The Relative Biopotency of Fermentation Beta-Carotene, Crystalline Beta-Carotene and Vitamin A for Poultry1

PHOSPHATASE ISOZYMES AND PRODUCTION 1959. The Connecticut and Cornell random bred populations of chickens. World's Poultry Sci. J. 15: 139-159. Law, G. R., and S. S. Munro, 1965. Inheritance of two alkaline phosphatase variants in fowl plasma. Science, 149 : 1518. Wilcox, F. H., 1963. Genetic control of serum alkaline phosphatase in the chicken. J. Exp. Zool. 152: 195-204. Wilcox, F. H., 1966. A recessively inherited elec-

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trophoretic variant of alkaline phosphatase in chicken serum. Genetics, 5 3 : 799-805. Wilcox, F. H., L. D. Van Vleck and W. R. Harvey, 1963. Estimates of correlation between serum alkaline phosphatase level and productive traits. Poultry Sci. 42: 1457-1458. Wilcox, F. H., L. D. Van Vleck and C. S. Shaffner, 1962. Serum alkaline phosphatase and egg production. Proc. 12th World's Poultry Congress, Sydney, Australia: 19-22.

The Relative Biopotency of Fermentation Beta-Carotene, Crystalline Beta-Carotene and Vitamin A for Poultry1 AND H. H. HALL 4 Department of Poultry Science, Michigan State University, East Lansing, Michigan 43823 U.S. Department of Agriculture, Northern Research Laboratory, Peoria, Illinois (Received for publication July 23, 1969)

I

N 1957, research workers at the U. S. Department of Agriculture Northern Utilization Research and Development Division developed a procedure for producing beta-carotene (Hesseltine and Anderson, 1957; Anderson et al., 1958), by a fermentation process involving the mating of opposite types of the heterothallic mold, Blackeslea trispora. Beta-carotene is used as a source of vitamin A activity for poultry and thus the relative biopotency of this fermentation beta-carotene becomes of 1 Published with the approval of the Director of the Michigan State University Agricultural Experiment Station as Journal Article No. 4789. This research was conducted under contract No. 12-14-100-6850(71) with the Northern Utilization Research and Development Division, Agricultural Research Service, U.S. Department of Agriculture, Peoria, 111. 2 In partial fulfillment of the requirements for Ph.D. degree, Associate Professor, Department of Poultry Science. 3 Professor Emeritus. 4 U.S.D.A. Northern Regional Research Laboratory, Peoria, Illinois, (deceased).

economic importance. It is possible that this fermentation product may have an economic advantage in comparison with other sources of vitamin A. The following studies were designed and conducted to determine the relative biopotency of this newly-developed, beta-carotene product for growing broiler-type chicks. MATERIALS AND METHODS Experiment 1: This experiment was designed to determine a suitable depletion period for the chicks fed the vitamin A deficient diet and to compare three levels of dietary vitamin A activity for the following three products. Fermentation beta-carotene—This product was produced as dry fermentation solids and supplied by the United States Department of Agriculture Northern Regional Research Laboratory, Peoria, Illinois. Analysis by this laboratory showed that it contained 85 percent all-trans betacarotene, 6 percent neo-beta-carotene, and

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CAL J. FLEGAL2 AND PHILIP J. SCHAIBLE3

350

C. J . FLEGAL, P . J . SCHAIBLE AND H . H . HALL TABLE 1.—Composition of the basal ' 'et (Experiment 1)

Ingredient

Percent

White milo, ground Soybean meal, 44% protein Steamed bone meal Limestone, ground Salt, iodized Vitamin premix1

