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The Effects of a Pectic Enzyme on the Growth-Depressing and Rachitogenic Properties of Rye for Chicks1
The Effects of a Pectic Enzyme on the Growth-Depressing and Rachitogenic Properties of Rye for Chicks1 JEAN C. GRAMMER, JAMES McGINNIS, and MERT0N H. PUBOLS Department of Animal Sciences, Washington State University, Pullman, Washington 99164 (Received for publication November 23, 1981) ABSTRACT Rye has growth-depressing and rachitogenic properties when fed to chicks. Growth depression also is observed when chicks are fed certain polysaccharides, particularly pectin. Rye has about 8% pectin-like material. A commercially available pectic enzyme preparation was fed to chicks at .1% of the rye diet. This enzyme permitted better utilization of rye for growth and also alleviated the rachitogenic effects of rye. The supplemental enzyme permitted maximal bone mineralization with 200 to 300 ICU of vitamin D3/kg diet, whereas 500 ICU/kg otherwise were required with rye diets. (Key words: rye, chick growth, rickets, pectic enzyme) 1982 Poultry Science 61:1891-1896
INTRODUCTION Rye has not been used commonly as a feedstuff in poultry rations. Recently, several workers have reported the growth-depressing effect of rye when it replaced corn in the rations of poultry. Moran et al. (1970) observed growth depression when rye constituted more than 25% of the grain component of the ration. MacAuliffe and McGinnis (1971) noted growth depression when rye replaced wheat as 40% of the diet. Patel and McGinnis (1976) noted depression at levels as low as 25%. The factor(s) in rye that causes reduced growth in chicks has not been identified. One proposal is that pectin-like components are responsible. McNab and Shannon (1975) reported that rye contains approximately 7.9% pectic material, substantially more than other cereal grains. Vohra and Kratzer (1964) observed the growth-depressing effect of citrus pectin when fed to chicks. Wagner and Thomas (1977) and Day and Thomas (1980) also noted this depression and speculated that due to the correlation of high levels of pectin-like components in rye and the growth-depressing effects of citrus pectin, that pectin was the component in rye responsible for growth depression. Patel et al. (1980) reported that a
'Scientific Paper No. 6072. College of Agriculture Research Center, Washington State University, Pullman, Washington. Project No. 0311.
commercial pectic enzyme, Irgazyme 100, increased the growth rate and feed efficiency of chicks fed rye. Rye also has been shown to contain a factor responsible " for decreasing vitamin D 3 utilization in poults and chicks (MacAuliffe et al, 1976a,b). MacAuliffe et al. (1976a) observed that a simple substitution of rye for corn in the diets fed to turkey poults resulted in the development of rickets by 10 days of age. The vitamin D 3 level in the diets either met or exceeded the recommendation of the National Research Council (1971). MacAuliffe et al. (1976b) observed the same effect in chicks fed a diet containing 63% rye. The rachitogenic effect could be alleviated by feeding increased levels of vitamin D 3 . In further studies, these investigators concluded that a vitamin D 3 antagonist in rye interfered with the absorption of vitamin D 3 and not with its further metabolism (MacAuliffe and McGinnis, 1976). Chicks fed vitamin D3-deficient diets with 63% rye but exposed to ultraviolet light showed no difference in bone ash as compared to controls fed corn. The addition of fat to the diet also reduced the severity of the rickets. Both of these observations suggest that rye interferes with the absorption and not the further metabolism of vitamin D 3 . Whether this rachitogenic factor is the same as or different from the factor responsible for growth depression is not known, but it appears that they are different, because excess vitamin D 3 permits bone mineralization but does not overcome growth depression.
1891
GRAMME R ET AL.
1892
A series of experiments was conducted to determine the effect that supplementation with the pectic enzyme, Irgazyme 100, would have on vitamin D 3 utilization in chicks fed rye. MATERIALS AND METHODS
General. One to 3-day-old, commercial broiler-type chicks (Hubbard strain) were randomly assigned to pens of either 5 or 10, as specified for each experiment. Chicks were housed in electrically heated, wire floored battery brooders. The temperature was maintained at 35 C for the first week and 24 C for the second week. Treatments were randomly assigned to pens and feed and water were supplied ad libitum. The composition of the basal diet is listed in Table 1; vitamin D 3 is omitted. All chicks were group weighed at 1 and 2 weeks. Upon termination of the experiments at 2 weeks, 5 chicks from each pen were killed and the left tibia was removed for bone ash analysis. Bone ash was determined according to the methods of the Association of Official Analytical Chemists (1970). All results were
TABLE 1. Composition of the basal diet for Experiments 1, 2, and 3 Ingredients
(%)
Grain a Fishmeal Meat and bone meal Whey Dehydrated alfalfa Iodized salt Dicalcium phosphate Limestone Vitamin premix D Trace mineral mix c DL-Methionine Soybean meal Animal fat
62.55 3.01 3.00 1.00 1.10 .23 .77 .90 .25 .05 .15 25.00 2.00
Total
99.91
Corn or rye. Vitamin premix at .25% of the diet supplies the following per kilogram of the diet: vitamin A, 5,500 IU; vitamin E, 4.4 IU; riboflavin, 3.3 mg; calcium pantothenate, 4.4 mg (or panthotenic acid, 4.05 mg); niacin, 22 mg; choline chloride, 577 mg; vitamin B 1 2 , .011 mg; and ethoxyquin, 62.2 mg. Trace mineral mix supplies the following in mg/kg of the diet: Mn, 100; Fe, 100; Zn, 100; Cu, 10; I, 3.0; Ca, 120; Co, 1.0.
