effective use of phytase in poultry diets³n animal 2016...journal of nutrition, 101:1289-1293...
TRANSCRIPT
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David R. Ledoux
Professor Emeritus - Animal Sciences
University of Missouri, Columbia MO
Email: [email protected]
EFFECTIVE USE OF PHYTASE IN POULTRY DIETS
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GRAIN PHOSPHORUS CONTENT
~ 0.3% (BRAN & GERM)
..HOWEVER..
~ 50 - 70% AS PHYTATE
Cereals: In aleurone layer
Oil seed meals: Associated with
storage proteins throughout seed
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O
P
O
O O
H
H
O
H
H
H
O
OP
O
O O
H
PO
OH
OPO
O
O
OPO
O
O
OPO
O
H
O
STRUCTURE OF PHYTIC ACID
(Anderson, 1912)
1 2
3
45
6
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PHYTATE PHOSPHORUS CONTENT OF SELECTED FEEDSTUFFS FOR ANIMALS
(Ravindran, 1996)
Ingredient Phytate P, Phytate P,
g/100 as % of total P
Barley 0.23 63
Corn 0.21 68
Oats 0.22 61
Sorghum 0.22 71
Triticale 0.25 67
Wheat 0.23 65
Soybean meal 0.39 60
Cottonseed meal 0.84 70
Canola meal 0.54 52
Average 64
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-
+ +Ca
O
O
O O
-
H H
Protein
HOCH2
O O
Starch
O
NH3
+
CH2
Protein
Starch
+ +Ca
++Zn
H
O
P
O
O OH
-
O
P
O
O OH
-
H
H
H
H
P
O
O
O O
-
H
O
P
O
O OH
-
O
P
O
O O H-
O
P
O
O O
-1 4
32
56
O
+ +Ca
C O
-
Fattyacid
Phytate, P source and anti-nutrient in feed
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NUTRITIONAL IMPORTANCE OF P
Skeletal structure (80% - 85% in bones and teeth)
Nutrient metabolism (ATP)
Cell membranes (phospholipids)
Nucleic acids (DNA, RNA)
Intracellular buffer
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CONSEQUENCES OF A DEFICIENCY
Rickets, Osteomalacia
Pica
Decreased performance
Mortality
Loss of income
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RICKETS
Normal Vitamin D/Ca Phosphorus
deficiency deficiency
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ENVIRONMENTAL CONCERNS
Phytates in plant products
Unavailable to poultry
Requires non-phytate P (iP, oP)
Results in hi P in excreta
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ENVIRONMENTAL CONCERNS
Results in hi P in excreta
How is excreta P disposed of?
Applied to crop land and pastures
Source of both P and N
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Application of liquid manure
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Application of litter
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INITIAL AND FINAL SOIL ANALYSES FOR A BERMUDA GRASS PASTURE FERTILIZED WITH SWINE LAGOON
EFFLUENT (Kornegay, 1996)
P2O5 Zn Ca
Depth 1990 1992 1990 1992 1990 1992
(cm) kg/ha
0 - 15 540 973 2.6 10.6 0.9 5.3
15 - 30 180 869 0.8 4.8 1.0 3.3
30 - 61 20 209 0.4 2.8 0 3.5
61 - 91 16 64 0.5 2.0 0 2.4
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Algae growth due to N/P
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Fish Kill in Rivers, Lakes & Streams
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ENZYMES
CATALYSTS THAT FACILITATE FEED DIGESTION
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HOW DO ENZYMES WORK?
Enzymes are
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ENZYMES
Exogenous
Animal (pancreas, intestine))
Bacteria
Endogenous
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PHYTASE HISTORY
T. S. Nelson, T. R. Shieh, R. J. Wodzinski and J. H. Ware. 1968.
Poultry Science, 47:1842-1848.