60.0 34.0 2.0 1.5 0.5 2.0

1 The premix contained the following per kilogram of ration with sufficient soybean meal to make up the 2 percent.

mg. riboflavin mg. dl calcium pantothenate mg. niacin mg. choline chloride mg. vitamin Bi 2 I.C.U. vitamin D 8 mg. procaine penicillin gm. MnS0 4

the remainder unidentifiable material. Crystalline beta-carotene—This was a commercial product containing 100 percent all-trans beta-carotene.1 Vitamin A Reference Standard—This product from the Animal Nutrition Research Council was obtained from Dr. S. Ames, Distillation Products Industries, Rochester, New York. It is a gelatin-stabilized, oil solution of all-trans vitamin A acetate with a mixture of butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) as an antioxidant. All products were incorporated into premixes by diluting the various products to the desired premix concentration (based on I.U. of vitamin A activity) with soybean meal containing two percent of a 2:3 mixture of butylated hydroxyanisole and butylated hydroxytoluene. These premixes were chemically analyzed and packaged under refrigeration at - 2 3 ° C . (-10°F.) until they were used. Commercially-obtained, one-day-old 1 Crystalline beta-carotene, all-trans, Biochemicals, Chagrin Falls, Ohio.

General

FIG. 1. Plasma vitamin A concentration (chicks).

Cobb's strain White Rock cockerels were placed into heated chick batteries, with raised wire floors, and given the basal depletion diet (Table 1) and water ad libitum. The initial brooding temperature of 90° to 9S°F. was decreased five degrees each week until room temperature (about 70°F.) was reached. At intervals, blood samples were taken from several chicks to determine plasma vitamin A concentration. After 17 days (Figure 1), plasma levels were very low. The chicks were then placed in equalized weight groups, wing-banded and three replications of ten birds each randomly distributed in the raised-wire chick batteries. The chicks were then fed the test diets for four weeks. Feed and water were supplied ad libitum, The intended and assayed vitamin A activity2 per kilogram of the test diets are shown in Table 2. 2 Vitamin A activity for the beta-carotene products is expressed as I.U. per kilogram and was

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440 1,100 2,750 4,400 1.1 8,800 440 2.2

351

BlOPOTENCY OF CAROTENE AND VlTAMIN A

based on the conversion of 0.6 microgram of betacarotene to one I.U. of vitamin A activity. 3 At the end of two weeks, individual blood samples were taken from only one replicate. Pooled samples were obtained from the other replicates.

TABLE 2.—Intended and assayed vitamin A concentration of the nine test diets (Experiment 1) I.U. Vitamin A Activity/kg. Treatment of basal ration None Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard

[ntended

Assayed1

0 880 1,540 2,695 880 1,540 2,695 880

0 1,131 1,839 3,153 1,206 1,927 3,197 1,186

1,540

1,802

2,695

2,853

1 Average of analytical data obtained from feed samples analyzed at the Northern Utilization Research and Development Division, U. S. Department of Agriculture.

on individual livers according to the methods described by Carr-Price (1926) and by Gallup and Hoefer (1946), using a Baush and Lomb Spectronic-20 spectrophotometer. Individual body weights, vitamin A blood plasma and liver concentrations and survival time in days were subjected to analysis of variance (Snedecor, 1956). Duncan's multiple range test (1955) was used to determine which means were significantly different. Experiment 2: This experiment was conducted in the same manner as Experiment 1 with the following exceptions: The one-dayold chicks were placed on the basal depletion diet for 15 days. Also, 0.53 mg. per kg. vitamin K, 44 mg. per kg. zinc and l\ percent cottonseed oil (at the expense of milo) were added to the basal depletion diet shown in Table 1. Blood samples for vitamin A assay were taken only after the chicks had been on the test diets for four weeks. The intended and assayed vitamin A concentrations of the test diets are shown in Table 3.