expressed as percent ash of the fat-free, dry bone. Irgazyme 100 and Irgazyme 100-Special were obtained from the Food Industry Department of Ciba-Geigy Corporation, Greensboro, NC. They have about 90% diluent. The Irgazyme 100-Special contains amylase activity, which is not present in the Irgazyme 100. Data from all experiments were evaluated using analysis of variance as outlined by Steel and Torrie (1960). Means were compared by Duncan's new multiple range test (1955). Experiment 1. To establish the broad requirement for vitamin D 3 by chicks when fed rye or rye supplemented with pectin enzyme, Experiment 1 was conducted. Also, we wanted to learn if there was an advantage to supplementing with Irgazyme 100 or Irgazyme 100Special. Day-old chicks were used in this experiment. The experiment consisted of 16 treatments with 3 pens of 5 chicks, 3 females and 2 males per treatment. All chicks were killed for bone ash analysis. Vitamin D 3 was added to the basal diet containing either rye or corn as the grain component at levels of 0, 67, 133, or 200 ICU/kg. Irgazyme 100 or Irgazyme 100-Special was added to the rye diets at levels of 0 or .1%. Experiment 2. To determine the requirement for vitamin D 3 when chicks were fed rye and supplemental pectic enzyme, Experiment 2 was conducted. Three-day-old chicks were used in this experiment. The experiment consisted of 21 treatment with 3 pens of 10 chicks, 5 females and 5 males, per treatment. Vitamin D 3 was added to the basal diet containing either rye or corn at the level of 0, 100, 200, 400, 800, 1200, or 1600 ICU/kg. Irgazyme 100 was added at the level of .1% to diets containing rye. Experiment 3. To define the requirements of vitamin D 3 , within narrow limits, for chicks fed rye, with or without supplemental pectic enzyme, Experiment 3 was conducted. Day-old chicks were randomly assigned to groups of 10, 6 females and 4 males. There were 15 treatments and 3 pens per treatment. Vitamin D 3 was added to the basal diets containing rye at the level of 0, 200, 400, 500, 600, 700, or 800 ICU/kg. Diets containing rye plus . 1 % Irgazyme or corn received 0, 100, 200, or 300 ICU vitamin D 3 /kg diet.
PECTIN ENZYME AND RYE FOR CHICKS RESULTS Experiment 1, Chicks fed rye, as compared to those fed corn, had reduced growth and bone ash, even though 200 ICU vitamin D3/kg diet were added (Table 2). Supplementation of the rye diet with . 1 % Irgazyme 100 or Irgazyme 100-Special significantly increased the growth rate. However, neither enzyme supplement allowed chicks fed rye to attain the maximum bone ash of controls fed corn. This is especially apparent with chicks fed 200 ICU vitamin D3/ kg diet. Experiment 2. Chicks fed corn showed no significant increase in weight or bone ash above 400 ICU of vitamin D 3 (Table 3). Chicks fed rye showed no increase in weight (P>.05) until the vitamin D 3 level was 800 ICU/kg diet. There were no further significant increases in weight with higher levels, however, The addition of . 1 % Irgazyme 100 to rye diets did permit gradually increasing weights until 200 ICU vitamin Da/kg were added. There were no increases (P>.05) in body weight among those chicks fed this diet with Irgazyme 100 supplemented with as high as 1600 ICU vitamin D 3 /kg.
