Research Journal of Agriculture and Biological Sciences, 4(4): 273-277, 2008

2008, INSInet Publication

Dietary Inclusion of Enzyme Phytase in Egg Layer Diet on Retention of Nutrients,

Serum Biochemical Characters and Phosphorus Excretion
1

D. Kannan, 2K. Viswanathan, 3S.C. Edwin, 4R. Amutha and 5R. Ravi

Department of Laboratory Animal Medicine, Veterinary College and Research Institute,

Namakkal-637 002, Tamil Nadu, India.
2-4
Department of Poultry Science, Veterinary College and Research Institute,
Namakkal-637 002, Tamil Nadu, India.
5
Animal Feed Analytical and Quality Control Laboratory, Veterinary College and
Research Institute, Namakkal-637 002, Tamil Nadu, India.

Abstract: A study was carried out to find out the effect of enzyme phytase (One gram contained 2410
IU of phytase activity) supplemented at 300, 600, 900 and 1200 IU/kg in layer chicken diets containing
available phosphorus at 0.20, 0.25, and 0.30 per cent from 21 to 52 weeks of age. A control group fed
with diet containing 0.50 per cent available phosphorus alone was also maintained. The serum biochemical
characteristics viz. serum calcium, phosphorus and alkaline phosphatase did not differ significantly. Phytate
phosphorus, phosphorus and calcium retention revealed significant difference among treatment groups at
52 n d week of age. But retention of dry matter, nitrogen and crude protein did not exhibit any statistical
variation. Phosphorus excretion was significantly (P<0.01) reduced in phytase-supplemented groups than
0.5 per cent available phosphorus group without phytase.
Key words: Phytase, nutrients retention, serum biochemical, phosphorus excretion, layer chicken
of phosphorus from animal excreta to the environment,
the use of microbial phytase in the diets of broilers [4 ]
and laying hens [1 6 ] has received more attention in recent
years. Hence, an attempt was made to study the effect
of enyme phytase inclusion on layer chicken diet.

INTRODUCTION
Phosphorus is an essential nutrient in several
metabolic processes and it is one of the major mineral
elements required by poultry. Phosphorus along with
calcium plays a major role in the development and
maintenance of the skeletal system. About 50-80 per
cent of the total phosphorus naturally present in plant
feedstuffs is bound in the form of insoluble complex,
phytic acid or phytate, which is not available to birds.
About two-third of the total phosphorus in cereal seeds,
grain legumes and oil-bearing plants are present as
phytate phosphorus [1 2 ] and only one third of phytate
phosphorus is absorbed and the balance is excreted.
In areas with intensive poultry production, manure
nitrogen and phosphorus are considered potentially
detrimental to soil and water quality. The poor
bioavailability of phytate phosphorus from cereals,
seeds and their by-products is largely responsible for
phosphorus pollution. Phytase is an enzyme that
improves phosphorus availability of phytate phosphorus
as well as many other nutrients. It has also got the
potential impact to reduce environment pollution by
reducing the excretion of phosphorus and nitrogen in
manure. W ith the commercial availability of microbial
phytase and public concern surrounding the contribution

M ATERIALS AND M ETHODS

The biological experiment on the layer diets
was studied with three hundred and thirty six
commercial W hite Leghorn pullets of sixteen weeks
age from a single hatch were reared for adaptation up
to 20 weeks of age. T hen birds were weighed, leg
banded
and
randomly
allotted
into
sixteen
treatment groups with three replicates of seven birds
each. The experimental groups allotted for treatment
are given in Table 1. The layers were reared in cages
during their entire experimental period adopting
standard managmental practices. The layers were
subjected to ad libitum feeding and had free access to
wholesome water. Sixteen hours photo period was
provided daily through out the experimental period.
Layer diets were formulated (table 2) as per the
standards prescribed (BIS 1992) except for the
available phosphorus content, which was kept low at
the levels of 0.20, 0.25 and 0.30 per cent. Calcite and

Corresponding Author: D.Kannan, Assistant Professor, Dept. of Lab Animal Medicine, Veterinary College and Research
Institute, Namakkal- 637 002, Tamil Nadu, INDIA.
E-mail: kannan_kpalayam@yahoo.com
273

