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The utilization of fat by the growing chick Burdett, Michael Owen 1955

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THE UTILIZATION OF FAT BY THE GROWING- CHICK PART I FOLIO AOID AND FAT, TQT^ANCE IN THE OHICK PART II SURFACE ACTIVE AGENTS AND FAT IN OHICK NUTRITION by Michael 0; Burdett, B.S.A. A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN AGRICULTURE in the Department of Poultry Science We accept this thesis as conforming to. the standard required from candidates for the degree of Master of Science In Agriculture. Members of the Department of Poultry Science. THE UNIVERSITY OF BRITISH COLUMBIA May, 1955. ABSTRACT THE UTILIZATION OF FAT BY THE GROWING- CHICK PART I - FOLIC ACID AND FAT TOLERANCE IN THE CHICK Two p r a c t i c a l feeding experiments have been carr i e d out to investigate the supplementary value of f o l i c a c i d , When added to corn-fishmeal type chick s t a r t i n g rations containing d i f f e r e n t l e v e l s of herring o i l . Growth was depressed when herring o i l was added to rations containing sub-optimal amounts of f o l i o acid. The extent to which o i l depressed growth with a d i e t sub-optimal i n f o l i o acid was probably dependent on the quality of the o i l added to the d i e t . The addition of 0.36 mg. of f o l i c a c i d per pound of d i e t a l l e v i a t e d the f o l i c a c i d deficiency and counteracted the growth depressant aotlon of the o i l . PART I I - SURFACE ACTIVE AGENTS AND FAT IN CHICK NUTRITION Chicks on a p r a c t i c a l s t a r t e r r a t i o n were s i g n i f i c a n t l y heavier at the end of 8 weeks when t h e i r rations were supple-mented with 1 per cent of Tween-80 or 3.3 p.p.m. of procaine p e n i c i l l i n . The addition of 0.25 per cent Tween-80 to rations containing e i t h e r 2.5 or 5*0 per cent herring o i l f a i l e d to produce a growth response. Tween-80 enhanced the u t i l i z a t i o n of e i t h e r 5 per cent cottonseed o i l and tallow f o r growth promotion at 8 weeks but not at 4 weeks. Chicks were fed to 9 weeks of age on a series of rations each with and without the addition of 0.2 per cent of Santomerse-80. The addition of Santomerse-80 or procaine p e n i c i l l i n singly or i n combination improved the growth rate of the chicks. The addition of Santomerse-80 did not further stimulate the growth of chicks reoeiving rations containing both herring o i l and procaine p e n i c i l l i n . No significant effect on the rate of growth was observed as a result of replacing p e n i c i l l i n with aureomycin or replacing herring o i l with tallow. When chicks were fed to 4 or 5 weeks on a f o l i c acid deficient corn-flshmeal type of ration, Tween-80 did not alleviate f o l i o acid deficiency whereas Santomerse-80, aureomycin or p e n i c i l l i n did. Tween-80 in the presence of 5 per cent cottonseed o i l further aggravated f o l i o acid deficiency. Santomerse-80, when fed i n combination with f o l i c acid, increased the growth rate of chicks over that obtained with the combination of Tween-80 and f o l i c acid. Lecithin spared f o l i c acid when the rations did not contain additional o i l . In the presence of the additional o i l , l e c i t h i n aggravated f o l i c acid deficiency. ACKNOWLEDGMENT This study was undertaken with the guidance and counsel of Professor Jacob Biely of the Department of Poultry Science. I wish to express my sincere appreciation and gratitude to him for his kindliness and generous assistance during the course of this study. It i s also a privilege to acknowledge my debt of gratitude to Mrs. B. E. March for her kindness and invaluable help during the course of this work. The author gratefully acknowledges the assistance of Miss Freda Morel i n the care and weighing of the chicks. TABLE OF CONTENTS Page PART I I Introduction . , 1 II Review of Literature (a) F o l i c A d d 1 (b) Functions of F o l i c Acid . . . . . . . .6 I I I Experimental . . . . . . . . . . . . 8 Test 1 10 Test 2 11 IV Discussion 12 V Summary • • . • • 16 PART I I I Introduction l a I I Review of Literature (a) Fats .* 1 (b) Surface Active Agents 5 I I I Experimental 14 Experiment I • • 16 Test 1 . . . . . 17 Test 2 19 Test 3 20 Experiment I I 22 Test 1 22 Test 2 24 Test 3 27 Test 4 . 28 Test 5 31 TABLE OF CONTESTS Page Experiment I I I . . . » 33 Test 1 33 Test 2 3* IV D i s c u s s i o n 37 V Summary . . . . . 51 VI B i b l i o g r a p h y 53 INTRODUCTION Experiments conducted pertaining to the author's undergraduate essay indicated that the addition of either commercial herring o i l or herring o i l subjected to severe heating impaired the nutrit ive value of a corn-flshmeal type chick starter ration. Subsequent experiments indicated that such a ration was deficient i n f o l i c a d d . The addition of either type of o i l to the corn flshmeal rat ion adequately supplemented with f o l i c acid stimulated the growth of chicks to a greater extent than did the rations without additional o i l . In the present study further experimentation was conducted to study the effects of f o l i c acid on the tolerance of the chick for fat. REVIEW OF LITERATURE Fol ic acid i s the popular descriptive term for a group of closely related compounds of the Vitamin B. Complex. In early investigations, this vitamin was described under various names including Vitamin B c , Vitamin M, factor A, L.easel factor, S . lact is R factor, factor U, yeast norlt eluate factor depending upon the source of the material and the species of organisms used for test purposes. There was much evidence that the several factors a l l had a common chemical structure but differed from each other by: a) the presence of extra chemical groups attached to the parent molecule which made these forms available only to certain organisms, or b) the absence of c e r t a i n parts of the common structure which r e s t r i c t e d a v a i l a b i l i t y to those organisms which could synthesize the missing moiety. t h i s compound i s widely d i s t r i b u t e d i n nature, but i t i s also present i n modified forms especially i n compounds with added glutamic acid. F o l i o acid was i s o l a t e d by P f i f f n e r and associates (1943) i n o r y s t a l l i n e form from hog l i v e r and i d e n t i f i e d chemically as pteroylglutamlc acid. There are three d i f f e r e n t groups i n the f o l i c a c i d molecule, namely the pteridine part (a two-ringed nitrogen compound), para-aminobenzoic acid and glutamic acid. The unit consisting of pteridine and the para-aminobenzoic acid i s known as pterolo acid. This compound was shown to be highly active i n promoting the growth of L. helveticus and to antianemic a c t i v i t y i n the chick. Pteroylglutamlc acid was synthesized by Angler et a l (1945). Synthetic f o l i c a c i d functions not only f o r growth stimulation and feather production, but also f o r blood formation. HOOO-OH2 01% .0 -0 HG 0 II / OH HOO0-CH-HH-CO-C 0 = 0 glutamic acid p-amino-benzoio a c i d pteridine FOLIO AOID Following the establishment of the molecular constitu-tion of f o l i c or pteroylglutamlc acid, the nature of several of the molecular conjugates of this vitamin was demonstrated. The marked differences observed In physiological av a i l a b i l i t y of the conjugates to various species of animals and micro-organisms has been responsible for the apparent multiplicity of factors now recognized as variants of pteroylglutamlc acid. Pfiffner et a l (1945) prepared Vitamin Be conjugate from yeast and showed that this substance had a lower nitrogen content than had Vitamin B 0. Absorption spectra date of these two compounds indicate that the molecule of the conjugate i s nearly three times as big as that of Vitamin B c. It had practically no growth promoting action for S.lactis R. The conjugate known as fermentation L.easel faotor which differs from the l i v e r factor (l i v e r L.easel factor, Vitamin B 0) isolated by Pfiffner and associates, In i t s biological activity. It i s just as active, mole for mole, as pteroylglutamlc acif for L. easel, rats, chicks and monkey8, i s relatively inactive for Streptococcus faeoalls R; on the other hand, the degradation product pterolc aold, In which the glutamic acid portion of the pteroylglutamlc aeld molecule i a replaced by hydrogen Is active for the latter microorganisms but not for L. easel. They accordingly reserved the name Vitamin Be for the l i v e r factor and gave the name Vitamin B 0 conjugate to the factor present In yeast* Vitamin Bo conjugate is converted into pteroylglutamlc acid by the action of an enzyme, known as vitamin B 0 oonjugase. This enzyme has been reported by Bird et a l ( 1 9 4 6 ) to be present i n the l i v e r , pancreas and the intestinal mucosa of the chick. Keresztesy et a l ( 1 9 4 3 . ) obtained a factor from fumaric acid fermentation by Hhizapus nigricans whioh had no activity i n stimulating the growth of L.easel, although i t was effective for the growth of S.faecalls and S.lactis R. Accordingly i t was named the S.lactis R. Factor. Subsequent-ly , stokes et a l ( 1 9 4 4 ) found that f o l i c acid could replace the S. laotis R. factor for a l l bacteria that could u t i l i z e the latter and that f o l i c acid was produced when S. faecalls R. was grown on a f o l i c acid free medium containing the S. laotis R. factor. The experiments of Rlckes et a l ( 1 9 4 7 ) indicated that, unlike the L.easel faotor, S.Lactis R. factor did not stimulate growth or haemoglobin formation i n chicks. During a search for better organisms for the micro-biological determination for non-essential amlno-aclds, Sauberlich ( 1 9 4 8 ) found that Leuconostoc citrovorum failed to grow on a synthetic medium that was satisfactory for Leuconostoc mesenteroides and other assay organisms. Supple-mentation of the synthetic medium with l i v e r or yeast extract proved to be effective i n such small quantities In promoting rapid growth, that L.citrovorum could be used for the m i c r o b i o l o g i c a l determination of n o n - e s s e n t i a l amino a c i d s . In the presence of a l l the amino a c i d s , L.Citrovorum was used f o r the q u a n t i t a t i v e determination of the new growth f a o t o r , and a "citrovorum u n i t " was defined as the amount per m i l l l l i t r e of medium that produces h a l f - o p t i m a l growth of the organism. This f a c t o r Is considered t o be c l o s e l y r e l a t e d to pteroylglutamlc a c i d because when moderately l a r g e amounts of pteroylglutamlc a c i d were added to the medium and the c u l t u r e incubated f o r three days, a c i d production was almost o p t i m a l , even i n the absence of l i v e r o r yeast e x t r a c t . But whe)a the i n c u b a t i o n p e r i o d was s h o r t , moderately l a r g e amounts of pteroylglutamlc a o l d f a i l e d t o st imulate growth. In the presence of amlnopterln (hydroxyl group i n p o s i t i o n 4 o f the p t e r i d i n e r i n g r e p l a c e d by an amino group) growth of L . c i t r o v o r u m was i n h i b i t e d , and t h i s i n h i b i t i o n was reversed by the citrovorum f a o t o r but not by pteroylglutamlc a c i d . N i c h o l and Welch (1950) have shown that pteroylglutamlc a c i d Is converted by an enzymatic r e d u c t i o n t o the citrovorum f a o t o r , and they b e l i e v e that the f u n c t i o n of a s c o r b i c a c i d i s that of the reducing agent r a t h e r than of an e s s e n t i a l component of an enzyme system. Pteroylglutamlc a c i d may be regarded as a provitamin In much the same sense that carotene Is a precursor of Vitamin A. Both forms may e x i s t side by s ide i n c e r t a i n foods, and both may be present In the same t i s s u e , 6. but under ordinary conditions the efficiency of conversion i s such that the precursor effectively satisfies nutritional needs. FUNCTION OF FOLIC ACID The metabolic role of f o l i c acid i n the l i v i n g c e l l i s not as clear as some of the other B complex vitamins much as niacin and riboflavin, although i n animals and man i t i s necessary for the proper function of the haemopoietlc system and much evidence links i t to purine and pyrimldlne, (thymine) synthesis, to formate metabolism, and to general reactions involving the transfer of one carbon fragments. No satisfactory explanation has been presented in the literature to explain the mechanism whereby f o l i c acid stimulates red blood c e l l formation. Unconfirmed data by Davis (1946) Indicates that the action of f o l i c acid probably Is to Increase the choline esterase activity i n the body. He claimed that subcutaneous injection of acetylcholine produced hyperchromlc anemia i n dogs and that both l i v e r extract and f o l i c acid increased the number of reticulocytes and red blood c e l l s , at the same time increasing the choline esterase activity. The choline esterase activity of dog serum was increased i n vitro by Incubation with f o l i c acid or l i v e r extract, and oral administration of f o l i c acid to normal humans also increased the choline esterase activity. It has been suggested by Dinning (1949) that f o l i c acid may function i n a choline oxidase system, since the l i v e r and kidneys of aminopterin-treated monkeys were p r a c t i c a l l y devoid of choline oxidase, as were the l i v e r s and kidneys of monkeys fed a f o l i c a c i d d e f i c i e n t diet* The replacement of the methyl group of thiamine by an amino or hydroxyl group resulted i n the formation of compounds which i n h i b i t e d the growth of L. helvetieus. The addition of e i t h e r thiamine or f o l i c acid restored normal growth. This l e d to the hypothesis of Stokes (1944) that thiamine Is the product of an enzyme system of which f o l i c a c i d or a co-enzyme form of i t , functions In the synthesis of thiamine, Shlve et a l (1947) presented evidence that i n the presence of an antimetabolite of f o l i c aold, 4 amino-5-lmldazolecarboxamlde accumulated i n the media used f o r the growth of E. c o l i , These workers believed that the accumulated oompound functions as a precursor of purines by the organism and that para-aminobenzoic acid or a compound synthesized by the organism from para-aminobenzoic acid functions as a co-enzyme In converting such a precursor to purines (hypoxanthine). The accumulation of 4-amino-5-imldazolecarboxamlde has been confirmed by Woolley and Pringle (1950) who further showed that t h i s compound also accumulated i n the media used f o r the growth of E . c o l i In the presence of an anti-metabolite of para-amlnobenzoic acid. From t h i s work, these workers are i n agreement with the hypothesis of Woods (1948) that the i n h i b i t i o n of f o l i c 8 . acid synthesis i s due to a lack of available para-aminobenzoic acid, which i n turn Is responsible for the failure of purine formation. With the demonstration that f o l i o acid i s involved In the biosynthesis of thiamin and the biosynthesis of the purine ring i t follows that: a) a nutritional deficiency of f o l i c acid or related metabolite i n species requiring these factors should be accompanied by a reduced nucleic acid synthesis and, b) antimetabolites of f o l i c acid should inhibit nucleic acid synthesis. EXPERIMENTAL Previous work by Biely, March and Tarr (1951a), has shown that the nutritive value of a f o l i c acid deficient ration fed as a starting diet to chicks wa.s further reduced with the presence of added herring o i l . Available data on the mechanism of f o l i c acid i n metabolism are fragmentary and conflicting. Folic acid has been shown to be effective i n the treatment of sprue (faulty absorption of fats and carbohydrates) i n human beings by Darby et a l (1947), although Woodruff (1950) could not correlate the absorption of oleic acid i n the absence or presence of f o l i c acid In the diets of rats. According to Luckey et a l (1946) the response obtained from f o l i c acid by chicks depends on the type of diet used, the greater response occurring with lower levels of fat In the diet. The following experiments were conducted to ascertain the f o l i c acid requirements of chicks fed rations containing different types and amounts of fat. A series of two feeding t r i a l s has been conducted i n whloh the effect on growth rate of the addition of: ( l ) different levels of cold cleared herring o i l , (2) cold cleared herring o i l and heated herring o i l , to the basal rations containing the ingredients presented i n Table 1 was investigated. The basal ration used i n these experi-ments previously was shown to be deficient In f o l i c acid by Blely et a l (1951). The growth rates of the chicks whloh were fed the different supplements were compared to the growth rate of chicks fed