1893
Within each diet type — corn, rye, or rye supplemented with Irgazyme-100 — bone ash increased, although nonsignificantly, with increasing levels of vitamin D 3 . Bone ash was the same (P>.05) at 0 and 100 ICU/kg vitamin D 3 for each type of diet and at several higher levels of vitamin D 3 ; corn and rye with Irgazyme-100 showed no significant differences. Chicks fed corn or rye with Irgazyme-100 reached a maximum bone ash value at 400 ICU vitamin D 3 /kg, whereas chicks fed rye without Irgazyme-100 did not maximize bone ash until they were fed 800 ICU vitamin D 3 /kg. There were no differences (P>.05) among the three diet types at the 800, 1200, or 1600 ICU/kg vitamin D 3 . Experiment 3, Table 4 lists the average 2week weights and percentage bone ash values for Experiment 3. Chicks fed corn supplemented with either 100, 200, or 300 ICU vitamin D 3 /kg were equal in weight (P> .05) and all were heavier (P<.05) than chicks fed corn without vitamin D 3 . A similar trend was observed when chicks were fed rye with Irgazyme-100. Chicks fed rye never reached the maximum weight attained by chicks fed corn. However, chicks fed rye with Irgazyme-100 did not differ
TABLE 2. Average chick weight and percentage bone ash for Experiment 1 Treatment
Grain
Irgazyme 100 (.1%)
Irgazyme100 Special (.1%)
_ — — —
_ — — — — — -
Vitamin D 3 (ICU/kg)
Corn Corn Corn Corn Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye !i n c n p
+ + + +
_ —
+ + + +
0 67 133 200 0 67 133 200 0 67 133 200 0 67 133 200
Average chick weight1 (g) 198.5cde 211.0 d e 187.3 c d 224.3 e 139.1 a 148.7 a b 149.4 a b 147.8 a b 171.7 b c 176.9 b c 193.3cd 192.5 c d 169.4 b c 196.9cde 186.0 c d 194.5cde
' ' ' ' Means within the same column with different letters differ significantly (P<.05). Error mean squares are 266.3 and 1.87, respectively, for chick weight and bone ash. 2 Bone ash is expressed as percent of the dry, fat-free bone. 1
Bone ash1-2
(%) 29.52abcd 3 1 91bcd 36.60 e 40.50 f 29.37abc 30.14abcd 3 ! 8 4 abcd 30.52abcd 29.30 a b 30.69abcd 31.97 c d 34.40 e 29.20 a 31.47abcd 3164abcd 32.13 d
1894
GRAMMER ET AL. TABLE 3. Average chick weight and percentage bone ash for Experiment 2 Treatment Irgazyme Mitamin D 3
Average chick weight1
Bone ash 1 ' 2
(ICU/kg)
(g)
(%)
159.9 b c 1 8 3 2 cdef 202.9 d e % 212.S8 hi 219.3 h i 233. l h i 215.1g h i 124.7 a 132.3 a 147.5 a b 146.5 a b 176.8 c d 173.9 b c 1 8 7 2 cdefg 146.9 a b 168.8 b c 183.5cdef 181.0 c d e 208.4 e % h i 210.9% h l 210.0% h i
26.68 a 32.92 b 36.14" 1 41.30% 42.25% 43.23g 43.058 27.51 a 31.93 b 34.46 b c 37.02 d e 41.35% 41.85% 42.32% 28.71 a 33.56 b c 39.53 e f 40.97% 42.68% 43.33g 42.95g
100
Grain
Corn Corn Corn Corn Corn Corn Corn Rye Rye Rye Rye Rye Rye Rye
Rye Rye Rye
Rye Rye Rye Rye
(.1%)
— — — — — + + + + + + +
0 100 200 400 800
1200 1600 0 100 200 400 800
1200 1600 0 100 200 400 800
1200 1600
> i • i • >e> ' Means within the same column with different letters differ significantly (P<.05). 1
Error mean squares are 220.8 and 2.77, respectively, for chick weight and bone ash.
2
Bone ash is expressed as percent of the dry, fat-free bone.
significantly in weight as compared to chicks fed corn at the 300 ICU/kg level of vitamin D3. There was an increase in bone ash values with increasing vitamin D 3 levels for chicks fed either corn or rye supplemented with Irgazyme. There were no significant differences between these two diet types at comparable vitamin D3 levels except at the 200 ICU/kg level. There was a general increase in bone ash values for chicks fed rye and increasing vitamin D 3 levels up to the 500 ICU/kg level. Chicks fed either corn or rye with Irgazyme attained maximum and equal (P>.05) bone ash values at 300 ICU vitamin D 3 /kg. DISCUSSION These experiments clearly demonstrate the growth-depressing effect of rye when fed to chicks. The maximum weight of chicks fed rye was consistently lower, although in one case
not significantly lower, than that of chicks fed corn at the same level of vitamin D 3 . The rachitogenic effect of rye, as measured by reduced bone ash and reported by MacAuliffe and McGinnis (1976), was verified. High levels of vitamin D 3 , two to four times the National Research Council (1971) recommendation, overcame both the rachitogenic and the growthdepressing effects. Supplementation of rye diets with Irgazyme 100 or Irgazyme 100-Special overcame the growth-depressing effect of rye as reported by Patel et al. (1980). There were no significant differences in growth between groups fed these two preparations. Supplementation with Irgazyme 100 or 100-Special also alleviated the rachitogenic effect of rye, although chicks fed rye with Irgazyme 100 had significantly higher bone ash values as compared to chicks fed rye with Irgazyme 100-Special. The Irgazyme 100 apparently is degrading some component(s) in
PECTIN ENZYME AND RYE FOR CHICKS
1895
TABLE 4. Average chick weight and percentage bone ash for Experiment 3 Treatment
Grain
Corn Corn Corn Corn Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye
Irgazyme 100 (.1%)
—
+ + + +
— -
Vitamin D 3
Average chick weight1
(ICU/kg)
(g)
0 100 200 300 0 100 200 300 0 200 400 500 600 700 800
192.4 b c 220.8 e f '229.9 f 221.3 e f 184.1 b 199.6 c d 212.1de 210.2 d e 153.0 a 183.8 b 200.2 c d 191.4 b c 182.6 b 187.1bc 192.8 b c
Bone ash1'2 (%) 29.05° 34.50cd 38.74f 41.74" 26.96a 33.05c 36.77e 40.75gh 29.43b 35.76 de 38.75f 40.8lS h 41.23 h 39.08fg 42.12h
3.bcdcf?h
