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Effect of Gamma Irradiation, Fractionation, and Penicillin Supplementation on the Rachitogenic Activity of Rye for Chicks1
Effect of Gamma Irradiation, Fractionation, and Penicillin Supplementation on the Rachitogenic Activity of Rye for Chicks1 TOMAS Mac-AULIFFE, DOUGLAS ZAVIEZO, and JAMES McGINNIS Department of Animal Sciences, Washington State University, Pullman, Washington 99164 (Received for publication August 4, 1978) ABSTRACT Experiments were conducted to study the effect of gamma irradiation on the ryevitamin D antagonism in broiler chicks. In an initial study, the irradiated grain was exposed to gamma rays for 6 hr (2 Mrad) and was fed to rachitic chicks for only 12 hr before a single oral dose of vitamin D 3 . In another trial, birds were fed during the entire experiment (12 days) on the different experimental diets, and the irradiated grain was exposed to gamma rays for 20 hr. Results indicate that the factor in rye that interferes with vitamin D utilization was largely inactivated by exposing this grain to gamma rays for 20 hr. Additional information is reported in this paper related to the alleviation by extraction or antibiotic supplementation on the rachitogenic properties of rye. These studies were conducted by feeding to rachitic chicks the test materials for only a short period of time (12 hr) before a vitamin D 3 oral dose and measuring the bone mineralization 48 hr later. Results indicate that the rachitogenic factor in rye is not present in the ash portion of the grain, that it can be largely overcome by water extraction and penicillin supplementation, and that an organic solvent extraction has no effect. Cooked beans which depress growth and increase the growth response to antibiotics are not rachitogenic. 1979 Poultry Sci 58:329-332 INTRODUCTION At t h e present t i m e , all t h e irradiation of foods o r feedstuffs is believed t o be in t h e preservation b y sterilization of these. However, irradiation of food or feedstuffs has frequently been shown t o affect their nutritional value, and m o s t studies have been directed t o observe any possible harmful effect of it. Damage of a m i n o acids—especially cystine—by g a m m a irradiation has often been reported ( J o h n s o n and Metta, 1956). Based o n t h e fact t h a t c a r b o h y d r a t e s can b e affected by high-energy irradiation, Moran et al. ( 1 9 6 8 ) studied t h e effect of Cobalt-60 g a m m a irradiation on t h e utilization of energy of w h e a t bran b y t h e chicken. T h e y found t h a t t h e metabolizable energy was n o t affected b y irradiation of t h e b r a n . Recently, studies cond u c t e d in o u r laboratories (Patel a n d McGinnis, u n p u b l i s h e d ) have shown t h e effectiveness of g a m m a irradiation with Cobalt-60 in improving t h e p o o r nutritional value of rye for broiler chicks.
1 Scientific Paper No. 4825. College of Agriculture Research Center, Washington State University. Project 0107.
T h e presence of a c o m p o n e n t in r y e t h a t interferes with t h e utilization of vitamin D3 b y broiler chicks has b e e n reported (Mac-Auliffe et al, 1 9 7 6 ) . Moreover, Zaviezo et al. ( 1 9 7 6 ) rep o r t e d a short-term bioassay t o s t u d y t h e utilization of vitamin D 3 and also t h e r y e vitamin D 3 interference by broiler chicks. T h e latter offers an easy way to study t h e rachitogenic activity of rye b y using small a m o u n t s of test material, and a series of e x p e r i m e n t s were c o n d u c t e d t o s t u d y t h e effects of gamma-irradiation, fractionation, a n d antibiotic s u p p l e m e n t a t i o n of diets containing rye o n t h e b o n e mineralization of broiler chicks.