Res. J. Agric. & Biol. Sci., 4(4): 273-277, 2008

Table 1:
T1
T2
T3
T4
T5
T6
T7
T8
T9
T10
T11
T12
T13
T14
T15
T16

The experim ental groups allotted for treatm ent

0.2 % Available phosphorus
0.2 % Available phosphorus + 300 units of Phytase
0.2 % Available phosphorus + 600 units of Phytase
0.2 % Available phosphorus + 900 units of Phytase
0.2 % Available phosphorus + 1200 units of Phytase
0.25 % Available phosphorus
0.25 % Available phosphorus + 300 units of Phytase
0.25 % Available phosphorus + 600 units of Phytase
0.25 % Available phosphorus + 900 units of Phytase
0.25 % Available phosphorus + 1200 units of Phytase
0.3 % Available phosphorus
0.3 % Available phosphorus + 300 units of Phytase
0.3 % Available phosphorus + 600 units of Phytase
0.3 % Available phosphorus + 900 units of Phytase
0.3 % Available phosphorus + 1200 units of Phytase
0.5 % Available phosphorus (BIS 1992) Control

shell grit were added as calcium sources. Dicalcium

phosphate (DCP) was used to adjust three different
levels of available phosphorus in different experimental
diets.
Blood Parameter: Blood samples were collected from
the six birds in each treatment. Serum was separated
and analyzed for the following parameters.
C
C
C

Serum calcium [1 ]
Serum phosphorus [6 ]
Alkaline phosphatase [7 ]

Digestibility Studies: The digestibility studies were

conducted at the end of the experimental period by
selection of four birds from each treatment. The birds
were fed with the assay diets for 7 days, with the first
3 days serving as adaptation period. During the last 4
days, feed intake was monitored and the excreta was
collected daily at 08.00 h, dried for 24 hours at 80 0 C
in a forced-air oven, and pooled for analysis. Care
was taken to avoid contamination from feather scales
and debris.
The dried excreta were allowed to
equilibrate to atmospheric conditions before being
weighed.
Representative samples were taken and
ground to pass through a 0.5-mm sieve.
The feed and the excreta were analysed for dry
matter, crude protein, calcium, total phosphorus,
phytate phosphorus adopting standard procedures.
From the results obtained from the above estimations,
the retention of dry matter, nitrogen, protein, calcium,
phosphorus and phytate phosphorus were studied. All
the parametric data obtained in this study were
subjected to analysis of variance for statistical
significance as per the methods of Snedecor and
Cochran [1 3 ].

Table 2: Ingredient and nutrient com position of experim ental layer

diets from 21 to 52 weeks of age (on D M basis)
Available Phosphorus (per cent)
Ingredients (per cent)
-----------------------------------------------------0.20
0.25
0.30
0.50
M aize
48.10
48.10
48.10
48.10
D eoiled rice bran
10.20
10.20
10.20
10.20
Sunflower m eal
8.00
8.00
8.00
8.00
Soybean m eal
20.90
20.90
20.90
20.90
Fishm eal
3.00
3.00
3.00
3.00
Calcite
5.00
5.00
5.00
5.00
Shell Grit
4.80
4.46
4.30
3.00
D icalcium phosphate
0.00
0.34
0.50
1.80
Total
100.00
100.00
100.00
100.00
Supplements (g/100kg)
AB 2 D 3 K 1
10.00
10.00
10.00
10.00
B-com plex 2
20.00
20.00
20.00
20.00
Trace m ineral 3
100.00
100.00
100.00
100.00
Lysine
10.00
10.00
10.00
10.00
M ethionine
40.00
40.00
40.00
40.00
Available Phosphorus (per cent)
N utrients (per cent)
-----------------------------------------------------0.21
0.26
0.31
0.49
CP
18.04
18.03
18.03
18.03
D ry m atter
90.09
90.09
90.09
90.05
Crude fibre
6.75
6.75
6.75
6.79
Ether extract
4.92
4.92
4.92
4.95
M E(Kcal/kg)*
2530
2530
2530
2530
Calcium
3.75
3.71
3.69
3.69
Total phosphorus
0.52
0.58
0.63
0.83
Phytate phosphorus
0.35
0.39
0.42
0.55
M ethionine*
0.30
0.30
0.30
0.30
Lysine*
0.87
0.87
0.87
0.87
Total ash
12.46
12.51
12.55
12.67
* - Calculated values
1
O ne gram of Vitam in AB 2 D 3 K supplem ent contained 82500 IU
of Vitam in-A, 50 m g of V itam in-B 2 , 12000 IU of Vitam in-D 3 and
10 m g of Vitam in-K.
2
O ne
gram of B-Com plex supplem ent contained 8 m g of
Vitam in-B 1 , 16 m g of Vitam in-B 6 , 80 m cg of Vitam in B 1 2 ,
80 m g of Vitam in-E, 120 m g of N iacin, 8 m g of Folic acid, 80
m g of C alcium pantothenate, 120 m g of Calcium and 300 m g
of Phosphate.
3
One gram of Trace M inerals contained 54 m g of m anganese, 52
m g of zinc, 20 m g of iron, 2 m g of iodine and 1 m g of cobalt.
N ote O ne gram of phytase enzym e contained 2410 IU of phytase
activity.