' • ' ' • ' & ' Means within the same column with different letters differ significantly (P<.05). Error mean squares are 57.5 and 1.01, respectively, for chick weight and bone ash. 2 Bone ash is expressed as percent of the dry, fat-free bone. 1
rye and thus enables more efficient utilization of vitamin D 3 and possibly other nutrients. The manner in which the enzyme has its effect is still not clear. Possibly the enzyme reduces the viscosity of the gut contents, an effect demonstrated in vitro on rye water extract, which contains the factors responsible for the detrimental effects of rye. This explanation can be minimized, according to a recent report relating the nutritional qualities of barley varieties to the viscosity of the water extracts (Ullrich et al., 1981). These studies showed no correlation between chick growth and viscosity of barley extract and a low but positive correlation (r = .31) of extract viscosity and sticky feces. The viscosity of rye extract is lower than that of barley extract (M. H. Pubols, unpublished data). A second possible mechanism is that the enzyme alters a carbohydrate moiety and thus allows existence of a more beneficial microflora (MacAuliffe and McGinnis, 1971). The interaction of these dietary carbohydrates with other nutrients or cells of the intestinal lumen must be considered. For example, Ray et al. (1982) eliminated the growth-depressing effects of 2% guar gum in the diet by supplementation with a purified mannanase. Another
possibility is that Irgazyme-100 may be altering a complex carbohydrate or some other component and thus decreasing either specific or nonspecific binding of vitamin D 3 and, therefore, allowing increased absorption to occur. The supplemental enzyme clearly permits normal bone mineralization and greater chick growth when rye is fed. Maximal mineralization is attained with 200 to 300 IU of vitamin D 3 /kg diet when enzyme is added to the rye diet, but 500 IU/kg is required without enzyme (Table 4). The benefit of the enzyme is observed with bone mineralization before it is with maximal growth. It is uncertain if the factor(s) responsible for lower bone ash and growth rate can be attributed to the pectinlike compounds of rye, because the enzyme preparations are not pure. The enzyme preparation (Irgazyme 100) not only acts on pectin but also on guar gum (J. C. Grammer, unpublished results) and also may act on other substrates as well. Thus, this enzyme preparation may be degrading some other component(s) of rye, possibly not identified, which may be deleterious to the chick with respect to growth and bone formation. These experiments thus do not resolve the
GRAMMER ET AL.
1896
issue regarding the number and type of deleterious factors in rye, but we do show that an enzyme preparation can improve the utilization of rye by chicks with respect to both bone mineralization and growth. REFERENCES Association of Official Analytical Chemists, 1970. Official Methods of Analysis. 11th ed. Washington, DC. Day, R. M., and O. P. Thomas, 1980. Growth depression of chicks fed a crude rye extract containing pectic substances. Poultry Sci. 59:2754-2759. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics 11:1—42. MacAuliffe, T., and J. McGinnis, 1971. Effect of antibiotic supplements to diets containing rye on chick growth. Poultry Sci. 50:1130-1134. MacAuliffe, T., and J. McGinnis, 1976. Effect of ultraviolet light and oral vitamin D 3 on rachitic chicks fed diets containing either corn or rye. Poultry Sci. 55:2305-2309. MacAuliffe, T., A. Pietraszek, and J. McGinnis, 1976a. The effect of adding procaine penicillin to diets with different levels of vitamin D 3 on the performance of turkey poults. Poultry Sci. 55:183— 187. MacAuliffe, T., A. Pietraszek, and J. McGinnis, 1976b. Variable rachitogenic effects of grain and alleviation by extraction or supplementation with vitamin D 3 , fat and antibiotics. Poultry Sci. 55: 2142-2147. McNab, J. M., and D.W.F. Shannon, 1975. The nutritive value of triticale and rye for the laying hen. Br. Poultry Sci. 1 6 : 9 - 1 5 .
Moran, E. T., S. P. Lall, and J. D. Summers, 1970. Altering the proportion of rye to maize in the grain fraction of practical broiler rations: Effects on live performance, litter moisture, dressing yield and carcass quality. Br. Poultry Sci. 11: 147-152. National Research Council, 1971. Nutrient Requirements of Poultry. 6th rev. ed. Natl. Acad. Sci., Washington, DC. Patel, M. B., M. S. Jami, and J. McGinnis, 1980. Effect of gamma irradiation, penicillin, and/or pectic enzyme on chick growth depression and fecal stickiness caused by rye, citrus pectin and guar gum. Poultry Sci. 59:2105-2110. Patel, M. B., and J. McGinnis, 1976. The nutritive value of rye for chick growth: Effect of various levels, geographical area of production and procaine penicillin supplement. Poultry Sci. 55: 418-424. Ray, S., M. H. Pubols, and J. McGinnis, 1982. The effect of a purified guar degrading enzyme on chick growth. Poultry Sci. 61:488-494. Steel, R.G.D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Co., New York, NY. Ullrich, S. E., C. N. Coon, and J. S. Sever, 1981. Relationship of nutritional and malting quality traits of barley. Barley Genetics. IV. Proceedings of the Fourth International Barley and Genetics Symposium. Edinburgh, July 22—29. In Roger Whitehouse, ed. Vohra, P., and F. H. Kratzer, 1964. Growth inhibitory effect on certain polysaccharides for chickens. Poultry Sci. 43:1164-1170. Wagner, D. D., and O. P. Thomas, 1977. A rye type growth depression of chicks fed pectin. Poultry Sci. 56:615-619.