EXPERIMENTAL PROCEDURE All studies were c o n d u c t e d using day-old H u b b a r d (mixed sexes) chicks from stock k e p t at Washington State University. Birds were assigned a t r a n d o m t o different pens in an electrically heated b a t t e r y b r o o d e r . Feed and water were provided ad libitum and 14 h r of artificial light given daily. Composition of t h e basal diet w i t h o u t vitam i n D 3 is presented in Table 1. All fractions obtained were i n c o r p o r a t e d in diets for chicks at levels a p p r o x i m a t e l y equivalent t o 6 3 % of rye. T h o s e chicks given an oral dosage of vita329
330
MAC-AULIFFE ET AL. TABLE 1. Composition of the basal diet
Ingredient
(%)
Grain1 Soybean meal, dehulled Herring fish meal Meat and bone meal Dried whey Dehydrated alfalfa Limestone Salt, iodized Dicalcium phosphate Vitamin premix 2 Mineral premix 3 D, L-methionine
63.54 22.09
5.00 2.50 2.00 2.50 .73 .30 1.00 .25 .05 .04
1
The grain component in the diet was either corn or rye. 2 Supplied the following per kg of feed: vitamin A, 5,500 IU; riboflavin, 3.3 mg: choline CI, 580 mg; Ca pantothenate, 3.3 mg; niacin, 22 mg; ethoxyquin, 62.15 mg; B, 2 , .011 mg; and vitamin E, 4.4 IU. 'Supplied the following mg/kg of feed: Mn, 49.5; Fe, 49.5; Cu, 4.95; Zn, 49.5; 1, 1.5; and Co, .5.
min D 3 received 4 4 ICU each of a water suspension of this vitamin at 10 days of age using a c r o p i n t u b a t i o n t e c h n i q u e . T h e dose was administered 2 hr before t h e lights were t u r n e d off in t h e r o o m . Forty-eight h o u r s later, nine chicks from each t r e a t m e n t were sacrificed and their right tibia removed for b o n e ash analysis. T h r e e g r o u p s of t h r e e b o n e s were analyzed and t h e results expressed as p e r c e n t ash of t h e fatfree, d r y b o n e following t h e A.O.A.C. ( 1 9 7 0 ) m e t h o d of analysis. Data from all e x p e r i m e n t s were t r e a t e d b y analysis of variance as outlined b y Steel and Torrie ( 1 9 6 0 ) and t h e differences among treatments compared by Duncan's new multiple range test ( 1 9 5 5 ) . Experiment I. All chicks were fed t h e same c o r n - t y p e , vitamin D 3 free diet (Table 1). When t h e chicks were 10 days old, t h e y were fed t h e test material (Table 2) for 12 h r from t h e m o m e n t t h e lights came on ( 0 7 0 0 h r ) . Twelve h o u r s later, t h e y received t h e oral dose and were p u t b a c k o n t h e corn diet. T h e process of b e n z e n e e x t r a c t i o n of rye was as follows: r y e was finely g r o u n d , m i x e d with 2 volumes of b e n z e n e a n d stirred for half an h o u r . T h e m i x t u r e was air dried, r e g r o u n d , and mixed with t h e basal prem i x . R y e ash was o b t a i n e d b y simply b u r n i n g
Nuclear Radiation Center at Richland, WA.
TABLE 2. Experiment 1', effect of irradiation of the grain component of the diet on bone mineralization of rachitic chicks using a short-term assay (12 hr) Bone ash (%) 12 days 44 ICU D 3 dose at 10 days
Test material fed for 12 hr
+ bc
Corn Corn 2 Rye Rye 2 Rye 3 Corn + rye ash
36.5 a 36.9 a 33.9 b 34.0 b 33.4 b 37.l a
31.6 29.5 C 29.3 C 30.8 C 32.0b 29.9 C
1 Chicks were fed a vitamin D 3 deficient diet containing corn the first 10 days and this same diet after test materials were given. 2 Gamma-irradiated for 6 hr. 3 Benzene extracted.
' ,c Means having the same letter(s) do not differ significantly (P = .05 or less).
t h e ground grain at 6 0 0 C for 4 h r in a muffle furnace. T h e irradiated grain was ground and e x p o s e d t o C o b a l t - 6 0 2 for 6 h r (approximately 2 Mrad). Experiment 2. In this trial, birds were fed during t h e entire e x p e r i m e n t o n t h e different experimental diets (Tables 3 and 4 ) , and t h e irradiated grain was exposed t o C o b a l t - 6 0 2 for 2 0 hr ( a p p r o x i m a t e l y 10 M r a d ) . T h e rest of t h e p r o c e d u r e was t h e same used for E x p e r i m e n t 1. Experiment 3. In this trial, chicks were m a d e rachitic on a basal diet (Table 1) containing r y e .
TABLE 3. Experiment 2, effect of irradiation of the grain component of the diet on body weight of rachitic chicks using a long-term assay (12 days)
Diet (no vitamin D)
Body weight (g) 10 days
Body weight (g) 12 days 44 ICU D 3 dose at 10 days +
Corn Corn 1 Rye Rye 1
a
130.3 107.0 b 95.5 b 130.7 a
170.2 142.7 114.2 157.9
164.7 134.7 120.1 161.7
ab ' Means having the same letter do not differ significantly (P = .05 or less). 1 Gamma-irradiated for 20 hr.
