Animal Feeding Tests

phosphate, but equal response was not registered by the Sudan grass. ... feeding tests using rats and chicks as experimental animals to determine: the...
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INDUSTRIAL AND ENGINEERING CHEMISTRY

LIMESTONE

PULVERIZER

PULVERIZER

Vol. 41, No. 7

ment for the limestone and ferrophosphorus; a rotary kiln for roasting of the mixture; and pulverization, size classification, and bagging equipment. It is estimated that production would be economically attractive, and i t is believed that the process offers a profitable outlet for by-product ferrophosphorus. A patent has been obtaincd on this process and assigned to TVA (9). ACKhOWLEDGBIENT

The exploratory rat-feeding tests were carried out by R. B. L. Fleming of the TVA Health and Safety Department, DUST and the other animal-feeding tests were carried out by Dorothy E. TT’illiams, p”L>JERlzER$/ FlorenceL. MacLeod, and Homer Patrick of the University of Tennessee Agricultural Experiment Station. The greenPRODUCT zhouse tests xere carried out by IT7. H. BAGGING MACHINE McJntirc and S. H. Winterberg of the University of Tennessee Agricultural Figure 1. Flow Diagram of Proposed Process ExDeriment Stations. Assistance in the laboratory and pilot plant work was phosphate, but equal response was not registered by the Sudan received from G. M. Blouin, J. R. Gray, I. A. Hobbs, J. S. Lewis, grass. These exploratory results indicate that the limestoneR. Lockheed, C. E. Mumma, and J. R. Tusson. ferrophosphorus product has possibilities as a fertilizer. HowLITERATURE CITED ever, since the product appears less favorable in comparison with (1) Association of American Feed Control Officials, Inc., Official Pubstandard fertilizer materials than n-ith standard feed-supplement lication (1947). materials of higher commercial value, i t is believed that the (2) Assoc. Official Agr. Chemist,s, “Methods of Snalysie,” 5t,h ed., product is more promising as a feed supplement than as a fertiWashington, D. C., 1940. lizer, (3) Bridger, G. L., U. S.Patent 2,320,342 (June 1, 1943). (4) Mitchell, H. E€., ;Vat. Research Council (U.S.), Reprint and Circ.

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I

I

DESCRIPTION OF PROPOSED PROCESS

A flo~vdiagram of a process proposed for the manufacture of the limestone-ferrophosphorus product is shown in Figure 1. The process would consist of pulverizing and proportioning equip-

Ser. 113 (June 1942). ( 5 ) Revnolds, D. 9..Hill, W.L., arid Jacob, K. D., J . Assoc. Oficial Agr. Chemists, 27, 559-71 (1944). (6) Williams, D. E., et al., IKD. ER’G. CHEM.,41, 1396 (1949). RECEIWDFebruary 2 , 1948.

(Phosphatic Animal-Feed Supplement)

ANIMAL FEEDING TESTS D. E. WILLIAMS, F, L. NIAcLEOD, ELISE MORRELL, AND HOiCIER PATRICK The University of Tennessee Agricultural Experiment Station, Knoxville, Tenn., in cooperation w i t h the Tennessee Valley Authority, Wilson Dam, Ala.

D

URING the war the supply of bone meal was short due t o lack of transport facilities between the United States and the exporting countries. The need for substitute phosphate carriers for mineral feed supplements was acute. Although postwar conditions have rdieved the situation somewhat, there is still need for additional sources of such supplement in animal feeding. The roasting with limestone of a by-product of phosphorus production a t Wilson D a m ( 1 ) resulted in a product which contains approximately 20% phosphorus pentoxide, 40% calcium oxide, 35y0ferric oxide, 4% silicon dioxide, and less than 0.10% fluorine. The phosphorus pentoxide is 97 to 99% soluble in 0.4% hydrochloric acid. Tests with the crude unprocessed material showed that the phosphorus was practically unavailabIe to both plants

and animals. Preliminary r a t feeding experiments using the material roasted with limestone reported by Fleming, Swayne, and Patterson (3) gave evidence that some of its phosphorus was available to animals. It was decided to repeat and extend the feeding tests using rats and chicks as experimental animals to determine: the degree of availability of the phosphorus of the roasted product as compared to other phosphate supplements; and the increased level necessary to produce the same results as other supplements if a lower availability occurred. RAT FEEDING TESTS