INTRODUCTION Rye has not been used commonly as a feedstuff in poultry rations. Recently, several workers have reported the growth-depressing effect of rye when it replaced corn in the rations of poultry. Moran et al. (1970) observed growth depression when rye constituted more than 25% of the grain component of the ration. MacAuliffe and McGinnis (1971) noted growth depression when rye replaced wheat as 40% of the diet. Patel and McGinnis (1976) noted depression at levels as low as 25%. The factor(s) in rye that causes reduced growth in chicks has not been identified. One proposal is that pectin-like components are responsible. McNab and Shannon (1975) reported that rye contains approximately 7.9% pectic material, substantially more than other cereal grains. Vohra and Kratzer (1964) observed the growth-depressing effect of citrus pectin when fed to chicks. Wagner and Thomas (1977) and Day and Thomas (1980) also noted this depression and speculated that due to the correlation of high levels of pectin-like components in rye and the growth-depressing effects of citrus pectin, that pectin was the component in rye responsible for growth depression. Patel et al. (1980) reported that a
'Scientific Paper No. 6072. College of Agriculture Research Center, Washington State University, Pullman, Washington. Project No. 0311.
commercial pectic enzyme, Irgazyme 100, increased the growth rate and feed efficiency of chicks fed rye. Rye also has been shown to contain a factor responsible " for decreasing vitamin D 3 utilization in poults and chicks (MacAuliffe et al, 1976a,b). MacAuliffe et al. (1976a) observed that a simple substitution of rye for corn in the diets fed to turkey poults resulted in the development of rickets by 10 days of age. The vitamin D 3 level in the diets either met or exceeded the recommendation of the National Research Council (1971). MacAuliffe et al. (1976b) observed the same effect in chicks fed a diet containing 63% rye. The rachitogenic effect could be alleviated by feeding increased levels of vitamin D 3 . In further studies, these investigators concluded that a vitamin D 3 antagonist in rye interfered with the absorption of vitamin D 3 and not with its further metabolism (MacAuliffe and McGinnis, 1976). Chicks fed vitamin D3-deficient diets with 63% rye but exposed to ultraviolet light showed no difference in bone ash as compared to controls fed corn. The addition of fat to the diet also reduced the severity of the rickets. Both of these observations suggest that rye interferes with the absorption and not the further metabolism of vitamin D 3 . Whether this rachitogenic factor is the same as or different from the factor responsible for growth depression is not known, but it appears that they are different, because excess vitamin D 3 permits bone mineralization but does not overcome growth depression.
1891
GRAMME R ET AL.
1892
A series of experiments was conducted to determine the effect that supplementation with the pectic enzyme, Irgazyme 100, would have on vitamin D 3 utilization in chicks fed rye. MATERIALS AND METHODS
General. One to 3-day-old, commercial broiler-type chicks (Hubbard strain) were randomly assigned to pens of either 5 or 10, as specified for each experiment. Chicks were housed in electrically heated, wire floored battery brooders. The temperature was maintained at 35 C for the first week and 24 C for the second week. Treatments were randomly assigned to pens and feed and water were supplied ad libitum. The composition of the basal diet is listed in Table 1; vitamin D 3 is omitted. All chicks were group weighed at 1 and 2 weeks. Upon termination of the experiments at 2 weeks, 5 chicks from each pen were killed and the left tibia was removed for bone ash analysis. Bone ash was determined according to the methods of the Association of Official Analytical Chemists (1970). All results were
TABLE 1. Composition of the basal diet for Experiments 1, 2, and 3 Ingredients
(%)
Grain a Fishmeal Meat and bone meal Whey Dehydrated alfalfa Iodized salt Dicalcium phosphate Limestone Vitamin premix D Trace mineral mix c DL-Methionine Soybean meal Animal fat
62.55 3.01 3.00 1.00 1.10 .23 .77 .90 .25 .05 .15 25.00 2.00
Total
99.91
Corn or rye. Vitamin premix at .25% of the diet supplies the following per kilogram of the diet: vitamin A, 5,500 IU; vitamin E, 4.4 IU; riboflavin, 3.3 mg; calcium pantothenate, 4.4 mg (or panthotenic acid, 4.05 mg); niacin, 22 mg; choline chloride, 577 mg; vitamin B 1 2 , .011 mg; and ethoxyquin, 62.2 mg. Trace mineral mix supplies the following in mg/kg of the diet: Mn, 100; Fe, 100; Zn, 100; Cu, 10; I, 3.0; Ca, 120; Co, 1.0.