the basal diet and the basal diet f o r t i f i e d with added f o l i o acid. The o i l s were added to the rations at the expense of an equivalent amount of cornstarch In each instance. The experimental rations were fed to day-old White Leghorn cockerels In these tests. Randomization was followed In assigning the chicks to positions i n the electrica l l y heated battery brooders where the chloks were reared to 35 days In Test 1 and to 28 days In Test 2, Feed and water were offered ad libitum. A l l chicks were weighed individually at weekly intervals. The weights 10. reported are indicated i n the various tables. TEST 1 In t h i s t e s t , cold cleared herring o i l was added to the rations at l e v e l s of 2.5 and 5*0 per cent. F o l i o a c i d supplements were fed at a l e v e l of 0.36 mg. and 0.72 mg. per pound. At f i v e weeks of age the b i r d s fed the f o l i c a c i d d e f i c i e n t rations were showing symptoms of severe deficiency and as mortality was becoming heavy, i t was nec-essary therefore to terminate the t e s t . RESULTS The average weights of the chicks In Test 1, obtained by feeding the basal r a t i o n plus d i f f e r e n t percentages of o i l and amounts of f o l i c a c i d f o r the experimental period of f i v e weeks are shown i n Table 1. The data Indicate that 2.5 per cent o i l did not a l t e r the growth rate while 5.0 per oent o i l had a highly s i g n i f i c a n t depressing e f f e c t . Examination of the mortality data indicates that the b i r d s receiving 2.5 per cent o i l suffered more from mortality than d i d the basal. In every Instance, as was expected, the added supplements of f o l i c a c i d to the d e f i c i e n t d i e t s resulted i n an Increase i n growth. I t may also be noted that the addition of f o l i c a c i d completely overcame the depressing e f f e c t of o i l on the growth rate . The presence of f o l i c a c i d In the rations containing o i l permitted a s i g n i f i c a n t gain In weight by those chicks 11. over those receiving the basal r a t i o n s . The e f f i c i e n c y with which the b i r d s converted feed to body weight improved with the addition of f o l i c acid; the addition of o i l to these f o l i c a c i d f o r t i f i e d d i e t s Increased the e f f i c i e n c y of feed u t i l i z a t i o n , whereas, the addition of o i l to the f o l i c a c i d d e f i c i e n t d i e t s diminished the u t i l i z a t i o n of feed. TEST 2. Using the f o l i c acid d e f i c i e n t basal d i e t shown i n Table 1, a study was conducted to determine the e f f e c t s of the quantity and q u a l i t y of herring o i l upon early chick growth. The q u a l i t y of the herring o i l was altered by subjecting the o i l to a continuous flow of a i r f o r 30 hours. The o i l was maintained at a temperature of 110°C through-out the oxidative process. The o r i g i n a l and the oxidized o i l were each added to the d e f i c i e n t diets and to the diets supplemented with f o l i c acid at l e v e l s of 3 and 6 per cent. RESULTS From the data of t h i s Investigation (Table 1) i t i s apparent that the addition of unheated o i l to the rations e i t h e r d e f i c i e n t or supplemented with f o l i c acid had l i t t l e e f f ect on the growth of the chicks. The rate of growth of the chicks was not affeoted by the addition of 3 per cent heated herring o i l . On the other hand, the n u t r i t i v e content of the 12. r a t i o n was f u r t h e r Impaired through the a d d i t i o n of 6 per cent heated o i l t o the f o l i c a c i d d e f i c i e n t r a t i o n although t h i s d e t r i m e n t a l e f f e c t was overcome by the presence of adequate f o l i c a c i d . The c h i c k s r e c e i v i n g 6 per cent heated h e r r i n g o i l i n the presenoe of 0.36 mg. of f o l i c a c i d per pound of d i e t were 6 per cent h e a v i e r than the c o n t r o l s and were 59 per cent h e a v i e r than the l o t s r e c e i v i n g the f o l i c a c i d d e f i c i e n t r a t i o n s supplemented w i t h $ per cent heated o i l . DISCUSSION The r e s u l t s of the t e s t s i n v o l v i n g the corn-fishmeal type of r a t i o n s c o nfirm the e a r l i e r f i n d i n g s of B l e l y and March (1951a) th a t f o l i o a c i d d e f i c i e n c y i n ohicks Induced by feeding these r a t i o n s was f u r t h e r aggravated by the presence of h e r r i n g o i l . I t may a l s o be seen t h a t , w h i l e the d i e t a r y l a c k of f o l i c a c i d i n these r a t i o n s may be overcome by the a d d i t i o n of 0.36 mg. f o l i o a c i d per pound of d i e t , the a d d i t i o n of o i l t o these f o r t i f i e d d i e t s r e s u l t e d i n a f a s t e r r a t e of growth by the c h i c k s than when o i l was excluded from these d i e t s . There i s l i t t l e reference t o experiments concerning the r e l a t i o n s h i p between f a t u t i l i z a t i o n and f o l i o a c i d I d the l i t e r a t u r e . I t has been reporte d by Luokey et a l (1946) t h a t the response of c h i c k s t o a given l e v e l of f o l i c a c i d depends on the type of d i e t and t h a t the r e s -13. ponse I s l e s s w i t h a h i g h f a t d i e t than w i t h a low f a t d i e t . Results s i m i l a r t o these f i n d i n g s were a l s o obtained i n Test 1, where no advantage was obtained by i n c r e a s i n g the f a t l e v e l of the d i e t s beyond 2.5 per cent. I n Test 2, where two q u a l i t i e s of the same, o i l were compared, the commercial grade of h e r r i n g o i l d i d not aggravate f o l i c a c i d d e f i c i e n c y as d i d the heated o i l . The aggravated d e f i c i e n c y was d e c i d e d l y n o t i c e a b l e when the l e v e l of heated o i l was Increased from 3 to 6 per cent of the r a t i o n . Under the c o n d i t i o n s o f the present study i t was found t h a t a s i g n i f i c a n t growth d i f f e r e n c e between l o t s r e s u l t e d from the a d d i t i o n of h e r r i n g o i l t o the f o l i c a c i d d e f i c i e n t r a t i o n s . Growth d e f i c i e n c y was demonstrated not only t o be a s s o c i a t e d w i t h the o i l per se but a l s o w i t h i t ' s q u a l i t y , p o s s i b l y t o the chemical nature of the o i l . I t i s evident from the r e s u l t s t h a t the use of h e r r i n g o i l i n the r a t i o n s adequate i n f o l i c a c i d had a b e n e f i c i a l e f f e o t on average body weight and feed e f f i c i e n c y of the d i f f e r e n t l o t s . D i f f e r e n c e s were noted i n the r a t e of growth and feed e f f i c -i e n c i e s of the d i f f e r e n t l o t s r e c e i v i n g h e r r i n g o i l of d i f f e r e n t q u a l i t y . When heated h e r r i n g o i l was i n c l u d e d i n the b a s a l r a t i o n s , the growth response was d e f i n i t e l y dimin-ished f o r those c h i c k s reared on the d i e t s supplemented w i t h heated h e r r i n g o i l than f o r those re&red on the corresponding d i e t s adequate I n f o l i c a c i d . 14. I n connection w i t h the e f f e c t of o i l q u a l i t y on growth, Crampton and h i s a s s o c i a t e s (1951) observed t h a t I n r a t s growth was impaired when the d i e t s f e d contained heat p o l y -merized o i l s . From the r e s u l t s of t h e i r experiments, these I n v e s t i g a t o r s were of the o p i n i o n t h a t t o x i c compounds were developed i n the o i l s d u r i n g the p o l y m e r i z i n g process and i t was these compounds t h a t depressed growth. There i s a p o s s i -b i l i t y t h a t s i m i l a r t o x i c compounds are present I n commercial h e r r i n g o i l and these compounds exert t h e i r maximum i n f l u e n c e when c h i c k s are reared under s t r e s s . As an a l t e r n a t i v e , I t may be t h a t these t o x i c compounds are none other than peroxides formed i n the unsaturated o i l s and i t i s these peroxides t h a t I n a c t i v a t e o r destroy the f o l i c a c i d o c c u r r i n g i n the n a t u r a l Ingredients of the r a t i o n . I t i s b e l i e v e d t h a t the a c t i v e form of f o l i c a c i d I s l e u c o v o r i n and the f i r s t step i n the process of c o n v e r t i n g f o l i c a c i d i n t o t h i s a c t i v e form i s one of r e d u c t i o n . This r e d u c t i o n I s b e l i e v e d t o be I n i t i a t e d by a s c o r b i c a c i d . I t may be t h a t the peroxides present i n the added o i l s prevent t h i s r e d u c t i o n whereby a f u r t h e r d e f i c i e n c y of f o l i c a c i d would e x i s t i n the corn-fishmeal type of d i e t . Popp and T o t t e r (1952) found t h a t when c h i c k s were f e d a f o l i c a c i d d e f i c i e n t d i e t f o r a p e r i o d of three weeks, markedly low l e v e l s of Coenzyme A were found i n t h e i r l i v e r s . The reason f o r the apparent r e l a t i o n s h i p between Coenzyme A and f o l i o a c i d may not be d i f f i c u l t t o e x p l a i n . I t has been 15. r e p o r t e d by Gregory et a l (1952) t h a t Coenzyme A contains a d e n y l i c a c i d . Presumably normal s u p p l i e s of t h i s nucleo-t i d e would be necessary t o maintain normal Coenzyme A l e v e l s . According t o the hypothesis of Wood (1948) a n u t r i t i o n a l d e f i c i e n c y of f o l i o a c i d i n species m q u i r l n g t h i s f a c t o r should r e s u l t i n reduced n u c l e i c a c i d s y n t h e s i s . I t seems not unreasonable t h a t a r e d u c t i o n i n the b i o s y n t h e s i s of n u c l e i c a c i d would l e a d t o lowered l e v e l s of a d e n y l i c a c i d . This hypothesis has found support through the experiments of To t t e r (1953) who was able t o show t h a t t i s s u e a d e n y l i c a c i d was decreased i n f o l i c a o l d d e f i c i e n c y . The p r e l i m i n a r y n e c e s s i t y of f a t ab s o r p t i o n appears, except f o r small q u a n t i t i e s which may be absorbed as a f i n e emulsion of u n s p l i t f a t , t o be the h y d r o l y s i s of the f a t i n t o i t s c o n s t i t u e n t f a t t y a c i d s and g l y c e r i n e . The experiments of Lynen and Ochoa (1953) i n d i c a t e d t h a t the o x i d a t i o n of f a t t y a c i d s i s the reverse process of f a t t y a c i d s y n t h e s i s , furthermore, these i n v e s t -i g a t o r s and others b e l i e v e t h a t Coenzyme A i s an i n t e g r a l p a r t of t h i s system. Therefore, a f o l i c a c i d d e f i c i e n c y would l e a d to a reduced Coenzyme A content. The p o s s i b i l i t y e x i s t s t h a t w i t h a f o l i c a c i d d e f i c i e n t d i e t h i g h i n f a t , a s t r e s s i s placed upon a v a i l a b l e Coenzyme A. To meet t h i s s t r e s s , Coenzyme A i s m o b i l i z e d w i t h the r e s u l t a n t breakdown of other systems dependent on Coenzyme A and hence an aggravated d e f i c i e n c y . 16. SUMMARY Two p r a c t i c a l chick feeding experiments have been carried out to investigate the supplementary value of f o l i c acid and herring o i l when added to a corn-fishmeal type of chick s t a r t e r r a t i o n . 1) With such d i e t s , the f o l i c acid requirement f o r rapid growth was adequately met with a dietary supplement of 0.36 mg. f o l i c a c i d per pound of feed. 2) F o l i c a c i d deficiency i n chicks fed a corn-fishmeal type of s t a r t e r r a t i o n was aggravated by the addition of herring o i l . Incorporating oxidized herring o i l i n chick rations of t h i s type had a greater e f f e c t i n aggravating the deficiency. 3) A greater growth response was obtained with a corn-fish-meal type chick s t a r t e r r a t i o n adequate i n f o l i c acid i f the r a t i o n contained additional o i l . 4) The addition of heated o i l to the experimental rations adequate i n f o l i c acid increased the growth rate of chicks to a greater extent than did the fresh herring o i l when i t was added to a s i m i l a r r a t i o n . TABLE 1. Average^ weights of ohlck In Teat 1. Supplement to basal diet Ave. wt. Feed Mortal-• • • (gms. ) Gain l t v None 317 2.15 2 2.55^  herring o i l 318 2.46 4 5.0$ herring o i l 270 2.45 8 0.36 mg. f o l i c aold/lb. 340 2.11 0 0.36 mg. plus 2.5# herring o i l 381 2.06 1 0.36 mg. plus 5.0$ herring o i l 377 2.06 2 0.72 mg. f o l i c acid/lb. 325 2.24 0 0.72 mg. plus 2 . 5 # herring o i l 356 2.05 0 0.72 mg. plus 5.0$ herring o i l 354 2.06 1 * Twenty-five day-old White Leghorn cockerels chicks per l o t . Average weights at 5 weeks. MSD at 5% level 31, at 1% level 41. Average^ weights of chick i n Test 2. Supplement to basal diet Av. wt. (gms.) None 262 0.36 mg. f o l i c acid/lb. 306 3.0# herring o i l 267 0.36 mg. plus 3.0$ herring o i l 293 6,0% herring o i l 255 0.36 mg. plus 6,0% herring o i l 308 3.0# herring o i l * 253 0.36 mg. plus 3,0% herring o i l * 311 6,0% herring o i l " SQ4 0.36 mg. plus 6,0% herring o i l * 324 * Twenty-five White Leghorn day-old cockerel chicks per lo t . Average weights at 4 weeks. * Herring o i l heated at 110°C for 30 hours. Composition of basal rations used In tests (lbs./lOO lbs.) Ground yellow oorn 72.75 Cornstarch 6.00 Herring meal 19.00 Iodized salt 0.50 Limestone 1.25 Feeding o i l (2.250A-300D) 0.25 Manganese sulphate 10.00 gms. Riboflavin 0.16 gms. Calcium pantothenate 0.50 gms. Nicotinic acid 0.80 gms. INTRODUCTION During the past few years, considerable emphasis has been placed on the importance of compounding poultry , rations high In protein and vitamins to give accelerated growth along with minimum food intake. Through the application of modern methods i n the processing of protein supplements, rations are now lower i n fat than has hitherto been the case* Since the energy content contributes to the maximum u t i l i z a t i o n of the ration, i t might be advantageous to raise the energy level of these high protein, vltamln-rioh rations* With such rations, supplementation with various fats may offer a means of doing this* The successful use of animal and vegetable o i l s In feed depends to an important degree on the nutritive quality of the ration used. Furthermore, the increased growth rate obtained with high protein, vitamln-rich rations together with high efficiency of feed u t i l i z a t i o n might well increase the energy requirements of the chlok for maximum growth rate and feed u t i l i z a t i o n . Experiments oonducted i n various laboratories have demonstrated clearly that calories added i n the form of rendered fat are well u t i l i z e d by dogs and chicks* In view of the scarcity of information on the effects of fat i n chick starter rations, i t was considered desir-able to study the influence of varying amounts of fat i n rations commonly used for growing chicks* In the present study special attention i s given to the incorporation of various fats at levels that may be applied i n commercial feeds together with surface active agents and antibiotics i n the hope that increased u t i l i z a t i o n of the fat might be aohieved* REVIEW OF LITERATURE Evidence i n the literature for the advisability of adding fat to chick rations i s contradictory. Henderson (1940) found no variation i n the weight or feed consumption of chicks receiving rations supplemented with soybean o i l up to 10 per cent, however, above this level a negative regression of weight was observed* Reiser and Pearson's studies (1949a) revealed that lardfor commercial hydrogenated vegetable fat did not retard the growth of chicks when fed i n levels of twenty per cent. Pepper et a l (1953) presented evidence that broiler diets made up mainly of wheat and soybean o i l meal gave better growth with improved feed efficiency when supplemented with one or two per cent soybean o i l . Feeding tests conduoted by the United States Department of the Interior (1953) with menhaden o i l have indicated that the growth rate of chicks was satisfactory up to twelve weeks of age when the rations contained eight per cent o i l . Sledler and Sehweigert (1953) found that the addition of fat up to eight per cent did not alter the rates of gain of chicks fed a practical starter ration for nine weeks and that the added calories were ef f i c i e n t l y u t i l i z e d at the two and four per cent levels but not at the eight per cent l e v e l . There has been an increasing amount of experimental eyidenoe that fat may actually play a specific role i n nutrition other than as a concentrated source of calories and as a solvent for fat soluble vitamins. It has been repeatedly REVIEW OF LITERATURE Evidence i n the l i t e r a t u r e * f o r the a d v i s a b i l i t y of adding f a t t o c h i c k r a t i o n s i s c o n t r a d i c t o r y . Henderson (1940) found no v a r i a t i o n i n the