Aquaculture Nutrition 2002 8; 271^277

..............................................................................................

Effect of microbial phytase on apparent nutrient digestibility

of barley, canola meal, wheat and wheat middlings, measured
in vivo using rainbow trout (Oncorhynchus mykiss)
Z.J. CHENG & R.W. HARDY
Hagerman Fish Culture Experiment Station, University of Idaho, Hagerman, Idaho, USA

Introduction
Abstract
Aquaculture is one of the fastest growing food-producing
Microbial phytase (Natuphos 5000G) was supplemented
industries in the world. However, as its production system
into barley, canola meal, wheat and wheat middlings at 500
intensifies, it is also subjected to increasing environmental
FTU kg–1 diet to test the effectiveness of the phytase on
concerns and regulations. Phosphorus (P) is a critical
digestibility of crude protein (CP), gross energy (GE) and
pollutant in the aquatic environment; excessive excretion of
minerals for rainbow trout. A total of 180 rainbow trout
P into fresh water can stimulate the growth of algae and
(initial mean body weight 223.8 ± 17.9 g) were stocked into
phytoplankton, thus reducing dissolved oxygen and causing
ten 40-L fibreglass digestibility tanks with 18 fish per tank.
water pollution (Miller et al. 1974; Beveridge 1984; Boyd
Duplicate tanks were assigned to a reference diet and a single
1990; Sugiura et al. 1999). Excessive discharge of protein
tank was assigned randomly to each of the eight diets made
(nitrogen), macro and trace minerals also affects water
from these four ingredients supplemented with and without
quality. Therefore, reducing the amounts of these nutrients
phytase. The collection of faeces lasted for 2 weeks. Faeces
discharged into the aquatic environment is of great
collected in each week represented a replicate and they were
importance in intensive aquaculture operations. Data from
analysed separately. The mean apparent digestibility coeffi-
several studies with trout and salmon have shown that
cients (average of four ingredients) in phytase supplemented
only about 20% of dietary P in typical commercial feeds is
and nonsupplemented ingredients were: CP, 97.3% and
retained by the fish (Ketola 1982; Philips & Beveridge
96.5%; GE, 62.8% and 53.1%; calcium (Ca), 38.3% and –
1986; Ackefors & Enell 1990; Holby & Hall 1991; Ketola
5.3%; magnesium (Mg), 72.4% and 50.3%; manganese
& Harland 1993). This means that approximately 80% of
(Mn), 31.0% and –4.2%; total-phosphorus (total-P), 71.1%
dietary P is not available and is discharged into the aquatic
and 47.6%; phytate-phosphorus (phytate-P), 87.5% and
environment as soluble and faecal forms. The amount of P
22.3%; copper (Cu), 38.3% and 26.4%; potassium (K),
not retained by the fish is known to be influenced by the
97.1% and 97.0%; sulphur (S), 90.2% and 88.6%; zinc (Zn),
amount of P in fish feed and the bioavailability of the P
12.3% and –16.6%. A two-tailed t-test indicated that phytase
sources.
supplementation significantly (P < 0.05) improved the
Fish meal is a major feed ingredient in fish feeds. However,
digestibility of Ca, Mg, Mn, total-P, phytate-P, and GE.
fish meal contains relatively high levels of P and other
The efficacy of phytase on nutrient digestibility also depen-
minerals and the availability of P is relatively low for many
ded on the type of ingredient as measured in rainbow trout.
fish species including trout and salmon (NRC 1993; Sugiura
et al. 1998). The supply of fish meal is not growing worldwide
KEY WORDS: apparent nutrient digestibility, fish feed, phy-
and, it depends entirely on landings from the capture
tase, rainbow trout
fisheries. Peru and Chile are two countries that produce
about two-thirds of annual global fish meal production.
Received 21 May 2001, accepted 29 November 2001
However, fish meal production from these countries can
Correspondence: Zongjia J. Cheng, Hagerman Fish Culture Experiment fluctuate periodically by over 20% in El Nino years when
Station, 3059F National Fish Hatchery Road, Hagerman, ID, USA.
E-mail: [email protected]
ocean temperatures warm up and cause the fish stocks to