“The availability of phytate phosphorus in soybean meal
before and after treatment with a mold phytase”
T. S. Nelson, T. R. Shieh, R. J. Wodzinski and
J. H. Ware. 1971. Journal of Nutrition, 101:1289-1293
“Effect of supplemental phytase on the utilization of
phytate phosphorus by chicks”
■ Phytase a solution to an environmental problem
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Netherlands pollution issues - pigs (Simons et al., 1990; Brit. J. Nutrition 64:525-540)
Day old male broilers
CRD; 2 reps of 11 birds each
PC, 0.75% tP, 0.9% Ca
NC, 0.45% tP, 0.60% Ca
0, 250, 500, 750,1,000, and 1,500 U phytase/kg diet
Aspergillus ficuum - 3 phytase
Diets fed from day 1 to day 24
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Effect of phytase on BWG of broilers from hatch to day 24 (Simons et al., 1990)
200
300
400
500
600
700
PC NC 250 500 750 1000 1500
Bod
y w
eigh
t gain
(g)
-----------------------U/kg Phytase-----------------------
d
a
b
c
d
e
a-eP < 0.05 d
PC = 0.75% tP and 0.90% Ca; NC = 0.45% tP and 0.60% Ca
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Effect of phytase on FCR of broilers from hatch to day 24 (Simons et al., 1990)
1
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
PC NC 250 500 750 1000 1500
Fee
d c
on
ver
sio
n r
ati
o (
g:g
)
-----------------------U/kg Phytase-----------------------
bc
a
b bcc
c
abcP < 0.05
bc
PC = 0.75% tP and 0.90% Ca; NC = 0.45% tP and 0.60% Ca
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Effect of phytase on P digestibility by broilers from day 21-24 (Simons et al., 1990)
40
50
60
70
PC NC 250 500 750 1000 1500
P d
iges
tib
ilit
y (
%)
-----------------------U/kg Phytase-----------------------
b
a
c
cd
dee
a-eP < 0.05
cd
PC = 0.75% tP and 0.90% Ca; NC = 0.45% tP and 0.60% Ca
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Effect of phytase on Ca digestibility by broilers from day 21-24 (Simons et al., 1990)
40
50
60
70
PC NC 250 500 750 1000 1500
Ca d
iges
tib
ilit
y (
%)
-----------------------U/kg Phytase-----------------------
a a
b
bc
cd
da-dP < 0.05
bc
PC = 0.75% tP and 0.90% Ca; NC = 0.45% tP and 0.60% Ca
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Effect of phytase on excreta P of broilers from hatch to day 24 (Simons et al., 1990)
1.5
2
2.5
3
3.5
4
4.5
5
PC NC 250 500 750 1000 1500
Excr
eta
P (
%)
-----------------------U/kg Phytase-----------------------
c
a
d
de ee
a-eP < 0.05
de
PC = 0.75% tP and 0.90% Ca; NC = 0.45% tP and 0.60% Ca
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COMMERCIAL PHYTASE SOURCES
3 - phytases
6 - phytases
Fungal or Bacterial
Submerged liquid fermentation
Solid-state fermentation
1 26
5 34
Phytate
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Enzyme Submerged Surface
Phytase 1,500 PU/g 1,500 PU/g
Fungal Alpha Amylase ND 240 FAU/g
Beta Glucanase ND 2,160 BGU/g
Cellulase ND 310 CMCU/g
Fungal Protease ND 7,380 HUT/g
SUBMERGED VS SOLID SUBSTRATE FERMENTATION
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Commercially available microbial phytases (Lei et al., 2013)
Product Company Origin Subtype Expression
Allzyme® SSF Alltech Aspergillus niger 3 Aspergillus niger
Finase® EC AB Vista Escherichia coli 6 Trichoderma reesei
Finase® P/L AB Vista Aspergillus niger 3 Trichoderma reesei
Phyzyme® XP Dupont Escherichia coli 6 Schizosaccharomyces pombe
Ronozyme® P Novozyme/DSM Peniophora lycii 6 Aspergillus oryzae
Natuphos® BASF Aspergillus niger 3 Aspergillus niger
OptiPhos® Enzyvia (JBS) Escherichia coli 6 Pichia pastoris