weight or feed consumption of c h i c k s r e c e i v i n g r a t i o n s supplemented w i t h up t o 10 per cent of soybean o i l . Above t h i s l e v e l however, a negative r e g r e s s i o n of weight was observed. R e i s e r and Pearson rs s t u d i e s (1949a) revealed that l a r d or commercial hydrogenated vegetable f a t d i d not r e t a r d the growth of chicks when fed at l e v e l s of twenty per cent. Pepper et a l (1953) presented evidence that b r o i l e r d i e t s made up -mainly of wheat and soy-o bean o i l meal gave b e t t e r growth w i t h improved feed e f f i c i e n c y when supplemented w i t h one or two per cent soybean o i l . Feeding t e s t s conducted by the U n i t e d States Department of the I n t e r i o r (1953) w i t h menhaden o i l i n d i c a t e d that the growth r a t e of chicks was s a t i s f a c t o r y up to twelve weeks of age when the r a t i o n s contained e i g h t per cent o i l . S i e d l e r and Schweigert (1953) found that the a d d i t i o n of up to 8 per cent f a t d i d not a l t e r the r a t e s of gain of chicks fed a p r a c t i c a l s t a r t e r r a t i o n f o r nine weeks. They a l s o found th a t the added c a l o r i e s were e f f i c i e n t l y u t i l i z e d from f a t added at the two and four per cent l e v e l s but not at the eight per cent l e v e l . There i s i n c r e a s i n g evidence that f a t may a c t u a l l y play a s p e c i f i c r o l e i n n u t r i t i o n other than as a concentrated source of c a l o r i e s and as a solvent f o r f a t - s o l u b l e v i t a m i n s . I t has been repeatedly shown by Burr and Burr (1929) that r i g i d e x c l u s i o n of f a t from the d i e t s of r a t s and other animals r e s u l t s i n a c e s s a t i o n of growth and i n the appearance of s k i n symptoms which are prevented or cured by e i t h e r l i n o l e i c or arachidonic a c i d . The kind of p r o t e i n i n the d i e t appears to have some i n f l u e n c e on the onset and s e v e r i t y of symptoms of f a t d e f i c i e n c y . Working w i t h r a t s Holman (195D reported that symptoms of f a t d e f i c i e n c y appear e a r l i e r w i t h d i e t s compounded w i t h c e s e i n than w i t h egg albumin. R e i s e r (1950) was able to show that an acute d e f i c i e n c y syndrome developed i n c h i c k s fed a d i e t devoid of l i n o l e i c a c i d . When l a r d or cottonseed o i l was added to the r a t i o n the d e f i c i e n c y was remedied. Supplementation w i t h methyl p a l m i t a t e , s a t u r a t e d f a t s or bayberry t a l l o w , d i d not ameliorate d e f i c i e n c y sumptoms, i n d i c a t i n g that the e f f e c t was not due t o f a t alone. From h i s work, R e i s e r concluded that poly-unsaturated f a t t y acids are e s s e n t i a l n u t r i e n t s f o r the growth of c h i c k s . By studying the e f f e c t of adding vegetable o i l s and f a t t y a c i d concentrates to s e m i - p u r i f i e d d i e t s , Carver and Johnson (1954) improved the growth r a t e of chicks as much as t h i r t y per cent when four per cent corn or soybean o i l was included i n these d i e t s . They concluded t h a t chicks r e q u i r e some f a c t o r ( s ) present i n unsaturated f a t s f o r maximum growth. Normal growth i n c h i c k s was observed by Russel (1940) and by Davis (1941) using p r a c t i c a l c h ick s t a r t e r r a t i o n s which were extracted t o c o n t a i n no more than 0.1 per 3. f a t . There are a number of w e l l recognized f u n c t i o n s which f a t may play. Fat i s g e n e r a l l y regarded as an o p t i o n a l component of the d i e t which can be p a r t l y or w h o l l y replaced by other sources of c a l o r i e s although i t has been long known that f a t s are the f o o d s t u f f s which possess the highest c a l o r i c d e n s i t y . A few years ago only carbohydrate was b e l i e v e d to have an appreciable p r o t e i n - s p a r i n g a c t i v i t y . Forbes et a l (1946) working w i t h r a t s showed th a t a d i e t a r y f a t supplement "spared" p r o t e i n much more e f f e c t i v e l y than d i d a comparable amount of carbohydrate. The f a t was a l s o more e f f e c t i v e i n reducing the s p e c i f i c dynamic a c t i o n of ingested p r o t e i n . The work of W i l l i a m et a l (1947) i n d i c a t e d t h a t f a t i s able t o spare p r o t e i n under c o n d i t i o n s where carbohydrate i s completely i n e f f e c t i v e . Pearson and Panzer (1949b) a r r i v e d at s i m i l a r conclusions by another experimental approach. They reported that the l o s s of the e s s e n t i a l amino acids i n the u r i n e and the s t o o l was c o n s i d e r a b l y reduced when corn o i l was administered i n the d i e t of r a t s . I n s i m i l a r work w i t h r a t s , Hoover and Swanson (1950) observed that when f a t was omitted from a l o w - p r o t e i n low c a l o r i c d i e t , the r a t e of p r o t e i n catabolism was doubled. Ensuing work by Forbes and h i s a s s o c i a t e s (1946) brought forward evidence that the d i g e s t i b i l i t y of p r o t e i n and the r e t e n t i o n of n i t r o g e n were improved i n the order of the i n c r e a s i n g f a t content of the d i e t . This e f f e c t was noted not only i n growing r a t s but a l s o 4. with mature animals. Fat as a dietary component, i s characterized by several other unique capacities. In addition to being the nutrient of maximum energy value i t may also serve as a source of fa t soluble vitamins. Fish l i v e r o i l s , for example, contain Vitamin A as such. Fish which contain much body o i l such as salmon and herring are the richest natural sources of Vitamin D. Vegetable o i l s are regarded as the most s a t i s f a c t o r y foodstuff from which the tocopherols can be obtained. Another expression of the s p e c i f i c n u t r i t i v e power of fat i s i t s sparing action on some of the B. vitamins. The sparing effect of fa t on thiamine was noted by Evans and Lepkovsky (1929) and has been confirmed by Elvehjem (1939a, 1939b). Reports from the laboratories of Bi r c h (1938), Salmon (1941), and Qackenbush et a l (1942) have demonstrated that pyridoxime deficiency which i s s i m i l a r to a fa t deficiency i n rats can be re l i e v e d by the presence of d i e t -ary fat containing the ess e n t i a l f a t t y acid, l i n o l e i c a c i d . Many reports have appeared i n the l i t e r a t u r e with regard to the a b i l i t y of surface agents to improve the absorption of f a t . The h i s t o r y of emulsifying agents and surface active agents goes back to the discovery of soap, probably the f i r s t emulsifying agent which has been covered extensively i n the l i t e r a t u r e . In the past few decades and 5. p a r t i c u l a r l y i n the past few years, other c l a s s e s of surface a c t i v e agents have come i n t o prominence p r i m a r i l y f o r i n d u s t r i a l use but due to recent t e c h n o l o g i c a l developments, some of these compounds have found a p p l i c a -t i o n i n b i o l o g i c a l systems. When app l i e d t o such systems, phenomena such as p r e c i p i t a t i o n and denaturation of p r o t e i n , d e s t r u c t i o n of microorganisms, and growth promoting propert-i e s are some examples. Surface a c t i v e agents are defined as substances which c o n t a i n both a water s o l u b l e and o i l s o l u b l e group. The o i l s o l u b l e group of the surface a c t i v e molecule w i l l have l i t t l e a t t r a c t i o n f o r the water of an aqueous s o l u t i o n and there w i l l be a tendency f o r i t to accumulate i n a surface l a y e r . The molecules which are i n the surface l a y e r w i l l a l t e r the energy r e l a t i o n s h i p at i n t e r f a c e s . I n other words, a surface a c t i v e agent has the property of o r i e n t i n g between two i n t e r f a c e s i n such a way that i t becomes a coupling agent b r i n g i n g the i n t e r f a c e s i n t o more inti m a t e contact. The i n t e r f a c e may be between l i q u i d and gas, l i q u i d and s o l i d , or between l i q u i d and l i q u i d . When the two i n t e r f a c e s and immiscible l i q u i d s , the surfa c e a c t i v e agent lowers the i n t e r -f a c i a l t e n s i o n so that emulsions are formed, the agent i s then c h a r a c t e r i z e d as an e m u l s i f y i n g agent. The group r e c e i v i n g most recent and growing a t t e n t i o n i s that of the non-ionic surface a c t i v e agents. Non-ionics, 6. as the name implies do not ionize i n water solution as do the anionics and cationics. They depend on a proper balance i n the molecule between their hydrophilic groups (polar) and lypophilic groups (non-polar) for their effectiveness as surface active agents. Generally the hydrophilic portion of the molecule is represented by a grouping consisting of free hydroxyl groups, ether oxygen linkages or both. The lypophilic property is introduced into the molecule by the hydrocarbon chain of fatty acids or alcohols. Jones et a l (1948) reported improved absorption of fat and fat soluble substances with the addition of Tween-80 (polyoxyethylene sorbitan monoaleate) to the diet of patients having a poor fat d i g e s t i b i l i t y . Johnson et al £ l 9 5 0 ) reported that Tween-20 (polyoxyethylene sorbitan monolaurate) increased fat absorption to some extent i n the case of premature infants. The addition of 0 . 5 per cent glycerol monostearate to a diet containing 3 per cent hydrogenated cottonseed o i l was demonstrated by Huff et a l ( 1 9 5 D to significantly increase the plasma fat levels in the blood of calves. Working with young pigs, Luecke et a l ( 1 9 5 2 ) was able to present evidence that either 0 . 1 per cent Bthonoid^ C/15 or aureomycin i n their diets improved #-Ethonoids - substituted fatty acid amides, the substituents being polyoxyethylene groups. the growth rate of these animals as much as 1 5 per cent. 7. Shoshkes, Geyer and Stare (1950a) demonstrated that f a t i n the form of an emulsion was absorbed to a greater extent from r a t i n t e s t i n e than when the f a t was i n the unemulsified form. I n f u r t h e r experimentation these i n v e s t i g a t o r s (1950b) found that the a b s o r p t i o n of an e d i b l e vegetable o i l a f t e r i n t u b a t i o n i n t o the stomach was unaffected by the presence of l a r g e amounts of soybean phosphatide or Tween-80. I r w i n et a l (1936) have i n v e s t i g a t e d the e f f e c t s of the a d d i t i o n of various substances t o f a t d i e t s upon f a t absorption i n the r a t and found that they had l i t t l e or no p o s i t i v e e f f e c t , but l a r g e r amounts i n v a r i a b l y decreased the r a t e of f a t a b s o r p t i o n . H a r r i s and Sherman (195D found that the i n c l u s i o n of high l e v e l s of c e r t a i n surface a c t i v e agents c o n t a i n i n g polyoxyethylene and s o r b i t a n groups i n a s y n t h e t i c d i e t i n h i b i t e d the growth of r a t s . Chow et a l (1953) presented evidence that the feeding of the e m u l s i f i e r s Span-60 ( s o r b i t a n monostearate), Tween-60 (polyoxyethylene s o r b i t a n monosterate), or Marj -52 (polyoxyethylene monostearate) at l e v e l s of 5 per cent or 15 per cent of a b a s a l soybean d i e t t o young weanling or o l d r a t s of both sexes d i d r e s u l t i n growth r e t a r d a t i o n . Further experimentation w i t h these e m u l s i f i e r s by A l l i s o n et a l (1952) d i d not produce any d e l e t e r i o u s e f f e c t s on growth or maintenance of dogs, mice or hamsters. Schweigert and h i s associates (1950) reported 8. that feeding weanling hamsters polyoxyethylene monostearate at 5 or 15 per cent l e v e l s i n a d i e t c o n t a i n -i n g p u r i f i e d c a s e i n , produced a s i g n i f i c a n t l y lower r a t e of gain i n comparison w i t h t h a t observed i n animals fed the same co n c e n t r a t i o n of l a r d . Krantz (1949a) and co-workers have made extensive animal feeding s t u d i e s on the e m u l s i f i e r s designated Span, Tween and Marj and found that the i n g e s t i o n of these sub-stances by r a t s at l e v e l s of 2 to 5 per cent i n a commercial feed over a period of two years d i d not a f f e c t t h e i r growth r a t e s . I n a l a t e r i n v e s t i g a t i o n Krantz (1949b) reported t h a t the d a i l y o r a l a d m i n i s t r a t i o n to humans of Tween-80 i n doses of 4 .5 to 6 gm. f o r about 4 years d i d not r e s u l t i n any observable e f f e c t . According to T i d w e l l and Nagler (1952) the a d m i n i s t r a -t i o n of Tween-80 had no e f f e c t on the r a t e of f a t a b s o r p t i o n i n r a t s or human beings. These i n v e s t i g a t o r s suggest t h a t the " f a t a b s o r p t i o n r a t e i s not a f f e c t e d by exogenous e m u l s i f i e r s but may be increased by substances a f f e c t i n g the chemical mechanism of f a t a b s o r p t i o n " . The i n t r o d u c t i o n of surface a c t i v e agents t o p o u l t r y n u t r i t i o n was I n i t i a t e d by E l y (1951)• He supplemented an a l l - v e g e t a b l e c h i c k r a t i o n w i t h an ethylene condensate and obtained an inc r e a s e i n growth to the extent of about ten 9. per cent i n 10 - 12 weeks. The increase i n growth was of the same magnitude as obtained by the a d d i t i o n of a n t i b i o t i c s or v i t a m i n B 1 2 t o t h e s a m e b a s a l d i e t although the response obtained by the supplementary surface a c t i v e agent was not revealed u n t i l r a t h e r l a t e i n the growing p e r i o d . E l y f u r t h e r emphasized that no s y n e r g i s t i c e f f e c t occurred between the surface a c t i v e agent and the a n t i b i o t i c v i t a m i n B]jp supplements. Fo l l o w i n g t h i s o r i g i n a l p u b l i c a t i o n , S c o t t et a l (1952) found no evidence of growth s t i m u l a t i o n from d i f f e r e n t surface a c t i v e agents when added t o an a l l plant d i e t adequate i n v i t a m i n B ^ and known to give a p o s i t i v e response t o aureomycin. I n one case a combination of two agents sh a r p l y depressed growth. S t e r n and McGinnis (1953) obtained no s i g n i f i c a n t growth response from the a d d i t i o n of detergents t o a d i e t c a l c u l a t e d t o c o n t a i n ample amounts of known vit a m i n s . I n some cases a growth depression occurred when the detergent l e v e l was in c r e a s e d . The r e s u l t s obtained from the l a t t e r two i n v e s t i g a t i o n s were from experiments conducted over a period of four weeks, whereas E l y ' s o r i g i n a l r e p o r t was based on an experimental period of 10-12 weeks. Using 10 weeks as the basis of study, Snyder et a l (1953) found no increase i n growth w i t h the a d d i t i o n of a surface a c t i v e agent, i n an a l l - v e g e t a b l e d i e t adequate i n v i t a m i n Bi2» however, i n the absence of t h i s 1 0 . v i t a m i n , the surface a c t i v e agent alone or i n combination w i t h aureomycin increased the growth r a t e of chicks at nine weeks, but the f i n a l weight was not equal to the d i e t supplemented w i t h v i t a m i n Bi2« T n e growth s t i m u l a t i o n obtained from v i t a m i n B12 and the surface a c t i v e agent was not manifested u n t i l l a t e r i n the experiment, whereas, those b i r d s r e c e i v i n g aureomycin made r a p i d gains during the f i r s t f our weeks of the experimental p e r i o d . Using a c o n t r o l period of 12 weeks, Branion and H i l l (1954) found no evidence of growth s t i m u l a t i o n w i t h a b a s a l r a t i o n c o n t a i n i n g animal p r o t e i n supplements along w i t h v i t a m i n B12 when s y n t h e t i c detergents were added at 0 . 5 per cent of the d i e t . They f u r t h e r reported that procaine p e n i c i l l i n a l s o f a i l e d t o give a response to the same ba s a l d i e t i n d i c a t i n g t h a t , i f s u r f a c t a n t s as growth promoters f u n c t i o n through s e l e c t i v e b a c t e r i a l i n h i b i t i o n as proposed by E l y ( 1 9 5 2 ) , i t i s p o s s i b l e under c e r t a i n environmental c o n d i t i o n s t h a t b a c t e r i a which can be i n h i b i t e d are not always present. E l y ( 1 9 5 2 ) s t a t e d that growth response t o surface a c t i v e agents would not be detected u n t i l a f t e r the chicks were 5 t o 6 weeks of age unless a g e r m i c i d a l type of surface a c t i v e agent was f e d . I t i s p o s s i b l e growth response observed w i t h the surface a c t i v e agents may be mediated i n a s i m i l a r way to the growth s t i m u l a t i n g p r o p e r t i e s of a n t i -b i o t i c s . 