expressed as percent ash of the fat-free, dry bone. Irgazyme 100 and Irgazyme 100-Special were obtained from the Food Industry Department of Ciba-Geigy Corporation, Greensboro, NC. They have about 90% diluent. The Irgazyme 100-Special contains amylase activity, which is not present in the Irgazyme 100. Data from all experiments were evaluated using analysis of variance as outlined by Steel and Torrie (1960). Means were compared by Duncan's new multiple range test (1955). Experiment 1. To establish the broad requirement for vitamin D 3 by chicks when fed rye or rye supplemented with pectin enzyme, Experiment 1 was conducted. Also, we wanted to learn if there was an advantage to supplementing with Irgazyme 100 or Irgazyme 100Special. Day-old chicks were used in this experiment. The experiment consisted of 16 treatments with 3 pens of 5 chicks, 3 females and 2 males per treatment. All chicks were killed for bone ash analysis. Vitamin D 3 was added to the basal diet containing either rye or corn as the grain component at levels of 0, 67, 133, or 200 ICU/kg. Irgazyme 100 or Irgazyme 100-Special was added to the rye diets at levels of 0 or .1%. Experiment 2. To determine the requirement for vitamin D 3 when chicks were fed rye and supplemental pectic enzyme, Experiment 2 was conducted. Three-day-old chicks were used in this experiment. The experiment consisted of 21 treatment with 3 pens of 10 chicks, 5 females and 5 males, per treatment. Vitamin D 3 was added to the basal diet containing either rye or corn at the level of 0, 100, 200, 400, 800, 1200, or 1600 ICU/kg. Irgazyme 100 was added at the level of .1% to diets containing rye. Experiment 3. To define the requirements of vitamin D 3 , within narrow limits, for chicks fed rye, with or without supplemental pectic enzyme, Experiment 3 was conducted. Day-old chicks were randomly assigned to groups of 10, 6 females and 4 males. There were 15 treatments and 3 pens per treatment. Vitamin D 3 was added to the basal diets containing rye at the level of 0, 200, 400, 500, 600, 700, or 800 ICU/kg. Diets containing rye plus . 1 % Irgazyme or corn received 0, 100, 200, or 300 ICU vitamin D 3 /kg diet.
PECTIN ENZYME AND RYE FOR CHICKS RESULTS Experiment 1, Chicks fed rye, as compared to those fed corn, had reduced growth and bone ash, even though 200 ICU vitamin D3/kg diet were added (Table 2). Supplementation of the rye diet with . 1 % Irgazyme 100 or Irgazyme 100-Special significantly increased the growth rate. However, neither enzyme supplement allowed chicks fed rye to attain the maximum bone ash of controls fed corn. This is especially apparent with chicks fed 200 ICU vitamin D3/ kg diet. Experiment 2. Chicks fed corn showed no significant increase in weight or bone ash above 400 ICU of vitamin D 3 (Table 3). Chicks fed rye showed no increase in weight (P>.05) until the vitamin D 3 level was 800 ICU/kg diet. There were no further significant increases in weight with higher levels, however, The addition of . 1 % Irgazyme 100 to rye diets did permit gradually increasing weights until 200 ICU vitamin Da/kg were added. There were no increases (P>.05) in body weight among those chicks fed this diet with Irgazyme 100 supplemented with as high as 1600 ICU vitamin D 3 /kg.
1893
Within each diet type — corn, rye, or rye supplemented with Irgazyme-100 — bone ash increased, although nonsignificantly, with increasing levels of vitamin D 3 . Bone ash was the same (P>.05) at 0 and 100 ICU/kg vitamin D 3 for each type of diet and at several higher levels of vitamin D 3 ; corn and rye with Irgazyme-100 showed no significant differences. Chicks fed corn or rye with Irgazyme-100 reached a maximum bone ash value at 400 ICU vitamin D 3 /kg, whereas chicks fed rye without Irgazyme-100 did not maximize bone ash until they were fed 800 ICU vitamin D 3 /kg. There were no differences (P>.05) among the three diet types at the 800, 1200, or 1600 ICU/kg vitamin D 3 . Experiment 3, Table 4 lists the average 2week weights and percentage bone ash values for Experiment 3. Chicks fed corn supplemented with either 100, 200, or 300 ICU vitamin D 3 /kg were equal in weight (P> .05) and all were heavier (P<.05) than chicks fed corn without vitamin D 3 . A similar trend was observed when chicks were fed rye with Irgazyme-100. Chicks fed rye never reached the maximum weight attained by chicks fed corn. However, chicks fed rye with Irgazyme-100 did not differ
TABLE 2. Average chick weight and percentage bone ash for Experiment 1 Treatment
Grain
Irgazyme 100 (.1%)
Irgazyme100 Special (.1%)
_ — — —
_ — — — — — -
Vitamin D 3 (ICU/kg)
Corn Corn Corn Corn Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye !i n c n p
+ + + +
_ —
+ + + +
0 67 133 200 0 67 133 200 0 67 133 200 0 67 133 200