Quantum™ AB Vista Escherichia coli 6 Pichia pastoris
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Bedford (2014)
Phytate degradation by phytase
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Materials and Methods
750 week-old female turkeys
CRD; 5 reps of 25 turkeys each
NRC, 0.6% npP, 1.2% Ca
Negative control, 60% NRC npP, 84% NRC Ca
0, 125, 250, 500, or 10,000 U phytase/kg diet
Bacterial 6 phytase
Diets fed from week 1 to week 14
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Effect of dietary treatments on FI of turkeys(Ledoux et al., 2005)
14
15
16
17
18
19
20
NRC Basal 125 250 500 10000
Fee
d I
nta
ke
(Kg)
------------------U/kg Phytase-------------------
ab
c
bb
ab
a
abcP < .001
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Effect of dietary treatments on WG of turkeys(Ledoux et al., 2005)
5
6
7
8
9
NRC Basal 125 250 500 10000
Wei
gh
t G
ain
(K
g)
------------------U/kg Phytase-------------------
b bb
c
b
a
abcP < .0001
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Effect of dietary treatments on tibia ash of turkeys (Ledoux et al., 2005)
46
48
50
52
54
56
NRC Basal 125 250 500 10000
Tib
ia A
sh (
%)
------------------U/kg Phytase-------------------
aa
a
b
a
a
abP < .05
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Effect of dietary treatments on litter P (Ledoux et al., 2005)
1
1.2
1.4
1.6
NRC Basal 125 250 500 10000
Lit
ter
P (
%)
------------------U/kg Phytase-------------------
aa
a
ab
a
a
abcP < .05
bcbc
cc
Range 22% to 28%
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Treatment 25 30 35 42 70
-------------- Age (Weeks) ---------------
0.10% aP 95ab 84b 61b 55c ---
0.15% aP 96ab 95a 95a 84b 73a
0.20% aP 92b 96a 94a 90ab 75a
0.25% aP 93ab 96a 94a 88ab 78a
0.45% aP 94ab 93a 93a 90ab 74a
0.10% aP+Phyt 97a 96a 94a 87ab 79a
0.15% aP+Phyt 94ab 94a 95a 88ab 78a
0.20% aP+ Phyt 94ab 94a 95a 94a 80a
EFFECTS OF AVAILABLE P AND PHYTASE (300 U/kg) ON PERCENT HEN-DAY EGG PRODUCTION (Boling et al., 2000)
a-cWithin a column, values with no common superscript differ (P < 0.05)
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Treatment 21-30 31-40 41-50 51-60 61-70
-------------- Age (Weeks) ---------------
0.10% aP 53.6a 42.0b ---- ---- ---
0.15% aP 53.6a 58.0a 54.0a 52.7a 47.3a
0.20% aP 53.2a 57.5a 57.0a 55.7a 50.3a
0.25% aP 63.8a 58.5a 56.5a 53.0a 50.0a
0.45% aP 53.0a 56.2a 57.0a 52.7a 48.0a
0.10% aP+Phyt 54.4a 57.2a 56.0a 54.7a 52.0a
0.15% aP+Phyt 54.2a 58.5a 56.0a 55.3a 51.7a
0.20% aP+ Phyt 54.8a 58.0a 58.0a 53.0a 52.3a
a-bWithin a column, values with no common superscript differ (P<0.05)
EFFECTS OF AVAILABLE P AND PHYTASE (300 U/kg) ON EGG YIELD (g/hen/d; Boling et al., 2000)
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Effect of P level and phytase on performance of broiler breeders 29-64 weeks of age (Plumstead et al., 2007)
TRT NPP Phyt Eggs per Feed per Hen Day Fertility Chicks per
hen housed dozen eggs egg prod. hen housed
(%) (FTU/kg) (n) (g) (%) (%) (n)
A 0.37 0 158 2,999xy 62.1b 97.5a 144
B 0.27 500 165 3,038x 62.1b 95.0b 145
C 0.19 0 164 3,068x 61.2b 95.5b 145
D 0.09 500 171 2,875y 65.2a 95.3b 152
SEM 3.7 46.7 0.89 ----- 3.26
P Value 0.178 0.059 0.041 <0.001 0.269
Total P in manure (%): A = 3.23a ; B = 2.79a ; C = 1.97b ; D = 1.87b
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Plant phytase
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PHYTASE ACTIVITY (U/KG) OF SELECTED FEEDSTUFFS (Nys et al., 1996)
Ingredient Phytase activity (U/kg)
Cereals:
Wheat 508 - 1193
Rye 2700 - 5130
Triticale 880 - 1688
Barley 288 - 582
Oats 19 - 42
Corn 15 - 46
Cereal byproducts:
Wheat bran 1100 - 2857
Wheat bran middlings 1900 - 4381
Rye bran 6190 - 6410
Rice bran 122
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Biological Activity of Wheat Phytase Compared with Natuphos® in Pigs (Eeckhout and Paepe, 1990)
Digestibility, %
P Ca
Basal ration 34.9 40.7
+ wheat phytase 45.6 53.2
(500 FTU/kg)
+ Natuphos 53.5 62.6
(500 FTU/kg)
Relative activity 57.5 57.0
of wheat phytase
(Natuphos = 100)
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pH Profile of Microbial and Plant Phytase(Heinzl, 1996)
pH
0
20
40
60
80
100
2 3 4 5 6 7 8
microbialphytase
plant phytase
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OTHER BENEFITS OF PHYTASE
Increased Ca utilization
Improved AA utilization
Improved energy (AME) utilization
Increased trace mineral (Zn, Mn) utilization
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-
+ +Ca
O
O
O O
-
H H
Protein
HOCH2
O O
Starch
O
NH3
+
CH2
Protein
Starch
+ +Ca
++Zn
H
O
P
O
O OH
-
O
P
O
O OH
-
H
H
H
H
P
O
O
O O
-
H
O
P
O
O OH
-
O
P
O
O O H-
O
P
O
O O
-1 4
32
56
O
+ +Ca
C O
-
Fattyacid
Phytate, P source and anti-nutrient in feed
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Effect of phytase on ileal N digestibility in turkeys (Ledoux unpublished)
86
86.5
87
87.5
88
88.5
89
Basal 200 400 600
N d
iges
tib
ilit
y (
%)
b
c
bb
b
a
abP < 0.05
ab
Phytase (U/kg)
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Effect of phytase on N digestibility in broilers fed a lysine deficient diet
(Ravindran et al., 2001)
76
78
80
82
0 125 250 375 500 750 1000
N d
iges
tib
ilit
y(%
)
b
a
Phytase effect (Linear, P < 0.001)
Phytase (U/kg)
Average increase from 0 to 1000 FTU/kg = 5.2%
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0 1,000 FTU/kg Increase
AA1 (%) (%) (%)
Phe 73.7 83.4 13.1*
Leu 72.2 82.6 14.4*
Ile 72.1 82.1 13.8*
Met 88.0 92.8 5.5*
Lys 80.6 88.9 10.3
Val 66.9 77.4 15.7*
Arg 82.3 90.0 9.4*
Hist 73.0 82.1 12.4*
Thr 66.1 76.5 15.7*
1Averaged across 4 Ca:aP ratios (1.43 – 3.57; aP = 0.28%)
Effects of phytase (0 vs 1,000 FTU/kg) on ilealdigestibility of essential AA (Amerah et al., 2014)
*P < 0.0001
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0 1,000 FTU/kg Increase
AA1 (%) (%) (%)
Ser 68.4 80.9 18.2*
Gly 65.8 76.5 16.2*
Asp 75.6 85.0 12.4*
Glu 80.7 88.2 9.3*
Ala 69.9 80.7 15.4*
Pro 72.2 81.2 12.4*
Tyr 75.5 84.4 11.7*
Cyst 54.3 68.5 26.1*
1Averaged across 4 Ca:aP ratios (1.43 – 3.57; aP = 0.28%)
Effects of phytase (0 vs 1,000 FTU/kg) on ileal digestibility of non essential AA (Amerah et al., 2014)
*P < 0.000120 studies poultry & pigs 1996-2009: Adeola and Cowieson, 2011
Ranged from no effect to 6 to 7% improvement; TiO2 & AIA vs Cr2O3
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Effect of phytase on AA digestibility in broilers fed a lysine deficient diet
(Ravindran et al., 2001)
70
75
80
0 125 250 375 500 750 1000
Aver
age
AA
dig
esti
bil
ity(%
)
b
a
Phytase effect (Linear, P < 0.001)
Phytase (U/kg)
Average increase from 0 to 1000 FTU/kg = 5.6%
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Effect of phytase on AME in turkeys (Ledoux unpublished)
2900
3000
3100
Basal 200 400 600
AM
E (
Kca
l/k
g)
b
c
bb
ab
a
abP < .05
b
Phytase (U/kg)
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Effect of phytase on AME in broilers fed a lysine deficient diet (Ravindran et al., 2001)
3200
3300
3400
3500
0 125 250 375 500 750 1000
AM
E (
Kca
l/k
g)
b
a