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The diets were mixed weekly. They were prepared by adding appropriate amounts of the beta-carotene or vitamin A premixes and the soybean meal-antioxidant diluent to the basal depletion diet. Approximately one-half of the weekly needs was fed immediately; the remainder was put in covered metal containers and refrigerated ( —10°F.). After one-half week, the feed not consumed was removed and replaced by the portion which had been kept refrigerated. Each diet was sampled at the time of mixing, at mid-week, and at the end of each week for determination of vitamin A activity. The samples were packaged under nitrogen in sealed containers and kept refrigerated ( —10°F.) until analyzed. The chicks were individually weighed at weekly intervals and feed consumption per lot was recorded. Blood samples were obtained by heart puncture from each chick3 at the end of the second and fourth weeks on the experimental diets for plasma vitamin A determinations. After having been on the test diets four weeks, one-half of the birds fed each diet were randomly selected and sacrificed. Their livers were removed and wrapped in Saran Wrap and aluminum foil and refrigerated at about — 10°F. until assayed for vitamin A. The remaining onehalf of the chicks were placed on the basal depletion ration and survival in days was recorded. The plasma vitamin A assays were made according to the method using antimony trichloride outlined by Yudkin (1941). Each liver was sectioned approximately at the middle line of all lobes and a portion from this area was removed for analysis. Vitamin A concentrations were determined

352

C. J . FLEGAL, P . J . SCHAIBLE AND H . H . HALL

TABLE 3.—Intended and assayed vitamin A concentration of the twelve test diets (Experiment 2)

Treatment of basal ration

Intended

Assayed1

0 1,760 2,464 3,450 4,829 1,760 2,464 3,450 4,829 1,760

0 1,683 2,277 3,443 4,554 1,782 2,398 3,322 4,510 2,266

2,464

2,904

3,450

3,498

4,829

4,598

1 Average of analytical data from feed samples analyzed at the Northern Utilization Research and Development Division, United States Department of Agriculture.

Experiment 3: The purpose of this experiment was to determine the relative effectiveness of the fermentation beta-carotene and a vitamin A product to influence vitamin A liver concentration when fed for a one-week period at relatively high feed levels. The sources of vitamin A activity in this trial were: Fermentation beta-carotene, previously described, Vitamin A palmitate—this product, obtained commercially,4 contained 250,000 I.U. alltrans vitamin A palmitate per gram plus edible tallow, gelatin, glucose, BHT, BHA, and soybean feed. Commercially-obtained, day-old, Cobb's strain White Rock cockerels were placed into heated, raised-wire chick batteries (as in the two previous experiments) and given the same depletion diet used in Experiment 2. After seven days, the birds were placed in equalized weight groups, wing-banded and four replications of ten birds each were "PGB-250 Dry Vitamin A Feed Supplement, Distillation Products Ind., Rochester, N.Y.

RESULTS AND DISCUSSION Experiment 1: The effect on plasma vitamin A concentration when day-old chicks were fed the basal depletion diet is graphically shown in Figure 1. The concentration dropped very rapidly and almost in a straight line during the first seven days from an initial value of 194 to about 43 micrograms per 100 ml. of plasma. The decline was less rapid from 7 to 14 days when the concentration dropped from about 43 to 5 micrograms per 100 ml. of plasma. The initial (day-old) plasma vitamin A concentration was higher than that reported by Squibb (1961) and lower than that recorded by Castano et al. (1951),

TABLE 4.—Intended and assayed vitamin A concentration of the four test diets (Experiment 3)

Treatment of basal ration

None Fermentation beta-carotene Fermentation beta-carotene Vitamin A palmitate Vitamin A palmitate

I. U. Vitamin A Activity/kg. of diet Intended

Assayed1

0 22,000 55,000 22,000 55,000

0 19,690 46,970 22,880 47,740

1 Average of analytical data from feed samples analyzed at the U. S. Department of Agriculture Northern Utilization Research and Development Division.

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None Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard

I.U. Vitamin A Activity/kg.

randomly distributed in the chick batteries. The test diets were mixed once. Preparation, storage and sampling procedures were similar to those previously mentioned. The intended and assayed vitamin A activity of the test diets are shown in Table 4. After one week on the test diets, all birds were sacrificed; livers were removed and wrapped in Saran Wrap and aluminum foil. They were refrigerated at approximately — 10°F. until assayed for vitamin A. Assay procedures and analytical interpretations were identical to those used in Experiments 1 and 2.