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2002 Blackwell Science Ltd 271

272 Z.J. Cheng & R.W. Hardy

move offshore, out of reach of the fishery (Hardy 1995). Table 1 Composition of reference diet (as is basis)
Furthermore, fish meal is often more expensive than most Ingredients g kg)1
oilseed meals such as canola meal or grains and their by-
Casein1 443.9
product meals. Thus, using oilseed meals or grains and their Gelatine2 100.0
by-product meals is increasingly important in formulating Dextrin2 110.0
economic and environmentally friendly fish feeds. Carboxymethyl cellulose2 10.0
Alpha-cellulose2 45.0
Approximately two-thirds of total-P in oilseed meal or
Mineral mixture3 33.0
grains and their by-product meals is present as phytate and Vitamin mixture4 20.0
its bioavailability is very limited to fish (Ogino et al. 1979; Amino acid mixture5 41.0
NRC 1993; Raboy 1997). Sugiura et al. (1998) reported that Ascorbic acid6 2.0
Choline chloride1 10.0
22% of the P in soyabean meal was available for rainbow Yttrium oxide2 0.1
trout. Alternatively, Riche & Brown (1996) found that P in Herring oil7 170.0
soyabean meal was completely unavailable to rainbow trout. Finstim8 15.0
Trace mineral solution9 (100.0)
Phytase is an enzyme specific to phytate hydrolysis. This Water (300.0)
enzyme is present in the digestive tract of many animals, but
1
Purchased from ICN Biomedicals, Inc., Cleveland, OH, USA.
the amount is normally too small to digest dietary phytate to 2
Purchased from Sigma Chemical Co., St Louis, MO, USA.
a significant extent (Bitar & Reinhold 1972). By applying 3
Supplies the following per kg dry diet: KCl, 12.4 g; CaHPO4, 22 g; MgO,
phytase to fish feeds containing oilseed meals or grains and 3 g; NaCl, 2.7 g.
4
Supplies the following per kg dry diet: thiamin mononitrate, 62 mg;
their by-product meals, not only can the P level in fish feeds
ribo£avin, 71 mg; niacin, 294 mg; calcium pantothenate, 153 mg;
be reduced, but the P discharged into the aquatic environ- pyridoxine hydrochloride, 50 mg; folic acid, 22 mg; vitamin B12,
ment can be reduced as well. Commercial phytase is currently 0.08 mg; d-biotin, 0.8 mg; myoinositol, 176 mg; retinol acetate, 8818 IU;
available as a feed supplement for fish and other animal vitamin D3, 588 IU; a-tocopherol acetate, 670 mg; menadione sodium
bisulphate complex, 37 mg.
feeds. However, the potency of the phytase may be different 5
Supplies the following per kg dry diet: DL-methionine, 10 g; L-arginine,
depending on its origin as well as on the dietary formulation 10 g; L-histidine, 3 g; L-lysine, 10 g; L-glycine, 10 g; L-threonine, 2 g.
6
to which it is supplemented. Ascorbate-2-phosphate (Ho¡man La-Roche, Basel, Switzerland).
7
Purchased from Rangen Inc., Buhl, ID.
The objectives of the present study were to determine the 8
Palatability enhancer, containing 48% betaine, EWOS Canada, LTD,
effect of microbial phytase on apparent digestibility coeffi- Surrey, BC, Canada.
9
cients (ADCs) for P, crude protein (CP), calcium (Ca), Supplies the following per kg dry diet: KI,1.5 mg; MnSO4 Æ H2O, 20 mg;
ZnSO4 Æ 7H2O, 75 mg; Na2SeO3, 2 mg; CoCl3 Æ 6H2O, 1.0 mg;
copper (Cu), potassium (K), magnesium (Mg), manganese
CuSO4 Æ 5H2O, 3 mg; FeSO4 Æ 7H2O, 50 mg.
(Mn), sulphur (S), zinc (Zn) and gross energy (GE) in barley,
canola meal, wheat and wheat middlings, and to use this
information to develop recommendations for using phytase
Fish rearing and sampling
in rainbow trout feeds.
This experiment was conducted at the Hagerman Fish
Culture Experiment Station, University of Idaho, Hagerman,
Materials and methods
Idaho. A total of 180 rainbow trout, Oncorhynchus mykiss
(Walbaum) (initial mean body weight 223.8 ± 17.9 g), were
Experimental diets and diet preparation
stocked into ten 40-L fibreglass digestibility tanks with 18 fish
Four feed ingredients (barley, canola meal, wheat and wheat per tank. Duplicate tanks were assigned to a reference diet
middlings) were supplemented with and without phytase and a single tank was assigned to each test diet randomly.
(Natuphos-5000G, BASF Canada Inc., British Columbia, Each tank was supplied continuously with spring water
Canada). Each of the four ingredients was added at 30% into (15 C) at 5 L min–1.
a reference diet (70%, Table 1). Natuphos was mixed into The fish were subjected to a 14-h photo-period using
test diets at 500 FTU kg–1 diet. Thirty percent of water and fluorescent lighting. Fish were acclimated to experimental
10% of trace mineral solution were mixed with each diet conditions for 2 weeks, during which time all fish received a
before pelleting. The experimental diets were prepared as commercial trout feed (Silver Cup, Murray, UT, USA).
cold-extruded pellets using a noodle-making machine with a Experimental diets were fed once daily at 13.00 hours to
4-mm die. Pellets were dried at room temperature overnight apparent satiation for 1 week before sampling faeces. After
and stored at 0–5 C until use. the fish were fed, the tanks were completely cleaned and then