Phytase effect (linear, P < 0.001; qudratic, P < 0.001
Phytase (U/kg)
Phytase effect ranged 0 to 120 kcal/kg; Adeola and Cowieson, 2011
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Effect of phytase on tibia Mg concentration of broilers at day 41 (Ptak et al. 2013)
0.5
0.6
0.7
0.8
0.9
PC NC Phytase 1 Phytase 2 Phytase 3
Tib
ia M
g (
%)
a
b
aa
a-cP < 0.0001
c
Dietary treatments
NC diet formulated to meet mineral requirements except for Ca and P
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Effect of phytase on tibia K concentration of broilers at day 41 (Ptak et al. 2013)
0.5
0.6
0.7
0.8
PC NC Phytase 1 Phytase 2 Phytase 3
Tib
ia K
(%
)
a
b
aa
ab
a-cP < 0.0001
c
Dietary treatments
NC diet formulated to meet mineral requirements except for Ca and P
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Effect of phytase on tibia Fe concentration of broilers at day 41 (Ptak et al. 2013)
220
320
420
PC NC Phytase 1 Phytase 2 Phytase 3
Tib
ia F
e (m
g)
a
b
a
a
ab
a-bP = 0.0002
b
Dietary treatments
NC diet formulated to meet mineral requirements except for Ca and P
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Effect of phytase on tibia Zn concentration of broilers at day 41 (Ptak et al. 2013)
250
350
450
550
PC NC Phytase 1 Phytase 2 Phytase 3
Tib
ia Z
n (
mg)
bc
c
a
ab
ab
abP < 0.0001
d
Dietary treatments
NC diet formulated to meet mineral requirements except for Ca and P
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PHYTASE
Phosphorus
Excretion
(20-25%)
Carbohydrate
utilization
Mineral
utilization
Nitrogen (AA)
utilization
Endogenous
secretions
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Matrix Values for phytase
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Diet Control ME ME+AA SEM1
Final weight, g 2,219 2,200 2,188 13.2
ADG, g 51.7 51.3 51.0 0.30
AFI, g 94.1 92.8 92.0 1.0
Mortality, chicks/rep 0.60 1.00 0.80 0.25
Tibia ash, % 57.27 58.02 57.12 0.56
Litter P, % 1.43 1.29 1.24 0.023
Control = C-SBM 1Diet = NSME = C-SBM with 600 FTU/kg and matrix values for Ca, aP, and ME
ME+AA = C-SBM with 600 FTU/kg and matrix values for Ca, aP, ME, and AA
EFFECTS OF PHYTASE ON PERFORMANCE OF BROILERS FED DIETS DIFFERING IN NUTRIENT CONTENT (Shelton et al., 2004)
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Diet Control ME ME+AA SEM1
Live weight, kg 2.20 2.15 2.17 0.03
Eviscerated Wt., kg 1.58 1.53 1.55 0.02
Chill Wt., kg 1.62 1.57 1.58 0.02
Carcass Yield, % 72.1 71.8 71.3 0.30
Breast Wt. PLW, % 13.0 12.7 13.1 0.14
Breast Wt. PCW, % 17.6 17.4 17.9 0.18
Control = C-SBM 1Diet = NSME = C-SBM with 600 FTU/kg and matrix values for Ca, aP, and ME
ME+AA = C-SBM with 600 FTU/k and matrix values for Ca, aP, ME, and AA
EFFECTS OF PHYTASE ON CARCASS TRAITS OF BROILERS FED DIETS DIFFERING IN NUTRIENT CONTENT (Shelton et al., 2004)
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Treatment Egg Prod. Egg Wt. Egg Shell Wt.
(g) (%) (%)
Control 76.1 57.9 9.5
RN1 + Phytase (300 FTU) 77.2 58.3 9.5
RN2 + Phytase (300 FTU) 76.3 57.6 9.8
RN3 + Phytase (450 FTU) 78.7 57.2 9.5
RN1 = Reduction in aP by 0.115% and Ca by 0.1%
RN2 = Reduction in aP by 0.115%, Ca by 0.1% and ME by 32 kcal/kg
RN3 = Reduction in aP by 0.172%, Ca by 0.15%, and ME by 48 kcal/kg
All RN diets also reduced in AAs
EFFECTS OF PHYTASE ON PERFORMANCE OF LAYERS FED DIETS DIFFERING IN NUTRIENT CONTENT (Peron et al., 2010)
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Enzyme Cocktails
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Why enzyme cocktails?