11. The growth promoting a c t i o n of a n t i b i o t i c s i s a v a r i a b l e e f f e c t . The v a r i a b i l i t y holds true both f o r the d i f f e r e n t a n t i b i o t i c s and f o r d i f f e r e n t l e v e l s of the same a n t i b i o t i c . The v a r i a b i l i t y i n the response t o d i e t a r y a n t i b i o t i c s has been s t u d i e d e x t e n s i v e l y by many workers. From these s t u d i e s , the growth promoting a c t i o n of a n t i b i o t i c s i s most f r e q u e n t l y discussed as one ass o c i a t e d w i t h the f l o r a of the g a s t r o i n t e s t i n a l t r a c t . The method whereby a n t i b i o t i c s a c t may be one or a combination of the f o l l o w i n g : a) Favoring the absorption of n u t r i e n t s from the gastr o -i n t e s t i n a l t r a c t . b) Decreasing the growth of b a c t e r i a which would o r d i n a r i l y use la r g e amounts of some e s s e n t i a l d i e t -ary n u t r i e n t . c) S e l e c t i v e l y f a v o r i n g the growth of microorganisms that are capable of s y n t h e s i z i n g some growth f a c t o r . The f a c t o r i s then made a v a i l a b l e to the t e s t animal. d) I n h i b i t i n g the growth of microorganisms, t h a t might otherwise produce a d i s e a s e . The e f f i c i e n c y w i t h which chicks convert feed i n t o body weight when a n t i b i o t i c s are added to the r a t i o n has been i n v e s t i g a t e d by many workers. One of the most i n t r i g u i n g p r o p e r t i e s of the a n t i b i o t i c s used i n c h i c k r a t i o n s i s t h e i r a b i l i t y t o reduce the d i e t a r y requirements of various members 12. of the B-complex vi t a m i n s . S h e p i l e v s k i i (1928) studying experimental scurvey i n guinea pigs was of the o p i n i o n t h a t 0.015 per cent o l e a t e , which has no a n t i s c o r b u t i c e f f e c t , i n the d i e t of these animals somewhat retarded the l o s s i n body weight. H a l i c k (1953) reported that the a d d i t i o n to the d i e t of a surface a c t i v e agent (a polyoxyethylene o x i d e - s u b s t i t u t e d amine) enhanced the absorption of v i t a m i n B-^ and increased the amount of v i t a m i n B ^ appearing i n the egg y o l k . Aldersberg and Sabotka (1943) reported that v i t a m i n A was abs orbed at a f a s t e r r a t e by man i f l e c i t h i n , an e f f e c t i v e e m u l s i f y i n g agent, was added to the d i e t . Esh and Sutton (1948) a l s o reported t h a t l e c i t h i n improves the absorption of both carotene and v i t a m i n A i n the r a t . Burns, Hauge and Quackenbush (195D reported that the gr owth of r a t s f e d carotene e m u l s i f i e d i n a water suspension c o n t a i n i n g 10 per cent Tween-40 was somewhat b e t t e r than i n r a t s r e c e i v i n g the carotene i n an o i l s o l u t i o n . Eaton and co-workers (195D found that c a l v e s , depleted of vit a m i n A, had higher plasma l e v e l s of carotene and greater amounts of vit a m i n A i n the l i v e r and lung a f t e r an aquous suspension of carotene had been given i n t r a v e n o u s l y than f o l l o w i n g i t s o r a l a d m i n i s t r a t i o n . T o m a r e l l i and a s s o c i a t e s (1946) and more r e c e n t l y B i e r i and Sandman (195D» reported that -carotene can be r e a d i l y u t i l i z e d by r a t s when given p a r e n t e r a l l y , provided an aquous suspension i n Tween-80 or 13 Tween-40 (polyoxyethylene s o r b i t a n monopalmitate) was used. Halpern and B i e l y (1948) reported t h a t emulsif-i e d o i l s c o n t a i n i n g v i t a m i n A have a higher b i o l o g i c a l value f o r ch i c k s than i f the same o i l was d i s s o l v e d i n f r e s h vegetable o i l . Jones et a l (1948) obtained greater a b s o r p t i o n of v i t a m i n A by using Tween-80 as an e m u l s i f y i n g agent. 14. EXPERIMENTAL The r a t i o n s used i n the f o l l o w i n g experiments were made up of n a t u r a l i n g r e d i e n t s . Except where the r a t i o n s were formulated t o be d e f i c i e n t i n f o l i c a c i d they may be termed as p r a c t i c a l c h i c k s t a r t e r s . Except i n one instance a l l the c o n t r o l r a t i o n s contained co r n s t a r c h . The various o i l s were added at the expense of the c o r n s t a r c h . This procedure made i t p o s s i b l e to add o i l to the d i e t s without d i l u t i n g the components i n the r a t i o n or changing the proportions of the components. Some of the chicks used i n these experiments were obtained from a commercial hatchery and the remainder from the U n i v e r s i t y f l o c k . S i n g l e comb White Leghorn and New Hampshire males and females were used. For these experiments, day-old chicks were wing-banded f o r i d e n t i f i c a t i o n . They were d i s t r i b u t e d at random i n t o groups of 18-22 i n compartments of e l e c t r i c a l l y heated b a t t e r y brooders w i t h r a i s e d screen f l o o r s . Natural l i g h t was excluded and a r t i f i c i a l l i g h t was s u p p l i e d f o r a 12 hour day. Feed and water were kept before the b i r d s at a l l times. The chicks were i n d i v i d u a l l y weighed each week and the average weight at 4 or 5 weeks was used as a measure of growth response t o the d i e t f e d . Records of m o r t a l i t y and, i n some experiments, of feed consumption were kept. There were three analyses made during these experiments t o determine the r e l a t i o n s h i p between the amount of f a t 15. consumed and the amount of f a t excreted by ch i c k s on various d i e t s . I n order to determine the amount of f a t eaten, the q u a n t i t y of feed remaining a f t e r a weighted amount of feed had been o f f e r e d over a period of 24 hours was determined. I n a s c e r t a i n i n g the amount of f a t consumed, 10 gram samples of each d i e t under i n v e s t i g a t i o n was subjected t o ether e x t r a c t i o n by means of a Soxhlet f a t e x t r a c t o r and the per cent of f a t i n each d i e t c a l c u l a t e d . The feces of each experimental l o t of ch i c k s was allowed t o accumulate simultaneously as the known amount of feed o f f e r e d to the ch i c k s was consumed. At the end of 24 hours the feces was c o l l e c t e d and thoroughly ground. The procedure which was f o l l o w e d i n preparing the feces f o r the e x t r a c t i o n of f a t and the subsequent i s o l a t i o n of the f a t was as f o l l o w s : 5 grams of feces and approximately 15 grams of anhydrous sodium sulphate was placed i n a mortar and t r i t u r a t e d w i t h a p e s t l e u n t i l the mixture had a granular c o n s i s t e n c y . The samples were then t r a n s f e r r e d t o a Soxhlet f a t e x t r a c t o r and extracted w i t h d i - e t h y l ether f o r a pe r i o d of 16 hours. F o l l o w i n g e x t r a c t i o n , the ether e x t r a c t was t r a n s f e r r e d t o weighed beakers and the sol v e n t d i s t i l l e d on a steam bath. The per cent f a t i n the wet feces was then determined. I n a d d i t i o n t o the samples of feces obtained f o r f a t determinations, 2 gram samples were 16. ajlso taken and placed i n an e l e c t r i c a l l y heated oven maintained at a temperature of 110°C f o r 12 hours and at the end of t h i s time the dry weight of the feces was determined. I n a l l experiments the f a t e x c r e t i o n r a t i o was c a l c u l a t e d as f a t excreted: f a t consumed: dry matter excreted: dry matter consumed. EXPERIMENT I The e f f e c t of surface a c t i v e agent on the growth r a t e and e f f i c i e n c y of feed u t i l i z a t i o n of chicks fed d i e t s c o n t a i n i n g various l e v e l s of f a t . The use of surface a c t i v e agents as growth promotants f o r chicks was i n i t i a t e d by E l y ( 1 9 5 D . The a p p l i c a t i o n of an ethylene oxide condensate of coco f a t t y a c i d s , when added t o an a l l plant d i e t stimulated c h i c k growth t o the extent of about 10 per cent at 10 - 12 weeks. Later work by Sc o t t et a l (1952) S t e r n et al- (1952), and S t e r n and McGinnis (1953) i n d i c a t e d t h a t surface a c t i v e agents d i d not produce any s i g n i f i c a n t growth response, and i n some cases, when fed at high l e v e l s or i n the combination of two s u r f a c t a n t s , depressed growth. The experiments of these workers were of short d u r a t i o n (4 weeks) compared to the 10-12 week period of E l y . I n a l a t e r r e p o r t , Snyder et a l (1953) fed one s u r f a c t a n t f o r a period of 10 weeks w i t h -out i n c r e a s i n g the growth response. The l a t t e r i n v e s t i g a t o r s a l s o reported t h a t s e v e r a l other s u r f a c t a n t s were without e f f e c t over a 4 week p e r i o d . 17. Test l . As an i n t r o d u c t i o n to the f u r t h e r study of the p o s s i b i l i t y of adding s u r f a c t a n t s to ch i c k s t a r t e r r a t i o n s i n order t o increase growth r a t e and improve feed e f f i c i e n c y , i t was deemed d e s i r a b l e t o t e s t the e f f e c t of d i f f e r e n t l e v e l s of a s u r f a c t a n t i n a p r e l i m i n a r y experiment. The ba s a l d i e t chosen was one w i t h which p e h i c i l l i n was known t o give a response. Du p l i c a t e l o t s of White Leghorn co c k e r e l s used i n t h i s i n v e s t i g a t i o n were obtained as day o l d b i r d s . The c h i c k s were kept i n back warmer type b a t t e r y brooders under the usual management c o n d i t i o n s w i t h ad l i b i t u m feeding. The ba s a l d i e t used during the experimental period i s presented i n Table ( 5 ) . The surface a c t i v e agent, Tween-80 (polyoxy-ethylene s o r b i t a n monoleate) was added to the basal r a t i o n at l e v e l s of 0 .25, 0 .5 , 1.0 and 2.0 per cent. P e n i c i l l i n was added t o the bas a l r a t i o n at a l e v e l of 0.15 gm/100 l b s . RESULTS Table (2) shows the average weights and the e f f i c i e n c y of d i e t u t i l i z a t i o n at the end of four and eight weeks r e s p e c t i v e l y . At the end of four weeks of t h i s experiment, the b i r d s r e c e i v i n g p e n i c i l l i n were heavier than those r e c e i v i n g the b a s a l or the b a s a l supplemented w i t h Tween-80. The b i r d s r e c e i v i n g Tween-80 at l e v e l s greater than .25 per cent were a l l heavier than those r e c e i v i n g the b a s a l . The 18. same growth pattenn was maintained at 8 weeks but those b i r d s r e c e i v i n g .75 per cent to 2.0 per cent Tween-80 had weights almost equal to the l o t r e c e i v i n g p e n i c i l l i n . The feed e f f i c i e n c i e s of the d i f f e r e n t l o t s f o l l o w e d the same pa t t e r n as the growth r a t e s except that at four and e i g h t weeks r e s p e c t i v e l y , the l o t s r e c e i v i n g 2.0 per cent Tween-80 u t i l i z e d feed as w e l l as the l o t r e c e i v i n g p e n i c i l l i n . The body weight data show tha t the b i r d s r e c e i v i n g p e n i c i l l i n and Tween-80 f o l l o w e d d i f f e r e n t growth p a t t e r n s . The l o t s r e c e i v i n g .5 per cent and .75 per cent Tween-80 were s i g n i f i c a n t l y heavier at the 5 per cent l e v e l than the c o n t r o l at 4 weeks of age, whereas the l o t fed p e n i c i l l i n as a supplement were s i g n i f i c a n t l y h eavier at the 1 per cent l e v e l than the c o n t r o l at four weeks of age. These l o t s r e c e i v i n g supplements of Tween-80 greater than .75 per cent were heavier than the c o n t r o l but not s i g n i f i c a n t l y so. At 8 weeks of age, those b i r d s r e c e i v i n g .5 per cent and .75 per cent Tween-80 were now s i g n i f i c a n t l y heavier than the c o n t r o l as was the l o t r e c e i v i n g p e n i c i l l i n . The l o t s r e c e i v i n g the supplements of Tween-80 were no so heavy however as those r e c e i v i n g p e n i c i l l i n . The data provided by the f o l l o w i n g two experiments are an extension of the study of the e f f e c t of d i e t a r y f a t and s u r f a c t a n t s on the growth of c h i c k s . 1 9 . TEST 2. I n the second t e s t , l e v e l s of 0.25 and 0.50 per^ cent Tween-80 was added to the b a s a l , b a s a l plus 2.5 per cent h e r r i n g o i l and b a s a l plus 5.0 per cent h e r r i n g o i l . Three hundred and s i x t e e n day-old White Leghorn p u l l e t s were d i v i d e d i n t o nine d u p l i c a t e l o t s of seventeen c h i c k s each and fed the r a t i o n s o u t l i n e d . The composition of the ba s a l d i e t i s summarized i n Table 5. The chicks were reared under the usual management procedures and they were weighed i n d i v i d u a l l y at weekly i n t e r v a l s . The average weights at 4 and 8 weeks of age and the data w i t h regard t o feed u t i l i z a t i o n are shown i n Table 3 . RESULTS The presence of e i t h e r amount of the s u r f a c t a n t i n the bas a l r a t i o n had l i t t l e e f f e c t i n s t i m u l a t i n g growth of the chicks up to four weeks of age. The average weight of the bi r d s r e c e i v i n g o i l was only s l i g h t l y i n f e r i o r to that of the b i r d s r e c e i v i n g the b a s a l r a t i o n w i t h and without the s u r f a c t a n t . The growth response was somewhat d i f f e r e n t at 8 weeks of age. I n each i n s t a n c e , the added supplement of Tween-80 r e s u l t e d i n an inc r e a s e i n growth w i t h the b a s a l r a t i o n . This growth p a t t e r n was not present where o i l was incorporated i n the r a t i o n . The presence of the s u r f a c t a n t i n the d i e t s improved the e f f i c i e n c y of feed u t i l i z a t i o n at four weeks except i n 2 0 . the l o t r e c e i v i n g 0 . 2 5 per cent Tween - 8 0 i n the presence of 5 per cent h e r r i n g o i l . At 8 weeks of age, Tween - 8 0 had somewhat of the opposite e f f e c t that i t had at four weeks, that i s , poorer feed u t i l i z a t i o n was obtained as the l e v e l s of Tween - 8 0 i n c r e a s e d . I t i s a l s o evident t h a t at 8 weeks of age no group u t i l i z e d feed as w e l l as the c o n t r o l group which was not the case at the four week i n t e r v a l . TEST 3 . A d d i t i o n a l data on the growth r a t e u s i n g a surface a c t i v e agent i s derived from a study i n which chicks have re c e i v e d the experimental d i e t s f o r as long as 8 weeks, and where h e r r i n g o i l , cottonseed o i l , t a l l o w and Tween - 8 0 were the v a r ' i a b l e s u t i l i z e d . The b a s a l d i e t used f o r t h i s - study was the same as that reported i n a previous study w i t h the s u r f a c t a n t , Tween - 8 0. The composition of t h i s d i e t i s summarized i n Table 5. Management procedures were s i m i l a r to the other experiments of t h i s s e r i e s . The growth s t i m u l a t i n g e f f e c t s of the Tween - 8 0 and the f a t supplements are tabulated i n Table 4 . RESULTS The r e s u l t s obtained at four weeks i n d i c a t e that the chicks fed the d i e t s c o n t a i n i n g the added f a t grew at a f a s t e r r a t e than the c h i c k s fed the b a s a l r a t i o n although the presence of cottonseed o i l i n the d i e t d i d not improve • 21. the growth r a t e to the same extent as d i d the other two o i l s . The enhanced gain caused by the added f a t was not demonstrated i n the presence of Tween-80 i n the r a t i o n . I t may be noted that the i n c l u s i o n of Tween-80 to the b a s a l d i e t caused an improvement i n weight gain amounting to 22 grams or 5.8 per cent at four weeks. Data obtained at the end of 8 weeks fol l o w e d the same p a t t e r n as the four week data w i t h regard to the e f f e c t on growth of the a d d i t i o n of the d i f f e r e n t o i l s to the b a s a l r a t i o n . I n the presence of Tween-80, a greater response was obtained by the a d d i t i o n of the o i l t o these r a t i o n s as compared to the b a s a l , maximum growth appearing w i t h the oottonseed o i l and the t a l l o w . During the d i s t r i b u t i o n of the chicks i n t o t h e i r r e s p e c t i v e compartments at the commencement of the experiment, one hundred White Leghorn p u l l e t s were a c c i d e n t l y mixed w i t h the cockerels obtained f o r t h i s t r i a l . For t h i s reason, data regarding the u t i l i z a t i o n of feed at the eight week per i o d have been omitted from Table 4 . However, at four weeks of age the feed e f f i c i e n c -i e s were improved w i t h the a d d i t i o n of the o i l s to the b a s a l d i e t . When Tween-80 was added to the basal r a t i o n only cottonseed o i l improved the u t i l i z a t i o n of feed. Both h e r r i n g o i l and t a l l o w decreased feed e f f i c i e n c y when added to a r a t i o n supplemented w i t h Tween-80. TABLE 2 Average weight of chicks at 4 weeks i n Experiment I, Test 1. Supplement to Weight Feed  basal ration (gms.) Gain None A 3 4 2 « Q 2 1 0 B 3 1 8 5 5 0 2 # 1 ° 0.25% Tween-80 A 311 B 347 0.50% Tween-80 A 351 B 371 1.0% Tween-80 A 357 B 332 2.0% Tween-80 A 341 B 346 329 2.06 561 2.06 344 2.12 344 1.99 P e n i c i l l i n * A 3 8 2 B 3 8 6 384 1.95 Average weight of chicks at 3 weeks i n Experiment I, Test 1. Supplement to Weight Feed  basal ration (mas.) Gain None A 770 B 730 0.25% Tween-80 A 752 B 777 0.50% Tween-80 A 804 B 809 750 3.01 754 2.89 806 2.79 1.0% Tween-80 A 845 p B 803 2 2 » ° 5 2.0% Tween-80 A 829 B 820 Pe n i c i l l i n * A 826 844 824 2.74 835 2.70 0.15 gm. procaine p e n i c i l l i n G/100 lb. TABLE 3 . Average weight and feed u t i l i z a t i o n at 4 and 8 weeks of White Leghorn cockerels i n Experiment I, Test 2. 