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 Phytase and apparent nutrient digestibility 273

the faeces were collected the next morning at 08.00 hours by predigested in 3 mL of concentrated trace-metal grade nitric
the settling column technique (Cho et al. 1982; Hajen et al. acid at room temperature overnight followed by digestion at
1993). The collection of faeces lasted for 2 weeks. Faeces approximately 115 C for 3–4 h. Digestates were diluted to
collected in each week represented a replicate according to 10 mL with 18 MW Cm water and analysed for Ca, K, Mg,
the method used by Rawles & Gatlin (2000), and they were S, total-P, Cu, Mn, Zn and yttrium (Y) using an Optima 3200
analysed separately. During the entire experiment, no fish radial inductively coupled plasma atomic emission spectro-
mortality occurred. This experimental protocol followed the meter (Perkin-Elmer Corp., Norwalk, CT, USA).
guidelines approved by the Animal Care and Use Committee
of the University of Idaho.
Calculations and statistical methods
The ADCs of protein (% N · 6.25), GE, total-P, phytate-P
Chemical analyses
and other minerals were calculated as the fractional net
Feeds were dried in a convection oven at 105 C for 2 h, and absorption of nutrients from diets based on Y as a
faeces were dried overnight. The dried samples were finely nonabsorbable indicator (Marcus & Lengemann 1962; Hardy
ground by mortar and pestle and were analysed for total 1989; Sugiura et al. 1998). The digestibility data were
nitrogen using a LECO FP-428 nitrogen analyzer (Leco subjected to a two-tailed t-test to determine the effect of
Instruments, St Joseph, MI, USA). The GE was determined phytase on ADC values for protein, GE, total-P, phytate-P,
using an adiabatic bomb calorimeter (Parr Instrument Co., and other minerals in barley, canola meal, wheat and wheat
Moline, IL, USA). Phytate-P was determined according to the middlings. The ADC values for the reference and test diets
method of Latta & Eskin (1980). Briefly, 1 g of feed or 0.5 g were calculated as: ADC(%) ¼ 100 · [1 – (% marker in
faecal samples were acid extracted using 0.65 N HCl for 3 h at diets/% marker in faeces) · (% nutrient in faeces/% nutrient
room temperature with continuous shaking, centrifuged at in diets)] (Maynard & Loosli 1969; NRC 1993).
13 200 g for 5 min, and eluted through an anion exchange The ADC values for test ingredients were calculated using
column chromatograph (AG1-X4 resin, 100–200 mesh, Bio- the formula below (Cho & Slinger 1979; Hardy et al. 1984):
Rad Laboratory, Hercules, CA, USA) using the procedure of ADC (%) ¼ [ADCtest – (1 – i) · ADCref]/i, where ADCtest
Harland & Oberleas (1986). Phytate level was measured using and ADCref were the apparent nutrient (or energy) digest-
sodium phytate (dodecasodium salt, 99% purity, 12% water. ibility coefficients of the test and reference diets, respectively,
Sigma Chemical Co., St Louis, MO, USA) as a standard. and i is the percentage of ingredient included in the test diets;
Other mineral analyses were conducted by the University of i ¼ 30% or 0.3 in this experiment. Computer software,
Idaho Analytical Sciences Laboratory, Moscow, Idaho. Prism, version 2.0 (GraphPad, Inc., San Diego, CA, USA)
Briefly, samples were dried at 60 C for 48 h and ground to was used to perform statistical calculations; P < 0.05 was
pass through a 1-mm sieve. A 0.25-g sub-sample was considered to be statistically significant.