353

BlOPOTENCY OF CAROTENE AND VlTAMIN A TABLE 5.—Body weights and feed efficiency {Experiment 1)

Treatment of basal ration Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard

Vitamin A I.U./kg. (Assayed) 1,131 1.839 3,153 1,206 1,927 3,197 1,186 1,802 2,853

Mean body wts. (gms.)1 2 weeks 358.7 376.5 434.2 315.9 377.8 384.0 416.3 453.1 461.8

DEe Dde ABCbc Ef CDde CDde BCDcd ABabc ABabc

4 weeks 618.2 666.9 771.7 517.3 608.7 682.8 742.7 770.8 801.0

DEd CDd ABab Fe DEd BCDcd ABCbc ABab Aab

Four week feed efficiency ^Kg.reea/ kg. gain) 3.41 3.10 2.70 3.77 3.12 2.80 2.71 2.67 2.55

1 Any two means having the same letter are not significantly different; means not having the same letter are significantly different. Small letters indicate significance at the 0.05 level; large letters at the 0.01 level.

responding dietary level of the crystalline beta-carotene and the two lowest dietary levels (4 weeks) of the fermentation betacarotene. Feed efficiency was closely associated with rate of gain; the chicks with the faster rate of gain had the best feed efficiency. The use of the A.N.R.C. Vitamin A Reference Standard at all dietary levels resulted in a considerably improved feed conversion compared to that of either of the beta-carotene products. The effects of the test diets on the mean vitamin A plasma concentrations are shown in Table 6. In general, as the feed level of vitamin A activity was increased, plasma concentrations increased. In most instances, the plasma concentration was higher after four weeks on the test diets than after two weeks. After two weeks on the test diets, only the lowest levels of the fermentation beta-carotene and the crystalline beta-carotene failed to raise the plasma vitamin A concentration above the depleted value. However, after four weeks, all diets increased the plasma concentrations above the depleted value. Few significant plasma vitamin A concentrations were observed at two weeks. After four weeks on the test diets, the beta-carotene products resulted in similar vitamin A plasma con-

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while the concentration following depletion was about equal to that noted by Squibb (1961). These discrepancies were probably due to the difference in the amount of parental carry-over (Squibb, 1961). The effects of the test diets on weight gain and feed efficiency are shown in Table 5. Significant differences (P < 0.01) occurred in mean body weights after both 2 and 4 weeks. Both product and level of vitamin A activity had an influence on mean body weight. Mean body weights after both 2 and 4 weeks were larger as the vitamin A activity of the diet was increased for all of the products tested. The crystalline betacarotene fed at 1,206 I.U. per kilogram of ration resulted in mean body weights that were significantly smaller than those from all other diets. Both beta-carotene products resulted in weights that were not significantly different from each other at the two higher corresponding dietary levels of vitamin A activity. However, there was a trend toward larger mean body weights at all of the corresponding dietary levels of vitamin A activity when comparing the fermentation beta-carotene to the crystalline betacarotene. The A.N.R.C. Vitamin A Refence Standard resulted in significantly larger mean body weights (after both 2 and 4 weeks on the test diets) than at each cor-

354

C. J. FLEGAL, P . J . SCHAIBLE AND H . H . HALL TABLE 6.—Mean blood plasma vitamin A concentrations1 with different vitamin A sources and levels in the feed (Experiment 1) Blood plasma (Mcg./lOO ml.)

Treatment of basal ration

(Assayed I.U./kg.)

4 weeks

2 weeks Fermentation beta-carotene Fermentation beta-carotene F'ermentation beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard

1,131 1,839 3,153 1,206 1,927 3,197 1,186 1,802 2,853

16.0 Efg 18.9DEefg 30.5 Bb 15.4 Eg 18.2 DEefg 24.2 CDcd 21.1 CDEde 30.3 Bb 50.7 Aa

6.7 De 13.5 CDde 21.0BCDbcd 7.9 De 15.6 CDcd 14.2 CDde 21.9BCDbcd 28.1 ABCb 41.1 Aa

1 Any two means having the same letter are not significantly different; means not having the same letter are significantly different. Small letters indicate significance at the 0.05 level; large letters at the 0.01 level.