4 weeks of age. 8 weeks of afte. Supplement to Weight Feed Fat Weight Feed Fat basal ration. (gms.) Gain Exere (gms*) Gain Exere -tion -tion Ratio Ratio None 321 2.24 1.87 755 2.73 1*47 2.5% Herring o i l 302 2.25 1.77 734 2.81 1.20 • - . . . . . ) 5.0% Herring o i l 301 2.05 1.56 749 2.83 1.53 0.25JS Tween-80 322 2,20 1.24 772 2.75 ©•80 0,25% Tween-80 plus 2.5# Herring o i l 310 2.22 1.54 730 2.87 0.81 0.25% Tween-80 plus 5.0% Herring o i l 305 2.16 1.25 724 2.76 0.81 0.5% Tween-80 316 2.12 1.27 766 2.88 0.66 0*5% Tween-80 plus 2.5% Herring o i l 306 2.14 1.14 737 2.78 0.60 0.5# Tween-80 plus 5.0% Herring o i l 310 2.10 0.92 746 2.84 0.56 TABLE 4. Gain and feed utilization of White Leghorn cockerels fed a surfactant and various fats at 4 and 8 weeks of age. 4 weeks of age. 8 weeks of age. Fej Supplement to basal ration Average Gain (gms.) Average Gain (gms.} None 275 2^ 39 729 5.0* Herring o i l 296 2.27 762 5.0# Cottonseed o i l 278 2.15 743 5.0% Tallow 300 2.23 752 0.5% Tween-80 287 2.07 719 0i5% 5.0% Tween-80 plus Herring o i l 289 2.35 746 0.5% 5.0% Tween-80 plus Cottonseed o i l 283 2.00 774 0*5% 5.0% Tween-80 plus Tallow 287 2.10 775 Analysis of Variance - 4 weeks. Source of Error Sum of Squares d.f. Variance Total Treatment Error 442872 144348 332 15 298524 317 Minimum Significant Difference at P at p 0.05 ^  19 0.01= 25 9623 ** 942 TABLE 5. Composition of basal rations used i n Experiment I, (Weight of ingredients i n pounds unless otherwise stated.) Ingredients Ration 1* Ration 2. Ground yellow corn 32.60 35.10 Ground wheat 32.60 35.10 Soybean o i l meal 13*40 13.40 F i shmeal 6.70 6.70 Livermeal 3.00 3.00 Dried brewers' yeast 2.00 2.00 0 Dehydrated cereal grass 1.00 1.00 Iodized salt 0.50 0.50 Limestone 1.00 1.00 Bonemeal 1.50 1.50 Feeding o i l (2,250A-300D) 0.25 0.25 Choline chloride (25%) 0.40 0.40 Manganese sulphate 0.025 0.025 Nicotinic acid 0.80 gms. 0,80 gms. Ration 2 was used i n Tests 2 and 3. EXPERIMENT II A comparison of surface active agents and antibiotics as growth stimulants for the chick. In the previous experiments attempts were made to evaluatethe growth promoting properties of Tween-80 i n chick starter rations with and without added fat. The results of these t r i a l s have not been consistent. There remains the fact, however, that some of the added fats tended to promote increased growth. No improvement i n ^ growth could be attributed to the presenoe of the surfac-tant i n rations containing added fat. TEST 1. This investigation was undertaken i n an attempt to determine the supplemental value of herring o i l i n the presence of the detergent Santomerse-80 (an alkyl aryl sulfonate) and an antibiotic when they were added indiv-idually and i n combination i n the experimental rations. Gold cleared herring o i l was incorporated i n a starter ration as a replacement for the cornstarch. Partial replacement was made at the 2.5 and 5.0 per cent levels and complete replacement made at the 7.5 per cent le v e l . White Leghorn cockerels were used. The chicks were raised and managed under the same procedures as previously described. The composition of the individual diets i s shown i n Table 12. 23.. RESULTS The data presented i n Table 6 show tha t the a d d i t i o n of 0.15 gm. procaine p e n i c i l l i n G per 100 l b s . r e s u l t e d i n a ( s i g n i f i c a n t ) s t i m u l a t i o n i n growth, regardless of the o i l content of the d i e t f e d . The a d d i t i o n of o i l t o the bas a l r a t i o n r e s u l t e d i n an improve-ment i n growth at nine weeks. The response observed from the a d d i t i o n of p e n i c i l l i n t o each ba s a l r a t i o n was s i m i l a r . The growth response from the a d d i t i o n of the surface a c t i v e agent d i d not f o l l o w the same p a t t e r n as that obtained by the a n t i b i o t i c . The response t o Santomerse-80 i n the bas a l d i e t , without the a d d i t i o n of any h e r r i n g o i l , was s i m i l a r to the growth response produced by the a n t i -b i o t i c alone. The a n t i b i o t i c p e n i c i l l i n was the more e f f e c t i v e when added to the d i e t s c o n t a i n i n g o i l than was Santoraerse 80. The growth obtained w i t h the d i e t s c o n t a i n -i n g 5 per cent or 7.5 per cent o i l and Santomerse-80 caused a growth depression. The a d d i t i o n of Santomerse-80 to the b a s a l r a t i o n plus the a n t i b i o t i c r e s u l t e d i n no increase i n growth above that obtained by p e n i c i l l i n or Santomerse-80 alone. The c h i c k s r e c e i v i n g p e n i c i l l i n and Santomerse-80 w i t h d i e t s c o n t a i n i n g added h e r r i n g o i l showed a lower r a t e of growth at each l e v e l of o i l than was obtained w i t h the a n t i b i o t i c alone. 24. The feed e f f i c i e n c y f i g u r e s f o r a l l groups, i n most instances were s i m i l a r . The e f f i c i e n c y was improved by the a d d i t i o n of p e n i c i l l i n t o a l l d i e t s , which was not the case w i t h Santomerse-80. The combination of p e n i c i l l i n and Santomerse-80 w i t h no added o i l i n creased the e f f i c i e n c y of the u t i l i z a t i o n of the b a s a l d i e t but i n the presence of an o i l supplement no such r e s u l t was obtained. TEST 2. The c h i c k growth s t u d i e s conducted p r e v i o u s l y using o i l , s u r f a c t a n t s , and a n t i b i o t i c s , have, on the whole, given i n c o n c l u s i v e r e s u l t s . The data from the t e s t d e s c r i b e d h e r e i n are provided by the e f f e c t of the s u r f a c t a n t , Santomerse-80 and the a n t i b i o t i c , aureomycin, s i n g l y and together, on r a t i o n s c o n t a i n i n g 7.5 pei; cent h e r r i n g o i l and e d i b l e t a l l o w . I n the case of the o i l d i e t s , the experimental r a t i o n s were prepared by s u b s t i t u t i n g equal q u a n t i t i e s of the o i l i n place of corn s t a r c h i n the standard b a s a l d i e t . These d i e t s were f e d under comparable c o n d i t i o n s t o d u p l i c a t e l o t s of twenty New Hampshire p u l l e t s each f o r a period of nine weeks. The e f f e c t s of supplementing d i e t s c o n t a i n i n g 7.5 per cent h e r r i n g o i l or t a l l o w w i t h Santomerse-80 and aureomycin on growth and feed e f f i c i e n c i e s are summarized at f i v e and nine weeks i n Table 7. The r e s u l t s represent the combined data from d u p l i c a t e t r i a l s . 25. RESULTS During the f i r s t f i v e weeks of the experiment, there was a very s l i g h t decrease i n growth due t o the presence of the 7.5 per cent h e r r i n g o i l . With respect t o the 7.5 per cent t a l l o w , the unsupplemented d i e t was below that of the b a s a l , w h i l e the remainder were s l i g h t l y above. The a d d i t i o n of Santomerse-80 to the b a s a l r a t i o n and to the b a s a l plus t a l l o w r e s u l t e d i n an improvement i n growth. No such improvement was obtained w i t h the d i e t c o n t a i n i n g 7.5 per cent h e r r i n g o i l . The response observed from the a d d i t i o n of the aureomycin supplements showed, i n a l l cases, an improvement i n growth when fed alone. When Santomerse-80 was added w i t h aureomycin to the b a s a l d i e t s , there was again an improvement over the basal and the basa l supplemented w i t h the o i l s . This increase i n growth was approximately equal to that r e s u l t i n g from the a d d i t i o n of aureomycin alone except where the b a s a l contained 7.5 per cent t a l l o w , where growth was s l i g h t l y above that obtained from aureomycin by i t s e l f . At the age of nine weeks, moderately good growth had been obtained i n the group f e d the basa l d i e t . Comparably the d i e t c o n t a i n i n g 7.5 per cent h e r r i n g o i l and 0.25% aureomycin at the same time produced increased growth over the b a s a l . A l l the chicks r e c e i v i n g 7.5 per cent t a l l o w were heavier than the b i r d s fed corresponding d i e t s w i t h and without h e r r i n g o i l . The b a s a l d i e t w i t h t a l l o w added i n d i c a t e d an improvement i n c h i c k weight during the f i n a l 26. weeks of the experiment, because at f i v e weeks, i t w i l l be r e c a l l e d , t h i s l o t was l e s s heavy than the b a s a l , and now, four weeks l a t e r , i t was 3.4 per cent heavier than the b a s a l . There was no d i f f e r e n c e i n growth p a t t e r n r e s u l t i n g from the a d d i t i o n of Santomerse-80 at nine weeks than that obtained at f i v e . S i m i l a r l y , the same p a t t e r n was manifested w i t h aureomycin at nine weeks as at f i v e . The combination of Santomerse-80 and aureomycin at f i v e weeks showed an e f f e c t on improving growth w i t h the 7.5 per cent t a l l o w than w i t h aureomycin and t a l l o w alone. This e f f e c t had disappeared at the t e r m i n a t i o n of the experiment, and t h i s combination had no b e t t e r e f f e c t on growth r a t e than that obtained from aureomycin alone. The response observed from the a d d i t i o n of the above supplements t o the basal r a t i o n upon the u t i l i z a t i o n of feed should not be overlooked. At f i v e weeks, the b i r d s fed the h e r r i n g o i l supplements i n a l l cases, except w i t h a combination of aureomycin and Santomerse-80, u t i l i z e d feed b e t t e r than when these d i e t s contained c o r n s t a r c h . These b i r d s had feed e f f i c i e n c i e s of the same order of those r e c e i v i n g 7.5 per cent t a l l o w i n t h e i r r a t i o n s , except i n the case where aureomycin was i n c l u d e d which r e s u l t e d i n an increased e f f i c i e n c y . The presence of t a l l o w i n the r a t i o n s improved feed e f f i c i e n c i e s over that of the bas a l but where aureomycin was in c l u d e d alone, the e f f i c i e n c y dropped below that observed w i t h the c o r n s t a r c h . 27. At nine weeks there was l i t t l e change i n the p a t t e r n of feed e f f i c i e n c i e s except t h a t , i n the presence of aureomycin, feed e f f i c i e n c i e s were improved over s i m i l a r d i e t s c o n t a i n i n g t a l l o w . TEST ^ Since the i n c l u s i o n of o i l , s u r f a c t a n t s , and a n t i -b i o t i c s i n a b a s a l d i e t d e f i c i e n t and a b a s a l d i e t adequate i n f o l i c a c i d gave v a r i e d responses, a f u r t h e r study was made to determine the e f f e c t s of these substances on c h i c k growth. Twenty-four White Leghorn cockerels per l o t were used i n t h i s Test. F o l i c a c i d , aureomycin and the s u r f a c t a n t Tween-80 were the v a r i a b l e s added s i n g l y and together i n the b a s a l r a t i o n (Table 9) and the basal plus 5»° per cent h e r r i n g o i l . The u s u a l methods of management were adhered to throughout the experimental period. The experimental data are summarized i n Table 9. RESULTS Under the c o n d i t i o n s of t h i s Test i t was found again that the a d d i t i o n of 5»0 per cent h e r r i n g o i l r e s u l t e d i n a depression i n r a t e of growth when added to a f o l i c a c i d d e f i c i e n t d i e t . A d d i t i o n a l f o l i c a c i d added to these d e f i c i e n t d i e t s not only r e l i e v e d the d e f i c i e n c y but a l s o m u l l i f i e d the aggravated d e f i c i e n c y caused by the h e r r i n g o i l . 28. I t may be noted that at f i v e weeks of age aureomycin caused a weight increase of 6.2 per cent over the basal diet but 10.6 per cent less than when f o l i c acid was added to the basal d i e t . Aureomycin and f o l i c acid together did not improve the growth response over that obtained by f o l i c acid alone. Aureomycin and Tween-80 when added together to the basal diet increased the growth response but not as we l l as did f o l i c acid alone. The addition of Tween-80 did not improve the growth rate obtained with either the basal r a t i o n or the rations with added o i l . Tween-80 when fed with f o l i c acid did not augment the growth over that obtained with f o l i c acid alone, i n f a c t , chicks weighed less than those receiving f o l i c acid alone with either o i l - f r e e or o i l supplemented d i e t s . The addition of f o l i c acid had l i t t l e e f fect i n improving the f a t excretion r a t i o whereas i t was markedly decreased by the .presence of aureomycin or Tween-80. A combination of aureomycin and Tween-80 reduced the r a t i o to a greater extent than either supplement alone, either aureomycin or Tween-80 i n combination with f o l i c acid lowered the r a t i o to a greater extent than did f o l i c acid alone. TEST 4. Evidence obtained i n previous experiments indicates that with a f o l i c acid d e f i c i e n t d i e t s , the i n c l u s i o n of herring o i l i n such diets depressed growth. In the following two 29. t e s t s a t t e n t i o n i s d i r e c t e d towards the e f f e c t of d i f f e r e n t d i e t a r y o i l s , s u r f a c t a n t s and a n t i b i o t i c s and combinations of these substances when added to chick s t a r t e r r a t i o n s d e f i c i e n t and r a t i o n s adequate i n f o l i c a c i d . The b a s a l r.ation used f o r these t e s t s was compounded by the i n g r e d i e n t s appearing i n Table 12. I n t h i s t e s t , a comparison was made i n the e f f e c t of aureomycin, p e n i c i l l i n , Santomerse-80 and Tween-80 on t h i s type of r a t i o n w i t h and without the a d d i t i o n of .036 gms. of f o l i c a c i d per 100 l b s . The average weight of the chicks at four weeks of age are presented i n Table 10. RESULTS Again, as i n previous experiments, the data from t h i s i n v e s t i g a t i o n v e r i f y the observation that the a d d i t i o n of 5 per cent h e r r i n g o i l to a r a t i o n d e f i c i e n t i n f o l i c a c i d aggravates f o l i c a c i d d e f i c i e n c y . Aureo-mycin increased the growth r a t e of the c h i c k s fed the f o l i c a c i d d e f i c i e n t r a t i o n regardless whether there was o i l present or not. The growth stimulus from aureomycin was not, however, equal to that obtained by the a d d i t i o n of f o l i c a c i d , where the increase was h i g h l y evident. A combination of aureomycin and f o l i c a c i d gave no greater growth e f f e c t than d i d f o l i c a c i d alone on e i t h e r the basal or the b a s a l plus 5 per cent o i l . - 3 0 -I n every i n s t a n c e , the added supplements of Tween-80 t o the basal d i e t c o n t a i n i n g c o r n s t a r c h or o i l had no e f f e c t on improving the growth of the b i r d s . The combination of Tween-80 w i t h f o l i c a c i d d i d not improve c h i c k growth as w e l l as d i d f o l i c a c i d alone but the combination of Tween-80 and aureomycin gave b e t t e r growth than aureomycin alone regardless of whether the basa l d i e t contained o i l or not. No advantage was obtained by supplementing the bas a l r a t i o n w i t h Tween-80, aureomycin and f o l i c a c i d i n combination than by f o l i c a c i d supplementation alone. A l l groups r e c e i v i n g f o l i c a c i d , alone or i n combina-t i o n e x h i b i t e d a four week growth response that was i n some cases s i g n i f i c a n t due t o the a d d i t i o n of 5 per cent h e r r i n g o i l s . The a b i l i t y of aureomycin i n a m e l i o r a t i n g the f o l i c a c i d d e f i c i e n c y was i n h i b i t e d by the a d d i t i o n of 5 per cent h e r r i n g o i l to the r a t i o n . I t should be noted