GAMMA IRRADIATION AND RACHITOGENIC FACTORS IN RYE TABLE 4. Experiment 2, effect of irradiation of the grain component of the diet on bone mineralization of rachitic chicks, using a long-term assay (12 days)1
Diet (no vitamin D)
Corn Corn2 Rye Rye2 1,2
Bone ash (%) 10 days
34.7 37.3 31.7 32.4
Bone ash (%) 12 days 44ICU D3 dose at 10 days
33.7C 36.2° 31.0d 31.0d
38.8 ab 41.l a 33.6C 37.2b
See footnotes for Table 3.
At 14 days, they were fed the test material (Table 5) for 12 hr from the moment the lights came on (0700 hr). Twelve hours later, they received the oral dose and were put back on the rye diet. The process of water extraction of rye has been described by Fernandez et al. (1973). Experiment 4. In this last trial, birds were made rachitic on a basal diet (Table 1) containing corn. At 10 days, they were fed the test material (Table 5). The rest of the procedure was the same used for Experiment 3. RESULTS Experiment I. Chicks that were made rachitic on a corn diet during the first part of the trial (10 days) and were fed diets containing corn, gamma irradiated corn, or corn plus rye ash for 12 hr before a single oral dose of vitamin D 3 responded significantly better in bone mineralization to a dose of vitamin D3 when compared with chicks that received diets during the 12 hr period that contained rye, rye irradiated with Cobalt-60 for 6 hr, or benzene-extracted rye. Bone ash was measured 48 hr after the oral dose of vitamin D 3 was administered (Table 2). Experiment 2. Chicks that were made rachitic on different diets (Table 3) differed significantly in body weight at 10 days. The growth depression of chicks fed corn that had been irradiated is attributed to a rancidity of the fat content of this grain detected easily by a smelling test. However, a body weight increase was obtained by irradiating rye, and we were unable to detect any rancid odor, probably because this grain contains a relatively small amount of fat. No differences were observed in
331
TABLE 5. Experiment 3, vitamin D 3 oral dose on bone ash of chicks fed rye or fractionated rye diets' Test material fed for 12 hr
Bone ash (%) 16 days 44 ICU D3 dose at 14 days
Rye Corn+RWS2 Corn + WER3
29.3C 28.5C 27.7C
32.4b 31.6b 34.4a
1 Chicks were fed a vitamin D3 deficient diet containing rye the first 14 days and this same diet after test materials were given. 2 RWS = water-soluble fraction of rye (10% fraction). 3 WER = water-extracted rye (90% fraction). ' ' Means having the same letter do not differ significantly (P = .05 or less).
body weight due to the administration of a single dose of vitamin D 3 , and the differences among treatments observed at 10 days were evident at 12 days of age. Rachitic chicks that were given a single oral dose of vitamin D 3 at 10 days showed a highly significant increase in bone ash when they were fed a diet containing rye irradiated for 20 hr, corn, or irradiated corn. Rachitic chicks on a rye diet throughout the trial responded significantly less to the oral dose of vitamin D 3 than those on the above treatments. Experiment 3. Chicks that were made rachitic on a rye diet during the first part of the trial (14 days) and were fed diets containing rye, or the water soluble fraction of rye, for 12 hr before a single oral dose of vitamin D 3 responded significantly less in bone mineralization when compared with chicks that received the water-extracted rye fraction during this 12 hr period (Table 5). Experiment 4. Chicks that were made rachitic on a corn diet and were fed a diet containing rye for 12 hr before a D 3 oral dose responded significantly more in bone mineralization when 50 ppm of procaine penicillin was supplemented in comparison with the nonsupplemented rye treatment (Table 6). A numerical nonsignificant response was observed when the corn and cooked bean treatments were supplemented with the antibiotic.
DISCUSSION According to the results obtained in Experi-
332
MAC-AUL1FFE ET AL.