Table 2 Chemical analyses of

Barley Canola meal Wheat Wheat middlings
test ingredients (as is basis,
Item [4-00-549] [5-06-145] [4-05-268] [4-05-205]
n = 2)
Moisture (g kg)1) 88.4 98.3 75.6 109.0
Ash (g kg)1) 18.6 76.9 27.2 50.4
Lipid (g kg)1) 24.1 13.0 25.9 33.8
CP (g kg)1) 174.0 406.0 168.0 176.6
GE (KJ g)1) 18.2 19.5 18.3 19.2
Ca (g kg)1) 0.5 6.1 2.7 1.1
K (g kg)1) 4.5 16.5 5.1 12.0
Mg (g kg)1) 1.4 6.1 1.5 4.0
S (g kg)1) 1.4 5.8 1.7 1.6
Phytate-P (g kg)1) 2.5 8.3 3.6 8.2
Total-P (g kg)1) 4.1 11.0 4.5 8.6
Cu (mg kg)1) 5.2 5.2 4.4 9.6
Mn (mg kg)1) 21.5 73.5 43.5 120.0
Zn (mg kg)1) 34.0 70.0 34.5 79.0

[International feed no.].

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274 Z.J. Cheng & R.W. Hardy

improved the digestibility of GE, Ca, Mg, S, total-P, Cu, Mn

Results and discussion
and Zn in canola meal. For wheat, supplementing phytase
The results of chemical analyses of the test ingredients are only increased the digestibility of Mg and total-P, whereas
presented in Table 2. The nutrient contents in these ingre- the digestibilities of K, Cu and Zn were reduced. The reasons
dients were similar to those analysed by other workers for this negative effect of phytase were not clear. However,
(Church & Pond 1988; NRC 1993). Chemical analyses of for the wheat milling by-product, wheat middlings, the
reference and test diets are listed in Table 3. There were no negative effect of supplementing phytase did not occur. i.e.
significant differences in nutrient contents between diets supplementing wheat middlings with phytase significantly
containing the same test ingredient supplemented with and increased the digestibility of K, Mg, S and total-P. When
without phytase (P > 0.90), however, lipid and CP levels in the digestibility data for these ingredients were combined,
the reference diet were higher than those in the other test supplemental phytase was noted to significantly increase the
diets. This was expected because all other test diets had 30% ADCs of GE (P < 0.001), Ca (P < 0.05), Mg (P < 0.001),
of each ingredient included in the diets, and thus these two Mn (P < 0.05) and total-P (P < 0.05). It also improved the
nutrient concentrations were diluted in the test diets. bioavailabilities of other nutrients, but the effects were not
The ADCs of the test ingredients are presented in Table 4. statistically significant. i.e. CP (P ¼ 0.13), K (P ¼ 0.65), S
Supplementing phytase significantly increased the digestibil- (P ¼ 0.06), Cu (P ¼ 0.14) and Zn (P ¼ 0.08). The data in
ity of Ca, K, Mg, Cu, Mn and Zn in barley. Moreover, it also Table 4 also showed that the efficacy of phytase on nutrient