Phytate hydrolysis by phytase (max. 65%)
Limited access of phytase to phytate
Phytate occluded by protein, starch, lipids
Allowing access to phytate should increase phytate hydrolysis
In cereals: In aleurone layer
In oil seeds: associated with storage proteins throughout seed
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Approximately 1.5g feed sample was added to a 50 mL centrifuge tube, and was hydrated with 3 mL of 0.03 M HCL solution
Vortex
Sample incubated in 40 C water bath for 30 min
1 mL of 1.5 M HCL containing 6,000 units pepsin/mL added to sample, and sample incubated in 40 C water bath for 45 min
1.3 mL of 1 M NaHCO₃ containing 7.4 mg pancreatin/mL, added to sample and sample incubated for 2 h in 40 C water bath
At the end of incubation, sample centrifuged at 2,500 rpm for 10 min
Supernatant was collected and diluted with 2% nitric acid, and diluent was analyzed for P content
In vitro Procedure(Adapted from Zyla, et al. 1995; Brit. J. of Nutrition, 74, 3-17)
I step (crop)
II step (gizzard)
III step (small intestine)
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Validation of the In Vitro Procedure
(Zyla et al., 1995)
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Efficacy of an enzyme cocktail on phytate hydrolysis in vitro (Zyla et al., 1995)
Phytase activity Conversion Rate
( FTU/kg) (%)
0 57
231 84
347 91
462 93
694 100
Cocktail = Acid protease, Acid phosphatase, Pectinase, Citric acid
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Treatment aP BWG TBASH P RET
(%) (g) (%) (%)
PC 0.60 381b 47.6a 31.0c
Phytase (1000 U) 0.16 356c 42.7c 66.8b
Cocktail 0.16 393b 45.2b 77.0a
Mycelium (5%) 0.16 420a 46.8ab 79.5a
Cocktail: phytase, pectinase, acid phosphatase, acid protease, citric acid, Vit. C
Mycelium: phytase, acid phosphatase, protease, pectinase, PA, Biotin, myo-
inositol
EFFECTS OF PHYTASE AND ENZYME COCKTAIL ADDITIONS TO A CORN-SBM BASED DIET ON PERFORMANCE AND P RETENTION BY TURKEYS
(ZYLA et al., 1996)
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Treatment aP BWG TASH P RET
(%) (g) (%) (%)
PC 0.41 443b 11.94a 47.24c
NC 0.17 289d 8.73b 48.75c
Phytase (750 U) 0.17 389c 9.06b 58.75b
Cocktail 0.17 484a 11.96a 71.50a
Mycelium (4%) 0.17 500a 11.88a 73.00a
Cocktail: phytase, pectinase, acid phosphatase, citric acid
Mycelium: phytase, acid phosphatase, protease, pectinase, PA, Biotin, myo-inositol
EFFECTS OF PHYTASE AND ENZYME COCKTAIL ADDITIONS TO A WHEAT BASED DIET ON PERFORMANCE AND P RETENTION BY BROILERS
(Zyla et al., 2000)
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Why not 100% In Vivo?
Rapid transit time
pH of intestinal tract
Availability of substrate (Phytate)
Solubility of substrate
pH 4-6 pH 2-5
pH 2-5 pH 6-7.5
pH 5-5.7
pH 7
Duodenum
pH 6-6.5
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THE COCKTAIL STRATEGY IS A FEASIBLE APPROACH FOR IMPROVING PHOSPHORUS
AVAILABILITY IN POULTRY DIETS
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IS IT BENEFICIAL TO FEED OTHER ENZYMES IN COMBINATION
WITH PHYTASE INTHE DIET?
XYLANASE ALONE
PHYTASE + XYLANASE
BWG FCR
PHYTASE ALONE
*AVERAGE OF 17 PUBLISHED STUDIES AS REVIEWED BY ROSEN (2004)
15 wheat based diets and 2 corn/rice based diets
NEGATIVE CONTROL 1067 1.736
1079 (+1.12%) 1.690 (-2.66%)
1110 (+4.03%) 1.697 (-2.28%)
1144 (+7.18%) 1.648 (-5.09%)
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IS IT BENEFICIAL TO FEED OTHER ENZYMES IN COMBINATION
WITH PHYTASE IN THE DIET (NC = corn, rye, SBM)?
NC + XAP
NC + XAP + PHYTASE
BWG FCR
NC + PHYTASE
Cowieson and Adeola (2005)
NEGATIVE CONTROL 878 1.742
939 (+7.0%) 1.596 (-8.4%)
932 (+6.2%) 1.614 (-7.3%)
1000 (+14.0) 1.561 (-10.4%)
XAP = XYL+ AMYL+ PROTEASE
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IS IT BENEFICIAL TO FEED OTHER ENZYMES IN COMBINATION
WITH PHYTASE IN THE DIET (NC = Corn-SBM)?