RESULTS AND DISSCUSSION

Serum Biochemical Characteristics: Supplementations
of enzyme phytase on serum biochemical characteristics
are presented in Table 3. The analysis of variance
of data revealed no significant difference in
serum calcium, phosphorus and alkaline phosphatase
among treatment groups at 52 n d week of age.
Among the treatment groups, the serum obtained
from the birds with enzyme phytase supplemented
groups showed higher values of phosphorus than
their respective unsupplemented groups of 0.20 and
0.25 per cent available phosphorus. However, in
0.30 per cent available phosphorus groups with or
without enzyme did not show linear increase or
decrease in their values. Among all the groups 0.50
per
cent showed higher phosphorus (6.17 mg/dl)
value.

274

Res. J. Agric. & Biol. Sci., 4(4): 273-277, 2008

Table 3: Effect of enzym e phytase supplem entation on m ean (+ S.E.) serum calcium (m g/dl), phosphorus (m g/dl) and alkaline phosphatase
(KA U nits) and phosphorus excretion (g/bird/day) (+ S.E.) of layer chicken.
Treatm ent
Serum Calcium
Serum Phosphorus
Alkaline Phosphatase
Phosphorus excretion
groups
(m g/dl)
(m g/dl)
(K A U nits)
(g/bird/day)
T1
09.23 0.25
03.83 0.30
31.43 3.48
0.384 0.003 B C
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T2
09.00 0.40
04.10 0.73
38.64 5.00
0.362 0.001 A B
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T3
10.85 0.76
05.70 0.51
39.29 6.38
0.345 0.004 A
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T4
11.62 1.40
05.92 0.19
44.01 9.15
0.348 0.004 A
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T5
10.52 0.98
04.93 0.51
27.11 5.38
0.346 0.001 A
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T6
08.72 0.55
04.13 0.27
31.62 4.53
0.436 0.002 E F
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T7
10.15 0.93
05.23 0.53
26.34 4.27
0.406 0.003 C D
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T8
09.33 0.14
04.98 0.62
33.97 6.90
0.409 0.005 D
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T9
09.05 0.30
05.47 0.29
28.34 3.09
0.407 0.002 C D
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T10
08.60 0.25
05.20 0.43
42.52 4.46
0.415 0.001 D E
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T11
10.25 0.44
05.60 0.73
43.26 4.30
0.479 0.005 G
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T12
09.12 0.34
05.28 0.64
40.09 7.63
0.457 0.002 F G
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T13
09.13 0.21
05.07 0.47
24.45 4.34
0.449 0.005 F
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T14
10.20 0.66
05.00 0.19
35.78 4.67
0.452 0.003 F
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T15
09.55 0.27
04.93 0.26
29.62 4.24
0.459 0.003 F G
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T16
11.42 1.02
06.17 0.50
28.32 3.45
0.657 0.005 H
A -H
M eans within a colum n with no com m on superscript differ significantly (P<0.01
Effect of enzym e phytase supplem entation in layer chicken diets on m ean per cent (+ S.E.)retention of phytate phosphorus,
phosphorus, calcium , dry m atter, nitrogen and protein
Treatm ent groups
Phytate Phosphorus (% )
Phosphorus (% )
Calcium (% )
D ry m atter (% )
N itrogen (% )
Protein (% )
T1
32.22 0.54 B
36.69 0.34 D
40.42 0.77 a
61.14 0.15
53.91 0.65
54.23 0.79
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T2
35.54 0.15 A
38.64 0.16 B C D
43.13 0.95 a
60.58 0.76
55.08 1.38
54.97 0.09
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------A
AB
a
T3
36.55 0.20
40.29 0.37
42.53 0.25
60.70 0.84
55.11 0.98
55.28 0.61