Table 3 Chemical analyses of

Barley Canola meal Wheat Wheat middlings
reference and test diets supple-
Reference mented with and without phytase
diet Without With Without With Without With Without With (as is basis, n = 2)
Moisture (g kg)1) 66.9 72.7 78.7 79.4 77.2 81.4 76.4 79.9 73.9
Ash (g kg)1) 42.8 37.2 37.5 53.0 50.5 39.1 41.2 42.8 44.3
Lipid (g kg)1) 158.3 81.6 94.7 98.8 101.1 86.0 78.4 100.2 103.1
CP (g kg)1) 576.0 469.2 471.0 551.1 548.3 478.0 470.6 477.4 474.6
GE (KJ g)1) 23.7 22.1 24.4 22.9 22.5 22.1 22.3 22.3 24.1
Ca (g kg)1) 5.5 4.3 4.4 6.1 6.2 5.1 5.0 4.5 3.7
K (g kg)1) 6.1 5.5 5.8 9.4 9.3 5.9 5.8 7.6 7.2
Mg (g kg)1) 1.2 1.3 1.3 2.6 2.7 1.3 1.3 2.0 1.9
S (g kg)1) 5.1 4.0 4.0 5.4 5.3 4.2 4.1 4.1 4.3
Phytate-P (g kg)1) 0.0 0.5 0.6 2.8 2.7 0.4 0.4 2.4 2.1
Total-P (g kg)1) 7.5 6.0 6.1 8.2 8.1 6.3 6.2 7.0 8.5
Cu (mg kg)1) 34 34 27 35 35 36 32 34 33
Mn (mg kg)1) 275 275 265 285 280 280 280 305 255
Zn (mg kg)1) 540 535 505 525 515 525 500 525 435

Table 4 Apparent digestibility coefficients of ingredients supplemented with and without phytase, mean ± SD (%, n = 2)

Barley Canola meal Wheat Wheat middlings Average

Without With Without With Without With Without With Without With

CP 95.5 H 0.2 96.5 H 0.9 94.8 H 1.0 95.9 H 0.4 98.9 H 0.3 98.1 H 1.0 96.9 H 0.9 98.5 H 0.1 96.5 H 1.8 97.3 H 1.3
GE 38.5 H 5.2 50.1 H 0.5 64.8 H 1.8 74.8 H 1.2 55.1 H 1.7 62.1 H 1.8 54.0 H 4.9 63.9 H 1.2 53.1 H 10.5 62.8 H 9.5
Ca 35.3 H 3.1 70.4 H 9.8 )99.8 H 4.6 9.2 H 1.1 43.5 H 13.0 61.9 H 7.6 2.8 H 2.6 11.5 H 1.9 )5.3 H 62.9 38.3 H 30.4
K 96.5 H 0.2 97.4 H 0.1 97.3 H 0.1 97.4 H 0.2 96.6 H 0.3 95.6 H 0.0 97.6 H 0.0 98.2 H 0.2 97.0 H 0.5 97.1 H 1.0
Mg 70.9 H 2.1 86.5 H 0.2 2.3 H 5.3 39.7 H 5.2 75.3 H 1.4 86.4 H 2.7 52.7 H 2.2 76.9 H 0.8 50.3 H 31.1 72.4 H 20.7
S 88.9 H 0.8 90.1 H 0.2 84.1 H 0.4 86.8 H 0.4 92.0 H 0.8 90.8 H 0.7 89.3 H 0.4 93.2 H 0.8 88.6 H 3.1 90.2 H 2.5
Total-P 79.4 H 3.6 82.7 H 0.1 12.2 H 14.3 41.8 H 5.9 61.6 H 0.4 64.6 H 0.1 61.4 H 1.4 95.1 H 3.0 47.6 H 38.1 71.1 H 21.6
Cu 21.5 H 4.2 48.8 H 0.7 17.2 H 2.1 43.2 H 3.3 49.9 H 0.3 31.3 H 0.2 16.8 H 0.6 29.7 H 5.9 26.4 H 14.8 38.3 H 8.9
Mn 16.1 H 11.7 59.1 H 3.5 )62.3 H 17.0 10.4 H 0.9 35.7 H 4.6 56.3 H 10.7 )6.4 H 1.8 )2.1 H 11.1 )4.2 H 40.0 31.0 H 29.6
Zn )0.3 H 5.9 29.8 H 7.5 )70.5 H 9.3 18.5 H 2.9 21.2 H 0.1 18.2 H 0.9 )16.7 H 7.5 )17.1 H 11.7 )16.6 H 36.6 12.3 H 19.6