There was very little liver storage from any of the diets in this experiment (Table 7), although the highest level of the A.N.R.C. Vitamin A Reference Standard resulted in significantly higher liver vitamin A concentration than all other diets. There was also a trend toward longer survival (Table 7) as the level of the vitamin A activity in the diet increased from each of the products tested but few few significant differences were observed. Experiment 2: In this experiment, the plasma vitamin A concentrations (not shown) followed an almost identical pattern to that in Figure 1 and reached a low of about five micrograms per 100 ml. on

TABLE 7.—Mean liver vitamin A concentrations1 and survival periods with respect to different levels and types of vitamin A \ Treatment of basal ration Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard A.N.R.C. Vitamin A Reference Standard

Feed assaved (I.U./kg".)

Liver fresh basis (Mcg./gm.)

Survival (days until death)

1,131 1,839 3,153 1,206 1,927 3,197 1,186 1,802 2,853

.38 Dd .57CDd 1.14 BCbc .50CDd .50CDd .65 CDcd .47 CDd .86BCDbcd 3.32 Aa

26.2 ABCbcde 24.5 BCede 29.3 ABCabcd 21.6 Ce 23.4 BCde 28.9 ABCabcd 24.8BCcde 30.5 ABab 34.3 Aa

1 Any two means having the same letter are not significantly different; means not having the same letter are significantly different. Small letters indicate significance at the 0.05 level: large letters at the 0.01 level.

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centrations at the two lowest comparable levels. However, in comparing the beta-carotene products at the highest level, the crystalline beta-carotene produced significantly smaller (P < 0.01) plasma vitamin A concentrations. The A.N.R.C. Vitamin A Reference Standard caused significantly higher plasma concentrations at all dietary levels of vitamin A than either beta-carotene product. At 2,853 I.U. per kilogram of feed, the plasma vitamin A concentration was significantly higher than with any other diet. Gurcay et al. (1950) and Castano et al. (1951) obtained similar results in comparing vitamin A and beta-carotene in the capacity to influence vitamin A plasma levels in turkeys and chicks.

355

BlOPOTENCY OF CAROTENE AND VlTAMIN A and feed efficiency {Experiment 2)

TABLE 8.—Body -an

Feed assayed (I.U-Ag.) •

Treatment of basal ration

Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene A.N.R.C. Vitamin A Reference A.N.R.C. Vitamin A Reference A.N.R.C. Vitamin A Reference A.N.R.C. Vitamin A Reference

Standard Standard Standard Standard

1,683 2,277 3,443 4,554 1,782 ,398 3,322 4,510 2,266 2,904 3,498 4,598

Mean body weights (gms.) 2 weeks 427.3 ABabc 440.3 ABabc 435.4 ABabc 446.1 ABabc 406.8 Be 429.6 ABabc 422.8 ABbc 449.3 ABabc 457.6 Aab 458.8 Aab 464.3 Aa 453.4 ABab

4 weeks 743.6 788.5 807.9 807.6 707.2 747.4 761.6 833.2 818.6 846.0 814.9 835.2

CDcd ABCabc ABCab ABCab Dd CDcd BCDbcd ABa ABCab Aa ABCab ABa

Four week feed efficiency - (kg. feed/ kg. gain) 2.58 2.52 2.48 2.49 2.70 2.59 2.52 2.46 2.47 2.48 2.52 2.49

the fourteenth day after the chicks had been placed on the basal depletion ration. The effects of the test diets on weight gain and feed efficiency are evident from data in Table 8. After two weeks on the test diets, only the lowest level of the crystalline beta-carotene did not appear to support the rate of growth obtained with most of the other diets, but differences were not significant in most cases. After four weeks, the lowest level of the fermentation betacarotene and the three lowest levels of the crystalline beta-carotene did not support

the rate of gain obtained by the diet which supported the greatest weight gain (Basal + A.N.R.C. Vitamin A Reference Standard at 2,904 I.U. per kg.). All of the other diets resulted in mean body weights which were not significantly different. Feed efficiency was similar for chicks receiving all diets except that those receiving the lowest level of the crystalline beta-carotene had a somewhat poorer feed efficiency. The effects of the test diets on mean plasma vitamin A concentration are shown in Table 9 and Figure 2. All diets raised

TABLE 9.—Mean blood plasma vitamin A concentrations? liver vitamin A concentrations and survival of chicks {Experiment 2) Feed assayed (I.U./kg.)