t h a t r a t i o n s adequate i n f o l i c a c i d and c o n t a i n i n g 5 per cent h e r r i n g o i l produced the best r e s u l t s of the e n t i r e experiment w i t h regard to feed e f f i c i e n c y except i n the case where f o l i c a c i d and Tween-80 were i n combination. The presence of Tween-80 alone or w i t h f o l i c a c i d decreased feed e f f i c i e n c y when added t o the ba s a l r a t i o n c o n t a i n i n g c o r n s t a r c h . Aureomycin had l i t t l e e f f e c t on feed e f f i c i e n c y except when-in combination w i t h Tween-80 or f o l i c a c i d . When o i l replaced cornstarch i n the basa l r a t i o n , feed e f f i c i e n c i e s were improved i n a l l cases 31 regardless of the supplementation although the best efficiencies were obtained in the presence of adequate f o l i c acid. The combination of f o l i c acid and aureomycin improved feed u t i l i z a t i o n slightly more than either of these supplements alone i n the rations containing 5 per cent herring o i l . TEST 5. Evidence obtained i n the previous test indicates that with a f o l i c acid deficient diet the addition of 5 per cent herring o i l to the diet depressed growth. In Test 2 the M same level of supplementary cottonseed o i l was chosen for a further investigation on the relationship between o i l and a f o l i c acid deficient diet on chicks from day old through five weeks of age. The basal rations used i n this feeding t r i a l contained a l l the known nutrients for the chick with the exception of f o l i c acid. The cottonseed o i l substituted for an equal amount of cornstarch as before. The experimental diets were formulated according to Table 12, f o l i c acid, aureomycin, and Tween-80 were fed singly and in combination to various lots of chicks. RESULTS The efficiency of feed u t i l i z a t i o n , growth, and mortality are given i n Table II. It i s to be noted again that in a l l instances where the f o l i c acid deficient diet contained cottonseed o i l there was a decrease in growth rate in a l l but one instance. The deficient diet containing 32, aureomycin d i d not s u f f e r i n n u t r i t i o n a l value due to the presence of o i l . The a d d i t i o n of aureomycin to the b a s a l d i e t r e s u l t e d i n an increase i n the weight of the chicks from both the o i l and the o i l - f r e e d i e t s . I t did not, however, produce the growth obtained w i t h f o l i c a c i d alone. No a d d i t i o n a l growth stimulus was obtained from a combination of f o l i c a c i d and aureomycin. The presence of o i l had the e f f e c t of i n c r e a s i n g feed e f f i c i e n c i e s although i n the d i e t s c o n t a i n i n g Tween-io and a combination of aureomycin and f o l i c a c i d , feed u t i l i z a t i o n was impaired but i n the l a t t e r case, very good e f f i c i e n c y was maintained. The presence of Tween-80 i n the o i l - f r e e d e f i c i e n t d i e t aggravated the d e f i c i e n c y which i s i n d i c a t e d i n the m o r t a l i t y data appearing i n Table 11. TABLE 6. Average weight of ohloks at nine weeks resulting from the dietary supplements* Supplement to basal ration None 2.5% Herring o i l 5.0% Herring o i l 7.5% Herring o i l Weight (gms*) Weight (gms*) Weight (gms.) Weight (gms*) None 775 865 902 852 Penicillin** 850 912 952 900 Santomerse-80* 841 880 897 827 Santomerse-80 plus Penicillin 841 834 916 882 Feed efficiencies of ohloks at nine weeks resulting from the dietary supplements* Supplement to basal ration None 2.5% Herring Oil 5.0% Herring o i l 7.5% Herring o i l Feed Gain Feed. Gain Feed Gain Feed Gain None 3.28 3;02 2.89 2.92 Penicillin** 3.13 2.98 2.82 2.75 Santomerse-80* 2;97 3a4 2^ 95 2.-94 Santomerse-80 plus Penicillin 3*02 3ao 2i94 2.-87 .0*15 gm. procaine penicillin G added per 100 lbs. *Santomerse-80 added at 0*2% level. TABLE 7. Average weights of Hew Hampshire pullets fed the experimental diets at 5 weeks of age* Supplement to None 7.5% Herring 7.5% Tallow basal ration Oil Weight (mns*) Feed Gain Weight («ms.) Feed Gain Weight (gms.) Ga?n None 486 2.55 471 2.42 478 2.45 Santomerse-80 479 2.55 452 2.41 494 2.37 Aureomycin 516 2.46 508 2.37 522 2.49 Santomerse-80 plus Aureomycin 518 2.50 503 2.52 534 2.49 Average weights of New Hampshire pullets fed the experimental diets at 9 weeks,of age. Supplement to None 7.5% Herring 7.5% Tallow basal ration O i l Weight (ffms.) Feed Gain Weight (was.) Feed Gain Weight ( M I S . ) None 1,101 2.76 1,095 2.57 1,140 2.53 Santomerse-80 1,118 2.71 1,050 2.63 1,162 2.57 Aureomycin 1,110 2.91 1,154 2.47 1,194 2.58 Santomerse-80 plus Aureomycin 1,152 2.64 1,100 2.55 1,178 2.61 Santomerse-80 was added at the rate of 0.10^. Aureomycin was added at the rate of 4.5 mg./lb. (from Aurofao A.) TABLE 8 Analysis of Variance Experiment I I , Test 2 - 4 weeks. Source of Error Sum of Squares d.f. Variance Total 1 8 7 3 9 9 9 4 0 3 Treatment 2 3 3 7 2 4 2 3 1 0 1 6 2 Error 1 6 4 0 2 7 5 3 8 0 4 3 1 7 Minimum Significant Difference at p 0 . 0 5 4 4 • ** at p 0 . 0 1 5 7 Experiment I I , Test 2 - 9 weeks. Souroe of Error Sum of Squares d.f. Variance Total 6 4 2 7 3 3 1 3 6 8 Treatment 9 0 8 1 8 2 2 3 3 9 4 8 6 * * Error 5 5 1 9 1 4 9 3 4 5 1 5 9 9 8 Minimum Significant Difference at p 0 . 0 5 ^ 9 0 at p 0 . 0 1 = 1 1 8 TABLE 9. Average weight (gmsv) of White Leghorn pullets at four weeks* Supplement to basal ration With 5% With 5% Cornstarch Herring Fat Excretion o i l Ratio None 354 318 1.63 Folio acid 416 447 1.54 Aureomycin 376 377 1.14 Tween-80 351 315 1.27 Folic acid plus aureomycin 417 447 1.18 Folic acid plus Tween-80 369 408 1.24 Tween-80 plus aureomycin 399 384 1*05 Folic acid plus aureomycin plus Tween-80 407 443 1.24 Folic acid was added at the rate of 0.36 mg.Ab., aureomycin at the rate of 4.5 mg./lb. (from Aurofac A) and Tween-80 at the rate of 0. Source of Error Sum of Squares d.f. Variance Total Treatment Error 1740908 528803 1212105 357 15 342 35253 3544 4 * Minimum Significant Difference ^ at p 0.05 -35 ** at p 0.01 > 47 TABLE 10. Results of feeding f o l i o acid, detergents and antibiotics to New Hampshire cockerels for a period of four weeks. Supplement to Mean Weight Increase over basal diet at 4 weeks. basal (%) (gms,) None 229 -Folic acid J 356 55.5 Aureomycin • 365 60.0 P e n i c i l l i n * 359 57.0 Tween-80 • 248 8.3 Santomerse-80 296 29.3 Folic acid and Tween-80 364 60.0 Folic acid and Santomerse-80 384 68.0 Aureomycin and Tween-80 336 47.0 Aureomycin and Santomerse-80 364 60.0 P e n i c i l l i n and Tween-80 329 44.0 P e n i c i l l i n and Santomerse-80 373 63.0 Folic acid and aureomycin 425 86.0 Folic acid and p e n i o i l l i n 398 74.0 • Quantities of supplements added per 100 lbs. Folic acid 0.036 gms. Aurofao A 0.25 lbs. Procaine p e n i c i l l i n 0 1.5 gms. Tween-80 0.5 l b . Santomerse-80 0.1 l b . Analysis of Varlanoe Source of Error Sum of Squares d.f. Variance Total 1537967 298 Treatment 714050 13 54927 H Error 823917 285 2891 Minimum Significant Difference at p 0.05 =23 ** at p 0.01 *43 TABLE 11 Average weight (gms.) of New Hampshire cockerels at 5 weeks. Supplement to basal ration With 5% Mort- With 5% Mort-Comstareh a l i t y Cottonseed a l i t y O i l None Folic acid Aureomycin Tween-80 Folic acid plus aureomycin Folic acid plus Tween-80 Tween-80 plus aureomycin Folic acid plus aureomycin plus Tween-80 438 514 458 421 534 487 528 519 2 1 408 547 451 377 548 524 457 576 2 2 3 3 Analysis of Variance Source of Error Sum of Squares d.f. Variance Total Treatment Error 2324367 959691 1364676 309 15 294 63979 ** 4642 Minimum Significant Difference at p 0.05 ^43 at p 0.01s 57 TABLE 12. Composition of basal diets used in outlined Tests, (Weight of ingredients in pounds unless otherwise stated*) Ingredients Test 1 Test 2 Test 4 Test 345 Ground yellow oora 31.05 27.41 79.75 72.75 Ground wheat 31.05 27.41 Soybean o i l meal 13.90 20*00 Fishmeal 6.90 10.00 3.00 18.00 i9;oo Livermeal 3.00 Dried brewers1 yeast 2.00 1.00 Dehydrated oereal grass 1.00 Iodized salt 0.50 0.50 0.50 0.50 Limestone 1.00 1.00 1.25 1.25 Bonemeal 1.50 1.50 Feeding o i l (2,250A-300D) 0.25 0.25 0.25 0.25 Choline chloride 0.40 0.40 0.25 0.25 Manganese sulphate 0.025 0.025 10.00 gm* 10.00 gm. Riboflavin 0.16 gm. 0.10 gm* 0.16 gm* 0.16 gm. Calcium pantothenate 0.50 gm. - 0.50 gm. 0.50 gm. Nicotinic acid 0.80 gm. 7.50 0.90 gm. 0.80 gm. 0.80 gm. Cornstarch 7.50 - 6.00 Supplements were added to 100 pounds of basal ration at the following levels. Test 1. Test Test Test 4. Test Folio acid (gms.) 0.036 0.036 0.036 Aureomycin (lbs.) 0.25 0.25 0.25 Q.25 Penicillin (gms*) 1.50 1.50 1.50 Tween-80 (lbs.) 0.50 0.50 0.50 Santomerse-80 (lbs.) 0.20 0.10 0.10 0.10 Aureomycin added as "Aurofao A" Penicillin added as Procaine penicillin G. 33. EXPERIMENT I I I The e f f e c t of l e c i t h i n on the growth rate of chicks fed various d i e t s . TEST 1. The f i r s t feeding test of the series was conducted to determine the e f f e c t of l e c i t h i n at l e v e l s of 0.5 and 1.0 per cent on the growth of chicks fed the basal r a t i o n , the composition of which appears i n Table 15 with 5 and 10 per cent herring o i l substituted f o r equal amounts of cornstarch i n the basal r a t i o n . The l e c i t h i n was dissolved i n each o i l supplement and i n the feeding o i l of the o i l -free diets p r i o r to combining i t with the experimental rations. RESULTS The e f f e c t of l e v e l s of l e c i t h i n and o i l on weight and feed e f f i c i e n c y i s shown i n Table 13. The weight data were examined s t a t i s t i c a l l y according to the method of analysis of variance (Snedecor, 1946). They indicate that i n the absenoe of l e o i t h l n , the effect on growth of 5 per oent herring o i l was s i g n i f i c a n t . There was a s i g n i f i c a n t decrease on the growth of the chicks receiving the basal r a t i o n supple* mented with 10 per cent herring o i l . The addition of 5 per cent herring o i l to those die t s containing e i t h e r 0.5 or 1.0 per cent l e c i t h i n , did not Improve growth over that obtained i n the absence of o i l . The presence of 10 per cent o i l i n the diets containing e i t h e r l e v e l of l e c i t h i n resulted i n a lower rate of growth than that obtained without any o i l or with the 5 per oent l e v e l of o i l . A s l i g h t advantage i n growth was obtained by the addi-t i o n of 0.5 per cent l e c i t h i n to the basal r a t i o n and to the d i e t containing 10 per cent herring o i l , but no such advantage was evident with the r a t i o n containing 5 per cent herring o i l . The response of chicks to 1.0 per cent l e c i t h i n was no better than that obtained with the 0.5 per cent l e v e l , of l e c i t h i n , i n f a c t , there was a s l i g h t decrease i n growth i n the presence of e i t h e r 5 or 10 per cent herring o i l . The e f f i c i e n c y of feed u t i l i z a t i o n was affected benefi-c i a l l y by the presence of o i l whereas the presence of l e c i t h i n decreased the e f f i c i e n c y unless o i l was included. The best conversion of feed to body weight was obtained with 10 per cent herring o i l and 0.5 per oent l e c i t h i n , although the b i r d s fed the d i e t containing these supplements d i d not grow at the maximum rate observed i n t h i s t e s t . TEST 2 f Evidence obtained i n the previous test indicated that the presence of l e c i t h i n i n a chick s t a r t e r r a t i o n containing 10 per cent herring o i l reduced the growth of chicks. In t h i s i n v e s t i g a t i o n , the effects of l e c i t h i n , aureomycin and f o l i c a cid on the growth of the chicks fed a oorn-flshmeal type of r a t i o n with and without the addition of 7.5 per cent herring 011 were examined. 35. The composition of the basal diet i s shown i n Table 15. Day-old Hew Hampshire cockerels were used and d i s t r i b -uted at random i n heated brooders into twelve lots of 27 birds each. The usual methods of management were maintained throughout the experimental period of 5 weeks. The data obtained from the feeding t r a i l are summar-ized i n Table 14 . At the conclusion of the experiment, the hemoglobin content of the blood from chicks fed the basal diets with and without o i l , basal plus f o l i c with and without o i l and asal plus l e c i t h i n with and without o i l , was deter-mined by the copper sulphate specific gravity method. RESULTS As was anticipated the addition of f o l i c acid to the basal diet resulted i n a marked Increase i n growth. The addition of both f o l i o acid and aureomycin to the ration gave an Increase in growth over and above the basal containing f o l i c acid. Lecithin, when added to the basal ration defic-ient i n f o l i o acid improved the growth rate to the same extent as did f o l i c acid. However, the combination of f o l i c acid and lecithin did not increase the growth rate above that of f o l i c acid or l e c i t h i n alone. The response obtained from a combination of f o l i c acid, aureomycin and le c i t h i n was greater than f o l i c acid plus l e c i t h i n but less than f o l i c acid plUB aureomycin. Again as i n previous tests, the addition of herring o i l 36. to a fo l io acid deficient diet augmented the deficiency. The presence of f o l i c acid and o i l gave greater growth than did the fo l i c acid alone. The response observed with the combination of f o l i c acid and aureomycin was greater than with f o l i c acid alone with o i l but less than with f o l i c acid and aureomycin i n the oi l - free diet . When l e c i t h i n was added to the basal ration, a greater growth depression occurred than with the o i l alone. There was an additional response when fo l i c acid and l ec i th in i n these o i l supplemental diets was used than that obtained with fo l i c acid alone. Supplementation of the oil-containing diets with fo l io acid, aureomycin and l ec i th in resulted i n a considerable increase i n the rate of growth of the chicks. Shown i n Table 14 are the feed efficiencies of the different groups at the end of 5 weeks. These results indicate that the presence of l e c i th in i n the growing diet resulted i n better u t i l i z a t i o n of feed. The inclusion of supplementary herring o i l , on the other hand, tended to decrease the efficiency of u t i l i z a t i o n , with the exception of the groups containing f o l i c acid and l e c i t h i n . The u t i l i z a t i o n of the rations containing fo l io acid alone or i n combination with l e c i t h i n was further Improved when aureomycin was added to these diets . The greatest improve-ment i n feed efficiency occurred i n the ration containing fo l i c acid, l e c i t h i n and aureomycin supplemented with 7.5 per cent herring o i l . TABLE 13. Effect of l e c i t h i n and fat on the growth response of New Hampshire cockerels at four weeks of age. Supplement to None 5*0% Herring 10% Herring basal ration - o i l . o i l . Weights Feed Weight Feed Weight Feed (wna.) Gain (was.) Gain (was.) Gain None 357 1.96 382 1.88 338 2.00 0.5% Lecithin 370 2.01 372 1.96 355 1.81 1.0% Lecithin 371 2.00 364 1.99 349 1.84 Analysis of Variance Source of Error Sum of Squares d.f. Variance Total 434606 197 Treatment Error 41868 492738 8 189 5224 * 2607 Minimum significant Difference at p 0.05* 31 at p 0.01 TABLE 14. Effect of le c i t h i n , f o l i c acid and fat on growth and feed efficiencies o f chicks fed f o l i c acid deficient diets at five weeks* Supplements to Without 7.5% With 7.5% basal ration Herring o i l Herring o i l Weight Feed Weight Feed (gms.) Gain (gms.) Gain None 355 2.44 399 2.82 Folic aold * 392 2.14 424 2.15 Lecithin 395 1.91 299 2.65 Folic aold plus l e c i t h i n 399 1.91 447 1.84 Folic acid pluja aureomycin W 461 1.75 456 1.98 Folio acid plus leoithin plus aureomycin 424 1.68 509 1.57 •** Aureomycin added at level of 4.5 mg.Ab. (from Aurofao A) *+ Lecithin added at level of 6.5 per cent. 4 Folic aold added at level of 0.36 mg./lb. Analysis of Variance Source of Error Sum of Squares d.f. Variance Total 1617919 275 Treatment 761307 12 63442 ** Error 1056612 264 4002 Minimum Significant Difference at p 0.05 * 35 at p 0.012 47 TABLE 15* Composition of basal diets used i n studies on l e c i t h i n and fat* INGREDIENTS Test 1* Test 2. (percent) (percent) Ground yellov oorn 20*00 70*25 Ground wheat 26*00 Cornstarch 10*00 7*50 Soybean o i l meal 25*00 Fishmeal 8*00 20*00 Vacatone 2*00 Dehydrated cereal grass 2*50 Dried brewers' yeast 3*00 Bonemeal 1*50 Limestone 1*00 1*25 Iodized salt 0*50 0*50 Feeding o i l (2,250A-300D) 0*25 0*25 Choline chloride (25#) 0*25 0.25 Manganese sulphate 0.0125 10.00 gms. Riboflavin 0.16 gms. Calcium pantothenate 0.50 gms. Nicotinic aold 0.80 gms. Folic acid 0.036 gms. 37. DISCUSSION The above