TABLE 6. Experiment 4, bone mineralization of rachitic chicks fed different test material with and •without an antibiotic supplement1 Test material fed for 12 hr
Bone ash (%) 12 days 44 ICU D 3 dose at 10 days
Corn Corn + Procaine penicillin Rye Rye + Procaine penicillin Beans (cooked) Beans (cooked) + Procaine penicillin
27.0 d e
+
32.0°
de
27.2 26.3 e
32.9 a b 30.2 C
26.5 e 27.6 d e
32.5ab 33.4 a b
28.6 c d
34.0 a
Chicks were fed a vitamin D 3 deficient diet containing corn the first 10 days and this same diet after test materials were given. a,D,c, . e M e a n s having the same letter(s) do not differ significantly (P = .05 or less).
m e n t s 1 and 2, t h e factor in rye t h a t interferes with vitamin D3 utilization can be largely inactivated b y exposing this grain t o g a m m a rays for 2 0 hr. Moreover, it is entirely possible t h a t t h e rachitogenic factor in rye is directly related t o t h e growth-depressing factor in this grain. Even t h o u g h w e d o n o t k n o w h o w gamma-irradiation alleviates this effect, it is already k n o w n t h a t rye interferes with t h e absorption of vitamin D3 a t t h e gastro-intestinal site (Zaviezo et al, 1 9 7 6 ) . T h e rachitogenic factor in rye was largely removed b y water e x t r a c t i o n b u t n o t b y b e n z e n e . This result is in a g r e e m e n t w i t h F e r n a n d e z et al. ( 1 9 7 3 ) , whose results indicate t h a t a c e t o n e e x t r a c t i o n of rye had n o effect o n t h e growth-depressing properties of this grain.
T h e y also reported t h a t chicks receiving diets containing water e x t r a c t e d rye grew significantly b e t t e r , and t h e lyophilized r y e water extract was growth-depressing. T h e addition of procaine penicillin t o t h e rye-containing diet, fed for only 12 hr before a vitamin D3 oral dose was administered t o rachitic chicks, overcame t h e rachitogenic effect of rye and s u p p o r t e d b o n e mineralization as well as t h a t o b t a i n e d b y feeding c o r n . Finally, c o o k e d beans did n o t interfere with t h e vitamin D utilization b y t h e chick.
REFERENCES Association of Official Agricultural Chemists, 1970. Official methods of analysis. 11th ed. Washington, DC. Duncan, D. B., 1955. Multiple range and multiple F tests, 1955. Biometrics 11:1—42. Fernandez, R., E. Lucas, and J. McGinnis, 1973. Fractionation of a chick growth-depressing factor from rye. Poultry Sci. 52:2252-2259. Johnson, B. C , and V. C. Metta, 1956. The effect of radiation sterilization on the nutritive value of foods. 1. Biological value of milk and beef proteins. J. Nutr. 59:479-490. Mac-Auliffe, T., A. Pietraszek, and J. McGinnis, 1976. Variable rachitogenic effects of grain and alleviation by extraction or supplementation with vitamin D, fat and antibiotics. Poultry Sci. 55:2142— 2147. Moran, Jr., E. T., J. D. Summers, and H. S. Bayley, 1968. Effect of Cobalt-60 gamma-irradiation on the utilization of energy, protein and phosphorous from wheat bran by the chicken. Cereal Chem. 45; 469-479. Patel, M., and J. McGinnis, 1977. Effect of Cobalt-60 gamma irradiation of grains on growth performance of broiler chicks. (Unpublished). Steel, R. G. D., and J. H. Torrie, 1960. Principles and procedures of statistics. McGraw-Hill Book Co., New York, NY. Zaviezo, D., T. Mac-Auliffe, and J. McGinnis, 1976. Bioassay for vitamin D 3 antagonism and its site of interference using broiler chicks. (Unpublished).
1 Scientific Paper No. 4825. College of Agriculture Research Center, Washington State University. Project 0107.
T h e presence of a c o m p o n e n t in r y e t h a t interferes with t h e utilization of vitamin D3 b y broiler chicks has b e e n reported (Mac-Auliffe et al, 1 9 7 6 ) . Moreover, Zaviezo et al. ( 1 9 7 6 ) rep o r t e d a short-term bioassay t o s t u d y t h e utilization of vitamin D 3 and also t h e r y e vitamin D 3 interference by broiler chicks. T h e latter offers an easy way to study t h e rachitogenic activity of rye b y using small a m o u n t s of test material, and a series of e x p e r i m e n t s were c o n d u c t e d t o s t u d y t h e effects of gamma-irradiation, fractionation, a n d antibiotic s u p p l e m e n t a t i o n of diets containing rye o n t h e b o n e mineralization of broiler chicks.