PRELIMINARY GROTVTH TESTS. Five white rats of the same litter, sex, and approximate weight were put on each of three

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INDUSTRIAL AND ENGINEERING CHEMISTRY

July 1949

T h e preceding article describes how a waste material containing chiefly a compound of iron and phosphorus in a form almost completely unavailable to plants and animals has been converted to a Iimestone-ferrophosphorus product. In this form approximately three fourths of the phosphorus has been shown to be available for phosphorus retention to experimental animals as compared with the readily available phosphorus of a salt mixture. Feeding the product at increased levels effectively counteracts this partial unavailability. Since there is a need for additional sources of phosphatic feed supplements in animal feeding, this material may prove useful if it can be produced cheaply enough to compete with more concentrated sources of phosphorus.

diets in which the chief source of phosphorus was from the limestone-ferrophosphorus product, Osborne and Mendel salt mixture, and a 0,37&fluorine fused tricalcium phosphate, respectively. Each diet v-as fed a t three different phosphorus levels, 0.23, 0 35, and 0.46%. The level of 0.23% is just about minimal for normal growth and bone development. Data concerning the composition of the test materials are given in Table I. Food consumption for comparable animals fed a t any one phosphorus level was kept uniform by restricting the food fed to the amount consumed by the rat which ate the least. The diet containing the Osborne and Mendel salt mixture was the control diet since t h e phosphorus from this source is readily available to animals, T h e control diet contained the following percentage composition: starch 64, lactalbumin 18, Osborne and Mendel salt mixture 4,vegetable fat 8, cod liver oil 2, Vitab 4,and 1 mg. of riboflavin per 100 grams of diet.

TABLEI.

COMPOSITION OF MATERIALS

Limestone-ferrophosphorus Fused trioaloium phosphate (0.3% F) Osborne & lMendel salt mixtnre Control 0 23% P level Control 0 . 3 6 % & 0,4670P level Experimental

P, % 8.56 12.45

Ca, %

F. %

28.0 30.29

0 06

0.3

..

4.06 6.95 0

14.46 11.42

0.03 0.03

.. ..

o

0.03

% 35.5

Fe208,

In the experimental diets the Osborne and Mendel salt mixture was modified by leaving out both phosphorus and calcium. When the phosphorus was supplied by the limestone-ferrophosphorus, 1.9% was required t o bring the total phosphorus content of the diet to the 0.23% level; 3.3% for the 0.35% level; and 4.3% for the 0.4601, level. When the phosphorus was supplied by the fused tricalcium phosphate, 1.3% was required to bring the total phosphorus content to the 0.23% level; 2.2% for the 0.35% level; and 3.0% for the 0.46% level. Similar calcium t o phosphorus ratios were obtained by adding small amounts of calcium carbonate or potassium acid phosphate when necessary. Table I 1 gives the phosphorus, calcium, and fluorine contents of the diets. Growth and phosphorus storage during the 30-day experimental period were used as criteria for judging results. Student’s t-test was applied to all results. Table I11 presents data showing the relative effectiveness of limestone-ferrophosphorus, Osborne and Mendel salt mixture, and a fused tricalcium phosphate containing 0.3% fluorine. The limestone-ferrophosphorus proved to be 91 to 99% as effective for growth as the Osborne and Mendel salt mixture a t all phosphorus levels. Except for the 0.46% phosphorus level, the differences in effectiveness could be said to be due to individual variation since statistically they were not significant.