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T4
36.54 0.19 A
39.95 0.26 A B
42.71 0.39 a
61.24 0.29
55.08 0.44
55.52 0.34
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T5
36.30 0.15 A
39.85 0.32 A B
42.91 0.04 a
61.43 0.13
54.91 0.08
55.37 0.34
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------B
CD
a
T6
32.06 0.14
37.78 0.30
41.36 0.25
61.12 0.43
54.17 0.35
54.21 0.23
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T7
37.48 0.19 A
40.59 0.39 A B
42.14 0.24 a
61.44 0.54
54.64 1.18
55.42 0.25
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------A
ABC
a
T8
36.87 0.16
39.62 0.13
42.23 0.77
60.85 0.39
55.22 0.13
55.27 0.76
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------A
ABC
a
T9
36.73 0.19
39.68 0.07
42.39 0.54
61.54 0.46
55.28 0.21
55.57 0.54
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T10
36.49 0.16 A
39.68 0.22 A B C
42.56 0.75 a
61.06 0.44
55.20 0.67
55.48 0.41
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------B
D
a
T11
31.48 0.38
37.43 0.14
40.49 0.41
61.59 1.03
54.21 0.16
54.10 0.10
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------A
AB
a
T12
36.16 0.11
39.87 0.20
41.46 0.38
60.78 0.37
54.65 0.09
55.19 0.31
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T13
36.13 0.34 A
40.09 0.24 A B
40.95 0.28 a
61.73 0.21
55.13 0.30
55.51 0.21
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------A
ABC
a
T14
35.96 0.11
39.74 0.12
41.47 0.57
61.49 0.47
55.14 0.15
55.65 0.07
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T15
35.79 0.31 A
39.90 0.32 A B
41.14 0.65 a
61.64 0.07
55.13 0.58
55.31 0.17
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------T16
30.39 0.28 B
40.72 0.32 A
36.43 0.18 b
61.77 0.74
53.86 0.54
54.61 0.24
A -D
M eans within a colum n with no com m on superscript differ significantly (P<0.01)
a-b
M eans within a colum n with no com m on superscript differ significantly (P<0.05)
Table 4:

275

Res. J. Agric. & Biol. Sci., 4(4): 273-277, 2008

The inclusion of various levels of enzyme phytase
(One g contained 2410 IU of phytase activity) in layer
chicken diets showed that there was no significant
difference in serum calcium (mg/dl), phosphorus
(mg/dl) and alkaline phosphatase among treatment
groups at 52 n d week of age. But numerical decrease in
serum phosphorus was observed in low phosphorus diet
groups than phytase supplementation groups. Similar
findings were reported by Lan et al.[8 ] and Viveros et
al.[1 7 ].

Retention of Dry M atter, Nitrogen and Protein: The

effect of supplementation of various levels of enzyme
phytase in layer chicken diet on retention of dry
matter, nitrogen and protein is presented in Table 4.
The statistical analysis revealed no significant variation
among treatment groups. However, numerical increase
in per cent retention of nitrogen was found in enzyme
phytase supplemented groups than unsupplemented
groups. Among all the treatment groups, 0.5 per cent
available phosphorus without phytase showed lowest
(53.86 per cent) nitrogen retention.
The data on dry matter, nitrogen and protein
retention did not exhibit any statistical variation
between treatment groups by addition of enzyme
phytase (One g contained 2410 IU of phytase activity)
in layer chicken diet. But numerical increase in protein
retention was recorded in phytase supplemented groups
than other groups. Similar findings were observed by
Morz et al. [9 ] and Ravindran et al.[1 0 ]. This might be
due to liberation of protein from the phytate-protein
complex in case of phytase supplementation.