Average chick weight1 (g) 198.5cde 211.0 d e 187.3 c d 224.3 e 139.1 a 148.7 a b 149.4 a b 147.8 a b 171.7 b c 176.9 b c 193.3cd 192.5 c d 169.4 b c 196.9cde 186.0 c d 194.5cde
' ' ' ' Means within the same column with different letters differ significantly (P<.05). Error mean squares are 266.3 and 1.87, respectively, for chick weight and bone ash. 2 Bone ash is expressed as percent of the dry, fat-free bone. 1
Bone ash1-2
(%) 29.52abcd 3 1 91bcd 36.60 e 40.50 f 29.37abc 30.14abcd 3 ! 8 4 abcd 30.52abcd 29.30 a b 30.69abcd 31.97 c d 34.40 e 29.20 a 31.47abcd 3164abcd 32.13 d
1894
GRAMMER ET AL. TABLE 3. Average chick weight and percentage bone ash for Experiment 2 Treatment Irgazyme Mitamin D 3
Average chick weight1
Bone ash 1 ' 2
(ICU/kg)
(g)
(%)
159.9 b c 1 8 3 2 cdef 202.9 d e % 212.S8 hi 219.3 h i 233. l h i 215.1g h i 124.7 a 132.3 a 147.5 a b 146.5 a b 176.8 c d 173.9 b c 1 8 7 2 cdefg 146.9 a b 168.8 b c 183.5cdef 181.0 c d e 208.4 e % h i 210.9% h l 210.0% h i
26.68 a 32.92 b 36.14" 1 41.30% 42.25% 43.23g 43.058 27.51 a 31.93 b 34.46 b c 37.02 d e 41.35% 41.85% 42.32% 28.71 a 33.56 b c 39.53 e f 40.97% 42.68% 43.33g 42.95g
100
Grain
Corn Corn Corn Corn Corn Corn Corn Rye Rye Rye Rye Rye Rye Rye
Rye Rye Rye
Rye Rye Rye Rye
(.1%)
— — — — — + + + + + + +
0 100 200 400 800
1200 1600 0 100 200 400 800
1200 1600 0 100 200 400 800
1200 1600
> i • i • >e> ' Means within the same column with different letters differ significantly (P<.05). 1
Error mean squares are 220.8 and 2.77, respectively, for chick weight and bone ash.
2
Bone ash is expressed as percent of the dry, fat-free bone.
significantly in weight as compared to chicks fed corn at the 300 ICU/kg level of vitamin D3. There was an increase in bone ash values with increasing vitamin D 3 levels for chicks fed either corn or rye supplemented with Irgazyme. There were no significant differences between these two diet types at comparable vitamin D3 levels except at the 200 ICU/kg level. There was a general increase in bone ash values for chicks fed rye and increasing vitamin D 3 levels up to the 500 ICU/kg level. Chicks fed either corn or rye with Irgazyme attained maximum and equal (P>.05) bone ash values at 300 ICU vitamin D 3 /kg. DISCUSSION These experiments clearly demonstrate the growth-depressing effect of rye when fed to chicks. The maximum weight of chicks fed rye was consistently lower, although in one case
not significantly lower, than that of chicks fed corn at the same level of vitamin D 3 . The rachitogenic effect of rye, as measured by reduced bone ash and reported by MacAuliffe and McGinnis (1976), was verified. High levels of vitamin D 3 , two to four times the National Research Council (1971) recommendation, overcame both the rachitogenic and the growthdepressing effects. Supplementation of rye diets with Irgazyme 100 or Irgazyme 100-Special overcame the growth-depressing effect of rye as reported by Patel et al. (1980). There were no significant differences in growth between groups fed these two preparations. Supplementation with Irgazyme 100 or 100-Special also alleviated the rachitogenic effect of rye, although chicks fed rye with Irgazyme 100 had significantly higher bone ash values as compared to chicks fed rye with Irgazyme 100-Special. The Irgazyme 100 apparently is degrading some component(s) in
PECTIN ENZYME AND RYE FOR CHICKS
1895
TABLE 4. Average chick weight and percentage bone ash for Experiment 3 Treatment
Grain
Corn Corn Corn Corn Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye Rye
Irgazyme 100 (.1%)
—
+ + + +
— -
Vitamin D 3
Average chick weight1
(ICU/kg)
(g)
0 100 200 300 0 100 200 300 0 200 400 500 600 700 800
192.4 b c 220.8 e f '229.9 f 221.3 e f 184.1 b 199.6 c d 212.1de 210.2 d e 153.0 a 183.8 b 200.2 c d 191.4 b c 182.6 b 187.1bc 192.8 b c
Bone ash1'2 (%) 29.05° 34.50cd 38.74f 41.74" 26.96a 33.05c 36.77e 40.75gh 29.43b 35.76 de 38.75f 40.8lS h 41.23 h 39.08fg 42.12h
3.bcdcf?h
' • ' ' • ' & ' Means within the same column with different letters differ significantly (P<.05). Error mean squares are 57.5 and 1.01, respectively, for chick weight and bone ash. 2 Bone ash is expressed as percent of the dry, fat-free bone. 1