For phosphorus storage over the 30-day experimental period, the limestone-ferrophosphorus was found to be 77% a s effective as the Osborne and Mendel salt mixture a t the 0.23% phosphorus level; 90% a t the 0.35% level; and 87% a t the 0.46% phosphorus level, The. differences in this case cannot be considered to be due to chance since they are statistically significantthat is, the phosphorus of the test product apparently is not complete€y available for utilization by the rat. The extent of this lack of availability is indicated by the effectiveness of the material a t the minimal phosphorus level in comparison with the readily available phosphorus of the Osborne and Mendel salt mixture, When the total phosphorus level was raised to 0.35 or 0.46% the difference in availability was much less apparent. Although a t these levels the ferrophosphorus material was 90 and 87%, respectively, as effective as the Osborne and Mendel salt mixture, the small differences were statistically significant and the effectiveness was slightly less a t the higher phosphorus level. This raises the question of the possibility of some slightly adverse effect on phosphorus metabolism due to something besides the unavailability of the component phosphorus. Possibly some of the large amount of iron that is present is not bound up in an insoluble form and a t these higher levels of incorporation of the product is sufficient to interfere slightly with phosphorus utilization. This interpretation, however, is not borne out by results obtained by Ellis et al. (2) in their tests of this product (designated by them as phosphate slag No. 2282) when it was added a t the phosphorus levels of 0.26, 0.39, and 0.78%, respectively, to a basal diet containing 0.04% phosphorus. Comparing the effectiveness in terms of femur ash of the limestoneferrophosphorus product with their average results ( A ) with a salt mixture and ( B ) with steamed bone meal, the test product was 77 and 80% as effective a t the 0.26% phosphorus level; 85 and 88% a t the 0.39% level; and 102 and 106% a t the 0.78% level. The close agreement a t comparable levels of their results with these tests using a different criterion for judging results is noteworthy. Moreover, when statistical analyses are made comparing the authors’ results obtained a t the 0.35% level with those a t the 0.46% level, the differences which in most cases are in favor of the 0.35% level are found to be not significant. The conclusion of the authors, therefore, is that the apparent less effectiveness a t the 0.46% level is due to individual variations among the experimental animals and not t o the presence of some deleterious substance. Greenhouse studies made with similar products by MacIntire ( 4 ) are of distinct interest. He finds that the crude ferrophosphorus material is practically insoluble and its phosphorus only slightly available to plants. When roasted with limestone, however, the resultant product shows a high degree of solubility. It compares favorably in phosphorus pentoxide uptake in plants with superphosphate and in general is slightly superior t o a fused tricalcium phosphate of 100 mesh. On the basis of the results of the preliminary feeding experiments done a t Wilson Dam, the results of the extension of those studies in the Nutrition Laboratory a t Knoxville, and the supple-

TABLE 11. PHOSPHORUS, CALCIUM, AND FLUORINE CONTENT OF DIETS P level 0.23% Con&ol Ca FePa

P.T.P.b

P level, 0.35%

p, % 0.24 0.23 0.24

Ca, %

0.60 0.46 0.59

0.36 Control 1.08 Ca FeP 0.35 0.84 0.99 F.T.P. 0.37 P level 0.46% Conirol 0.44 1.20 0.43 Ca FeP 1.05 F.T.P. 0.46 1.33 a C a F e P = limestone-ferrophosphorus. b F.T.P. = fused tricaloium phosphate. ~

C a : P Ratio

F, P.P.M.

