.91
14.66
6.88
2.13
Amino Acids
Aspartic Acid
Glutamic Acid
Serine
Glycine
Histidine
Arginine
Threonine
Alanine
Proline
Tyrosine
Valine
Methionine
Cystine
Isoleucine
Leucine
Phenylalanine
Lysine
Total amino acids
%
2.32
4.59
1.62
7.05
0.79
3.43
1.24
3.60
4.11
0.93
1.61
0.88
0.46
1.10
2.31
1.31
2.27
39.85

12. Material and Methods
• Meat and Bone Meal
Composition
Amino Acids
Aspartic Acid
Glutamic Acid
Serine+Glycine
Histidine
Arginine
Threonine
Alanine
Proline
Tyrosine
Valine
Methionine
Cystine
Isoleucine
Leucine
Phenylalanine
Lysine
Total amino acids
Rostagno
et al., 2005
8.79
0.82
3.32
1.36
0.83
1.86
0.57
0.42
1.12
2.63
1.55
2.19
%
2.32
4.59
8.67
0.79
3.43
1.24
3.60
4.11
0.93
1.61
0.88
0.46
1.10
2.31
1.31
2.27
39.85

13. Material and Methods
•Broiler raised in floor pens until
be transferred to batteries
•Receiving the same feed w/out
enzyme

14. Material and Methods
Metabolism Room
• Room of 90 m2
• Controlled environment

15. Results
• Protease effect on Total Apparent Amino Acid
Digestibility of Meat Bone Meal / Essential Amino Acids
Amino Acids
Digestibility Coefficient, %
CV
P
Without Protease
With Protease
Arginine
48,12
55,00
1,55
0,0001
Threonine
66,04
73,09
2,16
0,0006
Valine
63,68
67,16
1,44
0,0019
Methionine
63,73
68,81
2,12
0,0022
Cystine
47,43
51,55
3,77
0,0205
Isoleucine
60,48
61,64
2,53
0,3260
Leucine
69,92
70,18
3,98
0,8975
Phenylalanine
67,45
69,05
1,24
0,0371
Lysine
76,86
82,41
1,61
0,0009
Histidine
70,53
73,50
2,19
0,0373
Total AA
63,25
67,25
1,71
0,0023

16. Results
• Protease improvement on Total Apparent Amino
Acid Digestibility of Meat and Bone Meal
Improvement (% Control)
– Essential Amino Acids
115%
*
*
110%
105%
Mean = 6,32% improvement
*
*
*
*
*
*
100%
His
* = P≤0.05
Arg
Thr
Val
Met
Cys
Ile
Leu
Phe
Lys

17. Results
• Protease effect on Total Apparent Amino Acid
Digestibility of Meat Bone Meal
– Non Essential Amino Acids
Amino Acids
Digestibility Coefficient, %
Without Protease
With Protease
CV
P
Aspartic Acid
67,70
76,44
1,65 0,0001
Glutamic Acid
64,00
66,84
1,99 0,0216
Serine
64,53
70,72
1,83 0,0004
Glycine
60,37
63,44
3,69 0,1062
Alanine
53,40
57,36
3,05 0,0160
Proline
52,96
57,71
4,64 0,0398
Tyrosine
77,89
79,81
2,27 0,1801

18. Results
• Protease improvement on Total Apparent Amino
Acid Digestibility of Meat and Bone Meal
Improvement (% Control)
– All Amino Acids
115%
*
*
105%
*
*
110%
*
Mean = 6,7% improvement
* *
*
*
* *
*
*
100%
Asp Glu Ser
* = P≤0.05
Gly
His Arg Thr
Ala Pro
Tyr
Val
Met Cys
Ile
Leu Phe Lys

19. Conclusion
• Using 200 ppm of RONOZYME® ProAct
effectively increased utilization of amino acids
in Meat and Bone Meal on an average of
6.32%.

20. Acknowledgment
• UFLA
• NECTA
• DSM Nutritional Products

21. Visit us on:
• DSM Animal Nutrition and Hea