rye and thus enables more efficient utilization of vitamin D 3 and possibly other nutrients. The manner in which the enzyme has its effect is still not clear. Possibly the enzyme reduces the viscosity of the gut contents, an effect demonstrated in vitro on rye water extract, which contains the factors responsible for the detrimental effects of rye. This explanation can be minimized, according to a recent report relating the nutritional qualities of barley varieties to the viscosity of the water extracts (Ullrich et al., 1981). These studies showed no correlation between chick growth and viscosity of barley extract and a low but positive correlation (r = .31) of extract viscosity and sticky feces. The viscosity of rye extract is lower than that of barley extract (M. H. Pubols, unpublished data). A second possible mechanism is that the enzyme alters a carbohydrate moiety and thus allows existence of a more beneficial microflora (MacAuliffe and McGinnis, 1971). The interaction of these dietary carbohydrates with other nutrients or cells of the intestinal lumen must be considered. For example, Ray et al. (1982) eliminated the growth-depressing effects of 2% guar gum in the diet by supplementation with a purified mannanase. Another
possibility is that Irgazyme-100 may be altering a complex carbohydrate or some other component and thus decreasing either specific or nonspecific binding of vitamin D 3 and, therefore, allowing increased absorption to occur. The supplemental enzyme clearly permits normal bone mineralization and greater chick growth when rye is fed. Maximal mineralization is attained with 200 to 300 IU of vitamin D 3 /kg diet when enzyme is added to the rye diet, but 500 IU/kg is required without enzyme (Table 4). The benefit of the enzyme is observed with bone mineralization before it is with maximal growth. It is uncertain if the factor(s) responsible for lower bone ash and growth rate can be attributed to the pectinlike compounds of rye, because the enzyme preparations are not pure. The enzyme preparation (Irgazyme 100) not only acts on pectin but also on guar gum (J. C. Grammer, unpublished results) and also may act on other substrates as well. Thus, this enzyme preparation may be degrading some other component(s) of rye, possibly not identified, which may be deleterious to the chick with respect to growth and bone formation. These experiments thus do not resolve the
GRAMMER ET AL.
1896
issue regarding the number and type of deleterious factors in rye, but we do show that an enzyme preparation can improve the utilization of rye by chicks with respect to both bone mineralization and growth. REFERENCES Association of Official Analytical Chemists, 1970. Official Methods of Analysis. 11th ed. Washington, DC. Day, R. M., and O. P. Thomas, 1980. Growth depression of chicks fed a crude rye extract containing pectic substances. Poultry Sci. 59:2754-2759. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics 11:1—42. MacAuliffe, T., and J. McGinnis, 1971. Effect of antibiotic supplements to diets containing rye on chick growth. Poultry Sci. 50:1130-1134. MacAuliffe, T., and J. McGinnis, 1976. Effect of ultraviolet light and oral vitamin D 3 on rachitic chicks fed diets containing either corn or rye. Poultry Sci. 55:2305-2309. MacAuliffe, T., A. Pietraszek, and J. McGinnis, 1976a. The effect of adding procaine penicillin to diets with different levels of vitamin D 3 on the performance of turkey poults. Poultry Sci. 55:183— 187. MacAuliffe, T., A. Pietraszek, and J. McGinnis, 1976b. Variable rachitogenic effects of grain and alleviation by extraction or supplementation with vitamin D 3 , fat and antibiotics. Poultry Sci. 55: 2142-2147. McNab, J. M., and D.W.F. Shannon, 1975. The nutritive value of triticale and rye for the laying hen. Br. Poultry Sci. 1 6 : 9 - 1 5 .
Moran, E. T., S. P. Lall, and J. D. Summers, 1970. Altering the proportion of rye to maize in the grain fraction of practical broiler rations: Effects on live performance, litter moisture, dressing yield and carcass quality. Br. Poultry Sci. 11: 147-152. National Research Council, 1971. Nutrient Requirements of Poultry. 6th rev. ed. Natl. Acad. Sci., Washington, DC. Patel, M. B., M. S. Jami, and J. McGinnis, 1980. Effect of gamma irradiation, penicillin, and/or pectic enzyme on chick growth depression and fecal stickiness caused by rye, citrus pectin and guar gum. Poultry Sci. 59:2105-2110. Patel, M. B., and J. McGinnis, 1976. The nutritive value of rye for chick growth: Effect of various levels, geographical area of production and procaine penicillin supplement. Poultry Sci. 55: 418-424. Ray, S., M. H. Pubols, and J. McGinnis, 1982. The effect of a purified guar degrading enzyme on chick growth. Poultry Sci. 61:488-494. Steel, R.G.D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Co., New York, NY. Ullrich, S. E., C. N. Coon, and J. S. Sever, 1981. Relationship of nutritional and malting quality traits of barley. Barley Genetics. IV. Proceedings of the Fourth International Barley and Genetics Symposium. Edinburgh, July 22—29. In Roger Whitehouse, ed. Vohra, P., and F. H. Kratzer, 1964. Growth inhibitory effect on certain polysaccharides for chickens. Poultry Sci. 43:1164-1170. Wagner, D. D., and O. P. Thomas, 1977. A rye type growth depression of chicks fed pectin. Poultry Sci. 56:615-619.