Treatment of basal ration Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Fermentation beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene Crystalline beta-carotene A.N.R.C. Vitamin A Reference A.N.R.C. Vitamin A Reference A.N.R.C. Vitamin A Reference A.N.R.C. Vitamin A Reference

Standard Standard Standard Standard

1,683 2,277 3,443 4,554 1,782 2,398 3,322 4,510 2,266 2,904 3,498 4,598

Blood plasma Liver fresh basis (Mcg./lOO ml.) (Mcg./gm.) 28.6 Ggh 38.1 EFef 51.2 CDc 53.9 Cc 20.9 Hi 26.4 GHh 33.1 FGfg 44.8 DEd 41.3 Ede 52.7 Cc 64.5 Bb 76.1 Aa

1.22 Efg 1.47 DEfg 3.38 CDde 4.97 Cc .79 Eg 1.08 Efg 1.84 DEfg 2.49 DEef 2.28DEefg 4.66 Ccd 9.47 Bb 21.32 Aa

Survival (days until death) 34.7 39.1 45.7 44.0 25.0 39.2 34.8 42.1 40.3 48.1 51.4 48.0

BCc ABbc ABabc ABabc Cd ABbc BCc ABabc ABabc ABab Aa ABabc

1 Any two means having the same letter are not significantly different; means not having the same letter are significantly different. Small letters indicate significance at the 0.05 level; large letters at the 0.01 level.

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1 Any two means having the same letter are not significantly different; means not having the same letter are significantly different. Small letters indicate significance at the 0.05 level; larger letters at the 0.01 level.

356

C. J. FLEGAL, P. J. SCHAIBLE AND H. H. HALL

The effects of the test diets on mean liver vitamin A concentrations are shown in Ta70 ble 9 and Figure 3. There was a large variation in liver vitamin A concentration in chicks consuming the same diet. This has also been observed by other researchers 50 (Nestler et al., 1948; Guilbert and Hin6 shaw, 1934). In general, the liver vitamin 9 40 A concentration increased as the level of the vitamin A activity in the feed increased for all products tested. At the lowest dietary level of vitamin A activity, there were A = ANRC Vitamin A Reference Standard no significant differences in liver concentraB = Fermentation beta-carotene C = Crystalline beta-carotene tions from any of the products tested. However, at all other comparable levels, the liver vitamin A concentration obtained 1,650 2200 2,750 3,300 3,850 4,400 was significantly higher (P < 0.01) in the IU vitamin A per kilogram feed chicks receiving the A.N.R.C. Vitamin A FIG. 2. Mean plasma vitamin A Reference Standard than from either of the concentration (chicks). beta-carotene products. At all comparable the plasma vitamin A concentration above dietary levels, liver concentrations were the value for the depleted diet and the similar but slightly higher from the fermenplasma concentration increased when the tation product than from the crystalline dietary level of vitamin A activity in- beta-carotene. creased regardless of the product in the Using liver vitamin A concentrations affeed. At all comparable levels tested, the ter four weeks on the test diets as the critefermentation beta-carotene resulted in a ria, the relative biopotency of the fermensignificantly (P < 0.01) higher mean tation and the crystalline beta-carotenes plasma vitamin A concentration than did varied with the dietary levels when comthe crystalline beta-carotene. Each dietary increase in the crystalline beta-carotene reA = ANRC Vitamin A Reference Standard sulted in a corresponding increase in the B = Fermentation beta-carotene C = Crystalline beta-carotene plasma vitamin A concentration. This was not true with regards to the fermentation beta-carotene which did not result in such a straight line increase in plasma vitamin A S 15 concentration. At all comparable levels, the A.N.R.C. Vitamin A Reference Standard E" resulted in higher, although not always sig- £ 10 nificantly higher, vitamin A plasma values than did either of the beta-carotene products. When the dietary A.N.R.C. Vitamin A Reference Standard was increased from 3,498 I.U. to 4,598 LIT. per kilogram, the 1,650 2,200 2,750 3,300 3,850 4,400 IU vitamin A per kilogram feed plasma concentration did not increase correspondingly. FIG. 3. Vitamin A liver concentration (chicks). 80