EXPERIMENTAL PROCEDURE All studies were c o n d u c t e d using day-old H u b b a r d (mixed sexes) chicks from stock k e p t at Washington State University. Birds were assigned a t r a n d o m t o different pens in an electrically heated b a t t e r y b r o o d e r . Feed and water were provided ad libitum and 14 h r of artificial light given daily. Composition of t h e basal diet w i t h o u t vitam i n D 3 is presented in Table 1. All fractions obtained were i n c o r p o r a t e d in diets for chicks at levels a p p r o x i m a t e l y equivalent t o 6 3 % of rye. T h o s e chicks given an oral dosage of vita329
330
MAC-AULIFFE ET AL. TABLE 1. Composition of the basal diet
Ingredient
(%)
Grain1 Soybean meal, dehulled Herring fish meal Meat and bone meal Dried whey Dehydrated alfalfa Limestone Salt, iodized Dicalcium phosphate Vitamin premix 2 Mineral premix 3 D, L-methionine
63.54 22.09
5.00 2.50 2.00 2.50 .73 .30 1.00 .25 .05 .04
1
The grain component in the diet was either corn or rye. 2 Supplied the following per kg of feed: vitamin A, 5,500 IU; riboflavin, 3.3 mg: choline CI, 580 mg; Ca pantothenate, 3.3 mg; niacin, 22 mg; ethoxyquin, 62.15 mg; B, 2 , .011 mg; and vitamin E, 4.4 IU. 'Supplied the following mg/kg of feed: Mn, 49.5; Fe, 49.5; Cu, 4.95; Zn, 49.5; 1, 1.5; and Co, .5.
min D 3 received 4 4 ICU each of a water suspension of this vitamin at 10 days of age using a c r o p i n t u b a t i o n t e c h n i q u e . T h e dose was administered 2 hr before t h e lights were t u r n e d off in t h e r o o m . Forty-eight h o u r s later, nine chicks from each t r e a t m e n t were sacrificed and their right tibia removed for b o n e ash analysis. T h r e e g r o u p s of t h r e e b o n e s were analyzed and t h e results expressed as p e r c e n t ash of t h e fatfree, d r y b o n e following t h e A.O.A.C. ( 1 9 7 0 ) m e t h o d of analysis. Data from all e x p e r i m e n t s were t r e a t e d b y analysis of variance as outlined b y Steel and Torrie ( 1 9 6 0 ) and t h e differences among treatments compared by Duncan's new multiple range test ( 1 9 5 5 ) . Experiment I. All chicks were fed t h e same c o r n - t y p e , vitamin D 3 free diet (Table 1). When t h e chicks were 10 days old, t h e y were fed t h e test material (Table 2) for 12 h r from t h e m o m e n t t h e lights came on ( 0 7 0 0 h r ) . Twelve h o u r s later, t h e y received t h e oral dose and were p u t b a c k o n t h e corn diet. T h e process of b e n z e n e e x t r a c t i o n of rye was as follows: r y e was finely g r o u n d , m i x e d with 2 volumes of b e n z e n e a n d stirred for half an h o u r . T h e m i x t u r e was air dried, r e g r o u n d , and mixed with t h e basal prem i x . R y e ash was o b t a i n e d b y simply b u r n i n g
Nuclear Radiation Center at Richland, WA.
TABLE 2. Experiment 1', effect of irradiation of the grain component of the diet on bone mineralization of rachitic chicks using a short-term assay (12 hr) Bone ash (%) 12 days 44 ICU D 3 dose at 10 days
Test material fed for 12 hr
+ bc
Corn Corn 2 Rye Rye 2 Rye 3 Corn + rye ash
36.5 a 36.9 a 33.9 b 34.0 b 33.4 b 37.l a
31.6 29.5 C 29.3 C 30.8 C 32.0b 29.9 C
1 Chicks were fed a vitamin D 3 deficient diet containing corn the first 10 days and this same diet after test materials were given. 2 Gamma-irradiated for 6 hr. 3 Benzene extracted.