experiments regarding surface a c t i v e agents gave a d d i t i o n a l evidence i n c o n f i r m a t i o n of the previous f i n d i n g s of E l y and Schott (1952) and Synder et a l (1953) t h a t the growth response of c h i c k s i s i n f l u e n c e d by the composition of the d i e t f e d . R e s u l t s from these experiments show d i s s i m i l a r i t y i n the responses obtained by the a d d i t i o n o f the surface a c t i v e agents, Tween-80 and Santomerse-80 to the b a s a l d i e t s . P u b l i s h e d experiments on the growth of c h i c k s f e d d i e t s supplemented w i t h v a r i o u s s u r f a c t a n t s have f a i l e d t o show a growth s t i m u l a t i n g e f f e c t at 4 weeks. The growth response obtained when c h i c k s were f e d the corn-f l s h m e a l b a s a l d i e t , adequately f o r t i f i e d w i t h f o l i c a c i d , and supplemented w i t h surface a c t i v e agents showed co n s i d -erable v a r i a t i o n . I n Experiment I I , Test 1, 3 and 5, i t was demonstrated t h a t when the surface a o t i v e agents were added to the corn-flshmeal b a s a l d i e t s c o n t a i n i n g adequate f o l i c a c i d there was an improvement i n the r a t e of growth a t 4 weeks. I t may be noted t h a t , i n a l l t e s t s where an a n t i b i o t i c was a v a r i a b l e , s t i m u l a t i o n of growth occurred a t 4 weeks. The r e s u l t s obtained at 8 or 9 weeks tend t o confirm the r e s u l t s of E l y (1952) t h a t the weight responses obtained a t 10 weeks were d i f f e r e n t from those a t 4 weeks. These 38. results do not agree with the findings of Branion and H i l l (1954) or Snyder et a l (1953) who did not obtain any growth response over the 10-12 week feeding period with the addition of detergent to the bgsal diet. The data obtained from Experiment III, Test 4 indicate a similarity i n the action of detergents to that of anti-biotics since the addition of Santomerse-80 to a f o l i o acid deficient ration improved the rate of growth obtained with this ration. The growth promoting properties of detergents obtained by Ely (1951) has been reported by Stern and McGinn!s (1953) to be due to a deficiency i n the basal diet of vitamin &12* Equal growth responses resulted from the addition to this basal diet of vitamin B12, detergents or aureomycin, Whitehall et a l (1950) and Mariakulandai et a l (1952) suggests that under certain conditions antibiotics spare the require-ment of chicks for vitamin Bi2» possibly by increasing intest-i n a l synthesis or by increasing intestinal absorption of the vitamin. In the present feeding tests, the basal diets were considered an ample source of vitamin B12; fishmeal and l i v e r -meal supply unidentified factors as well. The a b i l i t y of the detergents to stimulate growth on these basal diets may be explained, therefore, by increasing the intestinal absorption of vitamins or unknown growth faetor(s). However, i n those rations where no additional growth response occurred with the presence of detergents, the possibility exists that more 39. rapid absorption of the nutrients from a r a t i o n , containing such nutrients i n excess of maximum requirements, would not re s u l t i n Increased growth. Examination of the data presented by the d i f f e r e n t tables show that a d i s s i m i l a r i t y i n the action of the deter-gents and a n t i b i o t i c s existed when the basal d i e t s contained o i l . The growth obtained with each of these supplements being s i m i l a r when the d i e t contained no a d d i t i o n a l o i l and the response to the surface active agent f a l l i n g o f f as o i l was added to the r a t i o n . In some of the experiments reported above, i t may be observed that a combination of a surface active agent and an a n t i b i o t i c stimulated growth greater than the a n t i b i o t i c alone while on the other hand, such combinations had the e f f e c t of reducing the growth below that acquired by the additi o n of an a n t i b i o t i c alone. The presence of o i l with such combinations reduced the growth response to a greater extent than when an a n t i b i o t i c and a surface active agent were present i n the o i l - f r e e diets.* I t i s apparent from the data presented that any growth stimulating e f f e c t by Incorporating the p a r t i c u l a r surface active agents i n the d i f f e r e n t rations could not be a t t r i b u t e d to an e f f e c t on the u t i l i z a t i o n of f a t . There appears to be no adequate explanation of the varying growth responses to the surface active agents observed i n these experiments. In speculating on the various responses, several explanations may 4 0 . be considered. If the stimulation of growth of chicks fed a complete ration containing an antibiotic i s due to an alteration of the bacterial flora, the inclusion of a sur-faoe active agent may also influence the intestinal flora i n a similar manner. Thus the effect of a surface active agent might be said to be due to an antibiotic effect. Ely and Schott (1952) suggested that selective bacterial inhib-i t i o n may also apply to surface active agents. As a second alternative theory these investigators proposed that more rapid absorption of nutritional factors from an altered intestinal surface tension. If such a phenomenon occurs, then the change i n the intestinal surface tension may also effect the intestinal microflora. It i s possible that one or a l l of these phenomena may be functioning simultaneously within the intestine of the chick and hence the variable responses. It therefore seems possible that variable growth responses to detergents may be mediated through intestinal bacteria and (or) altered surface tension. In the light of the data presented, and Inasmuch as other investigations have not yielded conclusive evidence on the mechanism of surface active agents within the intestine of the chick, the indiscriminate addition of surfactants to chick starter rations i s contraindicated. Biely and March (1951b) obtained an improvement i n the growth of chicks when aureomycin was added to a f o l i c acid deficient diet. They concluded that sub-optimal dietary levels of f o l i c aold are adequate when the antibiotic i s fed. The results of these tests show that the addition of aureo-mycin to a corn-flshmeal type chick starter ration deficient i n f o l i o acid resulted i n increased growth, i n agreement with the rsults of the above mentioned investigators. Furthermore, the data obtained i n Experiment II, Test 4 and Experiment III, Test 2 indicate that both p e n i c i l l i n and le c i t h i n lessened the degree of fo l i o acid deficiency when these compounds were added to the f o l i c aold deficient basal diets. With the f o l i c acid deficient rations supplemented with fat, i t w i l l be seen i n Tables 10, 11 that aureomycin or le c i t h i n did not spare the retirement for f o l i o acid as well as when these rations did not contain additional.fat. With the basal diets contain-ing optimal levels of f o l i c acid there was a slight improve-ment i n growth with the presence of leci t h i n whereas with the antibiotics no greater effect occurred than with f o l i c acid alone. The addition of f o l i c acid and aureomycin to the basal diet deficient i n f o l i c acid and containing 7.5% herring o i l resulted i n a slightly greater growth response than the addi-tion of f o l i c acid or f o l i c acid and le c i t h i n . From the results reported herein i t does not appear that the inclusion of l e c i t h i n i n diets containing added fat enhanced the u t i l i z a t i o n of the fat. The influence of l e c i -thin on the growth promoting a b i l i t y of the starting diets may 42. be seen i n Table 14 . It may be noted that, at five weeks of age, l e c i t h i n when added to the basal diet containing 7»5% herring o i l further augmented the f o l i c acid deficiency. Evidence i s presented i n the same table that a combination of f o l i c acid, aureomycin and le c i t h i n to the basal diet with included f a t Increased the growth response of the chicks to a greater extent than any other supplement or combination of supplements. Since aureomycin and le c i t h i n increased the growth rate of chicks fed the f o l i c acid deficient diets, their effect i n growth may have been indirect through an action on the intestinal microflora. The a b i l i t y of le c i t h i n to ameliorate f o l i c acid deficiency i n rations without additional fat and to exacerbate f o l i c acid deficiency when the diets contained additional fat cannot be associated with a similar mechanism as occurs with aureomycin. Why marginal intakes of f o l i c acid w i l l cause chicks to respond favorably to the addition of aureomycin to their diets has been postulated by Blely and March (1951b) by either reducing the intestinal microflora which compete with the'chick for B-complex vitamins, and/or by permitting the proliferation of microorganisms which syn-thesize the vitamins. A search of the literature reveals no data indicating that l e c i t h i n may have antibiotic properties although due to i t s emulsifying properties, the surface tension within the gastrointestinal tract of the chick may 43. been altered thereby affecting the microflora. It seems unreasonable to assume that any ohange i n surface tension within the gastrointestinal tract of the chick by l e c i t h i n would only Inhibit those micro-organisms that compete with the chick for certain vitamins and-or only permit the multiplication of others whioh synthesize certain vitamins, but that altered surface tension would affect a l l members of the f l o r a equally. A tentative explanation of the differences i n the growth promoting values of l e c i t h i n i n f o l i c acid deficient diets can be offered by considering that l e c i t h i n Increases the speed and degree of emulsification. Such an effect could be explained by an increased rate of absorption of nutrients from the gastrointestinal traot. On the other hand, i f f o l i c acid i s essential for the proper metabolism of fat, increasing the speed and degree of fat emulsifica-tion would heighten the demand for f o l i c acid In the basal diets containing supplemental fat. However, i t should be mentioned that i f growth i s influenced by changes i n surface tension within the gastrointestinal tract, the responses obtained by the presence of emulsifying agents i n the diet would be extremely variable. Analysis of the feces for fat content (Tables 3 and 9) indicated that the presence of a surface active agent i n the diet did affect the u t i l i z a t i o n of fat. 44. These findings are not i n agreement with the hypothesis of Tidwell and Nagler (1952) i n which they suggest that fat absorption i s not affected by exogenous emulsifiers* It i s interesting i n this connection that Auger et a l (1947) improved the d i g e s t i b i l i t y of hydrogenated cottonseed o i l by incorporating 3 per cent l e c i t h i n with the o i l . In these experiments a combination of Tween-80 and aureomycin had a greater effect on fat absorption than did each supplement singly. The fat excretion data also indicate that birds at 8 weeks of age are able to. u t i l i z e more fat than at 4 weeks. Tween-80 had a greater effect at 3 weeks i n lowering this ratio than i t did at 4 weeks. 45. The effeets of the addition of the oi l s i n various ehiok starter rations upon the growth of the chicks to four and i n some cases five weeks of age may be seen i n the data summarized i n Table 16. These data show that the addition of various levels of herring o i l , cottonseed o i l and tallow to the basal diets containing 18 or 20 per cent fishmeal resulted i n a stimu-lation to growth. Concentrations of 2.5» 5*0 and 7.5 per cent herring o i l produced heavier chicks at the end of the experimental period than did the basal rations. In those experiments containing either cottonseed o i l or tallow, increased growth responses also occurred. The u t i l i z a t i o n of feed by these experimental chicks followed the same pattern as the growth response, that i s , with the better growth more efficient assimilation of feed occurred. With the diets containing 8-9 per cent fishmeal, the results varied with the type of fat present. The chicks receiving the herring o i l supplement were only 3 grams heav-i e r i n one experiment (C) while in the other experiment (D) at the end of four weeks they weighed less than the control groups. On the other hand, those chicks receiving the diets with the supplementary tallow had greater weight at the conclusion of the experimental period than tfce basal diets. The weight data obtained from the experiments conducted with 6.7% fishmeal i n the rations indicate that a general 46, decrease In f i n a l weights occurred with the presence of herring o i l , whereas with tallow and oottonseed o i l , weights corresponding to or slightly greater than the controls occurred. The presence of fat i n these diets, i n a l l oases, regardless of the type, reduced the amount of feed reqired to produce a unit gain i n weight by the birds. Furthermore, the efficiency of feed u t i l i z a t i o n was affected beneficially as the fat level in the ration was increased. An inspection of the average weight records of the groups of chicks at the end of their respective experimental period reveals a strong trend of greater weight among the chicks fed a basal diet high in productive energy supple-mented with fat than that among chicks whose diet contained approximately the recommended productive energy less supplemental fat. The extent to which fat stimulated the growth rate i n chicks varied with the type of fat and the composition of the diet. Herring o i l showed a tendenoy to decrease growth i n chicks using rations low i n productive energy as compared to i t s use with the high productive energy of the com-fishmeal type of ration. No such growth depression accompanied the inclusion of cottonseed o i l or tallow i n the diets low i n productive energy although these fats increase the weight of the chicks when added to the corn-fishmeal type of basal ration. It i s apparent that 47. herring o i l , cottonseed o i l and tallow are worthwhile additions to an a l l fishmeal protein diet, whereas with a fishmeal-vegetable protein diet only cottonseed o i l or tallow are worthwhile additions. It i s d i f f i c u l t to offer a definite explanation why one fat should decrease the rate of growth with one type of ration while increasing growth on another, and furthermore, why one fat should increase growth regardless of the basal ration used. Some possible reasons for the discrepancy can be advanced. It may be possible that some distinctive properties of oi l s might account for this increased growth. It has been reported by Reiser (1950) that unsaturated fatty acids are essential for the proper nutrition of chicks. Carver and Johnson (1954) presented results which Indicate that unsat-urated fats contain some unidentified faotor(s) for maximum growth. However, the growth stimulation observed from the addition of tallow would not tend to support this evidence due to a relative low content of unsaturated fatty acids. Numerous investigators have reported that fishmeal contains an unknown factor(s) necessary i n obtaining maximum growth for chicks and poults. The possibility that such factor(s) may be more eff i c i e n t l y absorbed i n the presence of o i l i n the diet cannot be overlooked. This view stems from the finding of Saxena (1953) that the maximum effect 48. of the faotor(s) occurred when the ration contained 5 per cent herring meal. Whether fat i n such a ration supplies unidentified factor(s) or acts as a growth BpromotantM i n an indirect manner could not be determined from the experiments conducted. However, the "promotant" action of the fat may act by increasing the palatability of the ration, i n which case the ingredients of the ration would be more closely bound to each other and hence the chick would not be able to pick out certain ingredients of the ration. Comparatively few kinds of bacteria have the a b i l i t y to s p l i t fat, and probably small numbers of those normally present i n the Intestinal tract of the chick possess this property. Therefore the addition of more fat to the diet than i s naturally present i n the ingredients i s not expected to induce increases i n numbers of intestinal bacteria, but on the contrary tends to suppress some of those already present. This theory i s supported by the evidence put forward by Slinger, Pepper and H i l l (1952) i n feeding turkeys diets of relatively high fat content. These investigations showed that the increase i n growth may be accounted for by applying one of the