NC + XAP
NC + XAP + PHYTASE
BWG FCR
NC + PHYTASE
(Olukosi et al., 2007)
NEGATIVE CONTROL 553 1.45
566 (+2.4%) 1.44 (- 0.7%)
618 (+11.8%) 1.40 (- 3.4%)
629 (+13.9) 1.37 (- 5.5%)
XAP = XYL+ AMYL+ PROTEASE
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Factors affecting the efficacy of phytase
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Phytase Activity at Different Pelleting Temperatures (Coelho, 1996)
Pelleting temperature Phytase activity
ºF ºC %
140 60 91
158 70 66
176 80 24
194 90 7
212 100 0
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Approaches for addressing phytase degradation issues
Post pelleting applications
Coatings to improve heat stability
Newer phytases more thermostable
More pepsin resistance
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EFFECT OF Ca LEVEL ON THEEFFICACY OF PHYTASE
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Effect of Ca:tP Ratio on Efficacy of Phytase in Broilers and Turkeys (Qian et al., 1996, 1997)
400
450
500
550
600
Broilers Turkeys
1.1:1
1.4:1
1.7:1
2.0:1
Ca:tP ratio
Weig
ht
gain
(g
)
Phytase: 0 - 900 U/kg
Broilers:
0.27% nP; 0.5% tP
Turkeys:
0.36% nP; 0.63% tP
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Effect of Ca:tP Ratio on Efficacy of Phytase in Broilers and Turkeys (Qian et al., 1996, 1997)
10
10.5
11
11.5
12
12.5
Broilers Turkeys
1.1:1 1.4:1
1.7:1 2.0:1
Ca:tP ratio
To
e a
sh
(%
)
Phytase: 0 - 900 U/kg
Broilers:
0.27% nP; 0.5% tP
Turkeys:
0.36% nP; 0.63% tP
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Effect of Ca:tP Ratio on Efficacy of Phytase in Broilers and Turkeys (Qian et al., 1996, 1997)
40
45
50
55
60
Broilers Turkeys
1.1:1 1.4:1
1.7:1 2.0:1
Ca:tP ratio
P r
ete
nti
on
(%
)
Phytase: 0 - 900 U/kg
Broilers:
0.27% nP; 0.5% tP
Turkeys:
0.36% nP; 0.63% tP
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Effects of Formaldehyde on Phytase Efficacy
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Diet FOR Phytase Activity
Expected Analyzed
PC + 0 0
NC + 0 0
E.coli - 500 799
E.coli + 500 116
Fungal - 500 462
Fungal + 500 352
Effect of formaldehyde treatment of recovered phytase activity (Dos Santos et al., 2013)
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Diet FOR FI BWG FCR
PC - 1,163 850 1.370
PC + 1,183 842 1.405
NC - 1,045 740 1.412
NC + 1,087 753 1.445
E.coli - 1,127 816 1.338
E.coli + 1,133 805 1.408
Fungal - 1,175 833 1.411
Fungal + 1,118 800 1.398
FOR Effect NS NS S1
11.362 (- FOR) vs 1.408 (+ FOR) FOR added at 100% higher than recommended
Effect of formaldehyde treatment of on performance of broilers (Dos Santos et al., 2013)
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Superdosing
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Super-dosing
Enough phytase to completely degrade phytate
Less residual phytate (IP6) and other esters (IP5-IP3)
Less anti-nutritional effects
Less endogenous loss of nutrients
Generation of myo-inositol
Vitamin like effects
Lipotropic effects
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Superdosing of phytase
1) Complete break down of phytate
Using enough phytase to prevent build up of IP4 and IP3
IP6 to IP3 can all have anti-nutritional effects
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TRT FI BWG FCR
PC 1101ab 826bc 1.332a
NC 1066b 791c 1.348a
NC+500 U/kg 1132a 856ab 1.323a
NC+1,000 U/kg 1122a 865a 1.298b
NC+1,500 U/kg 1132a 876a 1.293b
Effects of high concentrations of phytase on performance of broiler chicks from hatch to
day 21 (Walk et al., 2014)
a-cMeans in a column with no common superscripts differ P < 0.05
NC = 0.30% aP and 0.82% Ca; PC = 0.45% aP and 1.08% Ca
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Treatment IP6 IP5 IP4 IP3 INOS
-------------- µmol/g of dry weight) ---------------
PC 2.85ab 0.91a 0.75 0.05c 0.91c
NC 1.91b 0.58a 0.67 0.07bc 1.21bc
NC+500 U/kg 0.04d 0.07b 0.72 0.13ab 1.57b
NC+1000 U/kg 0.00d 0.10b 0.55 0.02c 2.12a
NC+1500 U/kg 0.00d 0.11b 0.49 0.02c 2.22a
EFFECTS OF PHYTASE ON CONCENTRATIONS OF PHYTATE AND PHYTATE ESTERS IN THE GIZZARD OF BROILERS (Walk et al., 2014)
a-cMeans in a column with no common superscript differ P < 0.05