2.5 2.0 2.5

11 22

3.0 2.4 2.7

11 30 78

2.7 2.4 2.9

11 36 99

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Vol. 41, No. 7

food records were kept. Table Is' shows the reTABLE 111. RELATIVEEFFECTIVENESS OF LIMESTONE-FERROPHOSPHORUS, sults obtained using the same criteria as for the OSBORIiE AtiD MENDEL SlLT R'hXTURE (COXTROL), AND FUSED TRICALCIUM preliminary tests. PHOSPHATE AS SOURCE OF PHOSPHORUS FOR WHITE RATS These results indicate that when rats are t Mean for Five R a t s with Effecallowed to eat these diets ad libitum, the limestoneDifference Standard Error, Grains , tiveness, % Value ferrophosphorus is not so effective a source of GROWTHD U R I N G 3 0 - D a ~PERIOD phosphorus for growth and phosphorus retention Control os. C a PepQ Control Ca FeP 91 1 . 7 1 Not significant 46.0 t 9 . 8 0.23% P 60.4 i 8 . 5 as the inorganic phosphate of Osborne andMendeP 99 0 . 6 4 Kot significant 83.6 * 3 . 0 84.8 * 4.1 0.35% P 91 3 . 2 9 Significant 74.6 * 3 . 6 0.46% P 82.0 * 3.9 salt mixture, but is as effective as the commonly Controlus. F.T.P.b Control F.T.P. used feed supplements, bone meal and dicalcium 88 4 . 7 0 Significant 44.2 * 8.4 0.23% P 50.4 * 8 . 5 phosphate. 89 2 . 8 3 Significant 75.6 k 2 . 8 0.35% P 84.8 t 4 . 1 91 8 . 2 4 Significant 3.9 74.4 * 4 . 1 0.46% P 82.0 Since the diet containing the crudc ferrophosCa FePus. F.T.P. Ca F e P F.T.P. phorus material was not so effective for growth 96 0.76 Not significant 44.2 t 8 . 4 0.23% P 46.0 1 9 . 8 90 2 . 5 6 K o t significant 75.6 * 2 . 8 83.6 1 3.0 0.35% P nor phosphorus retention as the basal diet, it 99 0.11 h-ot significant 74.4 1 4 . 1 0.46% P 74.6 1 3 . 6 seemed that not only was its phosphorus practiNormal according to 85 cally unavailable to the animal but that it exerted Shcr&n B Quinn (6) a definitely adverse effect. It could not be deP H O S P H O R U S RETEKTIOK D U R I K G 30-D.4Y P E R I O D termined from these evperiments whether the Control os. Ca F e P Control Ca FeP 77 6 . 9 2 Significant 0 . 3 3 8 * 0.026 0.260 =t0.027 0.23% P adverse effects were due to toxicity of the crude 90 2 . 7 7 Significant 0.035 0.440 * 0 . 0 2 9 0.35% P 0.488 material or to secondary limiting factors due 87 6.74 Significant 0.46% P 0.494 * 0 . 0 2 3 0 . 4 3 1 * 0.016 to the marked decrease in food consumption on Control u s . F.T.P. Control F.T.P. 72 12.09 Significant 0.23% P 0.338 * 0.026 0 . 2 4 4 * 0 . 0 2 3 this diet. It seemed important t o test for the 87 2 . 9 7 Significant 0.488 * 0 . 0 3 6 0.426 * 0 . 0 2 0 0.3570 P 82 6.74 Bignificsnt 0 , 4 9 4 * 0.023 0.406 * 0.033 0.46% P possibility of toxicity of the crudc material in C a F e P cs. F.T.P. Ca F e P F.T.P. case unforeseen conditions occurrcd during the 0.244 t O . 0 2 3 94 1 . 2 1 Notsignificant 0.260 d O . 0 2 7 0,23% P processing which might result in the inadvertent 97 1.24 Not,significant 0.36% P 0.440 t 0 . 0 2 9 0.426 * 0 . 0 2 0 94 1 . 0 3 Not, significant 0 . 4 3 1 i.O.016 0.406 = 0 . 0 3 3 0.46% P inclusion of the crude product along with the Normal, according t o 0.513 roasted material. Sherman & Quinn (6) Forced feeding' of the crude material in a C R PeP = limestone-ferroghosphorus. daily doses, as much as eight times the amount b F.T.P. = fused trioalciuni phosphate. which caused the unfavorable results when included in the diet. failed to have a lethal effect. mentary evidence from the U.S.D.A., and from the greenhouse Table '5 gives the essential data in t v o trials undertaken to experiments a t t,he Knoxville Experiment Station, it was condetermine: the cffeci; of administering on alt,ernate days the cluded that the limestone-ferrophosphorus product showed sufcrude ferrophosphorus in capsules and the hone incal added as a ficient possibilities for use as an animal feed supplement t,o warseparate supplement to raise the phosphorus of t,he diet to 0.5%: and the effect of adding a readily available phosphate, potassium rant further experimentation. Studies, therefore, were initiated dihydrogen phosphat,e, t'o the basal diet in which crude ferroto compare the effectiveness for growth and reproduction of the phosphorus was incorporated. limestone-ferrophosphorus product with other phosphate carriers For purposes of comparison the data are given showing the low commonly used by stock producers when it was fed a t high enough food consumption, poor growth, and small phosphorus retention levels to counteract the unavailability of its phosphorus. for the fire animals which had previously been fed the basal FISALGROWTH TESTS. The basal diet, as described above for diet containing 0.14% phosphorus to which crude ferrophosphorus the experimental diets, mas adequate in all respects for ordinary had been added t o bring the hotal phosphorus level to 0.4%. growth except that the phosphorus was a t the level of 0.14%;--When crude ferrophosphorus was given by capsule on alternate that is, below the minimal requirement. The experimental diets, in this series, included a phosphate carrier in amounts days .ivith a sufficient bone meal supplement to raise the phosphorus of the basal diet to 0.5%, the animals ate and grcw almost sufficient to raise the total phosphorus level t o 0.4%. Bone meal, as much and stored more phosphorus than animal6 receiving the dicalcium phosphate, and the crude ferrophosphorus material same treatment except that an empty capsule was given in place were tested in addition to the Osborne and 11endel salt mixture and the roasted limestone-ferrophosphorus product of the preof one containing crude ferrophospliorus. When an ample supply of a readily available phosphate was liminary tests. The bone meal vas an Arniour product being used at the University of Tennessee as a cattle reed supplement added to the basal diet plus the crude ferrophospliorus, the average food consumption, gain in might, and phosphorus retention and containing 10.87, phosphorus and 23.6% calcium. It a-as were normal. ground t o pass an 80.mesh sieve for incorporation in the experimental diets. The dicalcium phosphate used contained 21.7% phosphorus and 27.370 calcium. A typical analysis of the crude ferrophozphorus TABLE I\r. EFFECTIVESESS F O R GROWTHAKD PHOSPHORCS RLTENTIUN was 22% phosphorus, 2 to 3% silicon, 3% O F OSBORKE AXD MENDEL SALT h f I X T U R E A A D EXPERrlfCXTAL 1'HOSPHATCS manganese, and the remainder an iron compound. -. 30-Day Experimental Period Groups of five animals each were fed as follon-s: n~~~~amt, Mean Gain in Weight-. hIean I' Retention ; i