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357

BlOPOTENCY OF CAROTENE AND VlTAMIN A

pared with the A.N.R.C. Vitamin A Reference Standard. The relative biopotency figured as described above was:

TABLE 10. Mean liver vitamin A concentrations1

{Experiment 3)

Treatment of basal ration Fermentation beta-carotene (I.U./kg. feed) Liver vitamin A concentrations: Values expressed as a percentage of those for similar feed concentrations of A.N.R.C. Vitamin A

3,443

4,554

64.5

35.7

23.3

Crystalline beta-carotene (I.U./kg.) 2,398 3,322 4,510

47.4

19.4

11.7

As the dietary vitamin A activity increased, the relative efficiency of the betacarotene products to influence liver vitamin A concentration was decreased. Table 9 shows the effects of placing the birds on the depletion diet after having been on the test diets for four weeks. There was a trend toward longer survival as the dietary vitamin A activity increased from all of the products tested. However, with the exception of the lowest level of the crystalline beta-carotene (1,782 I.U./kg.) which resulted in significantly shorter survival than all other diets, few significant differences in survival resulted. Experiment 3: The effects of the test diets on mean liver vitamin A concentration are shown in Table 10. Significant differences were observed between all diets. Using liver concentration of vitamin A as the criteria for comparison, the relative biopotency of the fermentation beta-carotene was 30.1 percent (19,690 I.U.) and 21.7 percent (46,970 I.U.) when compared to the alltrans vitamin A palmitate. These experiments confirm the observation of many researchers (Wilson et al., 1936; Record et al., 1937; Reynolds et al., 1947; Olsen et al., 1939; Marusich et al,

19,690 46,970 22,880 47,740

Liver, fresh basis (Mcg./gm.) 18.5 37.6 61.4 173.2

Dd Cc Bb Aa

1 Any two means having the same letter are not significantly different; means not having the same letter are significantly different. Small letters indicate significance at the 0.05 level; large letters at the 0.01 level.

1961) that as far as growth rate of chicks is concerned, carotene is as efficient as vitamin A but at higher levels of feeding, where liver storage of vitamin A is measurable, carotene performs less effectively than vitamin A. SUMMARY

Studies were conducted to determine the relative biopotency of a fermentation betacarotene product. Day-old broiler-type cockerels were successfully grown at an optimum rate of growth and feed efficiency when the fermentation beta-carotene supplied the only source of vitamin A activity. The fermentation beta-carotene compared favorably with a commercial crystalline all-trans beta-carotene in its influence on growth, feed efficiency and plasma and liver vitamin A concentration. At levels of vitamin A activity which support optimum chick growth, beta-carotene appeared to be as effective as the A.N.R.C. Vitamin A Reference Standard. However, both beta-carotene products, when fed at levels high enough to induce plasma concentration and liver storage, were inferior to the A.N.R.C. Vitamin A Reference Standard. A large variability in plasma and liver vitamin A concentration in chicks consuming the same diet was observed. Optimum growth was obtained without high levels of plasma and/or liver vitamin A concentra-

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Liver vitamin A concentrations: Values expressed as a percentage of those for similar feed concentrations of A.N.R.C. Vitamin A

2,277

Fermentation beta-carotene Fermentation beta-carotene Vitamin A palmitate Vitamin A palmitate

Feed assayed (I.U./kg.)