Asterisks show significant effect between same ingredients supplemented with and without phytase, or overall effect between average values of four
ingredients supplemented with and without phytase ( P < 0.05;  P < 0.01;  P < 0.001).

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 Phytase and apparent nutrient digestibility 275

digestibility depended on the type of ingredients tested in

rainbow trout.
The ADCs for CP, S and K for all ingredients were very
high (>80%) in the present experiment. These findings were
in agreement with those of Sugiura et al. (1998) who reported
that the ADCs for CP and K in soyabean meal, corn gluten,
wheat gluten, wheat middlings and wheat flour were also
high (>90%) for rainbow trout. By contrast, the ADCs of
Ca, Mg, total-P, Cu, Mn and Zn varied among the
ingredients that were not supplemented with phytase. Sugiura
et al. (1998) observed a similar trend for these elements
among different plant protein sources. Riche & Brown (1996)
also reported that the availability of P was more variable in
plant protein feedstuffs. In general, canola meal had lower Figure 1 Apparent digestibility coefficients (mean ± SD) of phytate-
digestibility of Ca, Mg, total-P, Cu, Mn and Zn than noted phosphorus in barley, canola meal, wheat and wheat middlings with
for these elements in barley, wheat and wheat middlings in and without phytase supplementation for rainbow trout (Oncorhyn-
the absence of supplemental phytase. However, with phytase chus mykiss) (P < 0.001 in all ingredients supplemented with and
without phytase).
supplementation, the digestibility of these nutrients was
increased significantly in canola meal. Our results are in
agreement with those of Riche & Brown (1996) who reported 74.8 ± 0.3 and 45.1 ± 3.2; 93.7 ± 1.7 and 12.1 ± 4.0;
that phytase supplementation increased apparent P availab- 81.4 ± 3.9 and 16.9 ± 3.8 for barley, canola meal, wheat
ility in cottonseed meal, peanut meal, soyabean meal and and wheat middlings that were supplemented with or without
corn gluten meal. phytase, respectively. The effectiveness of phytase supple-
The average ADCs of GE for the four test ingredients mentation on the availability of phytate-P was the highest in
analysed in this study was also increased significantly by barley, followed by wheat, wheat middlings and canola meal.
phytase supplementation. The degree of improvement This effect was negatively correlated with the phytate-P levels
depended on the type of ingredients. The bioavailability of in these ingredients. Thus, the higher the amount of phytate-
GE in canola meal, for example, was improved the most. The P in an ingredient, the less efficacy phytase had on the
digestible energy (DE) of each test ingredients were calcula- bioavailability of phytate-P. The phytate-P levels in barley,
ted as: DE ¼ GE · digestibility. The DE values for each of wheat, wheat middlings and canola meal were 2.6, 3.5, 8.2
the test ingredients with or without supplemental phytase and 8.3 g kg–1, respectively (Table 2). The effect of supple-
were: 9.14 and 7.02; 14.56 and 12.61; 11.38 and 10.10; and menting phytase on the utilization of phytate-P has been
12.27 and 10.37 KJ g–1, for barley, canola meal, wheat and demonstrated in rainbow trout and Atlantic salmon with
wheat middlings, respectively. Forster et al. (1999) also either positive effects or no effect whatsoever (Cain & Garling
reported that phytase supplementation of diets containing 1995; Rodehutscord & Pfeffer 1995; Lanari et al. 1998;
canola protein concentrate increased apparent energy digest- Storebakken et al. 1998; Vielma et al. 1998, 2000). Forster
ibility for rainbow trout, while other researchers did not find et al. (1999) showed that supplementing diets containing
a positive response (Lanari et al. 1998) for rainbow trout fed canola protein concentrate with phytase increased the avail-
soyabean meal based diets. This suggests that the effect of ability of phytate-P significantly for rainbow trout.
phytase supplementation on the utilization of energy may be Phytate has been shown to reduce the bioavailability of
dependent on ingredient source, fish size, phytase dosage, etc. various dietary minerals such as Ca, Mg and Zn (Mellanby
In this study, supplementing phytase improved DE in all 1949; Likuski & Forbes 1965; Taylor 1965; Oberleas et al.
ingredients with the effect on canola meal was the most 1966; House et al. 1982; Morris 1986; Zhou et al. 1992). In
significant. the present study, phytase supplementation increased the
The ADCs of phytate-P for all test ingredients with and bioavailability of Mg in all ingredients. Further, it also
without phytase supplementation are illustrated in Fig. 1. increased the bioavailability of Ca and Zn in barley and
Phytase supplementation increased the digestibility of phy- canola meal, but not in wheat and wheat middlings. In fact,
tate-P in all ingredients significantly (P < 0.001). The ADCs phytase supplementation reduced the bioavailability of Zn in
(%) for phytate-P were 100 ± 0.0 and 14.9 ± 2.8; wheat, indicating that phytate chelation with Ca or Zn may