4

1. Basal diet 2,

3. 4. 5. 6.

Basal diet ture Basal diet Basal diet Basal diet Basal diet

z i t h Standard Error Actual, Compargrams ative, c/o 97.8 b 6 . 4 100 62 60.6 - 6 . 1 79 77.4 * 5 . 4 12 12.2 1 6 . 8

with Standard Error Actual, Compargrams ative, "/c 0.585 * 0 . 0 3 2 100 0.168 * O . O l ; 29 0.429 *0.01,, 73 0.060 * 0 . 0 1 9 10

274

85.0

i2.7

87

0 434

282

85.6 t 3 . 9

88

0.430 i.O.015

food

+ Osborne and Mendel salt mix+ bone meal ++ crude ferrophosphorus limestone-ferrophosphorus + dicalcium phosphate

I n contrast to the preliminary t,ests the animals were fed the diets ad libitum and individual

+ +

Diet 0 & Id salt

Basal Basal Basal bone meal Basal +crudeferrophosphorus Basal limestone-ferrophosphorus Basal dlcalcium phosphate

+ +

eaten per r a t , grams 299 242 274 141

*0

027

74 77

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INDUSTRIAL AND ENGINEERING CHEMISTRY

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ferrophosphorus. Increasing the phosphorus level for both TABLE V. EFFECTS OF FEEDING CRUDEFERROPHOSPHORUS BY diets from below the minimal level up to about 0.4% CAPSULEOR WITH EXCESS OF READILY AVAILABLE PHOSPHATEtotal phosphorus brought about increased food consumption, Mean Mean Mean gain in weight, and phosphorus retention. After the 0.4% phosFood Gain in P No. of Eaten Weight, Stored, phorus level, additional amounts of phosphorus resulted in no Diet Rats G. per Rat G. G. further gains, either in weight or amount of phosphorus stored. 12 Basal (0.14% P) + crude ferro141 0.060 This experiment would seem to indicate that crude ferrophosphosphorus (total P 0.4%) 227 0.397 67 Basal (0.14% P) + 0.4 g. bone phorus included as 1% of the diet is not toxic to the white rat meal every other d a y total P 0.5%) + placebo (total 0.5%) when fed during the period of most rapid growth and, further59 218 0.408 (0.14% P) + 0.4 g. bone -Basal more, that its phosphorus is not assimilated by the rat. meal every other d a y (total P 0.5%) + 0.2 g. crude ferrophosBREEDINGTEsrs. Breeding experiments were set up to test phorus every other day b y capsule (total P 0.8%) the effectiveness of the limestone-ferrophosphorus for reproduc325 106 Basal (0.14% P) + crude ferro0.609 tion and lactation and also to determine the possibility of toxicity phosphorus (total P 0.4%) + KHzPO4 (total P 0.7%) of the crude material if fed continuously throughout the breeding period. Three breeding lots were made up of rats having identical hereditary histories. One lot was fed the basal laboratory diet These two feeding trials seemed to indicate that when crude used in the growth experiments t o which bone meal was added as ferrophosphorus was fed separately from an adequate diet or the chief source of phosphate. The total pposphorus content when phosphorus in a readily available form was added to the was 0.5%. This ration containing bone meal as the source of diet containing crude ferrophosphorus, the animals ate well and phosphorus was used as the control since bone meal is widely food consumption, growth, and phosphorus retention were satisused as a mineral supplement in livestock feeding. A second factory. breeding lot was fed the basal diet plus limestone-ferrophosphorus T o test these results further a series of rats was placed on diets added to raise the total phosphorus content to 0.5%, to test the containing successively higher amounts of phosphorus as potaseffectiveness of the experimental material for reproduction and sium dihydrogen phosphate beginning with the basal diet which lactation. The third breeding lot was placed on the same basal was designed to contain about 0.14% phosphorus. Five male plus bone meal diet as the controls but with crude ferrophosphorus rats were placed on diets a t each phosphorus level and mere added to supply the same amount of phosphorus as was supplied paired as to litter mates and food consumption with five male by the bone meal. Since the growth experiments had demonrats on the same diets plus 1.0% crude ferrophosphorus. Care strated that the phosphorus of this crude material wm unavailable was taken to randomize the rats in respect to the different diet8 to the animal the purpose of this breeding lot was to determine