Retention of Nutrients:
Retention of Phytate Phosphorus, Phosphorus and
Calcium: The effect of supplementation of various
levels of enzyme phytase in layer chicken diet on
phytatae phosphorus, phosphorus and calcium is
presented in Table 4. The retention of phytate
phosphorus was significantly (P<0.01) increased in
phytase supplemented groups than unsupplemented
groups. However, the addition of phytase in
different levels did not vary among themselves.
The lowest phytate phosphorus retention (30.39 per
cent) was noticed in 0.5 per cent available phosphorus
treatment groups.
The analysis of variance of data revealed a
significant influence (P<0.01) on retention of
phosphorus due to phytase supplementation, which
exhibited higher retention than the unsupplemented
groups. These values are comparable with that of the
group fed 0.5 per cent available phosphorus diet
without phytase.
The statistical analysis indicated highly significant
(P<0.01) difference in retention of calcium due to
supplementation of enzyme phytase in layer chicken
diet. A significant decrease in calcium retention was
noticed in 0.5 per cent available phosphorus without
phytase supplementation than other treatment groups.
The highest retention value (43.13 per cent) was found
in T2 group which had low available phosphorus
(0.2 per cent) with 300 units of enzyme.
The analysis of data on retention of phytate
phosphorus, and phosphorus revealed a highly
significant (P<0.01) difference among treatment groups
at 52 n d week of age. W here as the differences in
calcium retention was significant (P<0.05) only at 5
per cent level.
The retention of phytate phosphorus, phosphorus
and calcium was significantly increased in enzyme
phytase supplemented groups than their respective
unsupplemented groups. It is more pronounced at
low available phosphorus levels than at higher
levels. Similar results were obtained by Edwards [5 ],
Morz et al. [9 ] and Um et al.[1 5 ]. Increase in phytate
phosphorus retention by phytase enzyme may be due to
the possible breakdown and release of phytate bound
phosphorus from phytic acid.

Phosphorus Excretion: Incorporation of graded levels

of enzyme phytase on phosphorus excretion are
presented in Table 3. Incorporation of graded levels of
enzyme phytase fed to layer chicken revealed highly
significant (P<0.01) effect on phosphorus excretion.
The highest phosphorus excretion (0.657 g/bird/day)
was noted in 0.50 per cent phoshorus fed group.
However, in all groups addition of phytase decreased
the phosphorus excretion in droppings than that of
unsupplemented groups.
Similar results were observed by Simons et al.[1 1 ],
Broz et al. [3 ], Yi et al. [1 8 ], Um and Paik [1 4 ] and
Lan et al. [8 ].
ACKNOW LEDGM ENT
The authors are thankful to the Dean, Veterinary
College and Research Institute, Namakkal and Tamil
Nadu Veterinary and Animal Sciences University,
Chennai for providing necessary facilities to carry out
the work.
REFERENCES
1.

2.
3.
276

Baginski, E.S., S.S. Marie, W .L. Clark and B. Zak,

1973. Clin. Chem. Acta., 46:49. Cited by K.
W einer, 1988. Calcium, Magnesium and Phosphate.
In Varleys Practical Clinical Biochemistry, Ed. by
A.H. Gowenlock. J.L. M cM urray and D.M.
McLauchlan, 6 th Ed., pp: 601-621. Heinemann
Medical Books, London.
B.I.S., 1992. Nutrient requirement for poultry.
Bureau of Indian Standards, I.S. 13574: 1992.
Broz, J., P. Oldale, A.H. Perrein-Voltz, F. Rychen,

