USING ENZYMES TO INCREASE PHOSPHORUS AVAILABILITY IN POULTRY DIETS

AUSTIN H. CANTOR Sciences, University of Kentucky, Lexington,

Department of Animal Kentucky, USA

Introduction
There has been considerable interest in recent years in examining ways to decrease the amount of phosphorus excreted by animals in order to minimize water pollution. Much of the phosphorus in feed ingredients obtained from plants is in the form of phytic acid, a compound not degraded by the endogenous enzymes found in animals. Consequently, approximately two-thirds of this phosphorus is considered unavailable for monogastric animals. Thus, it is often necessary to add sources of inorganic phosphorus to diets for poultry and swine in order to meet the animals requirements for this element. One method of reducing the amount of phosphorus added to diets is to increase the availability of the phosphorus in the feed ingredients by the addition of microbially produced enzymes to the diet. This idea was introduced over 25 years ago, but was not applied in the feed industry for a variety of practical reasons. However, in the past several years, because of increasing environmental concerns and regulations, this concept has received considerable attention among researchers and application in the field. We recently conducted a study to compare the efficacy of acid phosphatases from yeast and Aspergillus niger with that of a commercial phytase (from A. niger) for improving availability of phosphorus in a corn/soybean meal broiler starter diet. The organisms used for production of acid phosphatase were genetically modified to increase their yield of the enzyme.

Methods
The study involved 468 day-old male broiler chicks housed in cages (61 x 51 cm). Four replicate groups of nine chicks were assigned to each of 13 dietary treatments. A basal broiler starter diet was formulated (Table 1) to meet all of the broilers nutrient requirements except for calcium and available phosphorus (calcium = 0.65%, available phosphorus = 349

Using enqmes

to increase phosphorus availability in poccltry diets

Table 1. Ingredient Composition of the basal diet. Per cent of diet 57.00 35.00 1.00 0.70 0.25 1.35 4.00 3.12 21.90 0.91 0.54 0.27 0.65

coin
Soybean meal Limestone Dicalcium phosphate Vitamin-mineral mix Cellulose VegetabIe oil Calculated nutrient composition ME, mcaVkg Protein Met f Cys P, total (analyzed) P, available Ca

0.27%). This diet was fed alone or with graded levels of calcium and phosphorus (providing 0.74, 0.83 and 0.92% calcium and 0.33. 0.39 and 0.45% available phosphorus) to establish standard dose response curves for the variables measured in the trial. The levels of calcium and phosphorus were adjusted by varying the amounts of limestone, dicalcium phosphate and cellulose in the diet. In addition to these four standard diets, there were nine experimental diets which consisted of the basal diet supplemented with 400, 800 or 1200 phytase units per kg provided by yeast acid phosphatase, A. niger acid phosphatase or Alfzyme Phytase. Chicks were fed the diets on a free-choice basis for 14 days.

Results
Variables measured in this study included weight gain, feed intake, feed efficiency (gain:feed ratio), plasma inorganic phosphorus, % toe ash, % tibia ash and tibia breaking strength. Compared with the basal diet (no added phosphorus or enzyme), weight gain to 14 days was significantly (P < 0.05) increased by 1200 units per kg of al1 three enzymes (Table 2). Feed intake was also increased by supplementing the diet with the three enzymes. There were no differences among these treatments in feed conversion. Adding inorganic phosphorus to the basal diet did not result in linear responses in plasma phosphorus and toe ash. Therefore, these parameters were not useful in evaluating the efficacy of the three enzymes. However, enzyme supplementation at 1200 units per kg resulted in plasma phosphorus levels equivalent to that obtained with the highest level of dietary phosphorus (Table 3). The addition of enzymes to the basal diet led to significant improvements in both breaking strength and per cent ash of the tibia (Table 4). 350

Austin H. Cantor
Table 2. Effect of dietary phosphorus growth performance. Dietary supplement Weight g (none)* 0.18% Pt YQS ANAPP PHYI SEM 320b 336=b 349= 348a 339a 6 gain, and enzymes on broikr

Feed intake, g 391b 4248 428a 422= 418a 7

Gain/feed

0.82 0.79 0.82 0.82 0.80 0.01

*Basal diet: 0.27% availabIe P, 0.54% total P, 0.65% Ca iProvided 0.45% available P, 0.70% total P, 0.92% Ca *Yeast acid phosphatase, 1200 units/kg A. niger phosphatase, 1200 units/kg IA. niger phytase, 1200 units/kg a,V < 0.05