to eliminate the factor of seasonal differences in growth. whether or not the crude material, if inadvertently present, Table VI gives phosphorus and calrium contrnt of each diet might have toxic effects. and growth and phosphorus retention in rats fed each diet. The criterion used for reproductive success was the number of In the control diets, as the phosphorus content was increased, young born; that for success in lactation, the number of young the calcium content was also increased by means of calcium carraised to weaning (21 days old). Records of weights a t birth and bonate to keep the calcium t o phosphorus ratios of all diets weaning were kept. approximately 2. For the diets containing crude ferrophosphorus, The total number of young born on the bone meal diet was the phosphorus content reported represents the actual analysis 359 and on the limestone-ferrophosphorus diet, 364; the numbers minus the phosphorus in the added crude ferrophosphorus and weaned were 121 and 178, respectively. Average weights at the calcium content was kept the same as in the corresponding birth were 4.7 and 4.8 grams and at weaning mere 24.6 and 24.3. control diet since previous results indicate that the additional The breeding performance of the rats receiving the bone meal plus phosphorus of the crude material is practically unavailable. crude ferrophosphorus was inferior to the controls receiving the The effective calcium to phosphorus ratio for the control plus bone meal both in regard to the number born, 149, and raised ferrophosphorus diets was considered to be approximately 2, the to weaning, 60. Birth weight was equal t o that of the controls same as for the controls. Food consumption, growth, and phosphorus retention records (4.8 grams) but weaning weight was lower than for the controls Qf the animals on the basal diet containing 0.15% phosphorus (20.9 grams). and the basal diet plus 1% crude ferrophosphorus, indicate that Thus, as in the preliminary growth experiments, rats fed a no adverse effect from the crude product occurred. The rats diet containing crude ferrophosphorus as well as bone meal on the basal diet plus ferrophosphorus in the later experiment ate showed inferior performance to those fed only bone meal. Also, more food than did those on the similar diet in the first experias in the growth experiments, food consumption on this diet waa ment; in fact, they ate approximately the same amount as did the rats on the basal TABLE VI. EFFECT ON GROWTH AND PHOSPHORUS RETENTION DURING 30-DAY PERIOD O F diet in the earlier experiment INCREASING TOTAL PHOSPHORUS OF DIETSCONTAINING CRUDEFERROPHOSPHORUS and their average gain in Control Diet Control + Ferrophosphorus Diet weight and phosphorus retenP Mean Ca Mean tion were essentially the same. in food Mean gain hlean P in food Mean gain Mean P diet, diet, consumpin ~ t . ~ , retentiona, diet, diet, consumpin wt.O, retentiona, Why the rats on the earlier % % tion, g. B. g. % % tion, 6. g. g. experiment failed to eat the 0.15 0.31 256 58.8 1 4 . 1 0.200 t 0.009 0.16 0.32 257 02.2 *4.2 0.206 *0.013 0.21 0.42 290 291 84.4 62.8 0,394 1 0.013 0.22 0.44 83.8 *2.3 0.377 1 0.008 diet is not evident. The later 0.26 0.51 290 290 91.0 -7.8 0,466 1 0.021 0.24 0.52 85.4 h7.5 0.458 1 0.032 experiment indicates quite 0.29 0.61 285 285 90.0 1 5.1 0.536 1 0,025 0.27 0.66 93.2 t 3 . 3 0.552 t0.014 t 0.026 0.36 322 0.38 322 0.68 0.76 5.5 0.591 10.021 104.0 *3.7 0.589 103.8 clearly that when food con0.41 0.82 324 112.6 * 5.4 0.636 * 0,025 0.41 0.82 325 107.4 1 8 . 2 0.607 10.037 1 0.45 t3.1 0.590 t 0 . 0 2 1 319 319 0.92 0.46 0.96 100.6 0.580 103.6 t 4.5 0.025 sumptions are equalized, rats 0.52 1.02 308 308 103.0 *2.2 0.596 =& 0,024 0.51 1.04 99.2 h4.9 0.596 =I 0.017 fed diets including 1% crude 0.59 1.24 310 308 104.0 1 5.1 0.545 f 0.044 0.65 1.27 98.0 1 3 . 4 0.593 *0.018 310 93.4 1 2 . 2 0.556 *0.019 310 0.67 1.43 97.2 12.9 0.559 t 0.028 0.65 1.47 ferrophosphorus show an equal 301 0.80 1.67 302 96.8 15.3 0.577 h 0,029 0.79 1.69 91.6 *2.5 0.538 *0.027 rate of gain and storage of a With standard error. phosphorus as animals fed the b Actual analysis minus oaloulated amount of phosphorus of added crude ferrophosphorus. same diet without the crude