' ,c Means having the same letter(s) do not differ significantly (P = .05 or less).
t h e ground grain at 6 0 0 C for 4 h r in a muffle furnace. T h e irradiated grain was ground and e x p o s e d t o C o b a l t - 6 0 2 for 6 h r (approximately 2 Mrad). Experiment 2. In this trial, birds were fed during t h e entire e x p e r i m e n t o n t h e different experimental diets (Tables 3 and 4 ) , and t h e irradiated grain was exposed t o C o b a l t - 6 0 2 for 2 0 hr ( a p p r o x i m a t e l y 10 M r a d ) . T h e rest of t h e p r o c e d u r e was t h e same used for E x p e r i m e n t 1. Experiment 3. In this trial, chicks were m a d e rachitic on a basal diet (Table 1) containing r y e .
TABLE 3. Experiment 2, effect of irradiation of the grain component of the diet on body weight of rachitic chicks using a long-term assay (12 days)
Diet (no vitamin D)
Body weight (g) 10 days
Body weight (g) 12 days 44 ICU D 3 dose at 10 days +
Corn Corn 1 Rye Rye 1
a
130.3 107.0 b 95.5 b 130.7 a
170.2 142.7 114.2 157.9
164.7 134.7 120.1 161.7
ab ' Means having the same letter do not differ significantly (P = .05 or less). 1 Gamma-irradiated for 20 hr.
GAMMA IRRADIATION AND RACHITOGENIC FACTORS IN RYE TABLE 4. Experiment 2, effect of irradiation of the grain component of the diet on bone mineralization of rachitic chicks, using a long-term assay (12 days)1
Diet (no vitamin D)
Corn Corn2 Rye Rye2 1,2
Bone ash (%) 10 days
34.7 37.3 31.7 32.4
Bone ash (%) 12 days 44ICU D3 dose at 10 days
33.7C 36.2° 31.0d 31.0d
38.8 ab 41.l a 33.6C 37.2b
See footnotes for Table 3.
At 14 days, they were fed the test material (Table 5) for 12 hr from the moment the lights came on (0700 hr). Twelve hours later, they received the oral dose and were put back on the rye diet. The process of water extraction of rye has been described by Fernandez et al. (1973). Experiment 4. In this last trial, birds were made rachitic on a basal diet (Table 1) containing corn. At 10 days, they were fed the test material (Table 5). The rest of the procedure was the same used for Experiment 3. RESULTS Experiment I. Chicks that were made rachitic on a corn diet during the first part of the trial (10 days) and were fed diets containing corn, gamma irradiated corn, or corn plus rye ash for 12 hr before a single oral dose of vitamin D 3 responded significantly better in bone mineralization to a dose of vitamin D3 when compared with chicks that received diets during the 12 hr period that contained rye, rye irradiated with Cobalt-60 for 6 hr, or benzene-extracted rye. Bone ash was measured 48 hr after the oral dose of vitamin D 3 was administered (Table 2). Experiment 2. Chicks that were made rachitic on different diets (Table 3) differed significantly in body weight at 10 days. The growth depression of chicks fed corn that had been irradiated is attributed to a rancidity of the fat content of this grain detected easily by a smelling test. However, a body weight increase was obtained by irradiating rye, and we were unable to detect any rancid odor, probably because this grain contains a relatively small amount of fat. No differences were observed in
331
TABLE 5. Experiment 3, vitamin D 3 oral dose on bone ash of chicks fed rye or fractionated rye diets' Test material fed for 12 hr
Bone ash (%) 16 days 44 ICU D3 dose at 14 days
Rye Corn+RWS2 Corn + WER3
29.3C 28.5C 27.7C
32.4b 31.6b 34.4a
1 Chicks were fed a vitamin D3 deficient diet containing rye the first 14 days and this same diet after test materials were given. 2 RWS = water-soluble fraction of rye (10% fraction). 3 WER = water-extracted rye (90% fraction). ' ' Means having the same letter do not differ significantly (P = .05 or less).
body weight due to the administration of a single dose of vitamin D 3 , and the differences among treatments observed at 10 days were evident at 12 days of age. Rachitic chicks that were given a single oral dose of vitamin D 3 at 10 days showed a highly significant increase in bone ash when they were fed a diet containing rye irradiated for 20 hr, corn, or irradiated corn. Rachitic chicks on a rye diet throughout the trial responded significantly less to the oral dose of vitamin D 3 than those on the above treatments. Experiment 3. Chicks that were made rachitic on a rye diet during the first part of the trial (14 days) and were fed diets containing rye, or the water soluble fraction of rye, for 12 hr before a single oral dose of vitamin D 3 responded significantly less in bone mineralization when compared with chicks that received the water-extracted rye fraction during this 12 hr period (Table 5). Experiment 4. Chicks that were made rachitic on a corn diet and were fed a diet containing rye for 12 hr before a D 3 oral dose responded significantly more in bone mineralization when 50 ppm of procaine penicillin was supplemented in comparison with the nonsupplemented rye treatment (Table 6). A numerical nonsignificant response was observed when the corn and cooked bean treatments were supplemented with the antibiotic.