many hypotheses proposed;, action,.by antibiotics as growth "promotants", w i l l deorease the growth of bacteria whioh would ordinarily use large amounts of some essential dietary nutrient. Dietary fat may act i n a similar manner. o 49. The l a c t i c acid bacteria which are believed to make up a large part of the Intestinal f l o r a of the chick require almost a l l of the known growth factors for their own growth and multiplication, and hence may be regarded as agents harmful to the chioks. The suppression of these bacteria i n the flora of the chick through the use of antibiotics has been shown by Biely and March (1951 )• If such bacteria are inhibited by the presence of dietary fat, then addition-a l vitamins and perhaps unknown factor(s) as well would then be available to the host. The influenoe of dextrin and other insoluble carbohy-drates i n maintaining a high oollform count i n chicks whloh are believed to be potent vitamin synthesizers i s f a i r l y well known. The high corn content may contribute the neo-essary carbohydrates i n order to produce a high coliform flora which through synthesis may contribute additional vitamins and unknown factor(s), providing that dietary fat does not decrease their numbers to a greater extent than the carbohydrates promote them. The failure of herring o i l to maintain or increase the growth over the basal or with the basal supplemented with tallow or cottonseed o i l cannot be readily accounted for. If the amino acids of f i s h meal are more completely u t i l i z e d and/or micro-biological synthesis favours the chick, i t does not seem probable that one o i l should favour these phenomena 50. while another i s adverse to i t . A more l i k e l y explanation would be the effect on the p a l a t i b i l i t y of the ration. Tallow as used i n these diets i s a solid at temperatures maintained i n the experimental rooms, whereas herring o i l i s a l i q u i d . When incorporated i n the rations, tallow tends to coalesce the ingredients of the ration more closely than the herring o i l , hence ingredients like wheat, corn and soybean o i l meal which tend to increase the bulklness of the ration are more closely adhered to the other ingredients of the ration by the presence of tallow. This reduces the a b i l i t y of the chick to select certain ingredients i n the ration with the overall effect that the chick ingests a complete diet continuously. TABLE 16. Effect of feeding different levels of the various fats on the average weight (gms.) and feed efficiencies of chicks at 4 weeks of age. (Unless otherwise indicated.) Ration Type Prod.*. Energy % Fish-meal Supplement Weight 4 wks. 5 wks.' % Increase Over Basal. Feed Gain A 888 20.00 Basal T.5% H.O. 392 * 424 * ^8.2 2.14 2.15 B 914 18.75 Basal 2.5% H.O. 5.0% H.O. 340 381 377 +12.0 +1.1..0 2.11 2.06 2.06 B 914 18.75 Basal 5.0% H.O. 416 * 447 * + 7.5 2.05 1.98 B 914 18.75 Basal 5.0% H.O. 5.0% Tallow 278 322 356 +15.8 t-28.0 B 914 18.75 Basal 5.0% C.S.O. 514 J 547 * + 6.8 1.97 1.88 C 744 9.20 Basal 6.0% H.O. 6.0% Tallow 260 263 281 +8.0 D 695 8.00 Basal 5.0% H.O. 5.0% Tallow 10.0% H.O. 10.0% Tallow 382 374 402 362 399 -2.1 +5.2 -5.5 +4.5 E 784 6.70 Basal 2.5% H.O. 5 . 0 * H.O. 7.5% H.O. 331 330 351 326 +6.0 -1.5 2.12 2.06 1.98 2.02 P 801 6.70 Basal 2.5% H.O. 5.0% H.O. 321 302 301 .15.8 -15.8 2.24 2.25 2.05 P 801 6.70 Basal 5.0% H.O. 5.0% C.S.O. 5.0% Tallow 358 331 369 362 ~8.2 +-3.0 +1.1 2.36 2.35 2.20 2.23 * The productive energy values calculated from Fraps' (1946) table on the caloric content of the various rations. (H.O. - Herring Oil C.S.O. - Cottonseed Oil) TABLE 17 Composition of the diets supplemented with the various oils, (Weight of ingredients i n pounds unless otherwise stated.) Ration Type C D E F Ingredient Ground Yellow corn 28.00 20.00 32.60 31.05 Ground wheat 28.00 26.00 32.60 31.05 Soyabean o i l meal 18.40 25.00 13.40 13.93 Flshmeal 9.20 8.00 6.70 6.97 Livermeal 3.00 3.00 Dried brewers' yeast 2.00 3.00 2.00 2.00 Dehydrated cereal grass 1.50 2.50 1.00 1.00 Vacatone 2.00 Iodized salt 0.50 0.50 0.50 0.50 Limestone 1.00 1.00 1.00 1.00 Bonemeal 1.75 1.50 1.50 1.50 Feeding o i l (2.250A-300D) 0.25 0.25 0.25 0.25 Choline chloride (25#) 0.40 0.25 0.40 0.40 Manganese Sulphate 0.025 0.025 0.025 0.025 Ration Type A B Ingredient Ground yellow corn 70.25 74.00 Fishmeal 20.00 18.75 Iodized salt 0.5© 0.50 Limestone 1.25 1.25 Feeding o i l (2,250A-300D) 0.25 0.25 Choline chloride (25%) 0.25 0.25 Manganese sulphate 10.00 gms. 10.00 gms. Riboflavin 0.16 gms. 0.16 gms. Calcium pantothenate 0.50 gms. 0.50 gms. Nicotinic acid 0.86 gms. 0.80 gms. Folic acid 0.036 mg. 0.036 mg. 51. SUMMARY A s e r i e s of three experiments i n v o l v i n g t e n t e s t s , was undertaken t o determine the e f f e c t s on c h i c k growth of v a r -i o u s surface a c t i v e agents and a n t i b i o t i c s when these substances were added t o c h i c k r a t i o n s . B a s a l r a t i o n s of d i f f e r e n t composition w i t h and without a d d i t i o n a l f a t were used t o determine whether the nature of the d i e t a f f e c t e d the growth of c h i c k s . Fat was in c o r p o r a t e d i n the b a s a l d i e t s a t the expense of eq u i v a l e n t amounts of c o r n s t a r c h . 1) Growth s t i m u l a t i o n has been obtained w i t h d i f f e r e n t surface a c t i v e agents and a n t i b i o t i c s . V a r i a b i l i t y was apparent f o r the same detergent i n d i f f e r e n t experiments. I n some cases a growth s t i m u l a t i o n of c h i c k s occurred w i t h the surface a c t i v e agents a t 4 weeks w h i l e i t was observed t h a t a n t i b i o t i c s i n eaoh case produced a growth stimulus a t 4 weeks. The t r i p l e combination of f o l i c a c i d , a n t i -b i o t i c s and surface a c t i v e agents i n c r e a s e d the growth r a t e of c h i c k s than the double combination of any of the supple-ments. The t r i p l e response was g r e a t e r i n the presence of a d d i t i o n a l f a t i n the b a s a l d i e t s . Growth s t i m u l a t i o n occur-r i n g w i t h the detergents at 8 weeks of age was not always apparent when the c h i c k s had reached the age of 4 weeks. Some of the d i f f e r e n c e s observed i n the response to surface a c t i v e agents seem t o be due to the composition of the ex-perimental d i e t s . 52. 2 ) An increase i n the weights of chicks and an accompanied improvement i n feed efficiencies was obtained by the addition of either herring o i l , cottonseed o i l or tallow to corn-fishmeal basal diets well f o r t i f i e d with a l l the known vitamins required for chick growth. On the other hand, with a fishmeal-soybean protein diet only cottonseed o i l or tallow are advantageous additions. 3) Supplementation of the fol i o acid deficient corn-fish-meal type of ration with l e c i t h i n ameliorated the deficiency while a combination of lecithin, aureomycin and f o l i c acid did not improve the growth of the chicks as much as f o l i c acid and aureomycin. 4^)i No enhanced u t i l i z a t i o n of fat as measured by the growth response of chicks could be attributed to the addition of lec i t h i n to diets containing relatively high levels of fat. 5) Analysis of the feces for far oontent indicated that fat was more effectively u t i l i z e d i n the presence of a surface active agent. The greater u t i l i z a t i o n of fat occurred with the higher level of the surface active agent and as the chicks became older. 53. BIBLIOGRAPHY Aldereberg, D. and H. Sobotka, 1943. Influence of l e c i t h i n on fat and vitamin A absorption i n man. J. Nutrition. 25: 255-263. Allison, J.B., H.L. Rosenthal and A.H. Mil l s , 1952. Effects of non-ionic surface active agents on the growth of animals. Fed. Proc. 11: 435. Angler, R.B., J.H. Booth, B.L. Hutchings, J.H. Howat, J. Semb, E.L.R. Stokatad, Y. Subbarow and C.W. Walker, 1945. Synthesis of a compound identical with the L. easel factor isolated from l i v e r . Science, 102: 227-228. Augur, V., H.S. Hollman and H.J. Deuel Jr., 1947. The effect of crude l e c i t h i n on the coefficient of di g e s t i b i l i t y and the rate of absorption of fat. J. Nutrition. 33: 177-186. Biely, J., B.E. March and H.L.A. Tarr, 1951. The nutritive value of f i s h meal and condensed f i s h solubles. III. Effect of heating fat - containing and hexane-extracted meal. Progress Reports, Pacific Coast Stations, Fisheries Research Board of Canada, 89: 79. Biely, J. and B. March, 1951. The effect of aureomycin and vitamins on the growth rate of chicks. Science 114: 330-331. B i e r i , J.G. and R.P. Sandman, 1951. Comparative u t i l i z a t i o n of carotene administered orally and parenterally. Proc. Soc. Exptl. B i o l . Med. 77: 617-619. Birch, T.W., 1938. The relation between vitamin B< and the unsaturated fatty acid faotor. J. Biol. Chem. 124: 775-793. Bird, O.D., M. Robblns, J.M. Vanderbett and J.J. Pfiffner, 1946. Observations on vitamin B c conjugase from Hog kidney. J. Bi o l . Chem. 163: 649-659. Branion, H.D. and D.C. H i l l , 1954. Detergents and chick growth. Poultry Sci. 33: 62-66. Burns, M.J., S.M. Hauge and F.W. Quaokenbush, 1951. U t i l i -zation of vitamin A and carotene by the rat. 1. Effects of tocopherol, Tween and dietary fat. 2. Effects of mineral o i l and fat content of diet. Arch. Biochem. 30: 341-346. 54 Burr, G-.0. and M.M. Burr, 1929* A new deficiency disease produced by the r i g i d exclusion of fat from the diet, J. B i o l . Ghem, 82: 345-367. Carver, D.S. and E,L, Johnston, 1954, Further studies of the unidentified chick growth factors i n unsaturated fats. Poultry Sci. 33: 543-548. Chow, B.F., J,M, Burnett, C,T. Long and L. Barrows, 1953* Effect of non-ionic surface active agents on the growth of animals. Fed. Proc. 2: 435. Commercial Fisheries Review, 1953. Fish and Wildlife Service, U.S.D.I. 15: No. 6, 13-15. Crampton, E.W., R.H, Common, F,A, Farmer, F.M. Berryhill and L. Wiseblatt, 1951. Studies to determine the nature of the damage to the nutritive value of some vegetable o i l s from heat treatment. I. The relation of autooxidation to decrease the nutritional value of heated linseed o i l . J. Nutrition 43: 533-539. Darby, W.J., M.M.- Kaser and E. Jones, 1947. The influence of pteroylglutamlo acid on the absorption of vitamin A and carotene by patients with sprue. J. Nutrition 33: 243-250. Davis, J.E., 1946. Hyperchronlc anemia produced by choline or acetylcholine and the induced remission of both by f o l i c acid or l i v e r Injection: The probable mechanism of action of l i v e r and f o l i c acid i n the treatment of anemia. Am. J. Physiol. 147: 404-411. Chow, B.F., J.M. Burnett, C.T. Ling and L. Barrows, 1953. Effects of basal diets on the response of rats to certain non-ionic surfaoe active agents. J. Nutrition. 49: 563-577. -Dinning, J.S., C.K. Keith and P,L. Day, 1949. A relation-ship of f o l i c acid to choline oxidase. Arch, Biochem, 24: 463-464, Eaton, H.D., L.D. Matterson, L. Decker, C.F. Helmboldt and E.L. Gungherr, 1951. Intravenous and oral administration of an aquous suspension of carotene to calves depleted of their vitamin A stores. J. Dairy Sci. 34: 1073-1080. Elvehjem, C.A. and A. Arnold, 1939a. Influence of the com-p position of the diet on the thiamin requirement of dogs. J. Physiol. 26: 289-292. 5 5 . Elvehjem, CA., A. Arnold and F.E. Stern, 1939b. The relation of dietary fat to the thiamin requirement of growing rate. J. Nutrition 17: 485-495. Ely, CM., 1951. Chick growth stimulation produced by sur-, factants. Science 114: 523-524. Ely, C.M., and S. Schott, 1952. Surface active agents as growth stimulators i n chick rations. Proc. 7th D i s t i l l e r s Feed Conference: 72-84. Esh, G.C. and T.S. Sutton, 1948. Lecithin i n vitamin A Nutrition. J. Nutrition. 36: 391-404. Evans, H.M. and S. Lepkovsky, 1929. Sparing action of fat on the antineuritic vitamin B. J. Biol. Chem. 83: 269-287. Forbes, E.B. and R.W. Swift, 1944. Associative dynamic effects of protein, carbohydrate and fat. J. Nut. 27: 453-468. Forbes, E.B., R.W. Swift and R.F. E l l i o t , 1946. Relation of fat to economy of food u t i l i z a t i o n by the mature albino rat. J. Nutrition 31: 213-227. Gregory, J.D., G.D. Novelli and F. Lipmann, 1952. The composition of Coenzyme A. J. Am. Chem. Soc. 74: 854. Haliok, J.V., B.L. Rsid, G.L. Brown and J.R. Couch, 1953. The vitamin B^o content of egg yolks as influenced by oral and parenteral administration of the vitamin. J. Nutrition. 5 0 : 330-340. Halpern, G.R. and J. Biely, 1948. The u t i l i z a t i o n of vitamin A i n various carriers. J. Biol. Chem. 174: 817-826. Harris, R.S. and H.S. Sherman, 1951. Nutritional and pathological effects of sorbitan monolaurate, polyoxy-ethylene monolaurate, and polyoxyethylene sorbitan monostearate when fed to rats. Arch. Biochem. and Biophys. 34: 249-258. Henderson, E.W. and W.E. Irwin, 1940. The tolerance of growing chicks for soybean o i l i n their ration. Poultry Sci. 19: 389-395. Holman, R.T., 1951. Fatty acids i n nutrition. Proc,of 3rd Conf. on Research. 56. Hoover, C. and P. Swanson, 1950, Role of fat In protein metabolism. Fed. Proc. 9 : 362. Huff, J.S., R.K. Waugh and Q.H. Wise, 1951. Effect of glycerol monosterate i n fat absorption, growth and health of calves. J. Dairy Sci. 34: 1056-1063. Irwin, M.H., J. Weber and H. Steenbock, 1936. The Influence of certain hydrotropic and other substances upon fat absorption. J. Nutrition. 12: 365-371. Johnson, A.L., R.B. Scott and L.H. Newman, 1950. Tween-20 and fecal fat i n premature infants. Am. J. Diseases Children. 80: 545-550. Jones, C.H., P.J. Culver, G.D. Drummer and A.E. Ryan, 1948. Modification of fat absorption i n the digestive tract by the use of an emulsifying agent. Am. Internal Med.29: 1-10. Keresztesy, J.C., E.L. Rickes and J.L. Stokes, 1943. Anew growth factor for streptococcus l a c t l s . Sci. 97: 465. Krantz, S.C., 1949. Unpublished observations cited i n Nutrition Revs. 7, 205-207. Luckey, T.D., P.R. Moore, C.A. Elvehjem and E.B. Hart, 1946. Effect of diet on the response of chicks to f o l i c acid. Proc. Soc. Exp. B i o l . Med. 62: 307-312. Leuoke, R.W., J.A. Haefer and F. Thorp Jr., 1952. The growth promoting effect on pigs of a surface active agent. Quarterly Bull, of the Mich. Agr. Exp. Stat. 34: 331-332. Lynen, F. and S. Oohoa, 1953. Enzymes and fatty acid.Meta-bolism Biochemica et Biophysics Acta 12: 299-314. Niekol, C.A. and A.D. Welch, 1950. Synthesis of citrovorum factor from f o l i c acid by l i v e r slices. Pro. Soc. Exper. Biol. Med. 74: 52-55. Pearson, P.B. and F. Panzer, 1949. Effect of fat in the diet of rats on their growth and their excretion of amino acids. J. Nutrition. 38: 257-265. Pepper, W.F., J.J. Slinger and E.S. Snyder, 1953. Value of low levels of soybean o i l i n Broiler diets containing a high percentage of wheat. Poultry Sci. 32: 1084-85. 57. Pfiffner, J.J., SfB. Blinkley, E.S. Bloom, R.A. Brown, O.D. Bird, A.D. Emmett, A.G. Hogan and B.L. 0'Dell, 1943. Isolation of the antianemia factor (vitamin B 0) i n orystalline form from l i v e r . Science 97: 404-405. Popp, E.M, and J.R. Totter, 1952. Studies on the relation-ship between f o l i c acid and Coenzyme A. J. B i o l . Chem. 199: 547-552. Quackenbush, F.W., F.A. Kummerov and H. Steenback, 1942. The effectiveness of l i n o l e i c , arachidonio and linolenic acid i n reproduction and lactation. J. Nut. 24: 213-224. Rickes, E.L., L. Chalet and J.C. Keresztesy, 1947. Rlzopterin, a new growth factor for Streptococcus Lactis R. J. Am. Chem.- Soc. 69: 2749. Reiser, R,, 1950. The essential role of fatty acids on growing chicks. J. Nutrition 42: 319-323. . Reiser, R. and P.B. Pearson, 1949. High levels of fat with suboptimum levels of riboflavin on the growth of the chick. J. Nutrition. 38: 247-256. Russel, W.G., M. Wright and L.J. Polskin, 1940. Fat require-ments of the growing chick. J. Nut. 19: 555-562. Salmon, W.D., 1941. The relationship of pantothenic acid, pyridoxime and l i n o l e i c acid to the cure of rat acrodynla. J. Biol. Chem. 140: CIX. Sauberlich, H.E. and C.A. Baumann, 1948. A factor required for the growth of leuconostoc citrovorum. J. B i o l . Chem. 176: 165-173. Saxena, H.G., L.G. Braylook, J.S. Carver and J. McGinnis, 1953. 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Proc. 6: 423. Woodruff, A.W., 1950. The absorption of oleic acid from the intestine of rats deficient i n fol i o acid and biotin. Brit. J. Exptl. Pathol. 31: 405-409. Woods, D.D., 1948. Les sulfamides en tant qu'antagonistes de 1' aeride p-aminobenzolque. Bull. Soc. Ghim. B i o l . 30: 730-747. Woolley, D.W. and R.B. Pringle, 1950. Formation of 4-amino-5-earboxamldoimidizole during the growth of E. co l i i n the presence of 4-amlnopteryl glutamic acid. J. Am. Chem. Soc. 72: 634-635. 

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