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INDUSTRIAL AND ENGINEERING CHEMISTRY

TABLE V1I. CHICKPHOSPHORUS DEFICIENT BASALDIET Feedstuff Yellow corn meal Soybean oil meal Casein Alfalfa leaf meal

Percentage 67.0 25.0 4.0 2.5

i:O

carbonate Vitamin Di (2000 b.O.A,C. units) Riboflavin (pure riboflavin) Phosphorus supplement Calcium carbonate

is otherwise adequate, the addition of crude ferrophosphorus has no adverse effect on reproduction and lactation. SUMMARY O F RESULTS FOR THE WHITE RAT

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0.03 0.0003 As indicated in Table VI11 Enough was added to maintain a level of 1.2y0calcium

markedly less and the same eventuality was possible that decreased palatability of the diet reduced food consumption to the extent that various essential nutrients were not present in adequate amounts for normal growth and reproduction. A litter of animals from the stock colony consisting of three males and three females were kept on the stock diet ( 2 parts ground whole wheat, 1part dried whole milk, 1% sodium chloride) with one male and one female receiving by capsule 0.2 gram of crude ferrophosphorus daily (6 days per week). The diets were continued until the two females not receiving the crude ferrophosphorus died from accidental causes. The female receiving the ferrophosphorus and the three males were then killed when they were 10.5 months old. At that time the male that had received ferrophosphorus weighed 416 grams; thc two which had had no ferrophosphorus weighed 420 and 380 grams, respectively. The female receiving the ferrophosphorus bore five litters ~ i t ah total of 34 young; 30 of these were raised to R-ehning. One of the sisters, that did not receive crude ferrophosphorus, bore four litters a i t h a total of 32 young of which 19 were raised; the corresponding numbers for the other sister were 2, 16, and 0. The female that received the crude material, therefore, had a record superior both for reproduction and lactation to either of her sisters for a period of repeated matings. For this breeding lot, then, the results indicate that when the crude ferrophosphorus material is fed separately from a diet adequate in other respects, no toxicity results as judged by reproduction and lactation. The breeding results for the control animals receiving the bone meal diet did not seem to be equal to those for our stock animals or for those that received Osborne and Mendel salt mixture as the chief source of phosphorus. It was therefore decided to set up four additional breeding lots to test further the possible toxicity of the crude material. Two lots which served as controls received the basal diet plus Osborne and Mendel salt mixture. Two other lots equally matched in size, sex, and hereditary background received the same diet plus the crude ferrophosphorus product, These breeding experiments are still under way. The results to date show that the control animals have borne 124 young of which 82 were raised to weaning, and that the rats receiving the crude ferrophosphorus have borne 152 rats of which 101 mere raised t o weaning. Average weights a t birth were 4.8 and 4.7 grams and a t weaning were 27.5 and 26.5 grams, respectively. These results indicate that when the diet contains a sufficient and readi1.v available source of phosphorus and