DISCUSSION According to the results obtained in Experi-
332
MAC-AUL1FFE ET AL.
TABLE 6. Experiment 4, bone mineralization of rachitic chicks fed different test material with and •without an antibiotic supplement1 Test material fed for 12 hr
Bone ash (%) 12 days 44 ICU D 3 dose at 10 days
Corn Corn + Procaine penicillin Rye Rye + Procaine penicillin Beans (cooked) Beans (cooked) + Procaine penicillin
27.0 d e
+
32.0°
de
27.2 26.3 e
32.9 a b 30.2 C
26.5 e 27.6 d e
32.5ab 33.4 a b
28.6 c d
34.0 a
Chicks were fed a vitamin D 3 deficient diet containing corn the first 10 days and this same diet after test materials were given. a,D,c, . e M e a n s having the same letter(s) do not differ significantly (P = .05 or less).
m e n t s 1 and 2, t h e factor in rye t h a t interferes with vitamin D3 utilization can be largely inactivated b y exposing this grain t o g a m m a rays for 2 0 hr. Moreover, it is entirely possible t h a t t h e rachitogenic factor in rye is directly related t o t h e growth-depressing factor in this grain. Even t h o u g h w e d o n o t k n o w h o w gamma-irradiation alleviates this effect, it is already k n o w n t h a t rye interferes with t h e absorption of vitamin D3 a t t h e gastro-intestinal site (Zaviezo et al, 1 9 7 6 ) . T h e rachitogenic factor in rye was largely removed b y water e x t r a c t i o n b u t n o t b y b e n z e n e . This result is in a g r e e m e n t w i t h F e r n a n d e z et al. ( 1 9 7 3 ) , whose results indicate t h a t a c e t o n e e x t r a c t i o n of rye had n o effect o n t h e growth-depressing properties of this grain.
T h e y also reported t h a t chicks receiving diets containing water e x t r a c t e d rye grew significantly b e t t e r , and t h e lyophilized r y e water extract was growth-depressing. T h e addition of procaine penicillin t o t h e rye-containing diet, fed for only 12 hr before a vitamin D3 oral dose was administered t o rachitic chicks, overcame t h e rachitogenic effect of rye and s u p p o r t e d b o n e mineralization as well as t h a t o b t a i n e d b y feeding c o r n . Finally, c o o k e d beans did n o t interfere with t h e vitamin D utilization b y t h e chick.
REFERENCES Association of Official Agricultural Chemists, 1970. Official methods of analysis. 11th ed. Washington, DC. Duncan, D. B., 1955. Multiple range and multiple F tests, 1955. Biometrics 11:1—42. Fernandez, R., E. Lucas, and J. McGinnis, 1973. Fractionation of a chick growth-depressing factor from rye. Poultry Sci. 52:2252-2259. Johnson, B. C , and V. C. Metta, 1956. The effect of radiation sterilization on the nutritive value of foods. 1. Biological value of milk and beef proteins. J. Nutr. 59:479-490. Mac-Auliffe, T., A. Pietraszek, and J. McGinnis, 1976. Variable rachitogenic effects of grain and alleviation by extraction or supplementation with vitamin D, fat and antibiotics. Poultry Sci. 55:2142— 2147. Moran, Jr., E. T., J. D. Summers, and H. S. Bayley, 1968. Effect of Cobalt-60 gamma-irradiation on the utilization of energy, protein and phosphorous from wheat bran by the chicken. Cereal Chem. 45; 469-479. Patel, M., and J. McGinnis, 1977. Effect of Cobalt-60 gamma irradiation of grains on growth performance of broiler chicks. (Unpublished). Steel, R. G. D., and J. H. Torrie, 1960. Principles and procedures of statistics. McGraw-Hill Book Co., New York, NY. Zaviezo, D., T. Mac-Auliffe, and J. McGinnis, 1976. Bioassay for vitamin D 3 antagonism and its site of interference using broiler chicks. (Unpublished).