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2002 Blackwell Science Ltd Aquaculture Nutrition 8; 271^277

276 Z.J. Cheng & R.W. Hardy

be ingredient specific. Sugiura et al. (1998) reported that net increasingly used in fish feeds, and the use of phytase may be
absorption of dietary Ca, P, Mg, iron, Mn, strontium and Zn justified because of its effectiveness in releasing P, Ca, Mg,
in rainbow trout was lower in grain by-product meal than Mn and dietary energy, and thus reducing water pollution.
casein–gelatine semi-purified diets. Richardson et al. (1985) Feeding trials aimed at reducing the amount of supplemental
found that a high phytate content in semi-purified diets minerals, especially P, are also needed to further elucidate the
(25.8 g kg–1) depressed growth and feed efficiency of chinook effect of phytase supplementation on fish growth and mineral
salmon. Phytate can also chelate with protein to form retention.
phytate–protein or phytate–mineral–protein complexes that
are resistant to proteolytic digestion (Cheryan 1980). In our
Acknowledgements
study, phytase supplementation did not improve protein
digestibility significantly (P ¼ 0.13), however, there was a Partial financial support from BASF Canada Inc., British
trend for improvement in barley, canola meal and wheat Columbia, Canada, is greatly appreciated. The authors thank
middlings, but not in wheat. This suggests that the supple- Dr Shozo Sugiura, Rudi Kampen, Mike Casten, Carol
mental phytase dosage used in the present study may not Hoffman, Jana Cole, David Brook, and Stefanie Weber for
have been high enough to cleave the bonds between protein their assistance, we also thank two anonymous reviewers for
and phytate. their critical reviews and helpful comments.
In the past, many authors used many different equations to
calculate ADCs of ingredients (Kleiber 1961; Cho & Slinger
References
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