1. The phosphorus of the crude ferrophosphorus material was not readily available for growth. 2. When the total dietary phosphorus was a t a minimal level, the phosphorus of the limestone-ferrophosphorus product was 91% as available for growth and 7’7% as available for phosphorus retention as the readily available phosphorus of Osborne and Mcndel salt mixture. I n other words, more ferrophosphorus product than other compounds with as readily available phosphorus as that in Osborne and hIendel salt mixture must be fed to furnish biologically equivalent amounts of phosphorus. 3. When included to raise the levels of total dietary phosphorus by 50 and 1007, over the minimal level, the limestoneferrophosphorus was 99 and 91%, respectively, as effective for growth and 90 and 87%, respectively, as effective for phosphorus retention as Osborne and Mendel salt mjxture. 4. Thc limestone-ferrophosphorus was as effective as bone meal and dicalcium phosphate for growth and phosphorus retention, and as effective as bone meal for reproduction and lactation when the total dietary phosphorus was 0.4% for growth and 0.5% for reproduction-that js, when the phosphorus level is sufficiently high to counteract the partial unavailability of the phosphorus of the limestone-ferrophosphorus product. 5. The crude ferrophosphorus showed no signs of toxicity for growth or reproduction when fed separately from a diet adequate for normal growth and reproduction or when sufficient phosphorus from a readily available source was present in the diet, in which the crude material was incorporated. 6. There was evidence that inclusion of the crude product decreased the palatability of the diet under some circumstances with consequent decrease in food consumption, growth, and reproduction. CHICK TESTS

PROCEDURE. Day-old S e w Hampshire and Barred Rock chicks were used to compare the availability of the phosphorus in steamed bone meal, limestone-ferrophosphorus, crude ferrophosphorus, and a defluorinated rock phosphate. Fifteen chicks were placed on each diet. At the end of 1 week, ten uniform chicks per diet were selected to complete the test. They were reared in battery brooders and giren feed and water ud Zibztzrnz. The basal diet which contained 0.3% total phosphorus shown in Table VI1 was supplemented as indicated in Table T?II. The supplemented diets were fed for a period of 21 days. The chicks were then killed, the left tibia removed and the percentage bone ash determined by the method outlined by the A.O.A.C. The results are summarized in Table VIII. RESULTS. (1) For bone development during early life in chicks, steamed bone meal, limestone-ferrophosphorus, and defluorinated rock phosphate have a similar biological value per unit of phosphorus. (2) The phosphorus of crude ferrophosphorus is not available for bone development in the chick. LITERATURE CITED

TABLE

Group 1 2

3 4

5 6

7 8 a

vm.

RESPONSE OF CHICKs

Supplementa None Steamed bone meal Steamed bone meal Steamed bone meal Limestone-ferrophosphorus Limestone-ferrophosphorus Crude ferrophosphorus Defluorinated rock phosphate

TO

DIFFERENT PHOSPHATES

Supplementary P, % 0 0.1 0 2

’

Kt. at

21 Days, Bone Ash,

0.3 0.1 0.2 0.2

0.2

G.

%

111 148 176 154 167

30 01 38.60 41.43 43.39 37 69 39.57 28.08 40.17

171

154

184

(1) Bridgcr, G. L., Moore, J. W., and McLeod. H. &I., JI., IND. EXG. CHEM.,41, 1391 (1949). (2) Ellis, N. R., Cabell, C. 9.Elmslie, , W. P., Fraps, G. S., Phillips, P. H., and Williams, D. E., J . Assoc. Oflciul A g r . Chemists, 28,

129-42 (1945). (3) FIeming, K. B. L., Swayne, V. R., Jr., and Patterson, IC. E., un-

published report. (4) MacIntire, W. H., unpublished report. ( 5 ) Sherman, H. C., and Quinn, E.J., J. B i d . Chem., 67, 667-77 (1926).

Basal diet contained approximately 0.3% phosphorus.

RECEIVED February 2 , 1948.