358

C. J . FLEGAL, P . J . SCHAIBLE AND H . H . HALL

tion(s). There was a different requirement for vitamin A for optimum growth, blood concentration, and liver concentration.

The Effects of Microwave Heating on the Properties of Raw Unextracted Soybeans for Utilization by the Chick1 M. A. GUSTAFSON, JR., 2 C. J. FLEGAL3 AND P. J. SCHAIBLE4 Department of Poultry Science, Michigan State University, East Lansing, Michigan 48823 (Received for publication August 28. 1969)

N

UMEROUS workers have reported on various methods of heating unextracted soybeans for use in chick diets. The methods investigated include steam cooking 1 Published with the approval of the Director of the Michigan State University Agricultural Experiment Station as Journal Article No. 4845. 2 Graduate Research Assistant. 3 Associate Professor. 4 Professor Emeritus.

at various pressures, extruding and infrared cooking. Combs (I960), Rogler and Carrick (1961) and Runnels (1961) reported on the utilization of steam cooking in the processing of unextracted soybeans. Other reports have been comprehensively reviewed by Featherston and Rogler (1966) and White et al. (1967). White et al. (1967) investigated infrared heating, autoclaving and extrusion as meth-

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REFERENCES Anderson, R. F., M. Arnold, G. E. N. Nelson and A. Ciegler, 1958. Microbial production of betacarotene in shaken flasks. J. Agr. Food Chem. 6: 543-545. Carr, F. H., and E. A. Price, 1926. Color reactions attributed to vitamin A. Biochem. J. 20: 497. Castano, F. F., R. V. Boucher and E. W. Callenbach, 1951. Utilization by the chick of vitamin A from different sources. I. Crystalline carotene, crystalline vitamin A acetate and "black cod" liver oil. J. Nutr. 45: 131-141. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 1 1 : 1-42. Gallup, W. D., and J. A. Hoefer, 1946. Determination of vitamin A in liver. Ind. Eng. Chem. Annal. 18: 288-298. Guilbert, H. R., and W. R. Hinshaw, 1934. Vitamin A storage in the livers of turkeys and chickens. J. Nutr. 8:45-56. Gurcay, R., R. V. Boucher and E. W. Callenbach, 1950. Utilization of vitamin A by turkey poults. J. Nutr. 4 1 : 565-582. Hesseltine, C. W., and R. F. Anderson, 1957. Microbiological production of carotenoids. I. Zygospores and carotene produced by intraspecific and interspecific crosses of choanephoraceal in

liquid media. Mycologia, 49: 449-452. Marusich, W. L., R. J. Krukar and J. C. Bauernfeind, 1961. |3-carotene as a vitamin A source of poultry. Poultry Sci. 40: 1428. Nestler, R. B., J. V. Derby and J. B. DeWitt, 1948. Storage by bobwhite quail of vitamin A fed in various forms. J. Nutr. 36: 323-329. Olsen, E. M., J. D. Harvey, D. C. Hill and H. D. Branion, 1959. Utilization and stability of commercial vitamin A supplements. Poultry Sci. 38: 929-942. Record, P. R., R. M. Bethke and O. H. M. Wilder, 1937. The vitamin A requirements of chicks with observations on the comparative efficiency of carotene and vitamin A. Poultry Sci. 16: 25-33. Reynolds, J. B., D. B. Parrish, H. L. Mitchell and L. F. Payne, 1947. The utilization of a stabilized carotene by chickens. Poultry Sci. 27: 182-185. Snedecor, G. W., 1956. Statistical Methods Applied to Experiments in Agriculture. 5th ed., Ames Iowa State College Press. Squibb, R. L., 1961. Relation of serum vitamin A activity levels of hens to reserves in the progeny. Poultry Sci. 40: 1197-1204. Wilson, W. O., C. H. Schroeder and W. A. Higgens, 1936. Further studies on the vitamin A requirements of chicks. Poultry Sci. 15: 426. Yudkin, S., 1941. Estimation of vitamin A and carotenes in human blood. Biochem. J. 35: 551-556.