Table 3. Effect of dietary phosphorus plasma phosphorus and toe ash. Dietary supplement Plasma

and enzymes on

inorganic mg/dl 5.61 6.61 7.01 6.48 7.49 0.45

P,

Toe ash, %

(none) * 0.18% Pt YAP+ ANAP PHYI SEM

10.7 11.9 11.0 10.9 12.2 0.5

*Basal diet: 0.27% available P, 0.54% totar P, 0.65% Ca j-Provided 0.45% available P, 0.70% total P, 0.92% Ca fYeast acid phosphatase, 1200 units/kg A. niger phosphatase, 1200 units/kg IA. niger phytase, 1200 units/kg

Table 4. Effect of dietary phosphorus tibia breaking strength and tibia ash. Dietary supplement Breaking

and enzymes on

strength, 5.53 10.06= 7.84b 7.98b 8.4P 0.34

kg Tibia ash, % 38.3 46.la 42.7b 42.7b 43.6b 0.5

(none)* 0.18% Pt YAP$ ANAPI PlWl SEM

*Basal diet: 0.27% available P, 0.54% total P, 0.65% Ca TProvided 0.45% available P, 0.70% total P, 0.92% Ca *Yeast acid phosphatase, 1200 units/kg A. niger phosphatase, 1200 units/kg flA. niger phytase, 1200 units/kg a.bG < 0.05.

351

Usipag enzynm

to increase

phosphorus

availability

in poultry

diets

However, the improvements noted with the highest levels of enzyme supplementation were not as great as those obtained with the addition of 0.18% inorganic phosphorus. We were interested in calcuiating the increase in available phosphorus due to using the enzymes. Per cent tibia ash was the parameter that had the most linear response to graded levels of supplemental calcium and phosphorus (Figure 1). Based on a linear regression equation for tibia ash vs. % available phosphorus in the diet (Y = 4.16X + 27.5, 1-2 = OK?), the equivalent available phosphorus was calculated for the various enzyme supplements. Supplementing the basal diet with 1200 units of yeast acid phosphatase, A. niger acid phosphatase and phytase increased the available phosphorus in the diet by 0.10, 0.10 and 0.12%,

50

45

3 g .l8

40

35 0.27 0.33
% Available P
Figure 1. Tibia ash as a function of dietary available phosphorus.

0.39

Table 5. Calculated increased available phosphorus based on tibia variabtes. Increased available Enzyme Breaking strength* 0.09 0.10 0.11 P(%) based on:

Per cent ash? 0.10 0.10 0.12

YAW
ANAPB -JWl

*Strength = 28.4 X (available P) - 2.4; P < 0.001, r = 0.89 t% ash = 41.6 X (availabIe P) + 27.5; P c 0.001, r = 0.90 *Yeast acid phosphatase, 1200 units/kg A. niger phosphatase, 1200 units/kg TA. niger phytase, 1200 unit&g

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Austin H. Cantor respectively (Table 5). A similar calculation was made using the results obtained with tibia breaking strength. Increases of 0.09, 0.10 and 0.11% available phosphorus were obtained from using 1200 units of yeast acid phosphatase, A. aiger acid phosphatase and phytase, which is in close agreement with the values calculated from the tibia ash data. The basal diet was calculated to contain 0.27% available phosphorus and 0.52% total phosphorus. Therefore, the unavailable phosphorus, which was presumably mostly in the form of phytic acid, was equal to 0.25%. Based on the assumption that the increases in available phosphorus were due to hydrolysis of phytate, we then calculated the percentage of phytate phosphorus liberated by the three enzymes using the values from Table 5. This value ranged from 36 to 44% and from 40 to 48% using breaking strength and ash data, respectively (Table 6).

Table 6. Percentage of phytate phosphorus hydrolyzed based on tibia variables. % Phytate phosphorus based on: strength 36 40 44 hydrolyzed

Enzyme YAP? ANAP+ PHYP

Breaking

Per cent ash 40 40 48

*Calculated phosphorus values: total = 0.52%, available = 0.27%, phytate = 0.25% TYeast acid phosphatase, 1200 units/kg $A. niger phosphatase, 1200 units/kg A. niger phytase, 1200 units/kg

Conclusions
The commercial phytase and the two experimental acid phosphatases were all effective in increasing phosphorus availability in the present study. The increases in available phosphorus due to enzyme supplementation were approximately O.lO%, which corresponds to roughly 40% of the phytate phosphorus. Thus, it appears that enzyme supplementation can be an effective method of replacing some of the supplemental phosphorus in poultry diets.

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