Res. J. Agric. & Biol. Sci., 4(4): 273-277, 2008

J. Schulza and C. Simues Nunes, 1994. Effects of
supplemental phytase on perform ance and
phosphorus utilization in broiler chickens fed a
low-phosphorus diet without addition of inorganic
phosphates. Br. Poult. Sci., 35: 273-280.
4. Denbow, D.M., V. Ravindran, E.T. Komegay, Z.
Yi and R.M. Hulet, 1995. Improving phosphorus
availability in soybean meal for broilers by
supplemental phytase. Poult. Sci. 74: 1831-1842.
5. Edwards H M
Jr., 1 9 93. D ietary 1,25dihydroxycholecalciferol supplementation increases
natural phytate phosphorus utilization in chickens.
J Nutr., 123: 567-577.
6. Gomorri, G., 1942. Lab. Clin. Med., 27: 955. Cited
by K. W einer. 1988. Calcium, Magnesium and
P hosphate. In V arleys P ractical Clinical
Biochemistry, Ed. by A.H. G owenlock. J.L.
McMurray and D .M. McLauchlan, 6 th Ed., pp:
601-621. Heinemann Medical Books, London.
7. King, E.J and A.R. Amstrong, 1934. Can. Med.
Ass. J., 31: 376. Cited by D.M. McLanchlan. 1988.
E n z ym e s . In V a rle ys P ra c tic a l C l in ic a l
Biochemistry, Ed. by A.H. G owenlock. J.L.
McMurray and D.M. McLauchlan, 6 th Ed., pp: 535.
Heinemann Medical Books, London.
8. Lan, G.Q, N. Abdullah, S. Jalaludin and Y.W . Ho,
2002. Efficacy of supplementation of a phytaseproducing bacterial culture on the performance and
nutrient use of broiler chickens fed corn-soybean
meal diets. Poult. Sci., 81: 1522-1532.
9. Morz, Z., A.W . Jongbloed and P.A. Kemme, 1994.
Apparent digestibility and retention of nutrients
bound to phytate complexes as influenced by
microbial phytase and feeding regimen in pigs.
J.Anim. Sci., 72: 126-132.
10. Ravindran, V., P.H. Selle, G. Ravindran, P.C.H.
Morel, A.K. Kies and W .L. Bryden, 2001.
Microbial phytase improves performance, apparent
metabolizable energy and ileal amino acid
digestibility of broilers fed a lysine-deficient diet.
Poult. Sci., 80: 338-344.

11. Simons, P.C.M ., A.W . Jongbloed and H.A.J.

Versteegh, 1992.
Improvement of phosphorus
availability by microbial phytase in poultry and
pigs. Br. J. Nutr., 66: 100-109.
12. Simons, P.C.M ., H .A.J. Versteegh, A.W .
Jongbloed, P.A. Kimme, P. Slump, K.D. Bos,
M .G .E. W olters, R.F. Beudeker and G .J.
Verschoor, 1990. Improvement of phosphorus
availability by microbial phytase in broiler and
pigs. Br. J. Nutr., 64: 525-540.
13. Snedecor, G.W . and W.C. Cochran, 1989.
Statistical methods. 8 th edn. Iowa State University
Press, Ames, Iowa.
14. Um, J.S. and I.K. Paik, 1999. Effects of microbial
phytase supplementation on egg production, egg
shell quality and mineral retention of laying hens
fed different levels of phosphorus. Poult. Sci.,
78: 75-79.
15. Um, J.S., H.S. Lim, S.H. Ahn and I.K. Paik, 2000.
Effects of microbial phytase supplementation to
low phosphorus diets on the performance and
utilization of nutrients in broiler chickens. AsianAustralasian J. Anim. Sci., 13: 824-829.
16. Van der klis, J.D., H.A.J. Versteegh, P.C.M.
Simons and A.K. Kies, 1997. The efficacy of
phytase in corn-soybean meal based diets for
laying hens. Poult. Sci., 76: 1535-1542.
17. Viveros, A., A. Brenes, I. Arija and C. Centeno,
2002. Effects of microbial phytase supplementation
on mineral utilization and serum enzyme activities
in broiler chicks fed different levels of phosphorus.
Poult. Sci., 81: 1172-1183.
18. Y i, Z., E.T. Kornegay, V. Ravindran and D.M.
Denbow, 1996a. Improving phytate phosphorus
availability in corn and soybean meal for broilers
using microbial phytase and calculation of
phosphorus equivalency values for phytase. Poult.
Sci., 75: 240-249.

277