NC = 0.30% aP and 0.82% Ca; PC = 0.45% aP and 1.08% Ca
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Effect of superdosing of phytase on performance of broilers from hatch to day 42
(Walk et al., 2013)
1.6
1.625
1.65
1.675
1.7
PC NC NC+500 NC+1,000 NC+1,500
Fee
d c
on
ver
sion
rati
o (
kg:k
g)
ab
bc
c
abcP < 0.05
Treatments
bcbc
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Effect of superdosing of phytase on performance of broilers from hatch to day 49
(Walk et al., 2013)
1.6
1.7
1.8
1.9
PC NC NC+500 NC+1,000 NC+1,500
Fee
d c
on
ver
sion
rati
o (
kg:k
g)
aba
c
abcP < 0.05
ab
Treatments
bcbc
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Superdosing of phytase
2) Release of inositol
Inositol shown to have growth promoting effects
Dietary requirement in fish and gerbils
May involve insulin-like mechanisms
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Inositol FI BWG FCR
0 838 537b 1.57a
0.1% 861 579a 1.47b
P Value NS 0.019 0.057
Effects of inositol on performance of broilers fed acorn-SBM based diet from hatch to day 19
(Zyla et al., 2004)
Avg of 2 NPP = 0.27% and 0.47%, and 2 Ca = 0.65% and 0.80%
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TRT FI BWG FCR
NC 565c 263c 2.195a
INOS (0.1%) 617b 321b 1.977b
Phy B 652ab 345ab 1.918b
Phy A+B 692a 371a 1.908b
P Values 0.001 0.001 0.007
Effects of inositol on performance of broiler chicks fed corn-SBM and corn-wheat diets from hatch to
day 21 (Zyla et al., 2013)
NC = 0.145% NPP and 0.516% Ca
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Effect of myo-inositol (0.15%) on feed intake of broilers (Cowieson et al., 2013)
2000
3000
4000
PC NC PC+IN NC+IN
Fee
d I
nta
ke
(g)
a
bb
abP = 0.003
ab
TreatmentsPC - 2590
NC - 2504
PC+IN - 2516
NC+IN - 2579
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Effect of myo-inositol (0.15%) on feed conversion ratio of broilers
(Cowieson et al., 2013)
1.5
1.6
1.7
1.8
PC NC PC+IN NC+IN
Fee
d c
on
ver
siom
rati
o (
g:g
)
a
b
b
abP < 0.0001
a
Treatments
PC - 2590
NC - 2504
PC+IN - 2516
NC+IN - 2579
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Efficacy of phytase and inositol(Bedford, 2014)
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Effect of phytase on plasma inositol concentrations in broilers fed a NC diet
(0.13% aP; Cowieson et al., 2014)
50
60
70
NC 1000 2000 3000
Pla
sma I
nosi
tol
(mg/L
)
ab
aa
abP < .001
ab
Phytase (U/kg)
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Effect of phytase on plasma inositol concentrations in broilers fed a PC diet
(0.40 % aP; Cowieson et al., 2014)
30
40
50
60
70
PC 1000 2000 3000
Pla
sma I
nosi
tol
(mg/L
)
c
ab
a
abcP < .001
bc
Phytase (U/kg)
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Hypothesized mechanism of inositol (Cowieson et al., 2014)
Phytase at high doses increased plasma inositol
Inositol has been found to be an insulin mimetic in several species
This suggests that the beneficial effect of high phytase doses
(increased inositol) may be due to insulin-like mechanisms
Such as insulin-like effects on glucose transport, gluconeogenesis,
and protein deposition
Jozefiak et al. (2010) observed no change in insulin in birds fed phytase (500 U) or phytase plus carbohydrase enzymes but did
see an increase in liver insulin receptor sensitivity by 9.3% and
12.3%, respectively
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Concluding Remarks - 1
Phytase shown to very effective in improving mineral
utilization and decreasing environmental concerns
Phytase shown to improve AA and energy utilization
90% market penetration in poultry diets
Cost effective with current ingredient prices
Phytase effective in combination with other enzymes
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Concluding Remarks - 2
Concerns include
- Dietary Ca concentrations
- Pelleting
- Formaldehyde treatment of feed
- Margins of safety (P equivalency ~ 0.10%)
Super-dosing appears to be effective
- Complete prevention of AN effects
- Inositol growth promoting effects
Will depend on economics
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